Biosurfactants and methods of uses and making

Abstract

Provided herein are biosurfactants or compositions comprising thereof. The biosurfactants provided exhibit various properties under various conditions, as described herein. Also provided herein are methods for using the biosurfactants in various applications, including but not limited to petroleum or oil applications, as described herein. Furthermore, the methods of making the biosurfactants is also provided.

Description

WSGR Docket No.: 69849-701.601BIOSURFACTANTS AND METHODS OF USES AND MAKING CROSS REFERENCE

[0001] This application claims the benefit of U.S. Provisional Pat. App. No. 63 / 775,717, filed on March 21, 2025, which is entirely incorporated by reference herein for all purposes.BACKGROUND

[0002] This disclosure is in the field of surfactants.SUMMARY

[0003] Provided herein, are biosurfactant compositions. In an aspect, a biosurfactant composition comprises: (i) at least one glycolipid, wherein the at least one glycolipid comprises at least one acetyl group; (ii) an enzyme; and (iii) an additive. In some embodiments, the at least one glycolipid comprises at least two acetyl groups. In some embodiments, the at least one glycolipid comprises at most five acetyl groups. In some embodiments, the at least oneHOv— ( glycolipid comprises a chemical structure of formula (I) or (II),0H(I),(II) wherein: each R1is independently selected from hydrogen, Ci-6 alkyl, CU,, alkenyl, C2-6 alkynyl, C1-6 haloalkyl, -C(=O)Rn, -CH2C(=O)Rn, -C(=O)N(Rn)2, and-CH2C(=O)N(Rn)2, wherein Ci.6 alkyl, C2-6 alkenyl, C2-ealkynyl, and C1-6 haloalkyl are optionally substituted with one or more substituents, each R11is independently selected from hydrogen, C1-6 alkyl, C2-6 alkenyl, and C2-6 alkynyl CH3; each R2is independently selected from hydrogen, halogen, -OH, C1-6 alkyl, C2-6 alkenyl, and C2-6 alkynyl, wherein C1-6 alkyl, C2-6 alkenyl, and C2-6 alkynyl are optionally substituted with one or more substituents; and each R3is independently selected from Cs-20 alkyl, C8-2o alkenyl, and C8.2o alkynyl wherein Cx-20 alkyl, C8.2o alkenyl, and C8.2o alkynyl are optionally substituted with one or more substituents. In some embodiments, R1is hydrogen. In some embodiments, R1is -C(=O)Rn. In some embodiments, R11is C1-6 alkyl. In some embodiments, R11is Ci alkyl. In some embodiments, R1is -C(=O)CH3.WSGR Docket No.: 69849-701.601In some embodiments, R2is optionally substituted C1-5 alkyl. In some embodiments, R2is optionally substituted Ci alkyl. In some embodiments, R3is optionally substituted Cx-20 alkyl. In some embodiments, R3is optionally substituted Cs-20 alkenyl. In some embodiments, R3is optionally substituted C12-19 alkenyl. In some embodiments, R3is optionally substituted C13 alkenyl or C15 alkenyl. In some embodiments, the optionally substituted Cs-20 alkenyl comprises one carboncarbon double bond. In some embodiments, the optionally substituted Cs-20 alkenyl comprises two carbon-carbon double bonds. In some embodiments, the one or more substituents are independently selected from halogen, -OH, -NO2, =0, =S, -CN, C1-6 aminoalkyl, C1-6 alkoxy, Ci-6 hydroxyalkyl, and Ci-ehaloalkyl. In some embodiments, the at least one glycolipid comprises achemical structure selected from the group consisting of:WSGR Docket No.: 69849-701.601WSGR Docket No.: 69849-701.601In some embodiments, the at least one glycolipidcomprises a chemical structureof In some embodiments, the atleast one glycolipid comprises a chemical structureof . In some embodiments, the at least one glycolipid comprises a chemical structure of. In some embodiments, the at least one glycolipid comprises achemical structure of . In some embodiments, the enzymeWSGR Docket No.: 69849-701.601comprises an enzyme from a fungus. In some embodiments, the fungus comprises a filamentous fungus. In some embodiments, the filamentous fungus comprises at least one species from the family Xylariaceae. In some embodiments, the enzyme is selected from the group consisting of alcohol dehydrogenase, monooxygenase, dioxygenase, peroxidase, and laccase. In some embodiments, the enzyme is present in an amount of at least about 0.01% w / w of the biosurfactant composition. In some embodiments, the enzyme is present in an amount of at least about 1 % w / w of the biosurfactant composition. In some embodiments, the enzyme is present in an amount of at most about 10 % w / w of the biosurfactant composition. In some embodiments, the enzyme is present in an amount of at most about 5 % w / w of the biosurfactant composition.

[0004] Provided herein, are biosurfactant compositions. In an aspect, a biosurfactant composition is configured to reduce surface tension between two phases under a condition, wherein the two phases comprise: (i) two different liquid phases, (ii) a first liquid phase and a solid phase, or (iii) a second liquid phase and a gaseous phase; wherein the condition is selected from the group consisting of: (a) a pH range that is: (1) from about 2 to about 4; (2) from about 10 to about 12; or (3) a combination of (1) and (2); (b) a temperature range that is: (1) from about -10°C to about 4°C; (2) from about 90°C to about 100°C; or (3) a combination of (1) and (2); (c) a salinity range that is: (1) from about 50000 total dissolved solids (TDS) to 70000 TDS; (2) from about 130000 TDS to about 150000 TDS or (3) a combination of (1) and (2); or (d) any combination of (a)-(c). In some embodiments, the first liquid phase and the second liquid phase are a same liquid phase. In some embodiments, the first liquid phase and the second liquid phase are different liquid phases.

[0005] Provided herein, are biosurfactant compositions. In an aspect, a biosurfactant composition is configured to remove or sequester a hydrocarbon from a mixture comprising an aqueous component and the hydrocarbon under a condition selected from the group consisting of: (a) a pH range that is: (1) from about 2 to about 4; (2) from about 10 to about 12; or (3) a combination of (1) and (2); (b) a temperature range that is: (1) from about -10°C to about 4°C; (2) from about 90°C to about 100°C; or (3) a combination of (1) and (2); (c) a salinity range that is: (1) from about 50000 Total dissolved solids (TDS) to 70000 TDS; (2) from about 130000 TDS to about 150000 TDS or (3) a combination of (1) and (2); or (d) any combination of (a)-(c).

[0006] Provided herein, are biosurfactant compositions. In an aspect, a biosurfactant composition is configured to facilitate emulsification of a mixture comprising two different liquids under a condition selected from the group consisting of: (a) a pH range that is: (1) from about 2 to about 4; (2) from about 10 to about 12; or (3) a combination of (1) and (2); (b) a temperature range that is: (1) from about -10°C to about 4°C; (2) from about 90°C to about 100°C; or (3) a combination of (1) and (2); (c) a salinity range that is: (1) from about 50000WSGR Docket No.: 69849-701.601Total dissolved solids (TDS) to 70000 TDS; (2) from about 130000 TDS to about 150000 TDS or (3) a combination of (1) and (2); or (d) any combination of (a)-(c).

[0007] Provided herein, are biosurfactant compositions. In an aspect, a biosurfactant composition is configured to not exhibit a cloud point formation under a condition selected from the group consisting of: (a) a pH range that is: (1) from about 2 to about 4; (2) from about 10 to about 12; or (3) a combination of (1) and (2); (b) a temperature range that is: (1) from about -10°C to about 4°C; (2) from about 90°C to about 100°C; or (3) a combination of (1) and (2); (c) a salinity range that is: (1) from about 50000 Total dissolved solids (TDS) to 70000 TDS; (2) from about 130000 TDS to about 150000 TDS or (3) a combination of (1) and (2); or (d) any combination of (a)-(c).

[0008] Provided herein, are biosurfactant compositions. In an aspect, a biosurfactant composition is configured to exhibit a surface tension of about 30 millinewton / meter (mN / m) under a condition selected from the group consisting of: (a) a pH range that is: (1) from about 2 to about 4; (2) from about 10 to about 12; or (3) a combination of (1) and (2); (b) a temperature range that is: (1) from about -10°C to about 4°C; (2) from about 90°C to about 100°C; or (3) a combination of (1) and (2); (c) a salinity range that is: (1) from about 50000 Total dissolved solids (TDS) to 70000 TDS; (2) from about 130000 TDS to about 150000 TDS or (3) a combination of (1) and (2); or (d) any combination of (a)-(c).

[0009] Provided herein, are biosurfactant compositions. In an aspect, a biosurfactant composition is configured to exhibit an interfacial tension of about 5 millinewton / meter (mN / m) under a condition selected from the group consisting of: (a) a pH range that is: (1) from about 2 to about 4; (2) from about 10 to about 12; or (3) a combination of (1) and (2); (b) a temperature range that is: (1) from about -10°C to about 4°C; (2) from about 90°C to about 100°C; or (3) a combination of (1) and (2); (c) a salinity range that is: (1) from about 50000 Total dissolved solids (TDS) to 70000 TDS; (2) from about 130000 TDS to about 150000 TDS or (3) a combination of (1) and (2); or (d) any combination of (a)-(c).

[0010] In some embodiments, the biosurfactant composition further comprises at least one glycolipid. In some embodiments, the biosurfactant composition further comprises an additive. In some embodiments, the at least one glycolipid comprises: sophorolipid, trehalose lipid, rhamnolipid, mannosylerythritol lipid, cellobiose lipid, polyol lipid, or a combination thereof. In some embodiments, the at least one glycolipid comprises: the sophorolipid, the trehalose lipid, the rhamnolipid, the mannosylerythritol lipid, or a combination thereof. In some embodiments, the at least one glycolipid comprises: the sophorolipid, the rhamnolipid, the mannosylerythritol lipid, or a combination thereof. In some embodiments, the at least one glycolipid comprises: the sophorolipid, the rhamnolipid, or a combination thereof. In some embodiments, the at least oneWSGR Docket No.: 69849-701.601glycolipid comprises the sophorolipid. In some embodiments, the sophorolipid comprises an acidic sophorolipid, a lactonic sophorolipid, or a combination thereof. In some embodiments, the at least one glycolipid is present in an amount of at least about 0.1% w / w of the biosurfactant composition. In some embodiments, the at least one glycolipid is present in an amount from about 20 % to about 30 % w / w of the biosurfactant composition. In some embodiments, the biosurfactant composition further comprises: a lipopeptide, a lipoprotein, or a combination thereof. In some embodiments, the lipopeptide is present in an amount of at least about 0.1 % by w / v of the biosurfactant composition. In some embodiments, the lipopeptide is present in an amount from about 10% to about 15% by w / v of the biosurfactant composition. In some embodiments, the lipoprotein is present in an amount of at least about 0.05 % by w / v of the biosurfactant composition. In some embodiments, the lipoprotein is present in an amount from about 5% to about 10% by w / v of the biosurfactant composition. In some embodiments, the additive is selected from the group consisting of antioxidants, preservatives, chelating agents, natural antioxidants, vegetable oil derivatives and any combination thereof. In some embodiments, the biosurfactant composition further comprises sodium benzoate. In some embodiments, the additive is present in an amount of at least about 0.001% w / v of the biosurfactant composition. In some embodiments, the additive is present in an amount from about 0.01% to about 2% w / v of the biosurfactant composition. In some embodiments, the biosurfactant composition exhibits: (a) a density of about 1 gram per cubic centimeter (g / cm3) at 25 °C; (b) a viscosity of about 2.5 Centipoise (cP) at 25 °C; (c) a hydrophilic-lipophilic balance of about 10; (d) a solids content of about 20 %; (e) a critical micelle concentration of about 9 at 25 °C, by w / v; (f) a surface tension of about 34 mN / m at 25 °C; (g) an interfacial tension of about 15 mN / m at 25 °C; (h) an emulsion index of about 100 %; (i) a hydrophilic contact angle of about 25 ° at 25 °C, when using glass as a reference substrate; (j) a hydrophobic contact angle of about 100 ° at 25 °C, when using polytetrafluoroethylene as a reference substrate; or (k) any combination of (a)-(j). In some embodiments, the biosurfactant composition is a lyophilizate.

[0011] Provided herein, are method. In an aspect, a method comprises using the biosurfactant composition as described herein (1) as an emulsifier, foaming control agent, cleaner, surfactant, wetting agent, oil recovery agent, or corrosion inhibitor; (2) to clean wells, facilitate enhanced oil recovery, disperse wax and parafilm facilitate flow assurance, facilitate high-salinity water disposal; or (3) for water treatment or polluted soil treatment; in a crude oil processing cycle.

[0012] Provided herein, are method. In an aspect, a method comprises using the biosurfactant composition as described herein to remove or sequester total petroleum hydrocarbons (TPH) from soil or water.WSGR Docket No.: 69849-701.601

[0013] Provided herein, are method. In an aspect, a method comprises using the biosurfactant composition as described herein to remove perfluoroalkyl and polyfluoroalkyl substances (PF AS) from the soil or the water.

[0014] Provided herein, are method. In an aspect, a method comprises using the biosurfactant composition as described herein as detergents, degreasers, fabric cleaners, or surface cleaners. In some embodiments, the biosurfactant composition is used to remove colored stains from a colored fabric or a colored hair.

[0015] Provided herein, are method. In an aspect, a method comprises using the biosurfactant composition as described herein to remove oil from a solid particle comprising rock. In some embodiments, the method further comprises using the biosurfactant composition and an acid to remove the oil from the solid particle.

[0016] Provided herein, are method. In an aspect, a method comprises using the biosurfactant composition as described herein as a dispersant, solubilizer, or anti-bacterial agent within agroindustrial products.

[0017] Provided herein, are method. In an aspect, a method comprises using the biosurfactant composition as described herein as a dispersant, solubilizer, or anti-bacterial agent within cosmetic products.

[0018] Provided herein, are method. In an aspect, a method comprises using the biosurfactant composition as described herein to facilitate emulsification in the cosmetic products, pharmaceutical products, petroleum products, or food products.

[0019] Provided herein, are method. In an aspect, a method comprises using the biosurfactant composition as described herein to facilitate stabilization of a vaccine.

[0020] Provided herein, are method. In an aspect, a method comprises (i) obtaining a crude extract from a microorganism; and (ii) generating the biosurfactant composition of any one as described herein using the crude extract. In some embodiments, the microorganism comprises a fungus. In some embodiments, the fungus comprises at least a filamentous fungus. In some embodiments, the fungus comprises at least one species from the family Xylariaceae. In some embodiments, the fungus comprises Entonaema liquescens. In some embodiments, the method further comprises fermenting the microorganism. In some embodiments, the fermenting comprises: (1) culturing the microorganism with a substrate comprising polysaccharides; (2) maintaining a culture of the microorganism: (i) from about 18 °C to about 30 °C, (ii) from about pH 3 to about pH 6, (iii) at an aeration rate from about 0.2 vessel volumes per minute (vvm) to about 1 vvm, or (iv) any combinations of (i)-(iii); (3) mechanically agitating the culture from about 100 rpm (revolution per minute) to about 200 rpm; or (4) a combination of (l)-(3). In some embodiments, the culturing comprises adding an inducer to the microorganism. In someWSGR Docket No.: 69849-701.601embodiments, the inducer is selected from the group consisting of hydrocarbons, synthetic lubricating oils, mineral base lubricating oils, vegetable fatty acids, and other sources of alkanes, alcohols, and aldehydes. In some embodiments, the inducer comprises an oil. In some embodiments, the oil comprises a vegetable oil. In some embodiments, the inducer is at a concentration range of 0.2% to 5% w / v of the culture. In some embodiments, the method further comprises concentrating the culture. In some embodiments, the concentrating comprises centrifugation. In some embodiments, the method further comprises purifying the crude extract from the culture. In some embodiments, the purifying comprises filtration. In some embodiments, the filtration comprises using a filter. In some embodiments, the filter has a pore size of about at most about 1 millimeter. In some embodiments, the filter has a pore size of about at least about 1 micrometer. In some embodiments, the filtration comprises vacuum filtration. In some embodiments, the substrate comprises agro-industrial waste or renewable sources. In some embodiments, the polysaccharides comprise starch, amylopectin, pectin, lignin, cellulose, or a derivative thereof, or a combination thereof. In some embodiments, the crude extract is substantially cell-free.

[0021] Additional aspects and advantages of the present disclosure will become readily apparent to those skilled in this art from the following detailed description, wherein only illustrative embodiments of the present disclosure are shown and described. As will be realized, the present disclosure is capable of other and different embodiments, and its several details are capable of modifications in various obvious respects, all without departing from the disclosure. Accordingly, the drawings and description are to be regarded as illustrative in nature, and not as restrictive.INCORPORATION BY REFERENCE

[0022] All publications, patents, and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference. To the extent publications and patents or patent applications incorporated by reference contradict the disclosure contained in the specification, the specification is intended to supersede and / or take precedence over any such contradictory material.BRIEF DESCRIPTION OF THE DRAWINGS

[0023] The novel features of the disclosure are set forth with particularity in the appended claims. A better understanding of the features and advantages of the present disclosure will be obtained by reference to the following detailed description that sets forth illustrativeWSGR Docket No.: 69849-701.601embodiments, in which the principles of the disclosure are utilized, and the accompanying drawings (“FIG.” or “FIGs.” herein), of which:

[0024] FIG. 1A depicts a schematic of the biosurfactant composition production process. FIG. IB depicts another schematic of the biosurfactant composition production process.

[0025] FIG. 2A depicts the growth of Entonaema liquescens on an agar plate. FIG. 2B depicts the growth of Entonaema liquescens in fermentation inoculum as descried herein.

[0026] FIG. 3A depicts the fermentation bioreactor. FIG. 3B depicts the aerated culture during fermentation. FIG. 3C depicts the aerated culture post-fermentation. FIG. 3D depicts a comparison of the crude extract (left vessel) and filtrate (right vessel) generated using the method described herein. FIG. 3E depicts a close-up view of the filtrated biosurfactants before lyophilization. FIG. 3F depicts a comparison between the initial state of the culture medium (pre-fermentation; left vessel) and the final state of the filtrate (right vessel). FIG. 3G depicts a close-up view of an example filtrated biosurfactants after lyophilization. FIG. 3H depicts a close-up view of an example reconstituted biosurfactant at 30% at 4 L fermentation. FIG. 31 depicts a close-up view of an example reconstituted biosurfactant at 30% at 40 L fermentation.FIG. 3J depicts an example cell-free enzymatic extract (ELC) after filtration that can be used for bioremediation. FIG. 3K depicts an example concentrated biosurfactant.

[0027] FIG. 4A depicts the various sophorolipid species generated in the biosurfactant using the methods as described herein. FIG. 4B depicts the chromatographic profile obtained by HPLC-ELS analysis of an example biosurfactant product, showing the correspondence between the chromatographic peaks of example biosurfactant #1 and those of a C18:l sophorolipid analytical standard, as generated using the methods described herein.

[0028] FIG. 5A depicts measurement of interfacial tension of the biosurfactant as described herein with lubricating oil at 27.5 °C. The left vessel shows the biosurfactant without an inducer. The right vessel shows the biosurfactant mixed with an inducer. FIG. 5B depicts measurement of interfacial tension of the biosurfactant as described herein with oil at 30.7 °C. The left vessel shows the biosurfactant without an inducer. The right vessel shows the biosurfactant mixed with an inducer. FIG. 5C depicts the pretreatment of an example biosurfactant formulation by centrifugation (left image) and its resulting appearance (right image) prior to physicochemical characterization. FIG. 5D depicts the surface tension-concentration curve of an example biosurfactant as described herein. FIG. 5E depicts the contact angle measurements of the example biosurfactant at CMC conditions. The left image shows a low contact angle on a hydrophilic glass surface (-26.79°), indicating high wettability, while the right image shows a high contact angle on a hydrophobic PTFE surface (-103.69°), indicating interaction with nonpolar substrates and balanced hydrophilic-lipophilic behavior.WSGR Docket No.: 69849-701.601

[0029] FIG. 6A depicts an image showing the formation of emulsions using fermentation product samples taken between days 0 and 5 (from the tube on the far left side to the far right side). The samples are contained in Falcon tubes with blue caps, placed in a blue rack. The emulsions exhibit different phases, indicating the interaction between the oil and the biosurfactant-rich extracts. FIG. 6B depicts a close-up of the formation and stability of the emulsion at days 0, 3, and 6. FIGs. 6C-6D depict a comparison of emulsion stability after 48 hours of resting, showing (FIG. 6C) the control sample without biosurfactant and (FIG. 6D) the sample containing the biosurfactant formulation. The image illustrates enhanced emulsion stability in the presence of the biosurfactant, as evidenced by reduced phase separation relative to the control. FIGs. 6E-6F depict a comparative foam formation assay recorded 60 seconds after air injection for 30 seconds. FIG. 6E shows a commercial synthetic surfactant used as a control, classified as high-foaming, exhibiting a tall and persistent foam column. FIG. 6F shows an example biosurfactant described herein, displaying substantially lower foam height and faster foam collapse under the same conditions. The reduced foam formation observed for the biosurfactant supports its suitability for applications requiring controlled or low foaming behavior. FIG. 6G depicts the emulsification assay of an example final biosurfactant product obtained at reactor scale, performed using a three-way stopcock mixing method. . Equal volumes of the biosurfactant formulation at its critical micelle concentration (CMC) and vegetable oil were transferred back and forth between two coupled syringes connected by a three-way valve, enabling controlled and reproducible mixing. The image illustrates the manual mixing setup and the formation of a homogeneous emulsion during repeated mixing cycles, prior to collection of the emulsion in Falcon tubes for subsequent stability evaluation immediately after preparation and after 24 and 48 hours of rest.

[0030] FIG. 7A depicts an image showing a laboratory test evaluating the activity of the biosurfactant in crude oil degradation at a concentration of 2%. The comparison was made between days 0 (left), 4 (middle), and 8 (right), demonstrating the progressive emulsification and biodegradation of crude oil over time. FIG. 7B depict a comparison between the initial day 0 (left container) and the final day 8 (right container) for the concentration of 2%. FIG. 7C shows the crude oil degradation results at the highest evaluated concentration (15%) on the initial day 0 (left container) and the final day 8 (middle container). The negative control (crude oil incubated without degradation) is shown in the container on the right. FIG. 7D shows the degradation results of different heavy hydrocarbon fractions (DRO - Diesel Range Organics, CIO to C25) using the initial concentration of 2% crude oil. The bar chart compares the treatment with the biosurfactant (blue) against the untreated control (gray), highlighting differences in hydrocarbon concentration after the degradation process.WSGR Docket No.: 69849-701.601

[0031] FIG. 8A depicts an image showing fungal growth in a culture medium without the addition of crude oil under low-temperature conditions (18°C), respectively. FIG. 8B depicts an image showing fungal growth in a culture medium without the addition of lubricant (growth control) under low-temperature conditions (18°C). FIG. 8C shows a close-up comparison between the two culture media of FIG. 8A (right vessel) and FIG. 8B (left vessel).

[0032] FIG. 9A depicts an image showing 2% crude oil sample treated with the biosurfactant composition for 8 days at low temperature (18°C). FIG. 9B depicts an image showing the untreated 2% crude oil control, incubated at low temperature (18°C) for 8 days. FIG. 9C depicts the removal of Diesel Range Organics (DRO) after 10 days of treatment in an aqueous matrix contaminated with 2% (v / v) crude oil. Treatments include live fungus, cell-free enzymatic extract (ELC), biosurfactant (BS) type 1 and type 2 applied alone or in combination with ELC, and an untreated control. Bars indicate residual DRO concentration (mg / L), while the line indicates DRO removal (%). The highest removal levels were observed for live fungus (about 97%) and ELC (about 93%), followed by biosurfactant type 1-based formulations, whereas biosurfactant type 2 showed lower removal efficiency. FIG. 9D depicts the visual appearance of an aqueous matrix contaminated with crude oil treated with live fungus at day 0 (left image) and after 10 days (right image), showing progressive emulsification and reduction of the oil phase.FIG. 9E depicts the visual appearance of an aqueous matrix contaminated with crude oil treated with biosurfactant type 1 combined with a cell-free enzymatic extract (ELC) at day 0 (left image a) and after 10 days (right image b), evidencing enhanced dispersion and homogenization of the hydrocarbon phase. FIG. 9F depicts the negative control (untreated system), in which no emulsification or hydrocarbon degradation is observed, showing persistent phase separation and no visible change after the incubation period. FIG. 9G depicts the removal of Diesel Range Organics (DRO) after 10 days of treatment in an aqueous matrix contaminated with 10% (w / w) petroleum sludge. Treatments include live fungus, biosurfactant type 1 applied alone or in combination with a cell-free enzymatic extract (ELC), biosurfactant type 2 combined with ELC, and an untreated control. Bars represent residual DRO concentration (mg / L), while the line indicates DRO removal (%). The highest removal efficiencies were observed for live fungus (about 99%) and biosurfactant type 1-based treatments (about 97%), followed by biosurfactant type 2 combined with ELC (about 94%), whereas no removal was detected in the untreated control. FIG. 9H depicts the visual appearance of an aqueous matrix contaminated with oil sludge treated with live fungus at day 0 (left image) and after 10 days (right image), showing progressive emulsification and reduction of the oil phase. FIG. 91 depicts the visual appearance of an aqueous matrix contaminated with oil sludge treated with biosurfactant type 1 combined with a cell-free enzymatic extract (ELC) at day 0 (left image a) and after 10 days (right imageWSGR Docket No.: 69849-701.601b), evidencing enhanced dispersion and homogenization of the hydrocarbon phase. FIG. 9J depicts the negative control (untreated system), in which no emulsification or hydrocarbon degradation is observed.

[0033] FIG. 10A depicts an image showing the fungal growth and biosurfactant production in a stirred-tank reactor (CSTR) at a 0.5-gallon scale (approximately 2L) in the presence of 1% crude oil as an inducer. Crude oil micelles can be seen integrating into the aqueous phase, indicating emulsification and biosurfactant activity. FIG. 10B depicts an image showing the crude oil was fully integrated into the aqueous phase after 48 hours, demonstrating biosurfactant production. FIG. 10C depicts an image showing the fermentation after 72 hours, with the crude oil fully degraded, the fungus growing in suspension, and biosurfactant formation.

[0034] FIG. 11A depicts an image showing water from fracture well (late return) contaminated with hydrocarbons (-0.22%) before treatment with the biosurfactant as described herein, obtained from oil wells in the Neuquen province, Argentina. FIG. 11B depicts an image showing water from fracture well (late return) contaminated with hydrocarbons (-0.22%) obtained from the Neuquen province, after 8 days of treatment with the biosurfactant formula. The visual change indicates a reduction in hydrocarbon content compared to the untreated sample. FIG. 11C depicts an image showing the experimental setup with 4L of fracture water from oil wells in Neuquen province, using oil-free compressors for aeration during the degradation process. FIG. 11D depicts the reduction of Total Petroleum Hydrocarbons (TPH) in fracture wastewater subjected to dilution treatments of 10%, 20%, and 50% and treated with the biosurfactant composition as described herein. Bars represent TPH concentration (mg / L) at day 0 and after 15 days of treatment, while the dashed line indicates the regulatory limit (10 mg / L). The results show TPH removals of approximately 97% for the 10% dilution, 89% for the 20% dilution, and 90% for the 50% dilution, demonstrating effective hydrocarbon reduction across all evaluated dilution levels and compliance with regulatory discharge standards.

[0035] FIG. 12A depicts an image showing hydrocarbon-contaminated soil (approximately 5.7% TPH) obtained from Neuquen province, before treatment with the biosurfactant formulation. The soil appears dark and compacted, indicating the presence of heavy hydrocarbons. FIG. 12B depicts an image showing hydrocarbon-contaminated soil (approximately 5.7% TPH) obtained from Neuquen province, after treatment with the biosurfactant formulation. The soil appears looser and with improved color, indicating enhanced physicochemical properties and reduced hydrocarbon content. FIG. 12C depicts the reduction of Total Petroleum Hydrocarbons (TPH) in two hydrocarbon-contaminated soil samples: Soil 1 (no previous treatment) and Soil 2 (diatomaceous earth pre-treatment) measured at day 0 and after 45 days of treatment with the biosurfactant formulation as described herein. Bars represent TPHWSGR Docket No.: 69849-701.601concentration (pg / g), while the dashed line indicates the applicable regulatory limit. A TPH reduction of approximately 51% was observed in Soil 1 and approximately 54% in Soil 2 after 45 days, evidencing the effectiveness of the biosurfactant treatment in promoting hydrocarbon removal in soil matrices under field-relevant conditions.

[0036] FIG. 13A depicts an image showing a first non-limiting example of filtered extract containing the biosurfactant generated without an inducer after the lyophilization process. FIG.13B depicts an image showing a second non-limiting example of filtered extract containing the biosurfactant generated with an inducer after the lyophilization process. In FIGs. 13A and 13B, the side view (top) and top view (bottom) of the containers containing the filtered extracts are shown.

[0037] FIG. 14A shows the stability of the filtered biosurfactant extracts (generated with an inducer vs. without an inducer) under low pH conditions. During the pH reduction process, no visible changes were observed in the samples' properties, such as color, suggesting stability under acidic conditions. FIG. 14B shows the stability of the filtered biosurfactant extracts (generated with an inducer vs. without an inducer) under high pH conditions.

[0038] FIG. 15 shows the biodegradability kinetics of the biosurfactant compositions as described herein (generated with an inducer vs. without an inducer).

[0039] FIGs. 16A-16B depict the emulsification properties of an example biosurfactant compared with two commercial surfactants. Emulsification tests were performed using vegetable oil as the oil phase and an aqueous phase containing either the example biosurfactant, Comparative surfactant A (synthetic), Comparative surfactant B (bio-based) or a negative control (water), each at 1% (w / v). In FIG. 16A, at 24 h (EI24), the example biosurfactant and Comparative surfactant A both exhibited an El of 100%, forming a dense, opaque emulsion layer, whereas Comparative surfactant B and the negative control showed El values ^2.5%, with almost complete phase separation. In FIG. 16B, the emulsified layer in the example biosurfactant samples remained macroscopically stable for more than 10 days at room temperature, indicating excellent emulsification capacity and long-term stability, with performance comparable to the synthetic surfactant. FIG. 16C depicts standardized washing test results obtained using a 1% (w / v) aqueous solution of an example biosurfactant formulation applied to two fabric types: 100% cotton (top panels) and a 65% polyester / 35% cotton blend (bottom panels). Images show fabric swatches stained with representative household soils (grass, chocolate drink, and red wine) before washing (“Before”) and after washing (“After”). The visual comparison illustrates effective stain removal across both fabric types while maintaining color stability in unstained controls, supporting the detergency performance of the biosurfactant formulation for laundry and Home Care applications. FIG. 16D depicts the color difference (AE,WSGR Docket No.: 69849-701.601CIELAB) measured after washing with a 1% (w / v) an example biosurfactant on two fabric types: 100% cotton and a 65% polyester / 35% cotton blend. Bars correspond to blank (unstained) fabrics and fabrics stained with grass, chocolate drink, and red wine. Low AE values for blanks indicate minimal background color alteration, while markedly higher AE values for stained samples demonstrate visually evident stain removal, with particularly strong performance on chocolate and wine stains, supporting the detergency efficacy of the biosurfactant formulation.

[0040] FIG. 17A depicts fungal growth and tolerance in the presence of per- and polyfluoroalkyl substances (PFAS). The left image (a) shows the macroscopic appearance of Enotonaema liquescens mycelium after 15 days of cultivation with PFOS, PFOA, or without PFAS (growth control). The right graph (b) shows the corresponding dry biomass (mg) and percentage of growth relative to the control. Reduced but measurable biomass formation is observed in PFOS and PFOA treatments compared to the growth control, indicating growth inhibition but sustained fungal tolerance to both PFAS compounds. FIG. 17B depicts the percentage degradation of per- and polyfluoroalkyl substances (PFAS) after 15 days of incubation with a fungal culture, compared with abiotic controls. Bars represent the degradation of branched PFOS (br-PFOS), linear PFOS, and PFOA by the fungus, while abiotic controls for PFOS and PFOA show negligible degradation. The results indicate partial biodegradation of PFOS isoforms (~25 - 35%) and substantially higher biodegradation of PFOA (~65 - 70%), demonstrating that biological activity rather than abiotic processes primarily drive PFAS removal.

[0041] FIG. 18 depicts images (upper panel) of representative carbonate rock discs coated with crude oil before and after treatment with different cleaning fluids and the graph summarizing the result (lower bar graph). The upper panels show oil-wet rock surfaces prior to treatment, while the lower panels show the same rocks after exposure to (i) brine / acid control, (ii) an acid formulation containing a commercial surfactant, and (iii) an acid formulation containing example biosurfactant #1. The lower bar graph summarizes the average percentage of residual oil coverage determined by image analysis after treatment. Rocks treated with the example biosurfactant #l-containing formulation exhibit a significantly lower oil-covered surface compared to both the brine control and the commercial surfactant, demonstrating enhanced oil removal and improved rock surface cleaning performance under simulated wellbore conditions.

[0042] FIG. 19 depicts representative images of a paraffin dispersal bench test comparing brine control, a commercial paraffin dispersant, and a biosurfactant formulation (example biosurfactant #1). The image shows paired transparent vials for each treatment, illustrating theWSGR Docket No.: 69849-701.601condition of paraffin deposits before and after 60 minutes of exposure at elevated temperature. In the brine control and commercial treatments, the paraffin remains largely adhered to the vial walls as a continuous dark film. In contrast, the example biosurfactant #l-treated vial exhibits a largely transparent glass surface with minimal residual deposits, indicating effective detachment and dispersion of paraffin into the bulk fluid. The visual differences demonstrate the superior ability of example biosurfactant #1 to mobilize and disperse paraffin deposits under simulated flow-assurance conditions.DETAILED DESCRIPTIONOverview

[0043] Surfactants have varieties of uses in different industries, such as but not limited to petroleum or oil industry, agro-industrial industry, environmental industry, cosmetic industry, food industry, and pharmaceutical industry. Commonly used surfactants can comprise synthetic surfactants. However, the use of synthetic surfactants in various industrial and domestic processes may cause environmental damage associated with their production and disposal. Another environmental issue can comprise the use of nonrenewable raw materials, such as petroleum derivatives. In addition to environmental issues, most of these synthetic compounds may be potentially associated with toxicological problems owing to their recalcitrant and persistent nature. There is a need to produce safe, biodegradable, and less toxic or non-toxic surfactants than those derived from petroleum.

[0044] Natural surfactants or biosurfactants can include amphiphilic biological compounds (including but not limited to those generated extracellularly), obtained from the secondary metabolism of microorganisms such as bacteria, yeasts, and filamentous fungi, and produced by biotechnological routes using various substances, including waste materials.

[0045] Provided herein are compositions comprising biosurfactants that can address the need for safe, biodegradable, and less toxic surfactants than those derived from petroleum. The biosurfactants provided herein can be biodegradable and low toxicity.

[0046] Biosurfactants can be classified by its microbial origin, chemical composition, or molecular weight. The molecular mass can range from 500 to 1500 daltons (Da). Low-molecular-weight biosurfactants can be more efficient in reducing the surface tension at the water-air interface and the water-oil interface (e.g., glycolipids or lipopeptides), while high-molecular-weight compounds can be more effective as stabilizers of oil-in-water emulsions (polymeric and particulate surfactants, such as lipoproteins). According to their chemical composition, biosurfactants can be classified into glycolipids, lipopeptides, lipoproteins, fatty acids, phospholipids, polymeric surfactants, and particulate surfactants.WSGR Docket No.: 69849-701.601

[0047] Biosurfactants can be produced by microorganisms in response to environmental conditions and can be associated with distinct growth stages. The use of different substrate sources by microorganisms can result in different structures with altered surfactant properties. Not only the species used but also the cultivation conditions can change the type of biosurfactant and its concentration in the medium. Physicochemical parameters such as the nature of the carbon and nitrogen source (the C:N ratio), as well as temperature, pH, and aeration rate, can result in products with different compositions.

[0048] For example, biosurfactants comprising sophorolipids can be associated with yeast or genetically modified bacteria. Sophorolipids may not be common at industrial scale.

[0049] Traditionally, metabolites, particularly biosurfactants produced by bacteria or yeasts (often GMO strains), lack stability under extreme conditions of pH, temperature, and salinity. This can limit their application to sectors such as cosmetics and personal care. To address this need, the biosurfactants provided herein can exhibit various properties under extreme environmental conditions and long-term physicochemical stability.

[0050] Provided herein are methods of generating the biosurfactants. The methods can comprise fermentation processes leveraging agro-industrial residues, particularly starch-rich byproducts such as potato peels, cassava, or other rejected harvests. Thus, the methods address the need of environmental issue such as uses of nonrenewable raw materials associated with petroleum derivatives.

[0051] Biosurfactants can be cost-prohibitive to produce as compared to synthetic surfactants. For example, the cost of raw materials can account for a large portion of production costs.Biosurfactant production yields can be low, especially from low-cost substrates. The methods provided herein can use renewable sources or waste materials generated from various industries and thus address the need of lowering the cost of the production of the biosurfactants.

[0052] Yeasts or filamentous fungi can generate biosurfactants with various beneficial properties, such as those as described herein. The industrial use of yeasts or filamentous fungi can be limited due to technical production challenges, including the reliance on non-commercial, custom-designed reactors and the need for particular and costly culture media. For example, yeasts or filamentous fungus may require the uses of solid growth media or conditions, preventing it from being grown in industrial scales. Additionally, various microbes for generating biosurfactants with beneficial properties may be pathogenic strains or generate virulent factors. Furthermore, the microbes may require genetic engineering for generating biosurfactants with beneficial properties.

[0053] The methods provided herein may comprise the use of low-cost culture medium and a production process that can enable the use of industrially viable systems, such as stirred-tankWSGR Docket No.: 69849-701.601reactors. The methods can effectively generate various constituents of the biosurfactant (such as but not limited to lipids, lipopeptides, lipoproteins, saccharides, enzymes, or pigments).Additionally, the methods provided herein may not require the use of genetically engineered microbes or pathogenic microbes.

[0054] Biosurfactants can be cost-prohibitive to recover or to be purified or further processed (such as the generation of excessive foaming). The methods provided her can comprise simplified downstream process with minimal steps, eliminating the need for solvents or complex equipment or minimizing foaming. For example, because the methods provided comprise generation of various constituents of the biosurfactant extracellularly, use of solvents or intracellular extraction or methods that generate foaming can be minimized.

[0055] Accordingly, the biosurfactants, methods of uses or manufacturing thereof can address various needs presented by synthetic surfactants or other biosurfactants, facilitating the effective uses and generation or wide-application of the biosurfactants.Biosurfactants

[0056] Provided herein are biosurfactants or compositions comprising the biosurfactants (biosurfactant compositions). In some cases, the biosurfactant compositions provided herein comprise at least one molecule as described herein. In some cases, the component of the biosurfactant composition may be a chemical (organic or inorganic chemical) or a biomolecule. In some cases, the biomolecule may be a metabolite or molecule synthesized by an organism as described herein. In some cases, the biosurfactant composition may comprise at least about: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 30, 40, 50 or more distinct molecules. A molecule distinct from another molecule may have a chemical structure that is different from that of the another molecule. In some cases, the biosurfactant composition may comprise at most about: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 30, 40, or 50 distinct molecules. In some cases, the biosurfactant compositions provided herein may exhibit various properties as described herein. The molecule or the biosurfactant composition may be generated extracellularly or secreted to the extracellular space by the microorganism.

[0057] The biosurfactant composition provided herein may comprise a lipid as described herein, an enzyme as described herein, a lipopeptide as described herein, a lipoprotein as described herein, a saccharide as described herein, an additive as described herein, a pigment as described herein, a derivative thereof, or any combinations thereof. As used herein when referring to a particular molecule of a biosurfactant composition, the particular molecule may comprise the derivative of the molecule.

[0058] In some cases, the biosurfactant composition may comprise a lipid as described herein, an enzyme as described herein, a lipopeptide as described herein, a lipoprotein as describedWSGR Docket No.: 69849-701.601herein, and an additive as described herein; or the derivatives thereof. In some cases, the biosurfactant composition may comprise a lipid as described herein, an enzyme as described herein, and an additive as described herein; or the derivatives thereof.

[0059] In some cases, the biosurfactant composition may comprise at least 2, 3, 4, or 5 of the lipids, enzyme, lipopeptide, lipoprotein, saccharide, pigment or additive, or the derivatives thereof. In some cases, the biosurfactant composition may comprise a lipid or a derivative thereof. In some cases, the biosurfactant composition may comprise an enzyme or a derivative thereof. In some cases, the biosurfactant composition may comprise a lipopeptide or a derivative thereof. In some cases, the biosurfactant composition may comprise a lipoprotein or a derivative thereof. In some cases, the biosurfactant composition may comprise an additive or a derivative thereof. In some cases, the biosurfactant composition may comprise a pigment or a derivative thereof. In some cases, the biosurfactant composition may comprise a saccharide or a derivative thereof.

[0060] The lipid may comprise a lipid synthesized by an organism as described herein. In some cases, the lipid may be one that is synthesized by the organism in vivo. In some cases, the lipid may be one that is synthesized in vitro. For example, the lipid may be synthesized by an enzyme (enzymatic pathway or biosynthetic pathway) of the organism.

[0061] In some cases, the biosurfactant composition may comprise a lipid. In some cases, the lipid may comprise a lipid from a fungus. In some cases, the lipid may comprise a lipid from a filamentous fungus. In some cases, the lipid may comprise a lipid from the family Xylariaceae. In some cases, the lipid may comprise a lipid from Entonaema liquescens.

[0062] In some cases, the lipid may be a glycolipid. In some cases, the glycolipid may comprise at least about: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 30, 40, 50 or more distinct glycolipids. In some cases, the glycolipid may comprise at most about: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 30, 40, or 50 distinct glycolipids.

[0063] In some cases, the glycolipid may comprise sophorolipid, trehalose lipid, rhamnolipid, mannosylerythritol lipid, cellobiose lipid, or polyol lipid, or any derivative thereof, or any combinations thereof. In some cases, the glycolipid may comprise sophorolipid, trehalose lipid, rhamnolipid, or mannosylerythritol lipid, or any derivative thereof, or any combinations thereof. In some cases, the glycolipid may comprise sophorolipid, rhamnolipid, or mannosylerythritol lipid, or any derivative thereof, or any combinations thereof. In some cases, the glycolipid may comprise sophorolipid, or rhamnolipid, or any derivative thereof, or any combinations thereof. In some cases, the glycolipid may comprise sophorolipid or a derivative thereof. In some cases, the glycolipid may comprise trehalose lipid or a derivative thereof. In some cases, the glycolipid may comprise rhamnolipid or a derivative thereof. In some cases, the glycolipid may compriseWSGR Docket No.: 69849-701.601mannosylerythritol lipid or a derivative thereof. In some cases, the glycolipid may comprise cellobiose lipid or a derivative thereof. In some cases, the glycolipid may comprise polyol lipid or a derivative thereof.

[0064] In some cases, the sophorolipid may comprise an acidic sophorolipid, a lactonic sophorolipid, a derivative thereof, or a combination thereof. In some cases, the sophorolipid may comprise an acidic sophorolipid and a lactonic sophorolipid, or a derivative thereof. In some cases, the sophorolipid may comprise an acidic sophorolipid or a derivative thereof. In some cases, the sophorolipid may comprise a lactonic sophorolipid or a derivative thereof.

[0065] In some cases, the lipid may be at least about: 0.001 %, 0.01 %, 0.1 %, 1 %, 2 %, 3 %, 4 %, 5 %, 6 %, 7 %, 8 %, 9 %, 10 %, 10 %, 11 %, 12 %, 13 %, 14 %, 15 %, 16 %, 17 %, 18 %, 19 %, 20 %, 21 %, 22 %, 23 %, 24 %, 25 %, 26 %, 27 %, 28 %, 29 %, 30 %, 31 %, 32 %, 33 %, 34 %, 35 %, 36 %, 37 %, 38 %, 39 %, 40 %, 41 %, 42 %, 43 %, 44 %, 45 %, 46 %, 47 %, 48 %, 49 %, 50 %, 51 %, 52 %, 53 %, 54 %, 55 %, 56 %, 57 %, 58 %, 59 %, 60 %, 61 %, 62 %, 63 %, 64 %, 65 %, 66 %, 67 %, 68 %, 69 %, 70 %, 71 %, 72 %, 73 %, 74 %, 75 %, 76 %, 77 %, 78 %, 79 %, 80 %, 81 %, 82 %, 83 %, 84 %, 85 %, 86 %, 87 %, 88 %, 89 %, 90 % or more, by weight / volume, of the biosurfactant composition. In some cases, the lipid may be at most about: 0.001 %, 0.01 %, 0.1 %, 1 %, 2 %, 3 %, 4 %, 5 %, 6 %, 7 %, 8 %, 9 %, 10 %, 10 %, 11 % 12 %, 13 %, 14 %, 15 %, 16 %, 17 %, 18 %, 19 %, 20 %, 21 %, 22 %, 23 %, 24 %, 25 %, 26 %, 27 %, 28 %, 29 %, 30 %, 31 %, 32 %, 33 %, 34 %, 35 %, 36 %, 37 %, 38 %, 39 %, 40 %, 41 %, 42 %, 43 %, 44 %, 45 %, 46 %, 47 %, 48 %, 49 %, 50 %, 51 %, 52 %, 53 %, 54 %, 55 %, 56 %, 57 %, 58 %, 59 %, 60 %, 61 %, 62 %, 63 %, 64 %, 65 %, 66 %, 67 %, 68 %, 69 %, 70 %, 71 %, 72 %, 73 %, 74 %, 75 %, 76 %, 77 %, 78 %, 79 %, 80 %, 81 %, 82 %, 83 %, 84 %, 85 %, 86 %, 87 %, 88 %, 89 %, or 90 %, by weight / volume, of the biosurfactant composition. In some cases, the lipid may be about: 20 %-30 %, 19 %-31 %, 18 %-32 %, 17 %-33 %, 16 %-34 %, 15 %- 35 %, 14 %-36 %, 13 %-37 %, 12 %-38 %, 11 %-39 %, 10 %-40 %, 9 %-41 %, 8 %-42 %, 7 %■ 43 %, 6 %-44 %, 5 %-45 %, 4 %-46 %, 3 %-47 %, 2 %-48 %, 1 %-49 %, 0.1 %-50 %, 0.01 %- 50 %, or 0.001 %-50 %, by weight / volume, of the biosurfactant composition. In some cases, the lipid may be at least about: 0.001 %, 0.01 %, 0.1 %, 1 %, 2 %, 3 %, 4 %, 5 %, 6 %, 7 %, 8 %, 9 %, 10 %, 10 %, 11 %, 12 %, 13 %, 14 %, 15 %, 16 %, 17 %, 18 %, 19 %, 20 %, 21 %, 22 % 23 %, 24 %, 25 %, 26 %, 27 %, 28 %, 29 %, 30 %, 31 %, 32 %, 33 %, 34 %, 35 %, 36 %, 37 %, 38 %, 39 %, 40 %, 41 %, 42 %, 43 %, 44 %, 45 %, 46 %, 47 %, 48 %, 49 %, 50 %, 51 %, 52 %, 53 %, 54 %, 55 %, 56 %, 57 %, 58 %, 59 %, 60 %, 61 %, 62 %, 63 %, 64 %, 65 %, 66 %, 67 %, 68 %, 69 %, 70 %, 71 %, 72 %, 73 %, 74 %, 75 %, 76 %, 77 %, 78 %, 79 %, 80 %, 81 %, 82 %, 83 %, 84 %, 85 %, 86 %, 87 %, 88 %, 89 %, 90 % or more, by weight / weight, of the biosurfactant composition. In some cases, the lipid may be at most about: 0.001 %, 0.01 %,WSGR Docket No.: 69849-701.6010.1 %, 1 %, 2 %, 3 %, 4 %, 5 %, 6 %, 7 %, 8 %, 9 %, 10 %, 10 %, 11 %, 12 %, 13 %, 14 %, 15 %, 16 %, 17 %, 18 %, 19 %, 20 %, 21 %, 22 %, 23 %, 24 %, 25 %, 26 %, 27 %, 28 %, 29 %, 30 %, 31 %, 32 %, 33 %, 34 %, 35 %, 36 %, 37 %, 38 %, 39 %, 40 %, 41 %, 42 %, 43 %, 44 %, 45 %, 46 %, 47 %, 48 %, 49 %, 50 %, 51 %, 52 %, 53 %, 54 %, 55 %, 56 %, 57 %, 58 %, 59 %, 60 %, 61 %, 62 %, 63 %, 64 %, 65 %, 66 %, 67 %, 68 %, 69 %, 70 %, 71 %, 72 %, 73 %, 74 %, 75 %, 76 %, 77 %, 78 %, 79 %, 80 %, 81 %, 82 %, 83 %, 84 %, 85 %, 86 %, 87 %, 88 %, 89 %, or 90 %, by weight / weight, of the biosurfactant composition. In some cases, the lipid may be about: 20 %-30 %, 19 %-31 %, 18 %-32 %, 17 %-33 %, 16 %-34 %, 15 %-35 %, 14 %-36 %, 13 %-37 %, 12 %-38 %, 11 %-39 %, 10 %-40 %, 9 %-41 %, 8 %-42 %, 7 %-43 %, 6 %-44 %, 5 %-45 %, 4 %-46 %, 3 %-47 %, 2 %-48 %, 1 %-49 %, 0.1 %-50 %, 0.01 %-50 %, or 0.001 %-50 %, by weight / weight, of the biosurfactant composition. In some cases, the lipid may be about 20 %-30 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lipid may be about 19 %-31 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lipid may be about 18 %-32 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lipid may be about 17 %-33 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lipid may be about 16 %-34 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lipid may be about 15 %-35 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lipid may be about 14 %-36 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lipid may be about 13 %-37 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lipid may be about 12 %-38 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lipid may be about 11 %-39 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lipid may be about 10 %-40 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lipid may be about 9 %-41 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lipid may be about 8 %-42 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lipid may be about 7 %-43 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lipid may be about 6 %-44 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lipid may be about 5 %-45 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lipid may be about 4 %-46 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lipid may be about 3 %-47 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lipid may be about 2 %-WSGR Docket No.: 69849-701.60148 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lipid may be about 1 %-49 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lipid may be about 0.1 %-50 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lipid may be about 0.01 %-50 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lipid may be about 0.001 %-50 %, by weight / weight or by weight / volume, of the biosurfactant composition.

[0066] In some cases, the glycolipid may be at least about: 0.001 %, 0.01 %, 0.1 %, 1 %, 2 %, 3 %, 4 %, 5 %, 6 %, 7 %, 8 %, 9 %, 10 %, 10 %, 11 %, 12 %, 13 %, 14 %, 15 %, 16 %, 17 %, 18 %, 19 %, 20 %, 21 %, 22 %, 23 %, 24 %, 25 %, 26 %, 27 %, 28 %, 29 %, 30 %, 31 %, 32 %, 33 %, 34 %, 35 %, 36 %, 37 %, 38 %, 39 %, 40 %, 41 %, 42 %, 43 %, 44 %, 45 %, 46 %, 47 %, 48 %, 49 %, 50 %, 51 %, 52 %, 53 %, 54 %, 55 %, 56 %, 57 %, 58 %, 59 %, 60 %, 61 %, 62 %, 63 %, 64 %, 65 %, 66 %, 67 %, 68 %, 69 %, 70 %, 71 %, 72 %, 73 %, 74 %, 75 %, 76 %, 77 %.78 %, 79 %, 80 %, 81 %, 82 %, 83 %, 84 %, 85 %, 86 %, 87 %, 88 %, 89 %, 90 % or more, by weight / volume, of the biosurfactant composition. In some cases, the glycolipid may be at most about: 0.001 %, 0.01 %, 0.1 %, 1 %, 2 %, 3 %, 4 %, 5 %, 6 %, 7 %, 8 %, 9 %, 10 %, 10 %, 11 %, 12 %, 13 %, 14 %, 15 %, 16 %, 17 %, 18 %, 19 %, 20 %, 21 %, 22 %, 23 %, 24 %, 25 % 26 %, 27 %, 28 %, 29 %, 30 %, 31 %, 32 %, 33 %, 34 %, 35 %, 36 %, 37 %, 38 %, 39 %, 40 %, 41 %, 42 %, 43 %, 44 %, 45 %, 46 %, 47 %, 48 %, 49 %, 50 %, 51 %, 52 %, 53 %, 54 %, 55 %, 56 %, 57 %, 58 %, 59 %, 60 %, 61 %, 62 %, 63 %, 64 %, 65 %, 66 %, 67 %, 68 %, 69 %, 70 %, 71 %, 72 %, 73 %, 74 %, 75 %, 76 %, 77 %, 78 %, 79 %, 80 %, 81 %, 82 %, 83 %, 84 %, 85 %, 86 %, 87 %, 88 %, 89 %, or 90 %, by weight / volume, of the biosurfactant composition. In some cases, the glycolipid may be about: 20 %-30 %, 19 %-31 %, 18 %-32 %, 17 %-33 %, 16 %- 34 %, 15 %-35 %, 14 %-36 %, 13 %-37 %, 12 %-38 %, 11 %-39 %, 10 %-40 %, 9 %-41 %, 8 %-42 %, 7 %-43 %, 6 %-44 %, 5 %-45 %, 4 %-46 %, 3 %-47 %, 2 %-48 %, 1 %-49 %, 0.1 %- 50 %, 0.01 %-50 %, or 0.001 %-50 %, by weight / volume, of the biosurfactant composition. In some cases, the glycolipid may be at least about: 0.001 %, 0.01 %, 0.1 %, 1 %, 2 %, 3 %, 4 %, 5 %, 6 %, 7 %, 8 %, 9 %, 10 %, 10 %, 11 %, 12 %, 13 %, 14 %, 15 %, 16 %, 17 %, 18 %, 19 %, 20 %, 21 %, 22 %, 23 %, 24 %, 25 %, 26 %, 27 %, 28 %, 29 %, 30 %, 31 %, 32 %, 33 %, 34 %, 35 %, 36 %, 37 %, 38 %, 39 %, 40 %, 41 %, 42 %, 43 %, 44 %, 45 %, 46 %, 47 %, 48 %, 49 %, 50 %, 51 %, 52 %, 53 %, 54 %, 55 %, 56 %, 57 %, 58 %, 59 %, 60 %, 61 %, 62 %, 63 %, 64 %, 65 %, 66 %, 67 %, 68 %, 69 %, 70 %, 71 %, 72 %, 73 %, 74 %, 75 %, 76 %, 77 %, 78 %, 79 % 80 %, 81 %, 82 %, 83 %, 84 %, 85 %, 86 %, 87 %, 88 %, 89 %, 90 % or more, by weight / weight, of the biosurfactant composition. In some cases, the glycolipid may be at most about: 0.001 %, 0.01 %, 0.1 %, 1 %, 2 %, 3 %, 4 %, 5 %, 6 %, 7 %, 8 %, 9 %, 10 %, 10 %,WSGR Docket No.: 69849-701.60111 %, 12 %, 13 %, 14 %, 15 %, 16 %, 17 %, 18 %, 19 %, 20 %, 21 %, 22 %, 23 %, 24 %, 25 %, 26 %, 27 %, 28 %, 29 %, 30 %, 31 %, 32 %, 33 %, 34 %, 35 %, 36 %, 37 %, 38 %, 39 %, 40 %, 41 %, 42 %, 43 %, 44 %, 45 %, 46 %, 47 %, 48 %, 49 %, 50 %, 51 %, 52 %, 53 %, 54 %, 55 %, 56 %, 57 %, 58 %, 59 %, 60 %, 61 %, 62 %, 63 %, 64 %, 65 %, 66 %, 67 %, 68 %, 69 %, 70 %, 71 %, 72 %, 73 %, 74 %, 75 %, 76 %, 77 %, 78 %, 79 %, 80 %, 81 %, 82 %, 83 %, 84 %, 85 %, 86 %, 87 %, 88 %, 89 %, or 90 %, by weight / weight, of the biosurfactant composition. In some cases, the glycolipid may be about: 20 %-30 %, 19 %-31 %, 18 %-32 %, 17 %-33 %, 16 %-34 %, 15 %-35 %, 14 %-36 %, 13 %-37 %, 12 %-38 %, 11 %-39 %, 10 %-40 %, 9 %-41 %, 8 %-42 %, 7 %-43 %, 6 %-44 %, 5 %-45 %, 4 %-46 %, 3 %-47 %, 2 %-48 %, 1 %-49 %, 0.1 %-50 %, 0.01 %-50 %, or 0.001 %-50 %, by weight / weight, of the biosurfactant composition. In some cases, the glycolipid may be about 20 %-30 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the glycolipid may be about 19 %-31 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the glycolipid may be about 18 %-32 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the glycolipid may be about 17 %-33 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the glycolipid may be about 16 %-34 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the glycolipid may be about 15 %-35 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the glycolipid may be about 14 %-36 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the glycolipid may be about 13 %-37 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the glycolipid may be about 12 %-38 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the glycolipid may be about 11 %-39 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the glycolipid may be about 10 %-40 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the glycolipid may be about 9 %-41 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the glycolipid may be about 8 %-42 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the glycolipid may be about 7 %-43 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the glycolipid may be about 6 %-44 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the glycolipid may be about 5 %-45 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the glycolipid may be about 4 %-46 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the glycolipid may be about 3 %-47 %, by weight / weight or by weight / volume, of the biosurfactantWSGR Docket No.: 69849-701.601composition. In some cases, the glycolipid may be about 2 %-48 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the glycolipid may be about 1 %-49 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the glycolipid may be about 0.1 %-50 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the glycolipid may be about 0.01 %-50 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the glycolipid may be about 0.001 %-50 %, by weight / weight or by weight / volume, of the biosurfactant composition.

[0067] In some cases, the sophorolipid may be at least about: 0.001 %, 0.01 %, 0.1 %, 1 %, 2 %, 3 %, 4 %, 5 %, 6 %, 7 %, 8 %, 9 %, 10 %, 10 %, 11 %, 12 %, 13 %, 14 %, 15 %, 16 %, 17 %, 18 %, 19 %, 20 %, 21 %, 22 %, 23 %, 24 %, 25 %, 26 %, 27 %, 28 %, 29 %, 30 %, 31 %, 32 %, 33 %, 34 %, 35 %, 36 %, 37 %, 38 %, 39 %, 40 %, 41 %, 42 %, 43 %, 44 %, 45 %, 46 %, 47 %, 48 %, 49 %, 50 %, 51 %, 52 %, 53 %, 54 %, 55 %, 56 %, 57 %, 58 %, 59 %, 60 %, 61 %, 62 %, 63 %, 64 %, 65 %, 66 %, 67 %, 68 %, 69 %, 70 %, 71 %, 72 %, 73 %, 74 %, 75 %, 76 %, 77 %, 78 %, 79 %, 80 %, 81 %, 82 %, 83 %, 84 %, 85 %, 86 %, 87 %, 88 %, 89 %, 90 % or more, by weight / volume, of the biosurfactant composition. In some cases, the sophorolipid may be at most about: 0.001 %, 0.01 %, 0.1 %, 1 %, 2 %, 3 %, 4 %, 5 %, 6 %, 7 %, 8 %, 9 %, 10 %, 10 %, 11 %, 12 %, 13 %, 14 %, 15 %, 16 %, 17 %, 18 %, 19 %, 20 %, 21 %, 22 %, 23 %, 24 % 25 %, 26 %, 27 %, 28 %, 29 %, 30 %, 31 %, 32 %, 33 %, 34 %, 35 %, 36 %, 37 %, 38 %, 39 %, 40 %, 41 %, 42 %, 43 %, 44 %, 45 %, 46 %, 47 %, 48 %, 49 %, 50 %, 51 %, 52 %, 53 %, 54 %, 55 %, 56 %, 57 %, 58 %, 59 %, 60 %, 61 %, 62 %, 63 %, 64 %, 65 %, 66 %, 67 %, 68 %, 69 %, 70 %, 71 %, 72 %, 73 %, 74 %, 75 %, 76 %, 77 %, 78 %, 79 %, 80 %, 81 %, 82 %, 83 %, 84 %, 85 %, 86 %, 87 %, 88 %, 89 %, or 90 %, by weight / volume, of the biosurfactant composition. In some cases, the sophorolipid may be about: 20 %-30 %, 19 %-31 %, 18 %-32 %, 17 %-33 %, 16 %-34 %, 15 %-35 %, 14 %-36 %, 13 %-37 %, 12 %-38 %, 11 %-39 %, 10 %-40 %, 9 %■ 41 %, 8 %-42 %, 7 %-43 %, 6 %-44 %, 5 %-45 %, 4 %-46 %, 3 %-47 %, 2 %-48 %, 1 %-49 %, 0.1 %-50 %, 0.01 %-50 %, or 0.001 %-50 %, by weight / volume, of the biosurfactant composition. In some cases, the sophorolipid may be at least about: 0.001 %, 0.01 %, 0.1 %, 1 %, 2 %, 3 %, 4 %, 5 %, 6 %, 7 %, 8 %, 9 %, 10 %, 10 %, 11 %, 12 %, 13 %, 14 %, 15 %, 16 %, 17 %, 18 %, 19 %, 20 %, 21 %, 22 %, 23 %, 24 %, 25 %, 26 %, 27 %, 28 %, 29 %, 30 %, 31 %, 32 %, 33 %, 34 %, 35 %, 36 %, 37 %, 38 %, 39 %, 40 %, 41 %, 42 %, 43 %, 44 %, 45 %, 46 %, 47 %, 48 %, 49 %, 50 %, 51 %, 52 %, 53 %, 54 %, 55 %, 56 %, 57 %, 58 %, 59 %, 60 %, 61 %, 62 %, 63 %, 64 %, 65 %, 66 %, 67 %, 68 %, 69 %, 70 %, 71 %, 72 %, 73 %, 74 %, 75 %, 76 %, 77 %, 78 %, 79 %, 80 %, 81 %, 82 %, 83 %, 84 %, 85 %, 86 %, 87 %, 88 %, 89 %, 90 % or more, by weight / weight, of the biosurfactant composition. In some cases, the sophorolipidWSGR Docket No.: 69849-701.601may be at most about: 0.001 %, 0.01 %, 0.1 %, 1 %, 2 %, 3 %, 4 %, 5 %, 6 %, 7 %, 8 %, 9 %, 10 %, 10 %, 11 %, 12 %, 13 %, 14 %, 15 %, 16 %, 17 %, 18 %, 19 %, 20 %, 21 %, 22 %, 23 %, 24 %, 25 %, 26 %, 27 %, 28 %, 29 %, 30 %, 31 %, 32 %, 33 %, 34 %, 35 %, 36 %, 37 %, 38 %, 39 %, 40 %, 41 %, 42 %, 43 %, 44 %, 45 %, 46 %, 47 %, 48 %, 49 %, 50 %, 51 %, 52 %, 53 %, 54 %, 55 %, 56 %, 57 %, 58 %, 59 %, 60 %, 61 %, 62 %, 63 %, 64 %, 65 %, 66 %, 67 %, 68 %, 69 %, 70 %, 71 %, 72 %, 73 %, 74 %, 75 %, 76 %, 77 %, 78 %, 79 %, 80 %, 81 %, 82 %, 83 %, 84 %, 85 %, 86 %, 87 %, 88 %, 89 %, or 90 %, by weight / weight, of the biosurfactant composition. In some cases, the sophorolipid may be about: 20 %-30 %, 19 %-31 %, 18 %-32 %, 17 %-33 %, 16 %-34 %, 15 %-35 %, 14 %-36 %, 13 %-37 %, 12 %-38 %, 11 %-39 %, 10 %-40 %, 9 %-41 %, 8 %-42 %, 7 %-43 %, 6 %-44 %, 5 %-45 %, 4 %-46 %, 3 %-47 %, 2 %-48 %, 1 %-49 %, 0.1 %-50 %, 0.01 %-50 %, or 0.001 %-50 %, by weight / weight, of the biosurfactant composition. In some cases, the sophorolipid may be about 20 %-30 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the sophorolipid may be about 19 %-31 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the sophorolipid may be about 18 %-32 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the sophorolipid may be about 17 %-33 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the sophorolipid may be about 16 %-34 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the sophorolipid may be about 15 %-35 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the sophorolipid may be about 14 %-36 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the sophorolipid may be about 13 %-37 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the sophorolipid may be about 12 %-38 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the sophorolipid may be about 11 %-39 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the sophorolipid may be about 10 %-40 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the sophorolipid may be about 9 %-41 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the sophorolipid may be about 8 %-42 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the sophorolipid may be about 7 %-43 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the sophorolipid may be about 6 %-44 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the sophorolipid may be about 5 %-45 %, by weight / weight or by weight / volume, of theWSGR Docket No.: 69849-701.601biosurfactant composition. In some cases, the sophorolipid may be about 4 %-46 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the sophorolipid may be about 3 %-47 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the sophorolipid may be about 2 %-48 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the sophorolipid may be about 1 %-49 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the sophorolipid may be about 0.1 %-50 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the sophorolipid may be about 0.01 %-50 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the sophorolipid may be about 0.001 %-50 %, by weight / weight or by weight / volume, of the biosurfactant composition.

[0068] In some cases, the acidic sophorolipid may be at least about: 0.001 %, 0.01 %, 0.1 %, 1 %, 2 %, 3 %, 4 %, 5 %, 6 %, 7 %, 8 %, 9 %, 10 %, 10 %, 11 %, 12 %, 13 %, 14 %, 15 %, 16 %, 17 %, 18 %, 19 %, 20 %, 21 %, 22 %, 23 %, 24 %, 25 %, 26 %, 27 %, 28 %, 29 %, 30 %, 31 %, 32 %, 33 %, 34 %, 35 %, 36 %, 37 %, 38 %, 39 %, 40 %, 41 %, 42 %, 43 %, 44 %, 45 %, 46 %, 47 %, 48 %, 49 %, 50 %, 51 %, 52 %, 53 %, 54 %, 55 %, 56 %, 57 %, 58 %, 59 %, 60 %, 61 %, 62 %, 63 %, 64 %, 65 %, 66 %, 67 %, 68 %, 69 %, 70 %, 71 %, 72 %, 73 %, 74 %, 75 %, 76 %, 77 %, 78 %, 79 %, 80 %, 81 %, 82 %, 83 %, 84 %, 85 %, 86 %, 87 %, 88 %, 89 %, 90 % or more, by weight / volume, of the biosurfactant composition. In some cases, the acidic sophorolipid may be at most about: 0.001 %, 0.01 %, 0.1 %, 1 %, 2 %, 3 %, 4 %, 5 %, 6 %, 7 %, 8 %, 9 %, 10 %, 10 %, 11 %, 12 %, 13 %, 14 %, 15 %, 16 %, 17 %, 18 %, 19 %, 20 %, 21 %, 22 %, 23 %, 24 %, 25 %, 26 %, 27 %, 28 %, 29 %, 30 %, 31 %, 32 %, 33 %, 34 %, 35 %, 36 %, 37 %, 38 %, 39 %, 40 %, 41 %, 42 %, 43 %, 44 %, 45 %, 46 %, 47 %, 48 %, 49 %, 50 %, 51 %, 52 %, 53 %, 54 %, 55 %, 56 %, 57 %, 58 %, 59 %, 60 %, 61 %, 62 %, 63 %, 64 %, 65 %, 66 %, 67 %, 68 %, 69 %, 70 %, 71 %, 72 %, 73 %, 74 %, 75 %, 76 %, 77 %, 78 %, 79 %, 80 %, 81 %, 82 %, 83 %, 84 %, 85 %, 86 %, 87 %, 88 %, 89 %, or 90 %, by weight / volume, of the biosurfactant composition. In some cases, the acidic sophorolipid may be about: 20 %-30 %, 19 %-31 %, 18 %-32 %, 17 %-33 %, 16 %-34 %, 15 %-35 %, 14 %-36 %, 13 %-37 %, 12 %■ 38 %, 11 %-39 %, 10 %-40 %, 9 %-41 %, 8 %-42 %, 7 %-43 %, 6 %-44 %, 5 %-45 %, 4 %- 46 %, 3 %-47 %, 2 %-48 %, 1 %-49 %, 0.1 %-50 %, 0.01 %-50 %, or 0.001 %-50 %, by weight / volume, of the biosurfactant composition. In some cases, the acidic sophorolipid may be at least about: 0.001 %, 0.01 %, 0.1 %, 1 %, 2 %, 3 %, 4 %, 5 %, 6 %, 7 %, 8 %, 9 %, 10 %, 10 %, 11 %, 12 %, 13 %, 14 %, 15 %, 16 %, 17 %, 18 %, 19 %, 20 %, 21 %, 22 %, 23 %, 24 % 25 %, 26 %, 27 %, 28 %, 29 %, 30 %, 31 %, 32 %, 33 %, 34 %, 35 %, 36 %, 37 %, 38 %, 39 %, 40 %, 41 %, 42 %, 43 %, 44 %, 45 %, 46 %, 47 %, 48 %, 49 %, 50 %, 51 %, 52 %, 53 %, 54 %,WSGR Docket No.: 69849-701.60155 %, 56 %, 57 %, 58 %, 59 %, 60 %, 61 %, 62 %, 63 %, 64 %, 65 %, 66 %, 67 %, 68 %, 69 %, 70 %, 71 %, 72 %, 73 %, 74 %, 75 %, 76 %, 77 %, 78 %, 79 %, 80 %, 81 %, 82 %, 83 %, 84 %, 85 %, 86 %, 87 %, 88 %, 89 %, 90 % or more, by weight / weight, of the biosurfactant composition. In some cases, the acidic sophorolipid may be at most about: 0.001 %, 0.01 %, 0.1 %, 1 %, 2 %, 3 %, 4 %, 5 %, 6 %, 7 %, 8 %, 9 %, 10 %, 10 %, 11 %, 12 %, 13 %, 14 %, 15 %, 16 %, 17 %, 18 %, 19 %, 20 %, 21 %, 22 %, 23 %, 24 %, 25 %, 26 %, 27 %, 28 %, 29 %, 30 %, 31 %, 32 %, 33 %, 34 %, 35 %, 36 %, 37 %, 38 %, 39 %, 40 %, 41 %, 42 %, 43 %, 44 %, 45 %, 46 %, 47 %, 48 %, 49 %, 50 %, 51 %, 52 %, 53 %, 54 %, 55 %, 56 %, 57 %, 58 %, 59 %, 60 %, 61 %, 62 %, 63 %, 64 %, 65 %, 66 %, 67 %, 68 %, 69 %, 70 %, 71 %, 72 %, 73 %, 74 %, 75 %, 76 %, 77 %, 78 %, 79 %, 80 %, 81 %, 82 %, 83 %, 84 %, 85 %, 86 %, 87 %, 88 %, 89 %, or 90 %, by weight / weight, of the biosurfactant composition. In some cases, the acidic sophorolipid may be about: 20 %-30 %, 19 %-31 %, 18 %-32 %, 17 %-33 %, 16 %-34 %, 15 %-35 %, 14 %-36 %, 13 %-37 %, 12 %-38 %, 11 %-39 %, 10 %-40 %, 9 %-41 %, 8 %-42 %, 7 %-43 %, 6 %-44 %, 5 %-45 %, 4 %-46 %, 3 %-47 %, 2 %-48 %, 1 %-49 %, 0.1 %-50 %, 0.01 %-50 %, or 0.001 %-50 %, by weight / weight, of the biosurfactant composition. In some cases, the acidic sophorolipid may be about 20 %-30 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the acidic sophorolipid may be about 19 %-31 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the acidic sophorolipid may be about 18 %-32 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the acidic sophorolipid may be about 17 %-33 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the acidic sophorolipid may be about 16 %-34 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the acidic sophorolipid may be about 15 %-35 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the acidic sophorolipid may be about 14 %-36 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the acidic sophorolipid may be about 13 %-37 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the acidic sophorolipid may be about 12 %-38 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the acidic sophorolipid may be about 11 %-39 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the acidic sophorolipid may be about 10 %-40 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the acidic sophorolipid may be about 9 %-41 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the acidic sophorolipid may be about 8 %-42 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the acidic sophorolipid may be about 7 %-43 %, byWSGR Docket No.: 69849-701.601weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the acidic sophorolipid may be about 6 %-44 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the acidic sophorolipid may be about 5 %-45 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the acidic sophorolipid may be about 4 %-46 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the acidic sophorolipid may be about 3 %-47 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the acidic sophorolipid may be about 2 %-48 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the acidic sophorolipid may be about 1 %-49 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the acidic sophorolipid may be about 0.1 %-50 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the acidic sophorolipid may be about 0.01 %-50 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the acidic sophorolipid may be about 0.001 %-50 %, by weight / weight or by weight / volume, of the biosurfactant composition.

[0069] In some cases, the lactonic sophorolipid may be at least about: 0.001 %, 0.01 %, 0.1 %, 1 %, 2 %, 3 %, 4 %, 5 %, 6 %, 7 %, 8 %, 9 %, 10 %, 10 %, 11 %, 12 %, 13 %, 14 %, 15 %, 16 %, 17 %, 18 %, 19 %, 20 %, 21 %, 22 %, 23 %, 24 %, 25 %, 26 %, 27 %, 28 %, 29 %, 30 %, 31 %, 32 %, 33 %, 34 %, 35 %, 36 %, 37 %, 38 %, 39 %, 40 %, 41 %, 42 %, 43 %, 44 %, 45 %, 46 %, 47 %, 48 %, 49 %, 50 %, 51 %, 52 %, 53 %, 54 %, 55 %, 56 %, 57 %, 58 %, 59 %, 60 %, 61 %, 62 %, 63 %, 64 %, 65 %, 66 %, 67 %, 68 %, 69 %, 70 %, 71 %, 72 %, 73 %, 74 %, 75 %, 76 %, 77 %, 78 %, 79 %, 80 %, 81 %, 82 %, 83 %, 84 %, 85 %, 86 %, 87 %, 88 %, 89 %, 90 % or more, by weight / volume, of the biosurfactant composition. In some cases, the lactonic sophorolipid may be at most about: 0.001 %, 0.01 %, 0.1 %, 1 %, 2 %, 3 %, 4 %, 5 %, 6 %, 7 %, 8 %, 9 %, 10 %, 10 %, 11 %, 12 %, 13 %, 14 %, 15 %, 16 %, 17 %, 18 %, 19 %, 20 %, 21 %, 22 %, 23 %, 24 %, 25 %, 26 %, 27 %, 28 %, 29 %, 30 %, 31 %, 32 %, 33 %, 34 %, 35 %, 36 %, 37 %, 38 %, 39 %, 40 %, 41 %, 42 %, 43 %, 44 %, 45 %, 46 %, 47 %, 48 %, 49 %, 50 %, 51 %, 52 %, 53 %, 54 %, 55 %, 56 %, 57 %, 58 %, 59 %, 60 %, 61 %, 62 %, 63 %, 64 %, 65 %, 66 %, 67 %, 68 %, 69 %, 70 %, 71 %, 72 %, 73 %, 74 %, 75 %, 76 %, 77 %, 78 %, 79 %, 80 %, 81 %, 82 %, 83 %, 84 %, 85 %, 86 %, 87 %, 88 %, 89 %, or 90 %, by weight / volume, of the biosurfactant composition. In some cases, the lactonic sophorolipid may be about: 20 %-30 %, 19 %-31 %, 18 %-32 %, 17 %-33 %, 16 %-34 %, 15 %-35 %, 14 %-36 %, 13 %-37 %, 12 %■ 38 %, 11 %-39 %, 10 %-40 %, 9 %-41 %, 8 %-42 %, 7 %-43 %, 6 %-44 %, 5 %-45 %, 4 %- 46 %, 3 %-47 %, 2 %-48 %, 1 %-49 %, 0.1 %-50 %, 0.01 %-50 %, or 0.001 %-50 %, by weight / volume, of the biosurfactant composition. In some cases, the lactonic sophorolipid mayWSGR Docket No.: 69849-701.601be at least about: 0.001 %, 0.01 %, 0.1 %, 1 %, 2 %, 3 %, 4 %, 5 %, 6 %, 7 %, 8 %, 9 %, 10 %, 10 %, 11 %, 12 %, 13 %, 14 %, 15 %, 16 %, 17 %, 18 %, 19 %, 20 %, 21 %, 22 %, 23 %, 24 %, 25 %, 26 %, 27 %, 28 %, 29 %, 30 %, 31 %, 32 %, 33 %, 34 %, 35 %, 36 %, 37 %, 38 %, 39 %, 40 %, 41 %, 42 %, 43 %, 44 %, 45 %, 46 %, 47 %, 48 %, 49 %, 50 %, 51 %, 52 %, 53 %, 54 %, 55 %, 56 %, 57 %, 58 %, 59 %, 60 %, 61 %, 62 %, 63 %, 64 %, 65 %, 66 %, 67 %, 68 %, 69 %, 70 %, 71 %, 72 %, 73 %, 74 %, 75 %, 76 %, 77 %, 78 %, 79 %, 80 %, 81 %, 82 %, 83 %, 84 %, 85 %, 86 %, 87 %, 88 %, 89 %, 90 % or more, by weight / weight, of the biosurfactant composition. In some cases, the lactonic sophorolipid may be at most about: 0.001 %, 0.01 %, 0.1 %, 1 %, 2 %, 3 %, 4 %, 5 %, 6 %, 7 %, 8 %, 9 %, 10 %, 10 %, 11 %, 12 %, 13 %, 14 %, 15 %, 16 %, 17 %, 18 %, 19 %, 20 %, 21 %, 22 %, 23 %, 24 %, 25 %, 26 %, 27 %, 28 %, 29 %, 30 %, 31 %, 32 %, 33 %, 34 %, 35 %, 36 %, 37 %, 38 %, 39 %, 40 %, 41 %, 42 %, 43 %, 44 %, 45 %, 46 %, 47 %, 48 %, 49 %, 50 %, 51 %, 52 %, 53 %, 54 %, 55 %, 56 %, 57 %, 58 %, 59 %, 60 %, 61 %, 62 %, 63 %, 64 %, 65 %, 66 %, 67 %, 68 %, 69 %, 70 %, 71 %, 72 %, 73 %, 74 %, 75 %, 76 %, 77 %, 78 %, 79 %, 80 %, 81 %, 82 %, 83 %, 84 %, 85 %, 86 %, 87 %, 88 %, 89 %, or 90 %, by weight / weight, of the biosurfactant composition. In some cases, the lactonic sophorolipid may be about: 20 %-30 %, 19 %-31 %, 18 %-32 %, 17 %-33 %, 16 %-34 %, 15 %-35 %, 14 %-36 %, 13 %-37 %, 12 %-38 %, 11 %-39 %, 10 %-40 %, 9 %-41 %, 8 %-42 %, 7 %-43 %, 6 %-44 %, 5 %-45 %, 4 %-46 %, 3 %-47 %, 2 %-48 %, 1 %-49 %, 0.1 %-50 %, 0.01 %-50 %, or 0.001 %-50 %, by weight / weight, of the biosurfactant composition. In some cases, the lactonic sophorolipid may be about 20 %-30 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lactonic sophorolipid may be about 19 %-31 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lactonic sophorolipid may be about 18 %-32 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lactonic sophorolipid may be about 17 %-33 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lactonic sophorolipid may be about 16 %-34 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lactonic sophorolipid may be about 15 %-35 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lactonic sophorolipid may be about 14 %-36 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lactonic sophorolipid may be about 13 %-37 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lactonic sophorolipid may be about 12 %-38 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lactonic sophorolipid may be about 11 %-39 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lactonic sophorolipid may be about 10 %-40 %, by weight / weight or by weight / volume, of theWSGR Docket No.: 69849-701.601biosurfactant composition. In some cases, the lactonic sophorolipid may be about 9 %-41 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lactonic sophorolipid may be about 8 %-42 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lactonic sophorolipid may be about 7 %-43 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lactonic sophorolipid may be about 6 %-44 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lactonic sophorolipid may be about 5 %-45 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lactonic sophorolipid may be about 4 %-46 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lactonic sophorolipid may be about 3 %-47 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lactonic sophorolipid may be about 2 %-48 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lactonic sophorolipid may be about 1 %-49 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lactonic sophorolipid may be about 0.1 %-50 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lactonic sophorolipid may be about 0.01 %-50 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lactonic sophorolipid may be about 0.001 %-50 %, by weight / weight or by weight / volume, of the biosurfactant composition.

[0070] In some cases, the lipid, glycolipid, or sophorolipid may acetylated. In some cases, the lipid, glycolipid, or sophorolipid may comprise at least: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or more acetyl groups. In some cases, the lipid, glycolipid, or sophorolipid may comprise at most about: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or acetyl groups. In some cases, the lipid, glycolipid, or sophorolipid may comprise 1 acetyl group. In some cases, the lipid, glycolipid, or sophorolipid may comprise 2 acetyl groups. In some cases, the lipid, glycolipid, or sophorolipid may comprise 3 acetyl groups. In some cases, the lipid, glycolipid, or sophorolipid may comprise 4 acetyl groups. In some cases, the lipid, glycolipid, or sophorolipid may comprise 5 acetyl groups. In some cases, the lipid, glycolipid, or sophorolipid may comprise 6 acetyl groups. In some cases, the lipid, glycolipid, or sophorolipid may comprise 7 acetyl groups. In some cases, the lipid, glycolipid, or sophorolipid may comprise 8 acetyl groups. In some cases, the lipid, glycolipid, or sophorolipid may comprise 9 acetyl groups. In some cases, the lipid, glycolipid, or sophorolipid may comprise 10 acetyl groups. In some cases, the lipid, glycolipid, or sophorolipid may comprise 11 acetyl groups. In some cases, the lipid, glycolipid, or sophorolipid may comprise 12 acetyl groups. In some cases, the lipid, glycolipid, or sophorolipid may comprise 13 acetyl groups. In some cases, the lipid,WSGR Docket No.: 69849-701.601glycolipid, or sophorolipid may comprise 14 acetyl groups. In some cases, the lipid, glycolipid, or sophorolipid may comprise 15 acetyl groups. In some cases, the lipid, glycolipid, or sophorolipid may comprise 16 acetyl groups. In some cases, the lipid, glycolipid, or sophorolipid may comprise 17 acetyl groups. In some cases, the lipid, glycolipid, or sophorolipid may comprise 18 acetyl groups. In some cases, the lipid, glycolipid, or sophorolipid may comprise 19 acetyl groups. In some cases, the lipid, glycolipid, or sophorolipid may comprise 20 acetyl groups.

[0071] In some embodiments, the glycolipid comprises sophorolipid. In some embodiments, the sophorolipid comprises a sophorolipidic compound of formula (I) or (II),wherein:each R1is independently selected from hydrogen, Ci.6alkyl, C2.6alkenyl, C2.6alkynyl, Ci-6 haloalkyl, -C(=O)Rn, -CH2C(=O)Rn, -C(=O)N(Rn)2, and-CH2C(=O)N(Rn)2, wherein Ci.6alkyl, C2.6 alkenyl, C2.6alkynyl, and Ci-6 haloalkyl are optionally substituted with one or more substituentseach R11is independently selected from hydrogen, Ci.6alkyl, C2.6alkenyl, and C2.6alkynyl CH3;each R2is independently selected from hydrogen, halogen, -OH, C1-6 alkyl, C2.„ alkenyl, and C2.6 alkynyl, wherein C1-6 alkyl, C2.6alkenyl, and C2.6alkynyl are optionally substituted with one or more substituents; andeach R3is independently selected from Cx-20 alkyl, Cs.2o alkenyl, and Cs.2o alkynyl wherein Cs-20 alkyl, C 8-2o alkenyl, and Cs-2o alkynyl are optionally substituted with one or more substituents.

[0072] In some embodiments, R1is hydrogen. In some embodiments, R1is Ci-6alkyl. In some embodiments, R1is C2-6alkenyl. In some embodiments, R1is C2-6alkynyl. In some embodiments, R1is -C(O)Rn. In some embodiments, R1is -CH2C(=O)Rn. In some embodiments, R1is -C(=O)N(Rn)2. In some embodiments, R1is -CH2C(=O)N(Rn)2.

[0073] In some embodiments, R11is hydrogen. In some embodiments, R11is C1-6 alkyl. In some embodiments, R11is Ci alkyl. In some embodiments, R11is C2alkyl. In some embodiments, R11isWSGR Docket No.: 69849-701.601C3alkyl. In some embodiments, R11is C4alkyl. In some embodiments, R11is C5alkyl. In some embodiments, R11is C6alkyl.

[0074] In some embodiments, R1is hydrogen. In some embodiments, R1is -C(=O)CH3. In some embodiments, R1is -C(=O)CH2CH3. In some embodiments, R1is -C(=O)CH2CH2CH3. In some embodiments, R1is -C(=O)CH2CH2CH2CH3. In some embodiments, R1is -C(=O)N(CH3)2. In some embodiments, R1is - CH2C(=O)N(CH3)2.

[0075] In some embodiments, R2is hydrogen. In some embodiments, R2is -OH. In some embodiments, R2is optionally substituted C1-5 alkyl. In some embodiments, R2is optionally substituted Ci alkyl. In some embodiments, R2is optionally substituted C2 alkyl. In some embodiments, R2is optionally substituted C3 alkyl.

[0076] In some embodiments, R3is optionally substituted Cs-20 alkyl. In some embodiments, R3is optionally substituted C5-20 alkenyl. In some embodiments, R3is optionally substituted C5-2oalkynyl. In some embodiments, R3is optionally substituted C12 alkyl. In some embodiments, R3is optionally substituted C13 alkyl. In some embodiments, R3is optionally substituted C14 alkyl. In some embodiments, R3is optionally substituted C15 alkyl. In some embodiments, R3is optionally substituted Ci6 alkyl. In some embodiments, R3is optionally substituted C17 alkyl. In some embodiments, R3is optionally substituted Cis alkyl. In some embodiments, R3is optionally substituted C19 alkyl. In some embodiments, R3is optionally substituted C20 alkyl. In some embodiments, R3is optionally substituted C12 alkenyl. In some embodiments, R3is optionally substituted C13 alkenyl. In some embodiments, R3is optionally substituted C14 alkenyl. In some embodiments, R3is optionally substituted C15 alkenyl. In some embodiments, R3is optionally substituted Ci6 alkenyl. In some embodiments, R3is optionally substituted C17 alkenyl. In some embodiments, R3is optionally substituted Cis alkenyl. In some embodiments, R3is optionally substituted C19 alkenyl. In some embodiments, R3is optionally substituted C20 alkenyl. In some embodiments, R3is optionally substituted Cnalkynyl. In some embodiments, R3is optionally substituted Cisalkynyl. In some embodiments, R3is optionally substituted Cualkynyl. In some embodiments, R3is optionally substituted Cisalkynyl. In some embodiments, R3is optionally substituted Ciealkynyl. In some embodiments, R3is optionally substituted Cnalkynyl. In some embodiments, R3is optionally substituted Cisalkynyl. In some embodiments, R3is optionally substituted Cisalkynyl. In some embodiments, R3is optionally substituted C2oalkynyl.

[0077] In some embodiments, R3is optionally substituted C12-19 alkenyl. In some embodiments, R3is optionally substituted C13 alkenyl or C15 alkenyl. In some embodiments, the optionally substituted Cs-20 alkenyl comprises one carbon-carbon double bond. In some embodiments, the optionally substituted Cs-20 alkenyl comprises two carbon-carbon double bonds.WSGR Docket No.: 69849-701.601

[0078] In some embodiments, the one or more substituents are independently selected from halogen, -OH, -NO2, =0, =S, -CN, C1-6 aminoalkyl, C1-6 alkoxy, C1-6 hydroxyalkyl, and C1-6 haloalkyl.

[0079] In some cases, a glycolipid may comprise a chemical structure ofWSGR Docket No.: 69849-701.601or any combinations thereof. In some cases, a glycolipid may comprise a chemical structure of. In some cases, a glycolipid may comprise a chemical structureWSGR Docket No.: 69849-701.601OHof . In some cases, a glycolipid may comprise a chemical O4a chemical structureof . In some cases, a glycolipid mayWSGR Docket No.: 69849-701.601OHcomprise a chemical structure of . In some cases, a glycolipidmay comprise a chemical structure of some cases,glycolipid may comprise a chemical structure ofcases, a glycolipid may comprise a chemical structure of some cases, a glycolipid may comprise a chemical structure ofWSGR Docket No.: 69849-701.601. In some cases, a glycolipid may comprise a chemicalstructure of chemical structureof . In some cases, a glycolipid mayWSGR Docket No.: 69849-701.601comprise a chemical structureof . In some cases, a glycolipidmay comprise a chemical structureof

[0080] In some cases, the glycolipid may comprise a chemical structure ofWSGR Docket No.: 69849-701.601WSGR Docket No.: 69849-701.601thereof, wherein each R is independently selected from -OH or -(C=O)-CH3. In some cases, theglycolipid may comprise a chemical structureof wherein each R is independently selected from -OH or -(C=O)-CH3. In some cases, the glycolipid may comprisea chemical structure of selected from -OH or -(C=O)-CH3. In some cases, the glycolipid may comprise a chemicalstructureof , wherein each R is independently selected from -OH or -(C=0)-CH3. In some cases, the glycolipid may comprise a chemical structure ofWSGR Docket No.: 69849-701.601, wherein each R is independently selected from -OH or -(C=O)- CH3. In some cases, the glycolipid may comprise a chemical structure of(C=0)-CH3.

[0081] In some cases, the biosurfactant composition may comprise an acetylated sophorolipid. In some cases, the biosurfactant composition may comprise a monoacetylated sophorolipid. In some cases, the biosurfactant composition may comprise a diacetylated sophorolipid. In some cases, the biosurfactant composition may comprise a lactonic diacetylated sophorolipid. In some cases, the biosurfactant composition may comprise an acidic diacetylated sophorolipid. In some cases, the sophorolipid may comprise a fatty acid chain. The fatty acid chain may comprise at least: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or more atoms. The fatty acid chain may comprise at most: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20. The fatty acid chain may comprise at least: 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or more carbon atoms. The fatty acid chain may comprise at most: 14, 15, 16, 17, 18, 19, 20, 22, 24, or 26 carbon atoms. The fatty acid chain may comprise 16 carbon atoms. The fatty acid chain may comprise 17 carbon atoms. The fatty acid chain may comprise 18 carbon atoms. The fatty acid chain may comprise 19 carbon atoms. The fatty acid chain may comprise 22 carbon atoms. The fatty acid chain may comprise at least: 1, 2, 3, 4, 5 or more double bonds. The fatty acid chain may comprise at most: 1, 2, 3, 4, 5 double bonds. The fatty acid chain may comprise one double bond. The fatty acid chain may comprise two double bonds. The fatty acid chain may comprise three double bonds. In some cases, the biosurfactant composition may comprise a C18:l sophorolipid. In some cases, the biosurfactant composition may comprise a C18:l diacetylatedWSGR Docket No.: 69849-701.601lactonic sophorolipid. In some cases, the biosurfactant composition may comprise a C18:l diacetylated acidic sophorolipid. In some cases, the sophorolipid may comprise an oleic acid chain. In some cases, the biosurfactant composition may comprise a C 16:1 sophorolipid. In some cases, the biosurfactant composition may comprise a C16:l diacetylated lactonic sophorolipid. In some cases, the biosurfactant composition may comprise a C16:l diacetylated acidic sophorolipid. In some cases, the sophorolipid may comprise a palmitoleic acid chain.

[0082] In some cases, the biosurfactant composition may comprise an enzyme. In some cases, the enzyme may comprise an enzyme from a fungus. In some cases, the enzyme may comprise an enzyme from a filamentous fungus. In some cases, the enzyme may comprise an enzyme from the family Xylariaceae. In some cases, the enzyme may comprise an enzyme from Entonaema liquescens. In some cases, the enzyme may comprise alcohol dehydrogenase, monooxygenase, dioxygenase, peroxidase, or laccase. In some cases, the enzyme may comprise alcohol dehydrogenase. In some cases, the enzyme may comprise monooxygenase. In some cases, the enzyme may comprise dioxygenases. In some cases, the enzyme may comprise peroxidase. In some cases, the enzyme may comprise laccase. In some cases, the enzyme may be at least about: 0.001 %, 0.01 %, 0.1 %, 1 %, 2 %, 3 %, 4 %, 5 %, 6 %, 7 %, 8 %, 9 %, 10 %, 10 %, 11 %, 12 %, 13 %, 14 %, 15 %, 16 %, 17 %, 18 %, 19 %, 20 %, 21 %, 22 %, 23 %, 24 %, 25 %, 26 %, 27 %, 28 %, 29 %, 30 %, 31 %, 32 %, 33 %, 34 %, 35 %, 36 %, 37 %, 38 %, 39 %, 40 %, 41 %, 42 %, 43 %, 44 %, 45 %, 46 %, 47 %, 48 %, 49 %, 50 %, 51 %, 52 %, 53 %, 54 %, 55 %, 56 %, 57 %, 58 %, 59 %, 60 %, 61 %, 62 %, 63 %, 64 %, 65 %, 66 %, 67 %, 68 %, 69 %, 70 %, 71 %, 72 %, 73 %, 74 %, 75 %, 76 %, 77 %, 78 %, 79 %, 80 %, 81 %, 82 %, 83 %, 84 %, 85 %, 86 %, 87 %, 88 %, 89 %, 90 % or more, by weight / volume, of the biosurfactant composition. In some cases, the enzyme may be at most about: 0.001 %, 0.01 %, 0.1 %, 1 %, 2 %, 3 %, 4 %, 5 %, 6 %, 7 %, 8 %, 9 %, 10 %, 10 %, 11 %, 12 %, 13 %, 14 %, 15 %, 16 %, 17 %, 18 %, 19 %, 20 %, 21 %, 22 %, 23 %, 24 %, 25 %, 26 %, 27 %, 28 %, 29 %, 30 %, 31 %, 32 %, 33 %, 34 %, 35 %, 36 %, 37 %, 38 %, 39 %, 40 %, 41 %, 42 %, 43 %, 44 %, 45 %, 46 %, 47 %, 48 %, 49 %, 50 %, 51 %, 52 %, 53 %, 54 %, 55 %, 56 %, 57 %, 58 %, 59 %, 60 %, 61 %, 62 %, 63 %, 64 %, 65 %, 66 %, 67 %, 68 %, 69 %, 70 %, 71 %, 72 %, 73 %, 74 %, 75 %, 76 %, 77 %, 78 %, 79 %, 80 %, 81 %, 82 %, 83 %, 84 %, 85 %, 86 %, 87 %, 88 %, 89 %, or 90 %, by weight / volume, of the biosurfactant composition. In some cases, the enzyme may be about: 1 %-5 %, 0.5 %-5.5 %, 0.5 %-6 %, 0.5 %-6.5 %, 0.5 %-7 %, 0.5 %-7.5 %, 0.5 %-8 %, 0.5 %-8.5 %, 0.5 %-9 %, 0.5 %-9.5 %, 0.5 %-10 %, by weight / volume, of the biosurfactant composition. In some cases, the enzyme may be at least about: 0.001 %, 0.01 %, 0.1 %, 1 %, 2 %, 3 %, 4 %, 5 %, 6 %, 7 %, 8 %, 9 %, 10 %, 10 %, 11 %, 12 %, 13 %, 14 %, 15 %, 16 %, 17 %, 18 %, 19 %, 20 %, 21 %, 22 %, 23 %, 24 %, 25 %, 26 %, 27 %, 28 %, 29 %, 30 %, 31 %, 32 %, 33 %, 34 %,WSGR Docket No.: 69849-701.60135 %, 36 %, 37 %, 38 %, 39 %, 40 %, 41 %, 42 %, 43 %, 44 %, 45 %, 46 %, 47 %, 48 %, 49 %, 50 %, 51 %, 52 %, 53 %, 54 %, 55 %, 56 %, 57 %, 58 %, 59 %, 60 %, 61 %, 62 %, 63 %, 64 %, 65 %, 66 %, 67 %, 68 %, 69 %, 70 %, 71 %, 72 %, 73 %, 74 %, 75 %, 76 %, 77 %, 78 %, 79 %.80 %, 81 %, 82 %, 83 %, 84 %, 85 %, 86 %, 87 %, 88 %, 89 %, 90 % or more, by weight / weight, of the biosurfactant composition. In some cases, the enzyme may be at most about: 0.001 %, 0.01 %, 0.1 %, 1 %, 2 %, 3 %, 4 %, 5 %, 6 %, 7 %, 8 %, 9 %, 10 %, 10 %, 11 %, 12 %, 13 %, 14 %, 15 %, 16 %, 17 %, 18 %, 19 %, 20 %, 21 %, 22 %, 23 %, 24 %, 25 % 26 %, 27 %, 28 %, 29 %, 30 %, 31 %, 32 %, 33 %, 34 %, 35 %, 36 %, 37 %, 38 %, 39 %, 40 %, 41 %, 42 %, 43 %, 44 %, 45 %, 46 %, 47 %, 48 %, 49 %, 50 %, 51 %, 52 %, 53 %, 54 %, 55 %, 56 %, 57 %, 58 %, 59 %, 60 %, 61 %, 62 %, 63 %, 64 %, 65 %, 66 %, 67 %, 68 %, 69 %, 70 %, 71 %, 72 %, 73 %, 74 %, 75 %, 76 %, 77 %, 78 %, 79 %, 80 %, 81 %, 82 %, 83 %, 84 %, 85 %, 86 %, 87 %, 88 %, 89 %, or 90 %, by weight / weight, of the biosurfactant composition. In some cases, the enzyme may be about: 1 %-5 %, 0.5 %-5.5 %, 0.5 %-6 %, 0.5 %-6.5 %, 0.5 %-7 %, 0.5 %-7.5 %, 0.5 %-8 %, 0.5 %-8.5 %, 0.5 %-9 %, 0.5 %-9.5 %, 0.5 %-10 %, by weight / weight, of the biosurfactant composition. In some cases, the enzyme may be about 1 %-5 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the enzyme may be about 0.5 %-5.5 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the enzyme may be about 0.5 %-6 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the enzyme may be about 0.5 %-6.5 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the enzyme may be about 0.5 %-7 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the enzyme may be about 0.5 %-7.5 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the enzyme may be about 0.5 %-8 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the enzyme may be about 0.5 %-8.5 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the enzyme may be about 0.5 %-9 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the enzyme may be about 0.5 %-9.5 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the enzyme may be about 0.5 %-10 %, by weight / weight or by weight / volume, of the biosurfactant composition.

[0083] In some cases, the biosurfactant composition may comprise an additive or a stabilizer. In some cases, the biosurfactant composition may comprise an additive. In some cases, the biosurfactant composition may comprise a stabilizer. In some cases, the biosurfactant composition may comprise an additive and a stabilizer. In some cases, an additive or a stabilizer of the biosurfactant composition may comprise antioxidants, preservatives, chelating agents,WSGR Docket No.: 69849-701.601natural antioxidants, vegetable oil derivatives, derivatives thereof, or any combinations thereof. In some cases, an additive or a stabilizer of the biosurfactant composition may comprise antioxidants. In some cases, an additive or a stabilizer of the biosurfactant composition may comprise preservatives. In some cases, an additive or a stabilizer of the biosurfactant composition may comprise chelating agents. In some cases, an additive or a stabilizer of the biosurfactant composition may comprise natural antioxidants. In some cases, an additive or a stabilizer of the biosurfactant composition may comprise vegetable oil derivatives. In some cases, an additive or a stabilizer of the biosurfactant composition may comprise sodium benzoate.

[0084] In some cases, the additive or stabilizer may be at least about: 0.0001 %, 0.0002 %, 0.0004 %, 0.0008 %, 0.001 %, 0.002 %, 0.003 %, 0.004 %, 0.005 %, 0.006 %, 0.007 %, 0.008 %, 0.009 %, 0.01 %, 0.011 %, 0.012 %, 0.013 %, 0.014 %, 0.015 %, 0.016 %, 0.017 %, 0.018 %, 0.019 %, 0.02 %, 0.021 %, 0.022 %, 0.023 %, 0.024 %, 0.025 %, 0.026 %, 0.027 %, 0.028 %, 0.029 %, 0.03 %, 0.031 %, 0.032 %, 0.033 %, 0.034 %, 0.035 %, 0.036 %, 0.037 %, 0.038 %, 0.039 %, 0.04 %, 0.041 %, 0.042 %, 0.043 %, 0.044 %, 0.045 %, 0.046 %, 0.047 %, 0.048 %, 0.049 %, 0.05 %, 0.055 %, 0.06 %, 0.065 %, 0.07 %, 0.075 %, 0.08 %, 0.085 %, 0.09 %, 0.095 %, 0.1 %, 0.105 %, 0.11 %, 0.115 %, 0.12 %, 0.125 %, 0.13 %, 0.135 %, 0.14 %, 0.145 %, 0.15 %, 0.155 %, 0.16 %, 0.165 %, 0.17 %, 0.175 %, 0.18 %, 0.185 %, 0.19 %, 0.195 %, 0.2 %, 0.3 %, 0.4 %, 0.5 %, 0.6 %, 0.7 %, 0.8 %, 0.9 %, 1 %, 1.1 %, 1.2 %, 1.3 %, 1.4 %, 1.5 %, 1.6 %, 1.7 %, 1.8 %, 1.9 %, 2 %, 3 %, 4 %, 5 %, 6 %, 7 %, 8 %, 9 %, 10 % or more, by weight / volume, of the biosurfactant composition. In some cases, the additive or stabilizer may be at most about: 0.0001 %, 0.0002 %, 0.0004 %, 0.0008 %, 0.001 %, 0.002 %, 0.003 %, 0.004 %, 0.005 %, 0.006 %, 0.007 %, 0.008 %, 0.009 %, 0.01 %, 0.011 %, 0.012 %, 0.013 %, 0.014 %, 0.015 %, 0.016 %, 0.017 %, 0.018 %, 0.019 %, 0.02 %, 0.021 %, 0.022 %, 0.023 %, 0.024 %, 0.025 %, 0.026 %, 0.027 %, 0.028 %, 0.029 %, 0.03 %, 0.031 %, 0.032 %, 0.033 %, 0.034 %, 0.035 %, 0.036 %, 0.037 %, 0.038 %, 0.039 %, 0.04 %, 0.041 %, 0.042 %, 0.043 %, 0.044 %, 0.045 %, 0.046 %, 0.047 %, 0.048 %, 0.049 %, 0.05 %, 0.055 %, 0.06 %, 0.065 %, 0.07 %, 0.075 %, 0.08 %, 0.085 %, 0.09 %, 0.095 %, 0.1 %, 0.105 %, 0.11 %, 0.115 %, 0.12 %, 0.125 %, 0.13 %, 0.135 %, 0.14 %, 0.145 %, 0.15 %, 0.155 %, 0.16 %, 0.165 %, 0.17 %, 0.175 %, 0.18 %, 0.185 %, 0.19 %, 0.195 %, 0.2 %, 0.3 %, 0.4 %, 0.5 %, 0.6 %, 0.7 %, 0.8 %, 0.9 %, 1 %, 1.1 %, 1.2 %, 1.3 %, 1.4 %, 1.5 %, 1.6 %, 1.7 %, 1.8 %, 1.9 %, 2 %, 3 %, 4 %, 5 %, 6 %, 7 %, 8 %, 9 %, or 10 %, by weight / volume, of the biosurfactant composition. In some cases, the additive or stabilizer may be about: 0.01 %-2 %, 0.009 %-2.25 %, 0.008 %-2.5 %, 0.007 %-2.75 %, 0.006 %-3 %, 0.005 %-3.25 %, 0.004 %-3.5 %, 0.003 %-3.75 %, 0.002 %-4 %, 0.001 %-4.25 %, 0.001 %-4.5 %, 0.001 %-4.75 %, 0.001 %-5 %,WSGR Docket No.: 69849-701.601by weight / volume, of the biosurfactant composition. In some cases, the additive or stabilizer may be at least about: 0.0001 %, 0.0002 %, 0.0004 %, 0.0008 %, 0.001 %, 0.002 %, 0.003 %, 0.004 %, 0.005 %, 0.006 %, 0.007 %, 0.008 %, 0.009 %, 0.01 %, 0.011 %, 0.012 %, 0.013 %, 0.014 %, 0.015 %, 0.016 %, 0.017 %, 0.018 %, 0.019 %, 0.02 %, 0.021 %, 0.022 %, 0.023 %, 0.024 %, 0.025 %, 0.026 %, 0.027 %, 0.028 %, 0.029 %, 0.03 %, 0.031 %, 0.032 %, 0.033 %, 0.034 %, 0.035 %, 0.036 %, 0.037 %, 0.038 %, 0.039 %, 0.04 %, 0.041 %, 0.042 %, 0.043 %, 0.044 %, 0.045 %, 0.046 %, 0.047 %, 0.048 %, 0.049 %, 0.05 %, 0.055 %, 0.06 %, 0.065 %, 0.07 %, 0.075 %, 0.08 %, 0.085 %, 0.09 %, 0.095 %, 0.1 %, 0.105 %, 0.11 %, 0.115 %, 0.12 %, 0.125 %, 0.13 %, 0.135 %, 0.14 %, 0.145 %, 0.15 %, 0.155 %, 0.16 %, 0.165 %, 0.17 % 0.175 %, 0.18 %, 0.185 %, 0.19 %, 0.195 %, 0.2 %, 0.3 %, 0.4 %, 0.5 %, 0.6 %, 0.7 %, 0.8 %, 0.9 %, 1 %, 1.1 %, 1.2 %, 1.3 %, 1.4 %, 1.5 %, 1.6 %, 1.7 %, 1.8 %, 1.9 %, 2 %, 3 %, 4 %, 5 %, 6 %, 7 %, 8 %, 9 %, 10 % or more, by weight / weight, of the biosurfactant composition. In some cases, the additive or stabilizer may be at most about: 0.0001 %, 0.0002 %, 0.0004 %, 0.0008 %, 0.001 %, 0.002 %, 0.003 %, 0.004 %, 0.005 %, 0.006 %, 0.007 %, 0.008 %, 0.009 %, 0.01 %, 0.011 %, 0.012 %, 0.013 %, 0.014 %, 0.015 %, 0.016 %, 0.017 %, 0.018 %, 0.019 %, 0.02 %, 0.021 %, 0.022 %, 0.023 %, 0.024 %, 0.025 %, 0.026 %, 0.027 %, 0.028 %, 0.029 %, 0.03 %, 0.031 %, 0.032 %, 0.033 %, 0.034 %, 0.035 %, 0.036 %, 0.037 %, 0.038 %, 0.039 %, 0.04 %, 0.041 %, 0.042 %, 0.043 %, 0.044 %, 0.045 %, 0.046 %, 0.047 %, 0.048 %, 0.049 %, 0.05 %, 0.055 %, 0.06 %, 0.065 %, 0.07 %, 0.075 %, 0.08 %, 0.085 %, 0.09 %, 0.095 %, 0.1 %, 0.105 %, 0.11 %, 0.115 %, 0.12 %, 0.125 %, 0.13 %, 0.135 %, 0.14 %, 0.145 %, 0.15 % 0.155 %, 0.16 %, 0.165 %, 0.17 %, 0.175 %, 0.18 %, 0.185 %, 0.19 %, 0.195 %, 0.2 %, 0.3 %, 0.4 %, 0.5 %, 0.6 %, 0.7 %, 0.8 %, 0.9 %, 1 %, 1.1 %, 1.2 %, 1.3 %, 1.4 %, 1.5 %, 1.6 %, 1.7 %, 1.8 %, 1.9 %, 2 %, 3 %, 4 %, 5 %, 6 %, 7 %, 8 %, 9 %, or 10 %, by weight / weight, of the biosurfactant composition. In some cases, the additive or stabilizer may be about 0.01 %-2 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the additive or stabilizer may be about 0.009 %-2.25 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the additive or stabilizer may be about 0.008 %- 2.5 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the additive or stabilizer may be about 0.007 %-2.75 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the additive or stabilizer may be about 0.006 %-3 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the additive or stabilizer may be about 0.005 %-3.25 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the additive or stabilizer may be about 0.004 %- 3.5 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the additive or stabilizer may be about 0.003 %-3.75 %, by weight / weight or by weight / volume,WSGR Docket No.: 69849-701.601of the biosurfactant composition. In some cases, the additive or stabilizer may be about 0.002 %- 4 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the additive or stabilizer may be about 0.001 %-4.25 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the additive or stabilizer may be about 0.001 %-4.5 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the additive or stabilizer may be about 0.001 %-4.75 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the additive or stabilizer may be about 0.001 %- 5 %, by weight / weight or by weight / volume, of the biosurfactant composition.

[0085] In some cases, the sodium benzoate may be at least about: 0.0001 %, 0.0002 %, 0.0004 %, 0.0008 %, 0.001 %, 0.002 %, 0.003 %, 0.004 %, 0.005 %, 0.006 %, 0.007 %, 0.008 %, 0.009 %, 0.01 %, 0.011 %, 0.012 %, 0.013 %, 0.014 %, 0.015 %, 0.016 %, 0.017 %, 0.018 %, 0.019 %, 0.02 %, 0.021 %, 0.022 %, 0.023 %, 0.024 %, 0.025 %, 0.026 %, 0.027 %, 0.028 %, 0.029 %, 0.03 %, 0.031 %, 0.032 %, 0.033 %, 0.034 %, 0.035 %, 0.036 %, 0.037 %, 0.038 %, 0.039 %, 0.04 %, 0.041 %, 0.042 %, 0.043 %, 0.044 %, 0.045 %, 0.046 %, 0.047 %, 0.048 %, 0.049 %, 0.05 %, 0.055 %, 0.06 %, 0.065 %, 0.07 %, 0.075 %, 0.08 %, 0.085 %, 0.09 %, 0.095 %, 0.1 %, 0.105 %, 0.11 %, 0.115 %, 0.12 %, 0.125 %, 0.13 %, 0.135 %, 0.14 %, 0.145 %, 0.15 %, 0.155 %, 0.16 %, 0.165 %, 0.17 %, 0.175 %, 0.18 %, 0.185 %, 0.19 %, 0.195 %, 0.2 %, 0.3 %, 0.4 %, 0.5 %, 0.6 %, 0.7 %, 0.8 %, 0.9 %, 1 %, 1.1 %, 1.2 %, 1.3 %. 1.4 %, 1.5 %, 1.6 %, 1.7 %, 1.8 %, 1.9 %, 2 %, 3 %, 4 %, 5 %, 6 %, 7 %, 8 %, 9 %, 10 % or more, by weight / volume, of the biosurfactant composition. In some cases, the sodium benzoate may be at most about: 0.0001 %, 0.0002 %, 0.0004 %, 0.0008 %, 0.001 %, 0.002 %, 0.003 %, 0.004 %, 0.005 %, 0.006 %, 0.007 %, 0.008 %, 0.009 %, 0.01 %, 0.011 %, 0.012 %, 0.013 %, 0.014 %, 0.015 %, 0.016 %, 0.017 %, 0.018 %, 0.019 %, 0.02 %, 0.021 %, 0.022 %, 0.023 %, 0.024 %, 0.025 %, 0.026 %, 0.027 %, 0.028 %, 0.029 %, 0.03 %, 0.031 %, 0.032 %, 0.033 %, 0.034 %, 0.035 %, 0.036 %, 0.037 %, 0.038 %, 0.039 %, 0.04 %, 0.041 %, 0.042 %, 0.043 %, 0.044 %, 0.045 %, 0.046 %, 0.047 %, 0.048 %, 0.049 %, 0.05 %, 0.055 %, 0.06 %, 0.065 %, 0.07 %, 0.075 %, 0.08 %, 0.085 %, 0.09 %, 0.095 %, 0.1 %, 0.105 %, 0.11 %, 0.115 %, 0.12 %, 0.125 %, 0.13 %, 0.135 %, 0.14 %, 0.145 %, 0.15 %, 0.155 %, 0.16 %, 0.165 %, 0.17 % 0.175 %, 0.18 %, 0.185 %, 0.19 %, 0.195 %, 0.2 %, 0.3 %, 0.4 %, 0.5 %, 0.6 %, 0.7 %, 0.8 %, 0.9 %, 1 %, 1.1 %, 1.2 %, 1. 3 %, 1.4 %, 1.5 %, 1.6 %, 1.7 %, 1.8 %, 1.9 %, 2 %, 3 %, 4 %, 5 %, 6 %, 7 %, 8 %, 9 %, or 10 %, by weight / volume, of the biosurfactant composition. In some cases, the sodium benzoate may be about: 0.01 %-2 %, 0.009 %-2.25 %, 0.008 %-2.5 %, 0.007 %-2.75 %, 0.006 %-3 %, 0.005 %-3.25 %, 0.004 %-3.5 %, 0.003 %-3.75 %, 0.002 %-4 %, 0.001 %-4.25 %, 0.001 %-4.5 %, 0.001 %-4.75 %, 0.001 %-5 %, by weight / volume, of the biosurfactant composition. In some cases, the sodium benzoate may be at least about: 0.0001 %,WSGR Docket No.: 69849-701.6010.0002 %, 0.0004 %, 0.0008 %, 0.001 %, 0.002 %, 0.003 %, 0.004 %, 0.005 %, 0.006 %, 0.007 %, 0.008 %, 0.009 %, 0.01 %, 0.011 %, 0.012 %, 0.013 %, 0.014 %, 0.015 %, 0.016 %, 0.017 %, 0.018 %, 0.019 %, 0.02 %, 0.021 %, 0.022 %, 0.023 %, 0.024 %, 0.025 %, 0.026 %, 0.027 %, 0.028 %, 0.029 %, 0.03 %, 0.031 %, 0.032 %, 0.033 %, 0.034 %, 0.035 %, 0.036 %, 0.037 %, 0.038 %, 0.039 %, 0.04 %, 0.041 %, 0.042 %, 0.043 %, 0.044 %, 0.045 %, 0.046 %, 0.047 %, 0.048 %, 0.049 %, 0.05 %, 0.055 %, 0.06 %, 0.065 %, 0.07 %, 0.075 %, 0.08 %, 0.085 %, 0.09 %, 0.095 %, 0.1 %, 0.105 %, 0.11 %, 0.115 %, 0.12 %, 0.125 %, 0.13 %, 0.135 %, 0.14 %, 0.145 %, 0.15 %, 0.155 %, 0.16 %, 0.165 %, 0.17 %, 0.175 %, 0.18 %, 0.185 %, 0.19 %, 0.195 %, 0.2 %, 0.3 %, 0.4 %, 0.5 %, 0.6 %, 0.7 %, 0.8 %, 0.9 %, 1 %, 1.1 %, 1.2 %, 1.3 %, 1.4 %, 1.5 %, 1.6 %, 1.7 %, 1.8 %, 1.9 %, 2 %, 3 %, 4 %, 5 %, 6 %, 7 %, 8 %, 9 %, 10 % or more, by weight / weight, of the biosurfactant composition. In some cases, the sodium benzoate may be at most about: 0.0001 %, 0.0002 %, 0.0004 %, 0.0008 %, 0.001 %, 0.002 %, 0.003 %, 0.004 %, 0.005 %, 0.006 %, 0.007 %, 0.008 %, 0.009 %, 0.01 %, 0.011 %, 0.012 %, 0.013 %, 0.014 %, 0.015 %, 0.016 %, 0.017 %, 0.018 %, 0.019 %, 0.02 %, 0.021 %, 0.022 %, 0.023 %, 0.024 %, 0.025 %, 0.026 %, 0.027 %, 0.028 %, 0.029 %, 0.03 %, 0.031 %, 0.032 %, 0.033 %, 0.034 %, 0.035 %, 0.036 %, 0.037 %, 0.038 %, 0.039 %, 0.04 %, 0.041 %, 0.042 %, 0.043 %, 0.044 %, 0.045 %, 0.046 %, 0.047 %, 0.048 %, 0.049 %, 0.05 %, 0.055 %, 0.06 %, 0.065 %, 0.07 %, 0.075 %, 0.08 %, 0.085 %, 0.09 %, 0.095 %, 0.1 %, 0.105 %, 0.11 %, 0.115 %, 0.12 %, 0.125 %, 0.13 %, 0.135 %, 0.14 %, 0.145 %, 0.15 %, 0.155 %, 0.16 %, 0.165 %, 0.17 %, 0.175 %, 0.18 %, 0.185 %, 0.19 %, 0.195 %, 0.2 %, 0.3 %, 0.4 %, 0.5 %, 0.6 %, 0.7 %, 0.8 %, 0.9 %, 1 %, 1.1 %, 1.2 %, 1.3 %, 1.4 %, 1.5 %, 1.6 %, 1.7 %, 1.8 %, 1.9 %, 2 %, 3 %, 4 %, 5 %, 6 %, 7 %, 8 %, 9 %, or 10 %, by weight / weight, of the biosurfactant composition. In some cases, the sodium benzoate may be about 0.01 %-2 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the sodium benzoate may be about 0.009 %-2.25 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the sodium benzoate may be about 0.008 %-2.5 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the sodium benzoate may be about 0.007 %-2.75 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the sodium benzoate may be about 0.006 %-3 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the sodium benzoate may be about 0.005 %-3.25 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the sodium benzoate may be about 0.004 %-3.5 %, by weight / weight or by weight / volume, of the biosurfactant. In some cases, the sodium benzoate may be about 0.003 %-3.75 %, by weight / weight or by weight / volume, of the biosurfactant. In some cases, the sodium benzoate may be about 0.002 %-4 %, by weight / weight or byWSGR Docket No.: 69849-701.601weight / volume, of the biosurfactant. In some cases, the sodium benzoate may be about 0.001 %-4.25 %, by weight / weight or by weight / volume, of the biosurfactant. In some cases, the sodium benzoate may be about 0.001 %-4.5 %, by weight / weight or by weight / volume, of the biosurfactant. In some cases, the sodium benzoate may be about 0.001 %-4.75 %, by weight / weight or by weight / volume, of the biosurfactant. In some cases, the sodium benzoate may be about 0.001 %-5 %, by weight / weight or by weight / volume, of the biosurfactant.

[0086] In some cases, the biosurfactant composition may comprise a lipopeptide. In some cases, the lipopeptide may comprise a lipopeptide from a fungus. In some cases, the lipopeptide may comprise a lipopeptide from a filamentous fungus. In some cases, the lipopeptide may comprise a lipopeptide from the family Xylariaceae. In some cases, the lipopeptide may comprise a lipopeptide from Entonaema liquescens.

[0087] In some cases, the lipopeptide may be at least about: 0.001 %, 0.01 %, 0.1 %, 1 %, 2 %, 3 %, 4 %, 5 %, 6 %, 7 %, 8 %, 9 %, 10 %, 10 %, 11 %, 12 %, 13 %, 14 %, 15 %, 16 %, 17 %, 18 %, 19 %, 20 %, 21 %, 22 %, 23 %, 24 %, 25 %, 26 %, 27 %, 28 %, 29 %, 30 %, 31 %, 32 %, 33 %, 34 %, 35 %, 36 %, 37 %, 38 %, 39 %, 40 %, 41 %, 42 %, 43 %, 44 %, 45 %, 46 %, 47 %, 48 %, 49 %, 50 %, 51 %, 52 %, 53 %, 54 %, 55 %, 56 %, 57 %, 58 %, 59 %, 60 %, 61 %, 62 %, 63 %, 64 %, 65 %, 66 %, 67 %, 68 %, 69 %, 70 %, 71 %, 72 %, 73 %, 74 %, 75 %, 76 %, 77 %, 78 %, 79 %, 80 %, 81 %, 82 %, 83 %, 84 %, 85 %, 86 %, 87 %, 88 %, 89 %, 90 % or more, by weight / volume, of the biosurfactant composition. In some cases, the lipopeptide may be at most about: 0.001 %, 0.01 %, 0.1 %, 1 %, 2 %, 3 %, 4 %, 5 %, 6 %, 7 %, 8 %, 9 %, 10 %, 10 %, 11 %, 12 %, 13 %, 14 %, 15 %, 16 %, 17 %, 18 %, 19 %, 20 %, 21 %, 22 %, 23 %, 24 %, 25 %, 26 %, 27 %, 28 %, 29 %, 30 %, 31 %, 32 %, 33 %, 34 %, 35 %, 36 %, 37 %, 38 %, 39 %, 40 %, 41 %, 42 %, 43 %, 44 %, 45 %, 46 %, 47 %, 48 %, 49 %, 50 %, 51 %, 52 %, 53 %, 54 %, 55 %, 56 %, 57 %, 58 %, 59 %, 60 %, 61 %, 62 %, 63 %, 64 %, 65 %, 66 %, 67 %, 68 %, 69 %, 70 %, 71 %, 72 %, 73 %, 74 %, 75 %, 76 %, 77 %, 78 %, 79 %, 80 %, 81 %, 82 %, 83 %, 84 %, 85 %, 86 %, 87 %, 88 %, 89 %, or 90 %, by weight / volume, of the biosurfactant composition. In some cases, the lipopeptide may be about: 10 %-15 %, 9.5 %-l 5.5 %, 9 %-16 %, 8.5 %- 16.5 %, 8 %- 17 %, 7.5 %-17.5 %, 7 %-18 %, 6.5 %-l 8.5 %, 6 %-19 %, 5.5 %-19.5 %, 5 %-20 %, 4.5 %- 20.5 %, 4 %-21 %, 3.5 %-21.5 %, 3 %-22 %, 2.5 %-22.5 %, 2 %-23 %, 1.5 %-23.5 %, 1 %- 24 %, by weight / volume, of the biosurfactant composition. In some cases, the lipopeptide may be at least about: 0.001 %, 0.01 %, 0.1 %, 1 %, 2 %, 3 %, 4 %, 5 %, 6 %, 7 %, 8 %, 9 %, 10 %, 10 %, 11 %, 12 %, 13 %, 14 %, 15 %, 16 %, 17 %, 18 %, 19 %, 20 %, 21 %, 22 %, 23 %, 24 % 25 %, 26 %, 27 %, 28 %, 29 %, 30 %, 31 %, 32 %, 33 %, 34 %, 35 %, 36 %, 37 %, 38 %, 39 %, 40 %, 41 %, 42 %, 43 %, 44 %, 45 %, 46 %, 47 %, 48 %, 49 %, 50 %, 51 %, 52 %, 53 %, 54 %, 55 %, 56 %, 57 %, 58 %, 59 %, 60 %, 61 %, 62 %, 63 %, 64 %, 65 %, 66 %, 67 %, 68 %, 69 %,WSGR Docket No.: 69849-701.60170 %, 71 %, 72 %, 73 %, 74 %, 75 %, 76 %, 77 %, 78 %, 79 %, 80 %, 81 %, 82 %, 83 %, 84 %, 85 %, 86 %, 87 %, 88 %, 89 %, 90 % or more, by weight / weight, of the biosurfactant composition. In some cases, the lipopeptide may be at most about: 0.001 %, 0.01 %, 0.1 %, 1 %, 2 %, 3 %, 4 %, 5 %, 6 %, 7 %, 8 %, 9 %, 10 %, 10 %, 11 %, 12 %, 13 %, 14 %, 15 %, 16 %, 17 %, 18 %, 19 %, 20 %, 21 %, 22 %, 23 %, 24 %, 25 %, 26 %, 27 %, 28 %, 29 %, 30 %, 31 %, 32 %, 33 %, 34 %, 35 %, 36 %, 37 %, 38 %, 39 %, 40 %, 41 %, 42 %, 43 %, 44 %, 45 %, 46 %, 47 %, 48 %, 49 %, 50 %, 51 %, 52 %, 53 %, 54 %, 55 %, 56 %, 57 %, 58 %, 59 %, 60 %, 61 %, 62 %, 63 %, 64 %, 65 %, 66 %, 67 %, 68 %, 69 %, 70 %, 71 %, 72 %, 73 %, 74 %, 75 %, 76 %, 77 %, 78 %, 79 %, 80 %, 81 %, 82 %, 83 %, 84 %, 85 %, 86 %, 87 %, 88 %, 89 %, or 90 %, by weight / weight, of the biosurfactant composition. In some cases, the lipopeptide may be about: 10 %-15 %, 9.5 %-l 5.5 %, 9 %-16 %, 8.5 %-16.5 %, 8 %-17 %, 7.5 %-17.5 %, 7 %-18 %, 6.5 %-l 8.5 %, 6 %-19 %, 5.5 %-19.5 %, 5 %-20 %, 4.5 %-20.5 %, 4 %-21 %, 3.5 %-21.5 %, 3 %-22 %, 2.5 %-22.5 %, 2 %-23 %, 1.5 %-23.5 %, 1 %-24 %, by weight / weight, of the biosurfactant composition. In some cases, the lipopeptide may be about 10 %-15 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lipopeptide may be about 9.5 %- 15.5 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lipopeptide may be about 9 %-16 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lipopeptide may be about 8.5 %- 16.5 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lipopeptide may be about 8 %-17 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lipopeptide may be about 7.5 %- 17.5 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lipopeptide may be about 7 %-18 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lipopeptide may be about 6.5 %- 18.5 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lipopeptide may be about 6 %-19 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lipopeptide may be about 5.5 %- 19.5 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lipopeptide may be about 5 %-20 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lipopeptide may be about 4.5 %-20.5 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lipopeptide may be about 4 %-21 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lipopeptide may be about 3.5 %-21.5 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lipopeptide may be about 3 %-22 %, byWSGR Docket No.: 69849-701.601weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lipopeptide may be about 2.5 %-22.5 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lipopeptide may be about 2 %-23 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lipopeptide may be about 1.5 %-23.5 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lipopeptide may be about 1 %-24 %, by weight / weight or by weight / volume, of the biosurfactant composition.

[0088] In some cases, the biosurfactant composition may comprise a lipoprotein. In some cases, the lipoprotein may comprise a lipoprotein from a fungus. In some cases, the lipoprotein may comprise a lipoprotein from a filamentous fungus. In some cases, the lipoprotein may comprise a lipoprotein from the family Xylariaceae. In some cases, the lipoprotein may comprise a lipoprotein from Entonaema liquescens.

[0089] In some cases, the lipoprotein may be at least about: 0.001 %, 0.01 %, 0.1 %, 1 %, 2 %, 3 %, 4 %, 5 %, 6 %, 7 %, 8 %, 9 %, 10 %, 10 %, 11 %, 12 %, 13 %, 14 %, 15 %, 16 %, 17 %, 18 %, 19 %, 20 %, 21 %, 22 %, 23 %, 24 %, 25 %, 26 %, 27 %, 28 %, 29 %, 30 %, 31 %, 32 %, 33 %, 34 %, 35 %, 36 %, 37 %, 38 %, 39 %, 40 %, 41 %, 42 %, 43 %, 44 %, 45 %, 46 %, 47 %, 48 %, 49 %, 50 %, 51 %, 52 %, 53 %, 54 %, 55 %, 56 %, 57 %, 58 %, 59 %, 60 %, 61 %, 62 %, 63 %, 64 %, 65 %, 66 %, 67 %, 68 %, 69 %, 70 %, 71 %, 72 %, 73 %, 74 %, 75 %, 76 %, 77 %.78 %, 79 %, 80 %, 81 %, 82 %, 83 %, 84 %, 85 %, 86 %, 87 %, 88 %, 89 %, 90 % or more, by weight / volume, of the biosurfactant composition. In some cases, the lipoprotein may be at most about: 0.001 %, 0.01 %, 0.1 %, 1 %, 2 %, 3 %, 4 %, 5 %, 6 %, 7 %, 8 %, 9 %, 10 %, 10 %, 11 %, 12 %, 13 %, 14 %, 15 %, 16 %, 17 %, 18 %, 19 %, 20 %, 21 %, 22 %, 23 %, 24 %, 25 % 26 %, 27 %, 28 %, 29 %, 30 %, 31 %, 32 %, 33 %, 34 %, 35 %, 36 %, 37 %, 38 %, 39 %, 40 %, 41 %, 42 %, 43 %, 44 %, 45 %, 46 %, 47 %, 48 %, 49 %, 50 %, 51 %, 52 %, 53 %, 54 %, 55 %, 56 %, 57 %, 58 %, 59 %, 60 %, 61 %, 62 %, 63 %, 64 %, 65 %, 66 %, 67 %, 68 %, 69 %, 70 %, 71 %, 72 %, 73 %, 74 %, 75 %, 76 %, 77 %, 78 %, 79 %, 80 %, 81 %, 82 %, 83 %, 84 %, 85 %, 86 %, 87 %, 88 %, 89 %, or 90 %, by weight / volume, of the biosurfactant composition. In some cases, the lipoprotein may be about: 5 %-10 %, 4.75 %-10.25 %, 4.5 %-10.5 %, 4.25 %- 10.75 %, 4 %-ll %, 3.75 %-l 1.25 %, 3.5 %-l 1.5 %, 3.25 %-l 1.75 %, 3 %-12 %, 2.75 %■ 12.25 %, 2.5 %-12.5 %, 2.25 %-12.75 %, 2 %-13 %, 1.75 %-I3.25 %, 1.5 %-13.5 %, 1.25 %■ 13.75 %, 1 %-14 %, 0.75 %-14.25 %, 0.5 %-14.5 %, 0.25 %-14.75 %, 0.25 %-15 %, by weight / volume, of the biosurfactant composition. In some cases, the lipoprotein may be at least about: 0.001 %, 0.01 %, 0.1 %, 1 %, 2 %, 3 %, 4 %, 5 %, 6 %, 7 %, 8 %, 9 %, 10 %, 10 %, 11 %, 12 %, 13 %, 14 %, 15 %, 16 %, 17 %, 18 %, 19 %, 20 %, 21 %, 22 %, 23 %, 24 %, 25 % 26 %, 27 %, 28 %, 29 %, 30 %, 31 %, 32 %, 33 %, 34 %, 35 %, 36 %, 37 %, 38 %, 39 %, 40 %,WSGR Docket No.: 69849-701.60141 %, 42 %, 43 %, 44 %, 45 %, 46 %, 47 %, 48 %, 49 %, 50 %, 51 %, 52 %, 53 %, 54 %, 55 %, 56 %, 57 %, 58 %, 59 %, 60 %, 61 %, 62 %, 63 %, 64 %, 65 %, 66 %, 67 %, 68 %, 69 %, 70 %, 71 %, 72 %, 73 %, 74 %, 75 %, 76 %, 77 %, 78 %, 79 %, 80 %, 81 %, 82 %, 83 %, 84 %, 85 %, 86 %, 87 %, 88 %, 89 %, 90 % or more, by weight / weight, of the biosurfactant composition. In some cases, the lipoprotein may be at most about: 0.001 %, 0.01 %, 0.1 %, 1 %, 2 %, 3 %, 4 %, 5 %, 6 %, 7 %, 8 %, 9 %, 10 %, 10 %, 11 %, 12 %, 13 %, 14 %, 15 %, 16 %, 17 %, 18 %, 19 %, 20 %, 21 %, 22 %, 23 %, 24 %, 25 %, 26 %, 27 %, 28 %, 29 %, 30 %, 31 %, 32 %, 33 %, 34 %, 35 %, 36 %, 37 %, 38 %, 39 %, 40 %, 41 %, 42 %, 43 %, 44 %, 45 %, 46 %, 47 %, 48 %, 49 %, 50 %, 51 %, 52 %, 53 %, 54 %, 55 %, 56 %, 57 %, 58 %, 59 %, 60 %, 61 %, 62 %, 63 %, 64 %, 65 %, 66 %, 67 %, 68 %, 69 %, 70 %, 71 %, 72 %, 73 %, 74 %, 75 %, 76 %, 77 %, 78 %, 79 %, 80 %, 81 %, 82 %, 83 %, 84 %, 85 %, 86 %, 87 %, 88 %, 89 %, or 90 %, by weight / weight, of the biosurfactant composition. In some cases, the lipoprotein may be about: 5 %-10 %, 4.75 %- 10.25 %, 4.5 %-10.5 %, 4.25 %-10.75 %, 4 %-ll %, 3.75 %-l 1.25 %, 3.5 %-l 1.5 %, 3.25 %- 11.75 %, 3 %-12 %, 2.75 %-12.25 %, 2.5 %-12.5 %, 2.25 %-12.75 %, 2 %-13 %, 1.75 %- 13.25 %, 1.5 %-13.5 %, 1.25 %-13.75 %, 1 %-14 %, 0.75 %-14.25 %, 0.5 %-14.5 %, 0.25 %- 14.75 %, 0.25 %-15 %, by weight / weight, of the biosurfactant composition. In some cases, the lipoprotein may be about 5 %-10 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lipoprotein may be about 4.75 %-10.25 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lipoprotein may be about 4.5 %- 10.5 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lipoprotein may be about 4.25 %-10.75 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lipoprotein may be about 4 %-l 1 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lipoprotein may be about 3.75 %-l 1.25 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lipoprotein may be about 3.5 %-l 1.5 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lipoprotein may be about 3.25 %-l 1.75 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lipoprotein may be about 3 %-12 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lipoprotein may be about 2.75 %-12.25 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lipoprotein may be about 2.5 %-12.5 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lipoprotein may be about 2.25 %-12.75 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lipoprotein may be about 2 %-13 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, theWSGR Docket No.: 69849-701.601lipoprotein may be about 1.75 %-13.25 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lipoprotein may be about 1.5 %- 13.5 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lipoprotein may be about 1.25 %-13.75 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lipoprotein may be about 1 %-14 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lipoprotein may be about 0.75 %-14.25 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lipoprotein may be about 0.5 %-14.5 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lipoprotein may be about 0.25 %-14.75 %, by weight / weight or by weight / volume, of the biosurfactant composition. In some cases, the lipoprotein may be about 0.25 %-15 %, by weight / weight or by weight / volume, of the biosurfactant composition.

[0090] In some cases, the biosurfactant composition may comprise a pigment. In some cases, the pigment may comprise a pigment from a fungus. In some cases, the pigment may comprise a pigment from a filamentous fungus. In some cases, the pigment may comprise a pigment from the family Xylariaceae. In some cases, the pigment may comprise a pigment from Entonaema liquescens. The pigment may comprise a pigment from Azaphilonas. The pigment may comprise mitroubin, orsellinic acid, or melanin, or a combination thereof.

[0091] In some cases, the pigment may be at least about: 0.001 %, 0.01 %, 0.1 %, 1 %, 2 %, 3 %, 4 %, 5 %, 6 %, 7 %, 8 %, 9 %, 10 %, 10 %, 11 %, 12 %, 13 %, 14 %, 15 %, 16 %, 17 %, 18 %, 19 %, 20 %, 21 %, 22 %, 23 %, 24 %, 25 %, 26 %, 27 %, 28 %, 29 %, 30 %, 31 %, 32 %, 33 %, 34 %, 35 %, 36 %, 37 %, 38 %, 39 %, 40 %, 41 %, 42 %, 43 %, 44 %, 45 %, 46 %, 47 %, 48 %, 49 %, 50 %, 51 %, 52 %, 53 %, 54 %, 55 %, 56 %, 57 %, 58 %, 59 %, 60 %, 61 %, 62 %, 63 %, 64 %, 65 %, 66 %, 67 %, 68 %, 69 %, 70 %, 71 %, 72 %, 73 %, 74 %, 75 %, 76 %, 77 %, 78 %, 79 %, 80 %, 81 %, 82 %, 83 %, 84 %, 85 %, 86 %, 87 %, 88 %, 89 %, 90 % or more, by weight / volume, of the biosurfactant composition. In some cases, the pigment may be at most about: 0.001 %, 0.01 %, 0.1 %, 1 %, 2 %, 3 %, 4 %, 5 %, 6 %, 7 %, 8 %, 9 %, 10 %, 10 %, 11 %, 12 %, 13 %, 14 %, 15 %, 16 %, 17 %, 18 %, 19 %, 20 %, 21 %, 22 %, 23 %, 24 %, 25 %, 26 %, 27 %, 28 %, 29 %, 30 %, 31 %, 32 %, 33 %, 34 %, 35 %, 36 %, 37 %, 38 %, 39 %, 40 %, 41 %, 42 %, 43 %, 44 %, 45 %, 46 %, 47 %, 48 %, 49 %, 50 %, 51 %, 52 %, 53 %, 54 %, 55 %, 56 %, 57 %, 58 %, 59 %, 60 %, 61 %, 62 %, 63 %, 64 %, 65 %, 66 %, 67 %, 68 %, 69 %, 70 %, 71 %, 72 %, 73 %, 74 %, 75 %, 76 %, 77 %, 78 %, 79 %, 80 %, 81 %, 82 %, 83 %, 84 %, 85 %, 86 %, 87 %, 88 %, 89 %, or 90 %, by weight / volume, of the biosurfactant composition.

[0092] In some cases, the biosurfactant composition may comprise a saccharide. The saccharide may comprise an exopolysaccharide. In some cases, the saccharide orWSGR Docket No.: 69849-701.601exopolysaccharide may be at least about: 0.001 %, 0.01 %, 0.1 %, 1 %, 2 %, 3 %, 4 %, 5 %, 6 %, 7 %, 8 %, 9 %, 10 %, 10 %, 11 %, 12 %, 13 %, 14 %, 15 %, 16 %, 17 %, 18 %, 19 % 20 %, 21 %, 22 %, 23 %, 24 %, 25 %, 26 %, 27 %, 28 %, 29 %, 30 %, 31 %, 32 %, 33 %, 34 %, 35 %, 36 %, 37 %, 38 %, 39 %, 40 %, 41 %, 42 %, 43 %, 44 %, 45 %, 46 %, 47 %, 48 %, 49 %, 50 %, 51 %, 52 %, 53 %, 54 %, 55 %, 56 %, 57 %, 58 %, 59 %, 60 %, 61 %, 62 %, 63 %, 64 %, 65 %, 66 %, 67 %, 68 %, 69 %, 70 %, 71 %, 72 %, 73 %, 74 %, 75 %, 76 %, 77 %, 78 %, 79 %, 80 %, 81 %, 82 %, 83 %, 84 %, 85 %, 86 %, 87 %, 88 %, 89 %, 90 % or more, by weight / volume or by weight / weight, of the biosurfactant composition. In some cases, the saccharide or exopolysaccharide may be at most about: 0.001 %, 0.01 %, 0.1 %, 1 %, 2 %, 3 %, 4 %, 5 %, 6 %, 7 %, 8 %, 9 %, 10 %, 10 %, 11 %, 12 %, 13 %, 14 %, 15 %, 16 %, 17 %, 18 %, 19 %, 20 %, 21 %, 22 %, 23 %, 24 %, 25 %, 26 %, 27 %, 28 %, 29 %, 30 %, 31 %, 32 %, 33 %, 34 %, 35 %, 36 %, 37 %, 38 %, 39 %, 40 %, 41 %, 42 %, 43 %, 44 %, 45 %, 46 %, 47 %, 48 %, 49 %, 50 %, 51 %, 52 %, 53 %, 54 %, 55 %, 56 %, 57 %, 58 %, 59 %, 60 %, 61 %, 62 %, 63 %, 64 %, 65 %, 66 %, 67 %, 68 %, 69 %, 70 %, 71 %, 72 %, 73 %, 74 %, 75 %, 76 %, 77 %, 78 %, 79 %, 80 %, 81 %, 82 %, 83 %, 84 %, 85 %, 86 %, 87 %, 88 %, 89 %, or 90 %, by weight / volume or by weight / weight, of the biosurfactant composition.

[0093] In some cases, the biosurfactant composition is frozen or lyophilized. In some cases, the biosurfactant composition is frozen. In some cases, the biosurfactant composition is lyophilized. In some cases, the biosurfactant composition is a powder. In some cases, the biosurfactant composition is a liquid. In some cases, the biosurfactant composition may not comprise a cryoprotectant. In some cases, the biosurfactant composition may comprise a cryoprotectant. In some cases, the cryoprotectant may comprise glycerol, dimethylsulfoxide (DMSO), ethylene glycol, propylene glycol, 2-methyl-2,4- pentanediol, trehalose, sucrose, diethyl glycol, triethylene glycol, polyvinylpyrrolidone (PVP), polyethylene glycol (PEG), saccharose, formamide, glycerol 3 -phosphate, proline, methyl alcohol, glucose, bovine serum albumin, polyvinyl alcohol, hydroxyethyl starch, sorbitol, an ice blocker (polyglycerol, polyvinyl alcohol, X-1000 or Z-100)., amino acids, or a combination thereof.

[0094] In some cases, the biosurfactant composition may comprise water. In some cases, the water may be at least about: 0.001 %, 0.01 %, 0.1 %, 1 %, 2 %, 3 %, 4 %, 5 %, 6 %, 7 %, 8 %, 9 %, 10 %, 10 %, 11 %, 12 %, 13 %, 14 %, 15 %, 16 %, 17 %, 18 %, 19 %, 20 %, 21 %, 22 %, 23 %, 24 %, 25 %, 26 %, 27 %, 28 %, 29 %, 30 %, 31 %, 32 %, 33 %, 34 %, 35 %, 36 %, 37 %, 38 %, 39 %, 40 %, 41 %, 42 %, 43 %, 44 %, 45 %, 46 %, 47 %, 48 %, 49 %, 50 %, 51 %, 52 %, 53 %, 54 %, 55 %, 56 %, 57 %, 58 %, 59 %, 60 %, 61 %, 62 %, 63 %, 64 %, 65 %, 66 %, 67 %, 68 %, 69 %, 70 %, 71 %, 72 %, 73 %, 74 %, 75 %, 76 %, 77 %, 78 %, 79 %, 80 %, 81 %, 82 %, 83 %, 84 %, 85 %, 86 %, 87 %, 88 %, 89 %, 90 % or more, by volume / volume, of theWSGR Docket No.: 69849-701.601biosurfactant composition. In some cases, the water may be at most about: 0.001 %, 0.01 %, 0.1 %, 1 %, 2 %, 3 %, 4 %, 5 %, 6 %, 7 %, 8 %, 9 %, 10 %, 10 %, 11 %, 12 %, 13 %, 14 %, 15 %, 16 %, 17 %, 18 %, 19 %, 20 %, 21 %, 22 %, 23 %, 24 %, 25 %, 26 %, 27 %, 28 %, 29 %, 30 %, 31 %, 32 %, 33 %, 34 %, 35 %, 36 %, 37 %, 38 %, 39 %, 40 %, 41 %, 42 %, 43 %, 44 %, 45 %, 46 %, 47 %, 48 %, 49 %, 50 %, 51 %, 52 %, 53 %, 54 %, 55 %, 56 %, 57 %, 58 %, 59 %, 60 %, 61 %, 62 %, 63 %, 64 %, 65 %, 66 %, 67 %, 68 %, 69 %, 70 %, 71 %, 72 %, 73 %, 74 %, 75 %, 76 %, 77 %, 78 %, 79 %, 80 %, 81 %, 82 %, 83 %, 84 %, 85 %, 86 %, 87 %, 88 %, 89 %, or 90 %, by volume / volume, of the biosurfactant composition.Various properties

[0095] The biosurfactant compositions described herein may exhibit various properties as described herein. These properties can be assayed using the methods as described herein (see, for example, Examples 1-4). In some cases, the biosurfactant composition may exhibit the various properties under a condition as described herein. In some cases, the condition may comprise a pH range as described herein, a temperature range as described herein, a salinity range as described herein, or a combination thereof.

[0096] In some cases, the biosurfactant composition may exhibit the various properties under a pH range. In some cases, the pH range may be at least about: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 or more. In some cases, the pH range may be at most about: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14. In some cases, the pH range may be about: 1-2, 1-3, 1-4, 1-5, 1-6, 1-7, 1-8, 1-9, 1-10, 1-11, 1-12, 1-13, 1-14, 2-3, 2-4, 2-5, 2-6, 2-7, 2-8, 2-9, 2-10, 2-11, 2-12, 2-13, 2-14, 3-4, 3-5, 3-6, 3-7, 3-8, 3-9, 3-10, 3-11, 3-12, 3-13, 3-14, 4-5, 4-6, 4-7, 4-8, 4-9, 4-10, 4-11, 4-12, 4-13, 4- 14, 5-6, 5-7, 5-8, 5-9, 5-10, 5-11, 5-12, 5-13, 5-14, 6-7, 6-8, 6-9, 6-10, 6-11, 6-12, 6-13, 6-14, 7-8, 7-9, 7-10, 7-11, 7-12, 7-13, 7-14, 8-9, 8-10, 8-11, 8-12, 8-13, 8-14, 9-10, 9-11, 9-12, 9-13, 9-14, 10-11, 10-12, 10-13, 10-14, 11-12, 11-13, 11-14, 12-13, 12-14, or 13-14.

[0097] In some cases, the biosurfactant composition may exhibit the various properties under a temperature range. In some cases, the temperature range may be at least about: -85 °C, -80 °C, - 75 °C -70 °C, -60 °C, -50 °C, -40 °C, -30 °C, -20 °C, -15 °C, -10 °C, -9 °C, -8 °C, -7 °C, -6 °C, - 5 °C, -4 °C, -3 °C, -2 °C, -1 °C, 0 °C, 1 °C, 2 °C, 3 °C, 4 °C, 5 °C, 6 °C, 7 °C, 8 °C, 9 °C, 10 °C, 11 °C, 12 °C, 13 °C, 14 °C, 15 °C, 16 °C, 17 °C, 18 °C, 19 °C, 20 °C, 21 °C, 22 °C, 23 °C, 24 °C, 25 °C, 26 °C, 27 °C, 28 °C, 29 °C, 30 °C, 31 °C, 32 °C, 33 °C, 34 °C, 35 °C, 36 °C, 37 °C, 38 °C, 39 °C, 40 °C, 41 °C, 42 °C, 43 °C, 44 °C, 45 °C, 46 °C, 47 °C, 48 °C, 49 °C, 50 °C, 51 °C, 52 °C, 53 °C, 54 °C, 55 °C, 56 °C, 57 °C, 58 °C, 59 °C, 60 °C, 61 °C, 62 °C, 63 °C, 64 °C, 65 °C, 66 °C, 67 °C, 68 °C, 69 °C, 70 °C, 71 °C, 72 °C, 73 °C, 74 °C, 75 °C, 76 °C, 77 °C, 78 °C, 79 °C, 80 °C, 81 °C, 82 °C, 83 °C, 84 °C, 85 °C, 86 °C, 87 °C, 88 °C, 89 °C, 90 °C, 91 °C, 92 °C, 93 °C, 94 °C, 95 °C, 96 °C, 97 °C, 98 °C, 99 °C, 100 °C, 101 °C,WSGR Docket No.: 69849-701.601102 °C, 103 °C, 104 °C, 105 °C, 105 °C, 106 °C, 107 °C„ 108 °C, 109 °C, 110 °C, 120 °C, 130 °C, 140 °C, 150 °C or more. In some cases, the temperature range may be at most about: -85 °C, -80 °C, -75 °C -70 °C, -60 °C, -50 °C, -40 °C, -30 °C, -20 °C, -15 °C, -10 °C, -9 °C, - 8 °C, -7 °C, -6 °C, -5 °C, -4 °C, -3 °C, -2 °C, -1 °C, 0 °C, 1 °C, 2 °C, 3 °C, 4 °C, 5 °C, 6 °C, 7 °C, 8 °C, 9 °C, 10 °C, 11 °C, 12 °C, 13 °C, 14 °C, 15 °C, 16 °C, 17 °C, 18 °C, 19 °C, 20 °C, 21 °C, 22 °C, 23 °C, 24 °C, 25 °C, 26 °C, 27 °C, 28 °C, 29 °C, 30 °C, 31 °C, 32 °C, 33 °C, 34 °C, 35 °C, 36 °C, 37 °C, 38 °C, 39 °C, 40 °C, 41 °C, 42 °C, 43 °C, 44 °C, 45 °C, 46 °C, 47 °C, 48 °C, 49 °C, 50 °C, 51 °C, 52 °C, 53 °C, 54 °C, 55 °C, 56 °C, 57 °C, 58 °C, 59 °C, 60 °C, 61 °C, 62 °C, 63 °C, 64 °C, 65 °C, 66 °C, 67 °C, 68 °C, 69 °C, 70 °C, 71 °C, 72 °C, 73 °C, 74 °C, 75 °C, 76 °C, 77 °C, 78 °C, 79 °C, 80 °C, 81 °C, 82 °C, 83 °C, 84 °C, 85 °C, 86 °C, 87 °C, 88 °C, 89 °C, 90 °C, 91 °C, 92 °C, 93 °C, 94 °C, 95 °C, 96 °C, 97 °C, 98 °C, 99 °C, 100 °C, 101 °C, 102 °C, 103 °C, 104 °C, 105 °C, 105 °C, 106 °C, 107 °C„ 108 °C, 109 °C, 110 °C, 120 °C, 130 °C, 140 °C, or 150 °C. In some cases, the temperature range may be about -20 °C to 4 °C, -20 °C to 5 °C, -20 °C to 6 °C, -20 °C to 7 °C, -20 °C to 8 °C, -20 °C to 9 °C, -20 °C to 10 °C, -20 °C to 11 °C, -20 °C to 12 °C, -20 °C to 13 °C, -20 °C to 14 °C, -20 °C to 15 °C, -10 °C to 4 °C, -10 °C to 5 °C, -10 °C to 6 °C, -10 °C to 7 °C, -10 °C to 8 °C, -10 °C to 9 °C, -10 °C to 10 °C, -10 °C to 11 °C, -10 °C to 12 °C, -10 °C to 13 °C, -10 °C to 14 °C, -10 °C to 15 °C, 0 °C to 4 °C, 0 °C to 5 °C, 0 °C to 6 °C, 0 °C to 7 °C, 0 °C to 8 °C, 0 °C to 9 °C, 0 °C to 10 °C, 0 °C to 11 °C, 0 °C to 12 °C, 0 °C to 13 °C, 0 °C to 14 °C, 0 °C to 15 °C, 85 °C to 95 °C, 85 °C to 96 °C, 85 °C to 97 °C, 85 °C to 98 °C, 85 °C to 99 °C, 85 °C to 100 °C, 85 °C to 101 °C, 85 °C to 102 °C, 85 °C to 103 °C, 85 °C to 104 °C, 85 °C to 105 °C, 85 °C to 106 °C, 90 °C to 95 °C, 90 °C to 96 °C, 90 °C to 97 °C, 90 °C to 98 °C, 90 °C to 99 °C, 90 °C to 100 °C, 90 °C to 101 °C, 90 °C to 102 °C, 90 °C to 103 °C, 90 °C to 104 °C, 90 °C to 105 °C, 90 °C to 106 °C, 95 °C to 96 °C, 95 °C to 97 °C, 95 °C to 98 °C, 95 °C to 99 °C, 95 °C to 100 °C, 95 °C to 101 °C, 95 °C to 102 °C, 95 °C to 103 °C, 95 °C to 104 °C, 95 °C to 105 °C, or 95 °C to 106 °C.

[0098] In some cases, the biosurfactant composition may exhibit the various properties under a salinity range. In some cases, the salinity range may be at least about: 25000 Total dissolved solids (TDS), 26000 TDS, 27000 TDS, 28000 TDS, 29000 TDS, 30000 TDS, 31000 TDS, 32000 TDS, 33000 TDS, 34000 TDS, 35000 TDS, 36000 TDS, 37000 TDS, 38000 TDS, 39000 TDS, 40000 TDS, 41000 TDS, 42000 TDS, 43000 TDS, 44000 TDS, 45000 TDS, 46000 TDS, 47000 TDS, 48000 TDS, 49000 TDS, 50000 TDS, 51000 TDS, 52000 TDS, 53000 TDS, 54000 TDS, 55000 TDS, 56000 TDS, 57000 TDS, 58000 TDS, 59000 TDS, 60000 TDS, 61000 TDS, 62000 TDS, 63000 TDS, 64000 TDS, 65000 TDS, 66000 TDS, 67000 TDS, 68000 TDS, 69000 TDS, 70000 TDS, 71000 TDS, 72000 TDS, 73000 TDS, 74000 TDS, 75000 TDS, 76000 TDS,WSGR Docket No.: 69849-701.60177000 TDS, 78000 TDS, 79000 TDS, 80000 TDS, 81000 TDS, 82000 TDS, 83000 TDS, 84000 TDS, 85000 TDS, 86000 TDS, 87000 TDS, 88000 TDS, 89000 TDS, 90000 TDS, 91000 TDS, 92000 TDS, 93000 TDS, 94000 TDS, 95000 TDS, 96000 TDS, 97000 TDS, 98000 TDS, 99000 TDS, 100000 TDS, 101000 TDS, 102000 TDS, 103000 TDS, 104000 TDS, 105000 TDS, 106000 TDS, 107000 TDS, 108000 TDS, 109000 TDS, 110000 TDS, 111000 TDS, 112000 TDS, 113000 TDS, 114000 TDS, 115000 TDS, 116000 TDS, 117000 TDS, 118000 TDS, 119000 TDS, 120000 TDS, 121000 TDS, 122000 TDS, 123000 TDS, 124000 TDS, 125000 TDS, 126000 TDS, 127000 TDS, 128000 TDS, 129000 TDS, 130000 TDS, 131000 TDS, 132000 TDS, 133000 TDS, 134000 TDS, 135000 TDS, 136000 TDS, 137000 TDS, 138000 TDS, 139000 TDS, 140000 TDS, 141000 TDS, 142000 TDS, 143000 TDS, 144000 TDS, 145000 TDS, 146000 TDS, 147000 TDS, 148000 TDS, 149000 TDS, 150000 TDS, 160000 TDS, 170000 TDS, 180000 TDS, 190000 TDS, 200000 TDS, 300000 TDS, 500000 TDS or more. In some cases, the biosurfactant composition may exhibit the various properties under a salinity range. In some cases, the salinity range may be at most about: 25000 Total dissolved solids (TDS), 26000 TDS, 27000 TDS, 28000 TDS, 29000 TDS, 30000 TDS, 31000 TDS, 32000 TDS, 33000 TDS, 34000 TDS, 35000 TDS, 36000 TDS, 37000 TDS, 38000 TDS, 39000 TDS, 40000 TDS, 41000 TDS, 42000 TDS, 43000 TDS, 44000 TDS, 45000 TDS, 46000 TDS, 47000 TDS, 48000 TDS, 49000 TDS, 50000 TDS, 51000 TDS, 52000 TDS, 53000 TDS, 54000 TDS, 55000 TDS, 56000 TDS, 57000 TDS, 58000 TDS, 59000 TDS, 60000 TDS, 61000 TDS, 62000 TDS, 63000 TDS, 64000 TDS, 65000 TDS, 66000 TDS, 67000 TDS, 68000 TDS, 69000 TDS, 70000 TDS, 71000 TDS, 72000 TDS, 73000 TDS, 74000 TDS, 75000 TDS, 76000 TDS, 77000 TDS, 78000 TDS, 79000 TDS, 80000 TDS, 81000 TDS, 82000 TDS, 83000 TDS, 84000 TDS, 85000 TDS, 86000 TDS, 87000 TDS, 88000 TDS, 89000 TDS, 90000 TDS, 91000 TDS, 92000 TDS, 93000 TDS, 94000 TDS, 95000 TDS, 96000 TDS, 97000 TDS, 98000 TDS, 99000 TDS, 100000 TDS, 101000 TDS, 102000 TDS, 103000 TDS, 104000 TDS, 105000 TDS, 106000 TDS, 107000 TDS, 108000 TDS, 109000 TDS, 110000 TDS, 111000 TDS, 112000 TDS, 113000 TDS, 114000 TDS, 115000 TDS, 116000 TDS, 117000 TDS, 118000 TDS, 119000 TDS, 120000 TDS, 121000 TDS, 122000 TDS, 123000 TDS, 124000 TDS, 125000 TDS, 126000 TDS, 127000 TDS, 128000 TDS, 129000 TDS, 130000 TDS, 131000 TDS, 132000 TDS, 133000 TDS, 134000 TDS, 135000 TDS, 136000 TDS, 137000 TDS, 138000 TDS, 139000 TDS, 140000 TDS, 141000 TDS, 142000 TDS, 143000 TDS, 144000 TDS, 145000 TDS, 146000 TDS, 147000 TDS, 148000 TDS, 149000 TDS, 150000 TDS, 160000 TDS, 170000 TDS, 180000 TDS, 190000 TDS, 200000 TDS, 300000 TDS, or 500000 TDS. In some cases, the salinity range, as measured in TDS, may be about 25000-30000, 25000-40000, 25000-50000, 25000-60000, 25000-70000, 25000-80000, 25000-90000, 25000-100000, 25000-WSGR Docket No.: 69849-701.601110000, 25000-120000, 50000-60000, 50000-70000, 50000-80000, 50000-90000, 50000-100000, 50000-110000, 50000-120000, 50000-130000, 50000-140000, 50000-150000, 75000-80000, 75000-90000, 75000-100000, 75000-110000, 75000-120000, 75000-130000, 75000-140000, 75000-150000, 75000-160000, 75000-170000, 100000-140000, 100000-150000, 100000-160000, 100000-170000, 100000-180000, 100000-190000, 100000-200000, 100000-210000, 100000-220000, 100000-230000, 115000-140000, 115000-150000, 115000-160000, 115000-170000, 115000-180000, 115000-190000, 115000-200000, 115000-210000, 115000-220000, 115000-230000, 130000-140000, 130000-150000, 130000-160000, 130000-170000, 130000-180000, 130000-190000, 130000-200000, 130000-210000, 130000-220000, or 130000-230000.

[0099] In some cases, the biosurfactant composition may not undergo phase transition at a temperature that is at least about: -85 °C, -80 °C, -75 °C -70 °C, -60 °C, -50 °C, -40 °C, -30 °C, -20 °C, -15 °C, -10 °C, -9 °C, -8 °C, -7 °C, -6 °C, -5 °C, -4 °C, -3 °C, -2 °C, -1 °C, 0 °C, 1 °C, 2 °C, 3 °C, 4 °C, 5 °C, 6 °C, 7 °C, 8 °C, 9 °C, 10 °C, 11 °C, 12 °C, 13 °C, 14 °C, 15 °C, 16 °C, 17 °C, 18 °C, 19 °C, 20 °C, 21 °C, 22 °C, 23 °C, 24 °C, 25 °C, 26 °C, 27 °C, 28 °C, 29 °C, 30 °C, 31 °C, 32 °C, 33 °C, 34 °C, 35 °C, 36 °C, 37 °C, 38 °C, 39 °C, 40 °C, 41 °C, 42 °C, 43 °C, 44 °C, 45 °C, 46 °C, 47 °C, 48 °C, 49 °C, 50 °C, 51 °C, 52 °C, 53 °C, 54 °C, 55 °C, 56 °C, 57 °C, 58 °C, 59 °C, 60 °C, 61 °C, 62 °C, 63 °C, 64 °C, 65 °C, 66 °C, 67 °C, 68 °C, 69 °C, 70 °C, 71 °C, 72 °C, 73 °C, 74 °C, 75 °C, 76 82 °C, 83 °C, 84 °C, 85 °C, 86 °C, 87 °C, 88 °C, 89 °C, 90 °C, 91 °C, 92 °C, 93 °C, 94 °C, 95 °C, 96 °C, 97 °C, 98 °C, 99 °C, 100 °C, 101 °C, 102 °C, 103 °C, 104 °C, 105 °C, 105 °C, 106 °C, 107 °C„ 108 °C, 109 °C, 110 °C, 120 °C, 130 °C, 140 °C, 150 °C or more, as measured by cloud point technique. When using the cloud point technique, the phase transition may comprise a solution being transitioned from being transparent to being turbid. In some cases, the cloud point technique may comprise subjecting a solution (such as biosurfactant composition) over various temperature points by gradually increasing and decreasing the temperature while assessing turbidity changes of the solution / biosurfactant composition. The absence of cloud points at a particular range of temperature points may indicate high solubility or thermal stability of the biosurfactant composition. In some cases, the biosurfactant composition may not undergo phase transition at a temperature that is at most about: -85 °C, -80 °C, -75 °C -70 °C, -60 °C, -50 °C, -40 °C, -30 °C, -20 °C, -15 °C, -10 °C, -9 °C, -8 °C, -7 °C, -6 °C, -5 °C, -4 °C, -3 °C, - 2 °C, -1 °C, 0 °C, 1 °C, 2 °C, 3 °C, 4 °C, 5 °C, 6 °C, 7 °C, 8 °C, 9 °C, 10 °C, 11 °C, 12 °C, 13 °C, 14 °C, 15 °C, 16 °C, 17 °C, 18 °C, 19 °C, 20 °C, 21 °C, 22 °C, 23 °C, 24 °C, 25 °C, 26 °C, 27 °C, 28 °C, 29 °C, 30 °C, 31 °C, 32 °C, 33 °C, 34 °C, 35 °C, 36 °C, 37 °C, 38 °C, 39 °C, 40 °C, 41 °C, 42 °C, 43 °C, 44 °C, 45 °C, 46 °C, 47 °C, 48 °C, 49 °C, 50 °C, 51 °C,WSGR Docket No.: 69849-701.60152 °C, 53 °C, 54 °C, 55 °C, 56 °C, 57 °C, 58 °C, 59 °C, 60 °C, 61 °C, 62 °C, 63 °C, 64 °C, 65 °C, 66 °C, 67 °C, 68 °C, 69 °C, 70 °C, 71 °C, 72 °C, 73 °C, 74 °C, 75 °C, 76 °C, 77 °C, 78 °C, 79 °C, 80 °C, 81 °C, 82 °C, 83 °C, 84 °C, 85 °C, 86 °C, 87 °C, 88 °C, 89 °C, 90 °C, 91 °C, 92 °C, 93 °C, 94 °C, 95 °C, 96 °C, 97 °C, 98 °C, 99 °C, 100 °C, 101 °C, 102 °C, 103 °C, 104 °C, 105 °C, 105 °C, 106 °C, 107 °C, 108 °C, 109 °C, 110 °C, 120 °C, 130 °C, 140 °C, or 150 °C. In some cases, the biosurfactant composition may not undergo phase transition: from about -10 °C to about 100 °C, from about -11 °C to about 101 °C, from about -12 °C to about 102 °C, from about -13 °C to about 103 °C, from about -14 °C to about 104 °C, from about -15 °C to about 105 °C, from about -16 °C to about 106 °C, from about -17 °C to about 107 °C, from about -18 °C to about 108 °C, from about -19 °C to about 109 °C, from about -20 °C to about 110 °C, from about -21 °C to about 111 °C, from about -22 °C to about 112 °C, from about -23 °C to about 113 °C, from about -24 °C to about 114 °C, from about -25 °C to about 115 °C, from about -26 °C to about 116 °C, from about -27 °C to about 117 °C, from about -28 °C to about 118 °C, from about -29 °C to about 119 °C, from about -30 °C to about 120 °C, from about -31 °C to about 121 °C, from about -32 °C to about 122 °C, from about -33 °C to about 123 °C, from about -34 °C to about 124 °C, from about -35 °C to about 125 °C, from about -36 °C to about 126 °C, from about -37 °C to about 127 °C, from about -38 °C to about 128 °C, from about -39 °C to about 129 °C, from about -40 °C to about 130 °C, from about -41 °C to about 131 °C, from about -42 °C to about 132 °C, from about -43 °C to about 133 °C, from about -44 °C to about 134 °C, from about -45 °C to about 135 °C, from about -46 °C to about 136 °C, from about -47 °C to about 137 °C, from about -48 °C to about 138 °C, from about -49 °C to about 139 °C, or from about -50 °C to about 140 °C.

[0100] In some cases, the biosurfactant composition may reduce interfacial tension between two phases. In some cases, the two phases may comprise two different liquid phases; a liquid phase and a solid phase; a liquid and a gaseous phase; or a combination thereof. In some cases, the two phases may comprise two different liquid phases. In some cases, the two phases may comprise a liquid phase and a solid phase. In some cases, the two phases may comprise a liquid and a gaseous phase. In some cases, a liquid phase may comprise a hydrophobic liquid. In some cases, the hydrophobic liquid may comprise oil or petroleum. In some cases, the two liquid phases may comprise a hydrophobic phase and a hydrophilic or aqueous phase. In some cases, the two liquid phases may comprise water and oil / petroleum. In some cases, the biosurfactant composition may reduce interfacial tension between two phases by at least about: 1 %, 2 %, 3 %, 4 %, 5 %, 10 %, 15 %, 20 %, 25 %, 30 %, 35 %, 40 %, 45 %, 50 %, 55 %, 60 %, 65 %, 70 %, 75 %, 80 %, 85 %, 90 %, 95 %, 96 %, 97 %, 98 %, 99 % or more, as compared to that of the two phases without the biosurfactant composition. In some cases, the biosurfactantWSGR Docket No.: 69849-701.601composition may reduce surface tension between two phases by at most about: 1 %, 2 %, 3 %, 4 %, 5 %, 10 %, 15 %, 20 %, 25 %, 30 %, 35 %, 40 %, 45 %, 50 %, 55 %, 60 %, 65 %, 70 %, 75 %, 80 %, 85 %, 90 %, 95 %, 96 %, 97 %, 98 %, or 99 %, as compared to that of the two phases without the biosurfactant composition. In some cases, the biosurfactant composition may reduce surface tension between two phases by at least about: 1 %, 2 %, 3 %, 4 %, 5 %, 10 %, 15 %, 20 %, 25 %, 30 %, 35 %, 40 %, 45 %, 50 %, 55 %, 60 %, 65 %, 70 %, 75 %, 80 %, 85 %, 90 %, 95 %, 96 %, 97 %, 98 %, 99 % or more, as compared to that of the two phases without the biosurfactant composition. . In some cases, the biosurfactant composition may reduce interfacial tension between two phases by at most about: 1 %, 2 %, 3 %, 4 %, 5 %, 10 %, 15 %, 20 %, 25 %, 30 %, 35 %, 40 %, 45 %, 50 %, 55 %, 60 %, 65 %, 70 %, 75 %, 80 %, 85 %, 90 %, 95 %, 96 %, 97 %, 98 %, or 99 %, as compared to that of the two phases without the biosurfactant composition.

[0101] In some cases, the biosurfactant composition may have a surface tension of at least about: 0.1 millinewton / meter (mN / m), 0.2 mN / m, 0.3 mN / m, 0.4 mN / m, 0.5 mN / m, 0.6 mN / m, 0.7 mN / m, 0.8 mN / m, 0.9 mN / m, 1 mN / m, 2 mN / m, 3 mN / m, 4 mN / m, 5 mN / m, 6 mN / m, 7 mN / m, 8 mN / m, 9 mN / m, 10 mN / m, 15 mN / m, 20 mN / m, 25 mN / m, 30 mN / m, 35 mN / m, 40 mN / m, 45 mN / m, 50 mN / m, 55 mN / m, 60 mN / m, 65 mN / m, 70 mN / m, 75 mN / m, 80 mN / m, 85 mN / m, 90 mN / m, 95 mN / m, 100 mN / m, 110 mN / m, 120 mN / m, 130 mN / m, 140 mN / m, 150 mN / m, 160 mN / m, 170 mN / m, 180 mN / m, 190 mN / m, 200 mN / m, 210 mN / m, 220 mN / m, 230 mN / m, 240 mN / m, 250 mN / m, 260 mN / m, 270 mN / m, 280 mN / m, 290 mN / m, 300 mN / m or more. In some cases, the biosurfactant composition may have a surface tension of at most about: 0.1 mN / m, 0.2 mN / m, 0.3 mN / m, 0.4 mN / m, 0.5 mN / m, 0.6 mN / m, 0.7 mN / m, 0.8 mN / m, 0.9 mN / m, 1 mN / m, 2 mN / m, 3 mN / m, 4 mN / m, 5 mN / m, 6 mN / m, 7 mN / m, 8 mN / m, 9 mN / m, 10 mN / m, 15 mN / m, 20 mN / m, 25 mN / m, 30 mN / m, 35 mN / m, 40 mN / m, 45 mN / m, 50 mN / m, 55 mN / m, 60 mN / m, 65 mN / m, 70 mN / m, 75 mN / m, 80 mN / m, 85 mN / m, 90 mN / m, 95 mN / m, 100 mN / m, 110 mN / m, 120 mN / m, 130 mN / m, 140 mN / m, 150 mN / m, 160 mN / m, 170 mN / m, 180 mN / m, 190 mN / m, 200 mN / m, 210 mN / m, 220 mN / m, 230 mN / m, 240 mN / m, 250 mN / m, 260 mN / m, 270 mN / m, 280 mN / m, 290 mN / m, or 300 mN / m. . In some cases, the biosurfactant composition may have a surface tension of about 25 mN / m-35 mN / m, 20 mN / m-40 mN / m, 15 mN / m-45 mN / m, 10 mN / m-50 mN / m, 5 mN / m-55 mN / m, 5 mN / m-60 mN / m, 5 mN / m-65 mN / m, 5 mN / m-70 mN / m, 5 mN / m-75 mN / m, 5 mN / m-80 mN / m, 5 mN / m-85 mN / m, 5 mN / m-90 mN / m, 5 mN / m-95 mN / m, or 5 mN / m- 100 mN / m.

[0102] In some cases, the biosurfactant composition may have an interfacial tension of at least about: 0.1 mN / m, 0.2 mN / m, 0.3 mN / m, 0.4 mN / m, 0.5 mN / m, 0.6 mN / m, 0.7 mN / m, 0.8 mN / m, 0.9 mN / m, 1 mN / m, 2 mN / m, 3 mN / m, 4 mN / m, 5 mN / m, 6 mN / m, 7 mN / m, 8 mN / m, 9WSGR Docket No.: 69849-701.601mN / m, 10 mN / m, 15 mN / m, 20 mN / m, 25 mN / m, 30 mN / m, 35 mN / m, 40 mN / m, 45 mN / m, 50 mN / m or more. In some cases, the biosurfactant composition may have an interfacial tension of at least about: 0.1 mN / m, 0.2 mN / m, 0.3 mN / m, 0.4 mN / m, 0.5 mN / m, 0.6 mN / m, 0.7 mN / m, 0.8 mN / m, 0.9 mN / m, 1 mN / m, 2 mN / m, 3 mN / m, 4 mN / m, 5 mN / m, 6 mN / m, 7 mN / m, 8 mN / m, 9 mN / m, 10 mN / m, 15 mN / m, 20 mN / m, 25 mN / m, 30 mN / m, 35 mN / m, 40 mN / m, 45 mN / m, or 50 mN / m. In some cases, the biosurfactant composition may have an interfacial tension of about: 4.5 mN / m-5.5 mN / m, 4 mN / m-6 mN / m, 3.5 mN / m-6.5 mN / m, 3 mN / m-7 mN / m, 2.5 mN / m-7.5 mN / m, 2 mN / m-8 mN / m, 1.5 mN / m-8.5 mN / m, 1 mN / m-9 mN / m, 0.5 mN / m-9.5 mN / m, 0.5 mN / m- 10 mN / m, 0.5 mN / m-20 mN / m, 0.5 mN / m-30 mN / m, 0.5 mN / m-40 mN / m, or 0.5 mN / m-50 mN / m.

[0103] In some cases, methods for measuring the surface tension and interfacial tension of the biosurfactant composition can comprise a static method, a dynamic method, or an indirect or complementary method. In some cases, methods for measuring the surface tension and interfacial tension of the biosurfactant composition can comprise a Kriiss K8 tensiometer and a Du Nouy ring (Pt-Ir); the Wilhelmy Plate Method; Pendant Drop Method; Sessile Drop Method; Bubble Drop Method; the Oscillating Drop / Bubble Method; Maximum Bubble Pressure Method; Spinning Drop Method; Interfacial Rheology; Contact Angle Microscopy; or Electrocapillarity.

[0104] In some cases, the biosurfactant composition may remove or sequester a hydrocarbon. Sequestering a hydrocarbon may comprise isolating the hydrocarbon within a mixture from the other components of the mixture. Sequestering a hydrocarbon may comprise isolating the hydrocarbon from a physical location to another physical location. Sequestering a hydrocarbon may comprise isolating the hydrocarbon from a physical compartment to another physical compartment. The ability of the biosurfactant composition for sequestering or removing a hydrocarbon may be used in various applications as described herein. In some cases, the biosurfactant composition may remove or sequester at least about: 1 %, 2 %, 3 %, 4 %, 5 %, 10 %, 15 %, 20 %, 25 %, 30 %, 35 %, 40 %, 45 %, 50 %, 55 %, 60 %, 65 %, 70 %, 75 %, 80 %, 85 %, 90 %, 95 %, 96 %, 97 %, 98 %, 99 % or more hydrocarbon. In some cases, the biosurfactant composition may remove or sequester at most about: 1 %, 2 %, 3 %, 4 %, 5 %, 10 %, 15 %, 20 %, 25 %, 30 %, 35 %, 40 %, 45 %, 50 %, 55 %, 60 %, 65 %, 70 %, 75 %, 80 %, 85 %, 90 %, 95 %, 96 %, 97 %, 98 %, or 99 % hydrocarbon. In some cases, the biosurfactant composition may remove or sequester a hydrocarbon from a mixture comprising an aqueous component and the hydrocarbon. In some cases, the biosurfactant composition may remove or sequester at least about: 1 %, 2 %, 3 %, 4 %, 5 %, 10 %, 15 %, 20 %, 25 %, 30 %, 35 %, 40 %, 45 %, 50 %, 55 %, 60 %, 65 %, 70 %, 75 %, 80 %, 85 %, 90 %, 95 %, 96 %, 97 %, 98 %, 99 %WSGR Docket No.: 69849-701.601or more hydrocarbon from a mixture comprising an aqueous component and the hydrocarbon. In some cases, the biosurfactant composition may remove or sequester at most about: 1 %, 2 %, 3 %, 4 %, 5 %, 10 %, 15 %, 20 %, 25 %, 30 %, 35 %, 40 %, 45 %, 50 %, 55 %, 60 %, 65 %, 70 %, 75 %, 80 %, 85 %, 90 %, 95 %, 96 %, 97 %, 98 %, or 99 % hydrocarbon from a mixture comprising an aqueous component and the hydrocarbon.

[0105] In some cases, the biosurfactant composition may remove or sequester a hydrocarbon from the surface of a solid particle. In some cases, the biosurfactant composition may remove or sequester at least about: 1 %, 2 %, 3 %, 4 %, 5 %, 10 %, 15 %, 20 %, 25 %, 30 %, 35 %, 40 %, 45 %, 50 %, 55 %, 60 %, 65 %, 70 %, 75 %, 80 %, 85 %, 90 %, 95 %, 96 %, 97 %, 98 %, 99 % or more hydrocarbon from the surface of a solid particle. In some cases, the biosurfactant composition may remove or sequester at most about: 1 %, 2 %, 3 %, 4 %, 5 %, 10 %, 15 %, 20 %, 25 %, 30 %, 35 %, 40 %, 45 %, 50 %, 55 %, 60 %, 65 %, 70 %, 75 %, 80 %, 85 %, 90 %, 95 %, 96 %, 97 %, 98 %, or 99 % hydrocarbon from the surface of a solid particle. The solid particle may have a volume of at least about: lxl0A0, lxl0Al, lxlOA2, lxl0A3, lxlOA4, lxl0A5, lxlOA6, lxlOA7, lxl0A8, lxlOA9, lxl0A10, lxl0All, lxlOA12, lxl0A13, lxlOA14, lxl0A15 or more cubic centimeter (cmA3). The solid particle may have a volume of at most about: lxl0A0, lxl0Al, lxlOA2, lxl0A3, lxlOA4, lxl0A5, lxlOA6, lxlOA7, lxl0A8, lxlOA9, lxl0A10, lxl0All, lxlOA12, lxl0A13, lxlOA14, or lxl0A15 cubic centimeter (cmA3). The solid particle may comprise a rock. The solid particle may comprise a carbonate rock. The solid particle may have a total surface area of at least about: lxl0A0, lxl0Al, lxlOA2, lxl0A3, lxlOA4, lxl0A5, lxlOA6, lxlOA7, lxl0A8, lxlOA9, lxl0A10, lxl0All, lxlOA12, lxl0A13, lxlOA14, lxl0A15 or more centimeter squared (cmA2). The solid particle may have a total surface area of at most about: lxl0A0, lxl0Al, lxlOA2, lxl0A3, lxlOA4, lxl0A5, lxlOA6, lxlOA7, lxl0A8, lxlOA9, lxl0A10, lxl0All, lxlOA12, lxl0A13, lxlOA14, or lxl0A15 centimeter squared (cmA2). The solid particle may have a diameter of at least about: lxl0A0, lxl0Al, lxlOA2, lxl0A3, lxlOA4, lxl0A5, lxlOA6, lxlOA7, lxl0A8, lxlOA9, lxl0A10, lxl0All, lxlOA12, lxl0A13, lxlOA14, lxl0A15 or more centimeter (cm). The solid particle may have a diameter of at most about: lxl0A0, lxl0Al, lxlOA2, lxl0A3, lxlOA4, lxl0A5, lxlOA6, lxlOA7, lxl0A8, lxlOA9, lxl0A10, lxl0All, lxlOA12, lxl0A13, lxlOA14, or lxl0A15 centimeter (cm). The solid particle may comprise a rock. The solid particle may comprise a carbonate rock.

[0106] The hydrocarbon removal or sequester ability of the biosurfactant is at least about: 0.01 %, 0.1 %, 1%, 2 %, 3 %, 4 %, 5 %, 6 %, 7 %, 8 %, 9 %, 10 %, 11 %, 12 %, 13 %, 14 %, 15 %, 20 %, 25 %, 30 %, 35 %, 40 %, 50 %, 60 %, 70 %, 80 %, 90 %, 95 %, 100 %, 150 %, 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, 100-fold, or 1000-fold, higher than that of a control biosurfactant composition. The control biosurfactant composition mayWSGR Docket No.: 69849-701.601comprise a biosurfactant composition that does not have a component of the biosurfactant composition. The control biosurfactant composition may comprise a biosurfactant composition that has different chemical make-up of the biosurfactant composition. The control biosurfactant composition may comprise a different biosurfactant composition from the biosurfactant composition. The control biosurfactant composition may comprise Biobased surfactant:BioLoop 84L from Lankem; Synthetic surfactant: STEPANBLEND or APB Stepan Blend from Stephan. The control biosurfactant composition may comprise the commercial synthetic surfactant described in this disclosure. The hydrocarbon removal or sequester ability of the biosurfactant is at most about: 0.01 %, 0.1 %, 1%, 2 %, 3 %, 4 %, 5 %, 6 %, 7 %, 8 %, 9 %, 10 %, 11 %, 12 %, 13 %, 14 %, 15 %, 20 %, 25 %, 30 %, 35 %, 40 %, 50 %, 60 %, 70 %, 80 %, 90 %, 95 %, 100 %, 150 %, 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, 100-fold, or 1000-fold, higher than that of a control biosurfactant composition.

[0107] In some cases, the biosurfactant composition may facilitate emulsification of a mixture comprising two different liquids. In some cases, the two liquids may comprise a hydrophobic phase and a hydrophilic or aqueous phase. In some cases, the two liquids may comprise water and oil / petroleum. In some cases, the biosurfactant composition may facilitate at least about: 1 %, 2 %, 3 %, 4 %, 5 %, 10 %, 15 %, 20 %, 25 %, 30 %, 35 %, 40 %, 45 %, 50 %, 55 %, 60 %, 65 %, 70 %, 75 %, 80 %, 85 %, 90 %, 95 %, 96 %, 97 %, 98 %, 99 % or more, by weigh or volume, of the two different liquids to become emulsions. In some cases, the two liquids may comprise water and oil / petroleum. In some cases, the biosurfactant composition may facilitate at most about: 1 %, 2 %, 3 %, 4 %, 5 %, 10 %, 15 %, 20 %, 25 %, 30 %, 35 %, 40 %, 45 %, 50 %, 55 %, 60 %, 65 %, 70 %, 75 %, 80 %, 85 %, 90 %, 95 %, 96 %, 97 %, 98 %, or 99 %, by weigh or volume, of the two different liquids to become emulsions. Emulsion formation can be measured using Dynamic light scattering (DLS) or Electrophoretic light scattering (ELS). The emulsion formation ability of the biosurfactant is at least about: 0.01 %, 0.1 %, 1%, 2 %, 3 %, 4 %, 5 %, 6 %, 7 %, 8 %, 9 %, 10 %, 11 %, 12 %, 13 %, 14 %, 15 %, 20 %, 25 %, 30 %, 35 %, 40 %, 50 %, 60 %, 70 %, 80 %, 90 %, 95 %, 100 %, 150 %, 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, 100-fold, or 1000-fold, higher than that of a control biosurfactant composition. The emulsion formation ability of the biosurfactant is at most about: 0.01 %, 0.1 %, 1%, 2 %, 3 %, 4 %, 5 %, 6 %, 7 %, 8 %, 9 %, 10 %, 11 %, 12 %, 13 %, 14 %, 15 %, 20 %, 25 %, 30 %, 35 %, 40 %, 50 %, 60 %, 70 %, 80 %, 90 %, 95 %, 100 %, 150 %, 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, 100-fold, or 1000-fold, higher than that of a control biosurfactant composition.

[0108] In some cases, the biosurfactant compositions provided herein are safe or do not negatively impact the environment. In some cases, the biosurfactant compositions providedWSGR Docket No.: 69849-701.601herein have low toxicity. In some cases, the Acute Toxicity category of biosurfactant composition described herein may be category V. In some cases, the Acute Toxicity category of biosurfactant composition described herein may be category IV. In some cases, the Acute Toxicity category of biosurfactant composition described herein may not be category I, II, or III. In some cases, the biosurfactant compositions provided herein are biodegradable or are capable of being decomposed by living organisms.

[0109] In some cases, the biosurfactant compositions provided herein has a density (at 25 °C) of at least about: 0.01 g / cmA3, 0.02 g / cmA3, 0.03 g / cmA3, 0.04 g / cmA3, 0.05 g / cmA3, 0.06 g / cmA3, 0.07 g / cmA3, 0.08 g / cmA3, 0.09 g / cmA3, 0.1 g / cmA3, 0.2 g / cmA3, 0.3 g / cmA3, 0.4 g / cmA3, 0.5 g / cmA3, 0.6 g / cmA3, 0.7 g / cmA3, 0.8 g / cmA3, 0.9 g / cmA3, 1 g / cmA3, 1.1 g / cmA3, 1.2 g / cmA3, 1.3 g / cmA3, 1.4 g / cmA3, 1.5 g / cmA3, 1.6 g / cmA3, 1.7 g / cmA3, 1.8 g / cmA3, 1.9 g / cmA3, 2 g / cmA3, 3 g / cmA3, 4 g / cmA3, 5 g / cmA3, 6 g / cmA3, 7 g / cmA3, 8 g / cmA3, 9 g / cmA3, 10 g / cmA3, 100 g / cmA3 or more. In some cases, the biosurfactant compositions provided herein has a density (at 25 °C) of at most about: 0.01 g / cmA3, 0.02 g / cmA3, 0.03 g / cmA3, 0.04 g / cmA3, 0.05 g / cmA3, 0.06 g / cmA3, 0.07 g / cmA3, 0.08 g / cmA3, 0.09 g / cmA3, 0.1 g / cmA3, 0.2 g / cmA3, 0.3 g / cmA3, 0.4 g / cmA3, 0.5 g / cmA3, 0.6 g / cmA3, 0.7 g / cmA3, 0.8 g / cmA3, 0.9 g / cmA3, 1 g / cmA3, 1.1 g / cmA3, 1.2 g / cmA3, 1.3 g / cmA3, 1.4 g / cmA3, 1.5 g / cmA3, 1.6 g / cmA3, 1.7 g / cmA3, 1.8 g / cmA3, 1.9 g / cmA3, 2 g / cmA3, 3 g / cmA3, 4 g / cmA3, 5 g / cmA3, 6 g / cmA3, 7 g / cmA3, 8 g / cmA3, 9 g / cmA3, 10 g / cmA3, or 100 g / cmA3.

[0110] In some cases, the biosurfactant compositions provided herein has a viscosity of at least about: 0.01 Centipoise (cP) or millipascal-seconds (mPa S) [cP / mPa S], 0.02 cP / mPa S, 0.03 cP / mPa S, 0.04 cP / mPa S, 0.05 cP / mPa S, 0.06 cP / mPa S, 0.07 cP / mPa S, 0.08 cP / mPa S, 0.09 cP / mPa S, 0.1 cP / mPa S, 0.2 cP / mPa S, 0.3 cP / mPa S, 0.4 cP / mPa S, 0.5 cP / mPa S, 0.6 cP / mPa S, 0.7 cP / mPa S, 0.8 cP / mPa S, 0.9 cP / mPa S, 1 cP / mPa S, 1.1 cP / mPa S, 1.2 cP / mPa S, 1.3 cP / mPa S, 1.4 cP / mPa S, 1.5 cP / mPa S, 1.6 cP / mPa S, 1.7 cP / mPa S, 1.8 cP / mPa S, 1.9 cP / mPa S, 2 cP / mPa S, 3 cP / mPa S, 4 cP / mPa S, 5 cP / mPa S, 6 cP / mPa S, 7 cP / mPa S, 8 cP / mPa S, 9 cP / mPa S, 10 cP / mPa S, or 100 cP / mPa S. In some cases, the biosurfactant compositions provided herein has a viscosity of at most about: 0.01 Centipoise (cP) and millipascal-seconds (mPa S), 0.02 cP / mPa S, 0.03 cP / mPa S, 0.04 cP / mPa S, 0.05 cP / mPa S, 0.06 cP / mPa S, 0.07 cP / mPa S, 0.08 cP / mPa S, 0.09 cP / mPa S, 0.1 cP / mPa S, 0.2 cP / mPa S, 0.3 cP / mPa S, 0.4 cP / mPa S, 0.5 cP / mPa S, 0.6 cP / mPa S, 0.7 cP / mPa S, 0.8 cP / mPa S, 0.9 cP / mPa S, 1 cP / mPa S, 1.1 cP / mPa S, 1.2 cP / mPa S, 1.3 cP / mPa S, 1.4 cP / mPa S, 1.5 cP / mPa S, 1.6 cP / mPa S, 1.7 cP / mPa S, 1.8 cP / mPa S, 1.9 cP / mPa S, 2 cP / mPa S, 3 cP / mPa S, 4 cP / mPa S, 5 cP / mPa S, 6 cP / mPa S, 7 cP / mPa S, 8 cP / mPa S, 9 cP / mPa S, 10 cP / mPa S, or 100 cP / mPa S.WSGR Docket No.: 69849-701.601

[0111] In some cases, the biosurfactant composition may have a solids content of at most about: 1 %, 2 %, 3 %, 4 %, 5 %, 10 %, 15 %, 20 %, 25 %, 30 %, 35 %, 40 %, 45 %, 50 %, 55 %, 60 %, 65 %, 70 %, 75 %, 80 %, 85 %, 90 %, 95 %, 96 %, 97 %, 98 %, or 99 %. In some cases, the biosurfactant composition may have a solids content of at least about: 1 %, 2 %, 3 %, 4 %, 5 %, 10 %, 15 %, 20 %, 25 %, 30 %, 35 %, 40 %, 45 %, 50 %, 55 %, 60 %, 65 %, 70 %, 75 %, 80 %, 85 %, 90 %, 95 %, 96 %, 97 %, 98 %, or 99 %.

[0112] In some cases, the biosurfactant composition may have critical micelle concentration / CMC (at 25 °C; by w / v), of at least about: 1 %, 2 %, 3 %, 4 %, 5 %, 10 %, 15 %, 20 %, 25 %, 30 %, 35 %, 40 %, 45 %, 50 %, 55 %, 60 %, 65 %, 70 %, 75 %, 80 %, 85 %, 90 %, 95 %, 96 %, 97 %, 98 %, or 99 %. In some cases, the biosurfactant composition may have CMC (at 25 °C; by w / v), of at most about: 1 %, 2 %, 3 %, 4 %, 5 %, 10 %, 15 %, 20 %, 25 %, 30 %, 35 %, 40 %, 45 %, 50 %, 55 %, 60 %, 65 %, 70 %, 75 %, 80 %, 85 %, 90 %, 95 %, 96 %, 97 %, 98 %, or 99 %.

[0113] In some cases, the biosurfactant composition at CMC at 9% and at 25 °C, may have a surface tension of at least about: 0.1 millinewton / meter (mN / m), 0.2 mN / m, 0.3 mN / m, 0.4 mN / m, 0.5 mN / m, 0.6 mN / m, 0.7 mN / m, 0.8 mN / m, 0.9 mN / m, 1 mN / m, 2 mN / m, 3 mN / m, 4 mN / m, 5 mN / m, 6 mN / m, 7 mN / m, 8 mN / m, 9 mN / m, 10 mN / m, 15 mN / m, 20 mN / m, 25 mN / m, 30 mN / m, 35 mN / m, 40 mN / m, 45 mN / m, 50 mN / m, 55 mN / m, 60 mN / m, 65 mN / m, 70 mN / m, 75 mN / m, 80 mN / m, 85 mN / m, 90 mN / m, 95 mN / m, 100 mN / m, 110 mN / m, 120 mN / m, 130 mN / m, 140 mN / m, 150 mN / m, 160 mN / m, 170 mN / m, 180 mN / m, 190 mN / m, 200 mN / m, 210 mN / m, 220 mN / m, 230 mN / m, 240 mN / m, 250 mN / m, 260 mN / m, 270 mN / m, 280 mN / m, 290 mN / m, 300 mN / m or more. In some cases, the biosurfactant composition at CMC at 9% and at 25 °C, may have a surface tension of at most about: 0.1 millinewton / meter (mN / m), 0.2 mN / m, 0.3 mN / m, 0.4 mN / m, 0.5 mN / m, 0.6 mN / m, 0.7 mN / m, 0.8 mN / m, 0.9 mN / m, 1 mN / m, 2 mN / m, 3 mN / m, 4 mN / m, 5 mN / m, 6 mN / m, 7 mN / m, 8 mN / m, 9 mN / m, 10 mN / m, 15 mN / m, 20 mN / m, 25 mN / m, 30 mN / m, 35 mN / m, 40 mN / m, 45 mN / m, 50 mN / m, 55 mN / m, 60 mN / m, 65 mN / m, 70 mN / m, 75 mN / m, 80 mN / m, 85 mN / m, 90 mN / m, 95 mN / m, 100 mN / m, 110 mN / m, 120 mN / m, 130 mN / m, 140 mN / m, 150 mN / m, 160 mN / m, 170 mN / m, 180 mN / m, 190 mN / m, 200 mN / m, 210 mN / m, 220 mN / m, 230 mN / m, 240 mN / m, 250 mN / m, 260 mN / m, 270 mN / m, 280 mN / m, 290 mN / m, or 300 mN / m.

[0114] In some cases, the biosurfactant composition at CMC at 9% and at 25 °C, using n-decane as a reference external liquid phase, may have an interfacial tension of at least about: 0.1 millinewton / meter (mN / m), 0.2 mN / m, 0.3 mN / m, 0.4 mN / m, 0.5 mN / m, 0.6 mN / m, 0.7 mN / m, 0.8 mN / m, 0.9 mN / m, 1 mN / m, 2 mN / m, 3 mN / m, 4 mN / m, 5 mN / m, 6 mN / m, 7 mN / m, 8 mN / m, 9 mN / m, 10 mN / m, 15 mN / m, 20 mN / m, 25 mN / m, 30 mN / m, 35 mN / m, 40 mN / m, 45WSGR Docket No.: 69849-701.601mN / m, 50 mN / m, 55 mN / m, 60 mN / m, 65 mN / m, 70 mN / m, 75 mN / m, 80 mN / m, 85 mN / m, 90 mN / m, 95 mN / m, 100 mN / m, 110 mN / m, 120 mN / m, 130 mN / m, 140 mN / m, 150 mN / m, 160 mN / m, 170 mN / m, 180 mN / m, 190 mN / m, 200 mN / m, 210 mN / m, 220 mN / m, 230 mN / m, 240 mN / m, 250 mN / m, 260 mN / m, 270 mN / m, 280 mN / m, 290 mN / m, 300 mN / m or more. In some cases, the biosurfactant composition at CMC at 9% and at 25 °C, using n-decane as a reference external liquid phase, may have an interfacial tension of at most about: 0.1 millinewton / meter (mN / m), 0.2 mN / m, 0.3 mN / m, 0.4 mN / m, 0.5 mN / m, 0.6 mN / m, 0.7 mN / m, 0.8 mN / m, 0.9 mN / m, 1 mN / m, 2 mN / m, 3 mN / m, 4 mN / m, 5 mN / m, 6 mN / m, 7 mN / m, 8 mN / m, 9 mN / m, 10 mN / m, 15 mN / m, 20 mN / m, 25 mN / m, 30 mN / m, 35 mN / m, 40 mN / m, 45 mN / m, 50 mN / m, 55 mN / m, 60 mN / m, 65 mN / m, 70 mN / m, 75 mN / m, 80 mN / m, 85 mN / m, 90 mN / m, 95 mN / m, 100 mN / m, 110 mN / m, 120 mN / m, 130 mN / m, 140 mN / m, 150 mN / m, 160 mN / m, 170 mN / m, 180 mN / m, 190 mN / m, 200 mN / m, 210 mN / m, 220 mN / m, 230 mN / m, 240 mN / m, 250 mN / m, 260 mN / m, 270 mN / m, 280 mN / m, 290 mN / m, or 300 mN / m.

[0115] In some cases, the salinity stability of the biosurfactant composition may be maintained at a concentration of sodium chloride that is at least about: 1 %, 2 %, 3 %, 4 %, 5 %, 10 %, 15 %, 20 %, 25 %, 30 %, 35 %, 40 %, 45 %, 50 %, 55 %, 60 %, 65 %, 70 %, 75 %, 80 %, 85 %, 90 %, 95 %, 96 %, 97 %, 98 %, or 99 %. In some cases, the salinity stability of the biosurfactant composition may be maintained at a concentration of sodium chloride that is at most about: 1 %, 2 %, 3 %, 4 %, 5 %, 10 %, 15 %, 20 %, 25 %, 30 %, 35 %, 40 %, 45 %, 50 %, 55 %, 60 %, 65 %, 70 %, 75 %, 80 %, 85 %, 90 %, 95 %, 96 %, 97 %, 98 %, or 99 %.

[0116] In some cases, at 20 °C, the biosurfactant composition may have a maximum sodium chloride solubility of at least about: 0.01, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, or 1000, gram per 100 mL. In some cases, at 20 °C, the biosurfactant composition may have a maximum sodium chloride solubility of at most about: 0.01, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, or 1000, gram per 100 mL.

[0117] In some cases, the biosurfactant composition may have an emulsion stability of at least about: 1 %, 2 %, 3 %, 4 %, 5 %, 10 %, 15 %, 20 %, 25 %, 30 %, 35 %, 40 %, 45 %, 50 %, 55 %,WSGR Docket No.: 69849-701.60160 %, 65 %, 70 %, 75 %, 80 %, 85 %, 90 %, 95 %, 96 %, 97 %, 98 %, or 99 %. In some cases, the biosurfactant composition may have an emulsion stability of at most about: 1 %, 2 %, 3 %, 4 %, 5 %, 10 %, 15 %, 20 %, 25 %, 30 %, 35 %, 40 %, 45 %, 50 %, 55 %, 60 %, 65 %, 70 %, 75 %, 80 %, 85 %, 90 %, 95 %, 96 %, 97 %, 98 %, or 99 %. The emulsion stability of the biosurfactant is at least about: 0.01 %, 0.1 %, 1%, 2 %, 3 %, 4 %, 5 %, 6 %, 7 %, 8 %, 9 %, 10 %, 11 %, 12 %, 13 %, 14 %, 15 %, 20 %, 25 %, 30 %, 35 %, 40 %, 50 %, 60 %, 70 %, 80 %, 90 %, 95 %, 100 %, 150 %, 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, 100-fold, or 1000-fold, higher than that of a control biosurfactant composition. The emulsion stability of the biosurfactant is at most about: 0.01 %, 0.1 %, 1%, 2 %, 3 %, 4 %, 5 %, 6 %, 7 %, 8 %, 9 %, 10 %, 11 %, 12 %, 13 %, 14 %, 15 %, 20 %, 25 %, 30 %, 35 %, 40 %, 50 %, 60 %, 70 %, 80 %, 90 %, 95 %, 100 %, 150 %, 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, 100-fold, or 1000-fold, higher than that of a control biosurfactant composition

[0118] In some cases, the biosurfactant composition may have an emulsion stability as described herein for at least about: 1 minute, 2 minutes, 3 minutes, 4 minutes, 5 minutes, 6 minutes, 7 minutes, 8 minutes, 9 minutes, 10 minutes, 11 minutes, 12 minutes, 13 minutes, 14 minutes, 15 minutes, 16 minutes, 17 minutes, 18 minutes, 19 minutes, 20 minutes, 21 minutes, 22 minutes, 23 minutes, 24 minutes, 25 minutes, 26 minutes, 27 minutes, 28 minutes, 29 minutes, 30 minutes, 31 minutes, 32 minutes, 33 minutes, 34 minutes, 35 minutes, 36 minutes, 37 minutes, 38 minutes, 39 minutes, 40 minutes, 41 minutes, 42 minutes, 43 minutes, 44 minutes, 45 minutes, 46 minutes, 47 minutes, 48 minutes, 49 minutes, 50 minutes, 51 minutes, 52 minutes, 53 minutes, 54 minutes, 55 minutes, 56 minutes, 57 minutes, 58 minutes, 59 minutes, 60 minutes, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, 18 hours, 19 hours, 20 hours, 21 hours, 22 hours, 23 hours, 24 hours, 25 hours, 26 hours, 27 hours, 28 hours, 29 hours, 30 hours, 31 hours, 32 hours, 33 hours, 34 hours, 35 hours, 36 hours, 37 hours, 38 hours, 39 hours, 40 hours, 41 hours, 42 hours, 43 hours, 44 hours, 45 hours, 46 hours, 47 hours, 48 hours, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months or more. In some cases, the biosurfactant composition may have an emulsion stability as described herein for at most about: 1 minute, 2 minutes, 3 minutes, 4 minutes, 5 minutes, 6 minutes, 7 minutes, 8 minutes, 9 minutes, 10 minutes, 11 minutes, 12 minutes, 13 minutes, 14 minutes, 15 minutes, 16 minutes, 17 minutes, 18 minutes, 19 minutes, 20 minutes, 21 minutes, 22 minutes, 23 minutes, 24 minutes, 25 minutes, 26 minutes, 27 minutes, 28 minutes, 29 minutes, 30 minutes, 31 minutes, 32 minutes, 33 minutes, 34 minutes, 35 minutes, 36 minutes, 37 minutes, 38 minutes, 39 minutes, 40 minutes, 41 minutes, 42 minutes, 43 minutes, 44 minutes, 45 minutes, 46WSGR Docket No.: 69849-701.601minutes, 47 minutes, 48 minutes, 49 minutes, 50 minutes, 51 minutes, 52 minutes, 53 minutes, 54 minutes, 55 minutes, 56 minutes, 57 minutes, 58 minutes, 59 minutes, 60 minutes, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, 18 hours, 19 hours, 20 hours, 21 hours, 22 hours, 23 hours, 24 hours, 25 hours, 26 hours, 27 hours, 28 hours, 29 hours, 30 hours, 31 hours, 32 hours, 33 hours, 34 hours, 35 hours, 36 hours, 37 hours, 38 hours, 39 hours, 40 hours, 41 hours, 42 hours, 43 hours, 44 hours, 45 hours, 46 hours, 47 hours, 48 hours, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, or 12 months.

[0119] In some cases, when using glass as the reference hydrophilic surface, the biosurfactant composition has a contact angle of at least about: 1 °, 2 °, 3 °, 4 °, 5 °, 6 °, 7 °, 8 °, 9 °, 10 °, 11 °, 12 °, 13 °, 14 °, 15 °, 16 °, 17 °, 18 °, 19 °, 20 °, 21 °, 22 °, 23 °, 24 °, 25 °, 26 °, 27 , 28 °, 29 °, 30 °, 31 °, 32 °, 33 °, 34 °, 35 °, 36 °, 37 °, 38 °, 39 °, 40 °, 41 °, 42 °, 43 °, 44 °, 45 °, 46 °, 47 °, 48 °, 49 °, 50 °, 51 °, 52 °, 53 °, 54 °, 55 °, 56 °, 57 °, 58 °, 59 °, 60 °, 61 °, 62 °, 63 , 64 °, 65 °, 66 °, 67 °, 68 °, 69 °, 70 °, 71 °, 72 °, 73 °, 74 °, 75 °, 76 °, 77 °, 78 °, 79 °, 80 °, 81 , 82 °, 83 °, 84 °, 85 °, 86 °, 87 °, 88 °, 89 °, 90 °, 91 °, 92 °, 93 °, 94 °, 95 °, 96 °, 97 °, 98 °, 99100 °, 101 °, 102 °, 103 °, 104 °, 105 °, 106 °, 107 °, 108 °, 109 °, 110 °, 111 °, 112 °, 113 °, 114 °, 115 °, 116 °, 117 °, 118 °, 119 °, 120 °, 121 °, 122 °, 123 °, 124 °, 125 °, 126 °, 127 °, 128 °, 129 °, 130 °, 131 °, 132 °, 133 °, 134 °, 135 °, 136 °, 137 °, 138 °, 139 °, 140 °, 141 °, 142 °, 143 °, 144 °, 145 °, 146 °, 147 °, 148 °, 149 °, 150 °, 151 °, 152 °, 153 °, 154 °, 155 °, 156 °, 157 °, 158 °, 159 °, 160 °, 161 °, 162 °, 163 °, 164 °, 165 °, 166 °, 167 °, 168 °, 169 °, 170 °, 171 °, 172 °, 173 °, 174 °, 175 °, 176 °, 177 °, 178 °, 179 °, 180 °, 181 °, 182 °, 183 °, 184 °, 185 °, 186 °, 187 °, 188 °, 189 °, 190 °, 191 °, 192 °, 193 °, 194 °, 195 °, 196 °, 197 °, 198 °, 199 °, 200 °, 201 °, 202 °, 203 °, 204 °, 205 °, 206 °, 207 °, 208 °, 209 °, 210 °, 211 °, 212 °, 213 °, 214 °, 215 °, 216 °, 217 °, 218 o 219 °, 220 o 221 °, 222 o 223 °, 224 °, 225 o 226 °, 227 o 228 °, 229 o 230 °, 231 °, 232 o 233 °, 234 °, 235 °, 236 °, 237 °, 238 o239o 240 °, 241 °, 242 °, 243 °, 244 °, 245 °, 246 °, 247 °, 248 °, 249 °, 250 °, 251 °, 252 °, 253 °, 254 °, 255 °, 256 °, 257 °, 258 °, 259 °, 260 °, 261 °, 262 °, 263 °, 264 °, 265 °, 266 °, 267 °, 268 °, 269 °, 270 °, 271 °, 272 °, 273 °, 274 °, 275 °, 276 °, 277 °, 278 °, 279 °, 280 °, 281 °, 282 °, 283 °, 284 °, 285 °, 286 °, 287 °, 288 °, 289 °, 290 °, 291 °, 292 °, 293 °, 294 °, 295 °, 296 °, 297 °, 298 °, 299 °, 300 °, 301 °, 302 °, 303 °, 304 °, 305 °, 306 °, 307 °, 308 °, 309 °, 310 °, 311 °, 312 °, 313 °, 314 °, 315 °, 316 °, 317 °, 318 °, 319 °, 320 °, 321 °, 322 °, 323 °, 324 °, 325 °, 326 °, 327 °, 328 °, 329 °, 330 °, 331 °, 332 °, 333 °, 334 °, 335 °, 336 °, 337 °, 338 °, 339 °, 340 °, 341 °, 342 °, 343 °, 344 °, 345 °, 346 °, 347 °, 348 °, 349 °, 350 °, 351 °, 352 °, 353 °, 354 °, 355 °, 356 °, 357 °, 358 °, 359 °, or 360 °. In some cases, when using glassWSGR Docket No.: 69849-701.601as the reference hydrophilic surface, the biosurfactant composition has a contact angle of at most about: 1 °, 2 °, 3 °, 4 °, 5 °, 6 °, 7 °, 8 °, 9 °, 10 °, 11 °, 12 °, 13 °, 14 °, 15 °, 16 °, 17 °, 18 °, 19 °, 20 °, 21 °, 22 °, 23 °, 24 °, 25 °, 26 °, 27 °, 28 °, 29 °, 30 °, 31 °, 32 °, 33 °, 34 °, 35 °, 36 °, 37 °, 38 °, 39 °, 40 °, 41 °, 42 °, 43 °, 44 °, 45 °, 46 °, 47 °, 48 °, 49 °, 50 °, 51 °, 52 °, 53 °, 54 °, 55 °, 56 °, 57 °, 58 °, 59 °, 60 °, 61 °, 62 °, 63 °, 64 °, 65 °, 66 °, 67 °, 68 °, 69 °, 70 °, 71 °, 72 °, 73 °, 74 °, 75 °, 76 °, 77 °, 78 °, 79 °, 80 °, 81 °, 82 °, 83 °, 84 °, 85 °, 86 °, 87 °, 88 °, 89 °, 90 °, 91 °, 92 °, 93 °, 94 °, 95 °, 96 °, 97 °, 98 °, 99 °, 100 °, b 31 °, 102 °, 103 °, 10' 1 °, 105 °, 106 °, 107 °, 108 °, 109 °, 110 °, 111 °, 112 °, 113 °, 114 °, 115 °, 116 °, 117 °, 118 °, 119 °, 120 °, 121 °, 122 °, 123 °, 124 °, 125 °, 126 °, 127 °, 128 °, 129 °, 130 °, 131 °, 132 °, 133 °, 134 °, 135 °, 136 °, 137 °, 138 °, 139 °, 140 °, 141 °, 142 °, 143 °, 144 °, 145 °, 146 °, 147 °, 148 °, 149 °, 150 °, 151 °, 152 °, 153 °, 154 °, 155 °, 156 °, 157 °, 158 °, 159 °, 160 °, 161 °, 162 °, 163 °, 164 °, 165 °, 166 °, 167 °, 168 °, 169 °, 170 °, 171 °, 172 °, 173 °, 174 °, 175 °, 176 °, 177 °, 178 °, 179 °, 180 °, 181 °, 182 °, 183 °, 184 °, 185 °, 186 °, 187 °, 188 °, 189 °, 190 °, 191 °, 192 °, 193 °, 194 °, 195 °, 196 °, 197 °, 198 °, 199 °, 200 °, 201 °, 202 °, 203 °, 204 °, 205 °, 206 °, 207 °, 208 °, 209 °, 210 °, 211 °, 212 °, 213 °, 214 °, 215 °, 216 °, 217 °, 218 °, 219 °, 220 221 °, 222 223 °, 224 °, 225 226 °, 227 228 °, 229 230 °, 231 o 232 °, 233 °, 234 °, 235 °, 236 °, 237 °, 238 °, 239 °, 240 °, 241 °, 242 °, 243 °, 244 °, 245 °, 246 °, 247 °, 248 249 °, 250 251 °, 252 °, 253 254 °, 255 256 °, 257 258 °, 259 260 261 °, 262 263 °, 264 265 °, 266 °, 267 268 °, 269 270 °, 271 272 °, 273 274 275 °, 276 277 °, 278 279 °, 280 °, 281 282 °, 283 284 °, 285 286 °, 287 288 289 °, 290 291 °, 292 293 °, 294 °, 295 296 °, 297 298 °, 299 300 °, 301 302 303 °, 304 305 °, 306 307 °, 308 °, 309 310 °, 311 312 °, 313 314 °, 315 316 317 °, 318 319 °, 320 321 °, 322 °, 323 324 °, 325 326 °, 327 328 °, 329 330 331 , 332 °, 333 °, 334 °, 335 °, 336 °, 337 °, 338 °, 339 °, 340 °, 341 °, 342 °, 343 ', 344 °, 345 , 346 °, 347 °, 348 °, 349 °, 350 °, 351 °, 352 °, 353 °, 354 °, 355 °, 356 °, 357 ', 358 °, 359 °, or 360 °.

[0120] In some cases, when using polytetrafluoroethylene / PTFE as the reference hydrophobic surface, the biosurfactant composition has a contact angle of at least about: 1 °, 2 °, 3 °, 4 °, 5 °, 6 °, 7 °, 8 °, 9 °, 10 °, 11 °, 12 °, 13 °, 14 °, 15 °, 16 °, 17 °, 18 °, 19 °, 20 °, 21 °, 22 °, 23 °, 24 °, 25 °, 26 °, 27 °, 28 °, 29 °, 30 °, 31 °, 32 °, 33 °, 34 °, 35 °, 36 °, 37 °, 38 °, 39 °, 40 °, 41 °, 42 °, 43 44 45 46 47 °, 48 °, 49 °, 50 °, 51 °, 52 °, 53 °, 54 °, 55 °, 56 °, 57 °, 58 °, 59 °, 60 °, 61 °, 62 °, 63 °, 64 °, 65 °, 66 °, 67 °, 68 °, 69 °, 70 °, 71 °, 72 °, 73 °, 74 °, 75 °, 76 °, 77 °, 78 °, 79 °, 80 °, 81 °, 82 °, 83 °, 84 °, 85 °, 86 °, 87 °, 88 °, 89 °, 90 °, 91 °, 92 °, 93 °, 94 °, 95 °, 96 °, 97 °, 98 °, 99 °, 100 °, 101 °, 102 °, 103 °, 104 °, 105 °, 106 °, 107 °, 108 °, 109 °, 110 °, 111 °, 112 °, 113 °, 114 °, 115 °, 116 °, 117 °, 118 °, 119 °, 120 °, 121 °, 122 °, 123 °, 124 °, 125 °,WSGR Docket No.: 69849-701.601126 °, 127 128 °, 129 130 °, 131 °, 132 133 °, 134 135 °, 136 137 °, 138 139 °, 140 °, 141 142 °, 143 144 °, 145 °, 146 147 °, 148 149 °, 150 151 °, 152 153 °, 154 °, 155 156 °, 157 158 °, 159 °, 160 161 °, 162 163 °, 164 165 °, 166 167 °, 168 °, 169 170 °, 171 172 °, 173 °, 174 175 °, 176 177 °, 178 179 °, 180 181 182 °, 183 184 °, 185 186 °, 187 °, 188 189 °, 190 191 °, 192 193 °, 194 195 196 °, 197 198 °, 199 200 °, 201 °, 202 203 °, 204 205 °, 206 207 °, 208 209 210 °, 211 212 °, 213 214 °, 215 °, 216 217 °, 218 219 °, 220 221 °, 222 223 224 °, 225 226 °, 227 228 °, 229 °, 230 231 °, 232 233 °, 234 235 °, 236 237 238 °, 239 240 °, 241 242 °, 243 °, 244 245 °, 246 247 °, 248 249 °, 250 251 252 °, 253 254 °, 255 256 °, 257 °, 258 259 °, 260 261 °, 262 263 °, 264 265 266 °, 267 268 °, 269 270 °, 271 °, 272 273 °, 274 275 °, 276 277 °, 278 279 280 °, 281 282 °, 283 284 °, 285 °, 286 287 °, 288 289 °, 290 291 °, 292 293 294 °, 295 296 °, 297 298 °, 299 °, 300 301 °, 302 303 °, 304 305 °, 306 307 308 °, 309 310 °, 311 312 °, 313 °, 314 315 °, 316 317 °, 318 319 °, 320 321 322 °, 323 324 °, 325 326 °, 327 °, 328 329 °, 330 331 °, 332 333 °, 334 335 336 °, 337 338 °, 339 340 °, 341 °, 342 343 °, 344 345 °, 346 347 °, 348 349 350 °, 351 352 °, 353 354 °, 355 °, 356 357 °, 358 359 °, or 360 °. In some cases, when using polytetrafluoroethylene / PTFE as the reference hydrophobic surface, the biosurfactant composition has a contact angle of at most about: 1 °, 2 °, 3 °, 4 °, 5 °, 6 °, 7 °, 8 °, 9 °, 10 °, 11 °, 12 13 °, 14 15 16 17 °, 18 °, 19 20 °, 21 22 °, 23 24 °, 25 26 °, 27 °, 28 29 °, 30 31 °, 32 33 34 35 36 °, 37 38 °, 39 40 °, 41 42 °, 43 44 °, 45 46 47 °, 48 49 °, 50 51 52 53 54 °, 55 56 °, 57 58 °, 59 60 °, 61 62 °, 63 64 65 °, 66 67 °, 68 69 °, 70 71 72 °, 73 74 °, 75 76 °, 77 78 °, 79 80 °, 81 82 83 °, 84 85 °, 86 87 °, 88 89 °, 90 °, 91 92 °, 93 94 °, 95 96 °, 97 98 °, 99 °, 100 °, 101 102 °, 103 104 °, 105 °, 106 107 °, 108 109 °, 110 111 °, 112 113 114 °, 115 116 °, 117 118 °, 119 °, 120 121 °, 122 123 °, 124 125 °, 126 127 128 °, 129 130 °, 131 132 °, 133 °, 134 135 °, 136 137 °, 138 139 °, 140 141 142 °, 143 144 °, 145 146 °, 147 °, 148 149 °, 150 151 °, 152 153 °, 154 155 156 °, 157 158 °, 159 160 °, 161 °, 162 163 °, 164 165 °, 166 167 °, 168 169 170 °, 171 172 °, 173 174 °, 175 °, 176 177 °, 178 179 °, 180 181 °, 182 183 184 °, 185 186 °, 187 188 °, 189 °, 190 191 °, 192 193 °, 194 195 °, 196 197 °, 198 °, 199 200 °, 201 202 °, 203 °, 204 205 °, 206 207 °, 208 209 °, 210 211 212 °, 213 214 °, 215 216 °, 217 °, 218 219 °, 220 221 °, 222 223 °, 224 225 226 °, 227 228 °, 229 230 °, 231 °, 232 233 °, 234 235 °, 236 237 °, 238 239 240 °, 241 242 °, 243 244 °, 245 °, 246 247 °, 248 249 °, 250 251 °, 252 253WSGR Docket No.: 69849-701.601254 °, 255 °, 256 °, 257 °, 258 °, 259 °, 260 °, 261 °, 262 °, 263 °, 264 °, 265 °, 266 °, 267 °, 268 °, 269 °, 270 °, 271 °, 272 °, 273 °, 274 °, 275 °, 276 °, 277 °, 278 °, 279 °, 280 °, 281 °, 282 °, 283 °, 284 °, 285 °, 286 °, 287 °, 288 °, 289 °, 290 °, 291 °, 292 °, 293 °, 294 °, 295 °, 296 °, 297 °, 298 °, 299 °, 300 °, 301 °, 302 °, 303 °, 304 °, 305 °, 306 °, 307 °, 308 °, 309 °, 310 °, 311 °, 312 °, 313 °, 314 °, 315 °, 316 °, 317 °, 318 °, 319 °, 320 °, 321 °, 322 °, 323 °, 324 °, 325 °, 326 °, 327 °, 328 °, 329 °, 330 °, 331 °, 332 °, 333 °, 334 °, 335 °, 336 °, 337 °, 338 °, 339 °, 340 °, 341 °, 342 °, 343 °, 344 °, 345 °, 346 °, 347 °, 348 °, 349 °, 350 °, 351 °, 352 °, 353 °, 354 °, 355 °, 356 °, 357 °, 358 °, 359 °, or 360 °.

[0121] In some cases, the hydrophilic-lipophilic balance (HLB) of the biosurfactant composition is at least about: 0.01, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 100. In some cases, the hydrophilic-lipophilic balance (HLB) of the biosurfactant composition is at most about: 0.01, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 100.

[0122] In some cases, the pour point of the biosurfactant composition is at least about: -20 °C, -15 °C, -10 °C, -9 °C, -8 °C, -7 °C, -6 °C, -5 °C, -4 °C, -3 °C, -2 °C, -1 °C, 0 °C, 1 °C, 2 °C, 3 °C, 4 °C, 5 °C, 6 °C, 7 °C, 8 °C, 9 °C, 10 °C, 11 °C, 12 °C, 13 °C, 14 °C, 15 °C, 16 °C, 17 °C, 18 °C, 19 °C, 20 °C, 21 °C, 22 °C, 23 °C, 24 °C, 25 °C, 26 °C, 27 °C, 28 °C, 29 °C, 30 °C, 31 °C, 32 °C, 33 °C, 34 °C, 35 °C, 36 °C, 37 °C, 38 °C, 39 °C, 40 °C, 41 °C, 42 °C, 43 °C, 44 °C, 45 °C, 46 °C, 47 °C, 48 °C, 49 °C, 50 °C or more. In some cases, the pour point of the biosurfactant composition is at most about: -20 °C, -15 °C, -10 °C, -9 °C, -8 °C, -7 °C, -6 °C, - 5 °C, -4 °C, -3 °C, -2 °C, -1 °C, 0 °C, 1 °C, 2 °C, 3 °C, 4 °C, 5 °C, 6 °C, 7 °C, 8 °C, 9 °C, 10 °C, 11 °C, 12 °C, 13 °C, 14 °C, 15 °C, 16 °C, 17 °C, 18 °C, 19 °C, 20 °C, 21 °C, 22 °C, 23 °C, 24 °C, 25 °C, 26 °C, 27 °C, 28 °C, 29 °C, 30 °C. 31 °C, 32 °C, 33 °C, 34 °C, 35 °C, 36 °C, 37 °C, 38 °C, 39 °C, 40 °C, 41 °C, 42 °C, 43 °C, 44 °C, 45 °C, 46 °C, 47 °C, 48 °C, 49 °C, or 50 °C.Methods of using the biosurfactant compositions

[0123] In some cases, the biosurfactant compositions may be used in various applications. In some cases, the application may comprise petroleum / gas or oil applications, home cleaning, industrial cleaning, agro-industrial applications, food applications, cosmetic applications, pharmacological applications, or a combination thereof. In some cases, the application may comprise petroleum / gas or oil applications. In some cases, the application may comprise home cleaning. In some cases, the application may comprise industrial cleaning. In some cases, the application may comprise agro-industrial applications. In some cases, the application mayWSGR Docket No.: 69849-701.601comprise food applications. In some cases, the application may comprise cosmetic applications. In some cases, the application may comprise pharmacological applications.

[0124] In petroleum / gas or oil applications, the biosurfactant composition may be used in different phases of the crude oil processing cycle. In some cases, the biosurfactant composition may be used in the upstream process, midstream process, or downstream, process, or a combination thereof. In some cases, the upstream process may comprise well preparation and crude extraction to the surface. In the upstream process, the biosurfactant composition may be used as an emulsifier, a defoamer agent, a spacer surfactant, or a wetting agent, or a combination thereof. In the upstream process, the biosurfactant composition may be used as an emulsifier. In the upstream process, the biosurfactant composition may be used a defoamer agent. In the upstream process, the biosurfactant composition may be used as a spacer surfactant. In the upstream process, the biosurfactant composition may be used as a wetting agent. In the upstream process, the biosurfactant composition may be used for foam control, cleaning wells, hydraulic fracturing, or enhanced oil recovery (EOR), or a combination thereof. In the upstream process, the biosurfactant composition may be used for foam control. In the upstream process, the biosurfactant composition may be used for cleaning wells. In the upstream process, the biosurfactant composition may be used for hydraulic fracturing. In the upstream process, the biosurfactant composition may be used for EOR. In some cases, the midstream process may comprise transportation and refining.

[0125] In the midstream process, the biosurfactant composition may be used as a dispersing wax, paraffin, or a corrosion inhibitor, or a combination thereof. In the midstream process, the biosurfactant composition may be used as a dispersing wax. In the midstream process, the biosurfactant composition may be used as a paraffin. In the midstream process, the biosurfactant composition may be used as a corrosion inhibitor. In the midstream process, the biosurfactant composition may be used for pipeline cleaning, flow assurance, or high salinity water disposal, or a combination thereof. In the midstream process, the biosurfactant composition may be used for pipeline cleaning. In the midstream process, the biosurfactant composition may be used for flow assurance. In the midstream process, the biosurfactant composition may be used for high salinity water disposal. In some cases, the downstream process may comprise the final stage of the lifecycle. In the downstream process, the biosurfactant composition can be used in water treatment, polluted soil treatment, or cleaning wells, or a combination thereof. In the downstream process, the biosurfactant composition can be used in water treatment. In the downstream process, the biosurfactant composition can be used in polluted soil treatment. In the downstream process, the biosurfactant composition can be used in cleaning wells. In the downstream process, the biosurfactant composition can be used for TPH (Total PetroleumWSGR Docket No.: 69849-701.601Hydrocarbons) removal. In the downstream process, the biosurfactant composition can be used in oil recovery, hydrocarbon remediation, or drilling fluids.

[0126] In some cases, the biosurfactant compositions may be an emulsifying agent, preservative, adjuvant, foaming agent, anti-bacterial agent (bactericide or bacteriostatic agent), anti-microbial agent, anti-fungal agent, detergent, dispersant, solubilizer, soil amendment, or a combination thereof, in the applications as described herein. In some cases, the biosurfactant compositions may be an emulsifying agent in the applications as described herein. In some cases, the biosurfactant compositions may be a preservative in the applications as described herein. In some cases, the biosurfactant compositions may be an adjuvant in the applications as described herein. In some cases, the biosurfactant compositions may be a foaming agent in the applications as described herein. In some cases, the biosurfactant compositions may be an antibacterial agent (bactericide or bacteriostatic agent) in the applications as described herein. In some cases, the biosurfactant compositions may be an anti-microbial agent in the applications as described herein. In some cases, the biosurfactant compositions may be an anti-fungal agent in the applications as described herein. In some cases, the biosurfactant compositions may be a detergent in the applications as described herein. In some cases, the biosurfactant compositions may be a dispersant in the applications as described herein. In some cases, the biosurfactant compositions may be a solubilizer in the applications as described herein. In some cases, the biosurfactant compositions may be a soil amendment in the applications as described herein.

[0127] In some cases, the biosurfactant composition may be used to remove total petroleum hydrocarbons (TPH) from soil or water; remove perfluoroalkyl and polyfluoroalkyl substances (PF AS) from the soil or the water; remove or kill bacteria present within cosmetic products; remove or kill bacteria present within agro-industrial products; facilitate emulsification in the cosmetic products; facilitate emulsification in the pharmaceutical products; facilitate emulsification in the petroleum products, facilitate emulsification in the food products; facilitate stabilization of a vaccine; or a combination thereof. In some cases, the biosurfactant composition may be used to remove total petroleum hydrocarbons (TPH) from soil or water. In some cases, the biosurfactant composition may be used to remove perfluoroalkyl and polyfluoroalkyl substances (PF AS) from the soil or the water. In some cases, the biosurfactant composition may be used to remove or kill bacteria present within cosmetic products. In some cases, the biosurfactant composition may be used to remove or kill bacteria present within agro-industrial products. In some cases, the biosurfactant composition may be used to facilitate emulsification in the cosmetic products. In some cases, the biosurfactant composition may be used to facilitate emulsification in the pharmaceutical products. In some cases, the biosurfactant composition mayWSGR Docket No.: 69849-701.601be used to facilitate emulsification in the petroleum products, facilitate emulsification in the food products.

[0128] In some cases, home cleaning applications may comprise removing a hydrocarbon- containing substance from a domestic item or household item. A domestic item or household item may comprise a furniture, cloth, clothing, appliances, a vessel for cooking, drinking, or eating; or a combination thereof. A domestic item or household item may comprise a furniture. A domestic item or household item may comprise a cloth. A domestic item or household item may comprise a clothing. A domestic item or household item may comprise an appliance. A domestic item or household item may comprise a cooking, drinking, or eating vessel. The hydrocarbon-containing substance may comprise domestic stains. Domestic stains may comprise any colored substances to be removed from a domestic or household item. The hydrocarbon- containing substance may comprise grass extract, red wine, chocolate milk solution, organic soils, or a combination thereof. The hydrocarbon-containing substance may comprise grass extract. The hydrocarbon-containing substance may comprise red wine. The hydrocarbon- containing substance may comprise chocolate solution. The hydrocarbon-containing substance may comprise organic soils.Methods of manufacturing the biosurfactants

[0129] Provided herein are methods of making the biosurfactants or the composition comprising the biosurfactants as described herein. In some cases, the method may comprise (a) obtaining a crude extract from a microorganism and (b) generating formulation or composition comprising the biosurfactant as described herein using the crude extract. In some cases, the microorganism may comprise a fungus. In some cases, the fungus may be a filamentous fungus. In some cases, the fungus may be a non-filamentous fungus. In some cases, the fungus may comprise at least a species from the family Xylariaceae. In some cases, the fungus may comprise Entonaema liquescens. In some cases, the microorganism may comprise Candida lipolytic, Candida tropicalis I JCP0996, Candida sphaerica, Candida antarctica, Starmerella bombicola, Starmerella bombicola, Pseudozyma tsukubaensis, Rhodotorula paludigena, Aureobasidium pullulans, Rhizopus arrhizus. Fusarium sp. BS-8, Mucor hiemalis, Aspergillus niger, Cunninghamella echinulata, or a combination thereof.

[0130] In some cases, the microorganism may not be pathogenic. In some cases, the microorganism may not generate virulence factors. In some cases, the microorganism may not be genetically engineered. In some cases, the microorganism may be genetically engineered.

[0131] In some cases, the method may further comprise preparing an initial inoculum. Prior to the inoculation, the inoculum culture may comprise yeast extracts or nutrients for culturing the microbes. The inoculum culture may comprise at least about: 1 %, 2 %, 3 %, 4 %, 5 %, 6 %,WSGR Docket No.: 69849-701.6017 %, 8 %, 9 %, 10 %, 11 %, 12 %, 13 %, 14 %, 15 %, 16 %, 17 %, 18 %, 19 %, 20 %, 21 %, 22 %, 23 %, 24 %, 25 %, 26 %, 27 %, 28 %, 29 %, 30 %, 31 %, 32 %, 33 %, 34 %, 35 %, 36 %, 37 %, 38 %, 39 %, 40 %, 41 %, 42 %, 43 %, 44 %, 45 %, 46 %, 47 %, 48 %, 49 %, 50 %, 51 %, 52 %, 53 %, 54 %, 55 %, 56 %, 57 %, 58 %, 59 %, 60 %, 61 %, 62 %, 63 %, 64 %, 65 %, 66 %, 67 %, 68 %, 69 %, 70 %, 71 %, 72 %, 73 %, 74 %, 75 %, 76 %, 77 %, 78 %, 79 %, 80 %, 81 %, 82 %, 83 %, 84 %, 85 %, 86 %, 87 %, 88 %, 89 %, 90 %, 91 %, 92 %, 93 %, 94 %, 95 %, 96 %, 97 %, 98 %, or 99 % nutrients, by volume / volume or weight / volume, of the inoculum culture. The inoculum culture may comprise at most about: 1 %, 2 %, 3 %, 4 %, 5 %, 6 %, 7 %, 8 %, 9 %, 10 %, 11 %, 12 %, 13 %, 14 %, 15 %, 16 %, 17 %, 18 %, 19 %, 20 %, 21 %, 22 %, 23 %, 24 %, 25 %, 26 %, 27 %, 28 %, 29 %, 30 %, 31 %, 32 %, 33 %, 34 %, 35 %, 36 %, 37 %, 38 %, 39 %, 40 %, 41 %, 42 %, 43 %, 44 %, 45 %, 46 %, 47 %, 48 %, 49 %, 50 %, 51 %, 52 %, 53 %, 54 %, 55 %, 56 %, 57 %, 58 %, 59 %, 60 %, 61 %, 62 %, 63 %, 64 %, 65 %, 66 %, 67 %, 68 %, 69 %, 70 %, 71 %, 72 %, 73 %, 74 %, 75 %, 76 %, 77 %, 78 %, 79 %, 80 %, 81 %, 82 %, 83 %, 84 %, 85 %, 86 %, 87 %, 88 %, 89 %, 90 %, 91 %, 92 %, 93 %, 94 %, 95 %, 96 %, 97 %, 98 %, or 99 % nutrients, by volume / volume or weight / volume, of the inoculum culture. The nutrient may comprise a carbon source, nitrogen source, saccharides, amino acids, peptides, lipids, vitamins, metabolites, organic chemicals, or inorganic chemicals for support the viability or growth of an organism or microbe present within the inoculum culture. At inoculation, the inoculum culture may comprise at least about: 1, lxl0Al, lxlOA2, lxl0A3, lxlOA4, lxl0A5, lxlOA6, lxlOA7, lxl0A8, lxlOA9 cells. At inoculation, the inoculum culture may comprise at most about: 1, lxl0Al, lxlOA2, lxl0A3, lxlOA4, lxl0A5, lxlOA6, lxlOA7, lxl0A8, lxlOA9 cells.

[0132] The method of preparing the initial inoculum may comprise those described herein, such as those in Example 1. In some cases, the method may comprise preparing the fermentation culture using the initial inoculum. In some cases, an initial inoculum may comprise at least about: 0.001 %, 0.002 %, 0.005 %, 0.01 %, 0.02 %, 0.05 %, 0.1 %, 0.2 %, 0.3 %, 0.4 %, 0.5 %, 0.6 %, 0.7 %, 0.8 %, 0.9 %, 1 %, 1.5 %, 2 %, 2.5 %, 3 %, 3.5 %, 4 %, 4.5 %, 5 %, 5.5 %, 6 %, 6.5 %, 7 %, 7.5 %, 8 %, 8.5 %, 9 %, 9.5 %, 10 %, 20 %, 50 % or more by volume of the fermentation culture. In some cases, an initial inoculum may comprise at most about: 0.001 %, 0.002 %, 0.005 %, 0.01 %, 0.02 %, 0.05 %, 0.1 %, 0.2 %, 0.3 %, 0.4 %, 0.5 %, 0.6 %, 0.7 %, 0.8 %, 0.9 %, 1 %, 1.5 %, 2 %, 2.5 %, 3 %, 3.5 %, 4 %, 4.5 %, 5 %, 5.5 %, 6 %, 6.5 %, 7 %, 7.5 %, 8 %, 8.5 %, 9 %, 9.5 %, 10 %, 20 %, or 50 % by volume of the fermentation culture. In some instances, the fermentation culture may be at least about: 1 L, 5 L, 10 L, 20 L, 30 L, 40 L, 50 L, 60 L, 70 L, 80 L, 90 L, 100 L, 150 L, 200 L, 250 L, 300 L, 350 L, 400 L, 450 L, 500 L, 1000 L, 1500 L, 2000 L, 2500 L, 3000 L, 3500 L, 4000 L, 4500 L, 5000 L, 7500 L, 10000 L,WSGR Docket No.: 69849-701.60120000 L, 30000 L, 40000 L, 50000 L, 60000 L, 70000 L, 80000 L, 90000 L, 100000 L, 110000 L, 120000 L, 130000 L, 140000 L, 150000 L, 160000 L, 170000 L, 180000 L, 190000 L, 200000 L, 300000 L, 400000 L, 500000 L, 600000 L, 700000 L, 800000 L, 900000 L, 1000000 L or more in volume. In some instances, the fermentation culture may be at most about 1 L, 5 L, 10 L, 20 L, 30 L, 40 L, 50 L, 60 L, 70 L, 80 L, 90 L, 100 L, 150 L, 200 L, 250 L, 300 L, 350 L, 400 L, 450 L, 500 L, 1000 L, 1500 L, 2000 L, 2500 L, 3000 L, 3500 L, 4000 L, 4500 L, 5000 L, 7500 L, 10000 L, 20000 L, 30000 L, 40000 L, 50000 L, 60000 L, 70000 L, 80000 L, 90000 L, 100000 L, 110000 L, 120000 L, 130000 L, 140000 L, 150000 L, 160000 L, 170000 L, 180000 L, 190000 L, 200000 L, 300000 L, 400000 L, 500000 L, 600000 L, 700000 L, 800000 L, 900000 L, or 1000000 L in volume. In some instances, the fermentation culture may comprise at least about: 1, lxl0Al, lxlOA2, lxl0A3, lxlOA4, lxl0A5, lxlOA6, lxlOA7, lxl0A8, or lxlOA9 cells / mL of the fermentation culture. In some instances, the fermentation culture may comprise at most about: 1, lxl0Al, lxlOA2, lxl0A3, lxlOA4, lxl0A5, lxlOA6, lxlOA7, lxl0A8, or lxlOA9 cells / mL of the fermentation culture. In some instances, the fermentation culture may comprise at least about: 1, lxl0Al, lxlOA2, lxl0A3, lxlOA4, lxl0A5, lxlOA6, lxlOA7, lxl0A8, lxlOA9, lxl0All, lxlOA12, lxl0A13, lxlOA14, lxl0A15, lxlOA16, lxlOA17, lxl0A18, lxlOA19, lxl0A20 or more cells. In some instances, the fermentation culture may comprise at most about: 1, lxl0Al, lxlOA2, lxl0A3, lxlOA4, lxl0A5, lxlOA6, lxlOA7, lxl0A8, lxlOA9, lxl0All, lxlOA12, lxl0A13, lxlOA14, lxl0A15, lxlOA16, lxlOA17, lxl0A18, lxlOA19, or lxl0A20 cells.

[0133] In some cases, the method may further comprise fermenting the microorganism or subjecting the microorganism to fermentation. In some cases, the fermentation may comprise culturing the microorganism with a substrate. In some cases, the substrate may comprise starch. In some cases, the substrate may comprise an amount of starch that is at least about 1 %, 2 %, 3 %, 4 %, 5 %, 6 %, 7 %, 8 %, 9 %, 10 %, 10 %, 11 %, 12 %, 13 %, 14 %, 15 %, 16 %, 17 %, 18 %, 19 %, 20 %, 21 %, 22 %, 23 %, 24 %, 25 %, 26 %, 27 %, 28 %, 29 %, 30 %, 31 %, 32 %, 33 %, 34 %, 35 %, 36 %, 37 %, 38 %, 39 %, 40 %, 41 %, 42 %, 43 %, 44 %, 45 %, 46 %, 47 %, 48 %, 49 %, 50 %, 51 %, 52 %, 53 %, 54 %, 55 %, 56 %, 57 %, 58 %, 59 %, 60 %, 61 %, 62 %, 63 %, 64 %, 65 %, 66 %, 67 %, 68 %, 69 %, 70 %, 71 %, 72 %, 73 %, 74 %, 75 %, 76 %, 77 %, 78 %, 79 %, 80 %, 81 %, 82 %, 83 %, 84 %, 85 %, 86 %, 87 %, 88 %, 89 %, 90 %, 95 %, 99 % or more, by weight / weight or weight / volume, of the substrate. In some cases, the substrate may comprise an amount of starch that is at most about 1 %, 2 %, 3 %, 4 %, 5 %, 6 %, 7 %, 8 %, 9 %, 10 %, 10 %, 11 %, 12 %, 13 %, 14 %, 15 %, 16 %, 17 %, 18 %, 19 %, 20 %, 21 %, 22 %, 23 %, 24 %, 25 %, 26 %, 27 %, 28 %, 29 %, 30 %, 31 %, 32 %, 33 %, 34 %, 35 %, 36 %, 37 %, 38 %, 39 %, 40 %, 41 %, 42 %, 43 %, 44 %, 45 %, 46 %, 47 %, 48 %, 49 %, 50 %, 51 %, 52 %, 53 %, 54 %, 55 %, 56 %, 57 %, 58 %, 59 %, 60 %, 61 %, 62 %, 63 %, 64 %, 65 %, 66 %, 67 %,WSGR Docket No.: 69849-701.60168 %, 69 %, 70 %, 71 %, 72 %, 73 %, 74 %, 75 %, 76 %, 77 %, 78 %, 79 %, 80 %, 81 %, 82 %, 83 %, 84 %, 85 %, 86 %, 87 %, 88 %, 89 %, 90 %, 95 %, or 99 %, by weight / weight or weight / volume, of the substrate. In some cases, the substrate may comprise agro-industrial waste or renewable source. In some cases, renewable source may comprise residues from crops. The crops may comprise peels, rejected harvests, or byproducts from processing. The residues may be generated from cassava, potato, sweet potato, corn or other rich in starch crops.

[0134] In some cases, the substrate may comprise agro-industrial waste. In some cases, the substrate may comprise agro-industrial waste. In some cases, the substrate may comprise starch, amylopectin, pectin, lignin, cellulose, or a combination thereof. In some cases, the substrate may comprise starch. In some cases, the substrate may comprise amylopectin. In some cases, the substrate may comprise pectin. In some cases, the substrate may comprise lignin. In some cases, the substrate may comprise cellulose. In some cases, the fermentation culture may be maintained at a temperature that is at least at about: 0 °C, 1 °C, 2 °C, 3 °C, 4 °C, 5 °C, 6 °C, 7 °C, 8 °C, 9 °C, 10 °C, 11 °C, 12 °C, 13 °C, 14 °C, 15 °C, 16 °C, 17 °C, 18 °C, 19 °C, 20 °C, 21 °C, 22 °C, 23 °C, 24 °C, 25 °C, 26 °C, 27 °C, 28 °C, 29 °C, 30 °C, 31 °C, 32 °C, 33 °C, 34 °C, 35 °C, 36 °C, 37 °C, 38 °C, 39 °C, 40 °C, 41 °C, 42 °C, 43 °C, 44 °C, 45 °C, 46 °C, 47 °C, 48 °C, 49 °C, 50 °C or more. In some cases, the fermentation culture may be maintained at a temperature that is at most at about: 0 °C, 1 °C, 2 °C, 3 °C, 4 °C, 5 °C, 6 °C, 7 °C, 8 °C, 9 °C, 10 °C, 11 °C, 12 °C, 13 °C, 14 °C, 15 °C, 16 °C, 17 °C, 18 °C, 19 °C, 20 °C, 21 °C, 22 °C, 23 °C, 24 °C, 25 °C, 26 °C, 27 °C, 28 °C, 29 °C, 30 °C, 31 °C, 32 °C, 33 °C, 34 °C, 35 °C, 36 °C, 37 °C, 38 °C, 39 °C, 40 °C, 41 °C, 42 °C, 43 °C, 44 °C, 45 °C, 46 °C, 47 °C, 48 °C, 49 °C, or 50 °C more. In some cases, the fermentation culture may be maintained at a temperature that is: from about 18°C to about 30°C, from about 17.5°C to about 30.5°C, from about 17°C to about 31°C, from about 16.5°C to about 31.5°C, from about 16°C to about 32°C, from about 15.5°C to about 32.5°C, from about 15°C to about 33°C, from about 14.5°C to about 33.5°C, from about 14°C to about 34°C, from about 13.5°C to about 34.5°C, from about 13°C to about 35°C, from about 12.5°C to about 35.5°C, from about 12°C to about 36°C, from about 11.5°C to about 36.5°C, from about 11°C to about 37°C, from about 10.5°C to about 37.5°C, from about 10°C to about 38°C, from about 9.5°C to about 38.5°C, from about 9°C to about 39°C, from about 8.5°C to about 39.5°C, from about 8°C to about 40°C, from about 18°C to about 30.5°C, from about 18°C to about 31°C, from about 18°C to about 31.5°C, from about 18°C to about 32°C, from about 18°C to about 32.5°C, from about 18°C to about 33°C, from about 18°C to about 33.5°C, from about 18°C to about 34°C, from about 18°C to about 34.5°C, from about 18°C to about 35°C, from about 18°C to about 35.5°C, from about 18°C to about 36°C, from about 18°C to about 36.5°C, from about 18°C to about 37°C, from about 18°C toWSGR Docket No.: 69849-701.601about 37.5°C, from about 18°C to about 38°C, from about 18°C to about 38.5°C, from about 18°C to about 39°C, from about 18°C to about 39.5°C, from about 18°C to about 40°C, from about 17.5°C to about 30°C, from about 17°C to about 30°C, from about 16.5°C to about 30°C, from about 16°C to about 30°C, from about 15.5°C to about 30°C, from about 15°C to about 30°C, from about 14.5°C to about 30°C, from about 14°C to about 30°C, from about 13.5°C to about 30°C, from about 13°C to about 30°C, from about 12.5°C to about 30°C, from about 12°C to about 30°C, from about 11.5°C to about 30°C, from about 11°C to about 30°C, from about 10.5°C to about 30°C, from about 10°C to about 30°C, from about 9.5°C to about 30°C, from about 9°C to about 30°C, from about 8.5°C to about 30°C, or from about 8°C to about 30°C. In some cases, the fermentation culture may be maintained at a pH that is at least about: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 or more. In some cases, the fermentation culture may be maintained at a pH that is at most about: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14. In some cases, the fermentation culture may be maintained at a pH that is: from about 3 to about 6, from about 2.9 to about 6.1, from about 2.8 to about 6.2, from about 2.7 to about 6.3, from about 2.6 to about 6.4, from about 2.5 to about 6.5, from about 2.4 to about 6.6, from about 2.3 to about 6.7, from about 2.2 to about 6.8, from about 2.1 to about 6.9, from about 2 to about 7, from about 1.9 to about 7.1, from about 1.8 to about 7.2, from about 1.7 to about 7.3, from about 1.6 to about 7.4, from about 1.5 to about 7.5, from about 3 to about 7.6, from about 3 to about 6.2, from about 3 to about 6.3, from about 3 to about 6.4, from about 3 to about 6.5, from about 3 to about 6.6, from about 3 to about 6.7, from about 3 to about 6.8, from about 3 to about 6.9, from about 3 to about 7, from about 3 to about 7.1, from about 3 to about 7.2, from about 3 to about 7.3, from about 3 to about 7.4, from about 3 to about 7.5, from about 2.9 to about 7.6, from about 2.8 to about 6, from about 2.7 to about 6, from about 2.6 to about 6, from about 2.5 to about 6, from about 2.4 to about 6, from about 2.3 to about 6, from about 2.2 to about 6, from about 2.1 to about 6, from about 2 to about 6, from about 1.9 to about 6, from about 1.8 to about 6, from about 1.7 to about 6, from about 1.6 to about 6, or from about 1.5 to about 6.

[0135] In some cases, the fermentation culture may be subjected to aeration that is at least about: 0.02 vessel volumes per minute (vvm), 0.021 vvm, 0.022 vvm, 0.023 vvm, 0.024 vvm, 0.025 vvm, 0.026 vvm, 0.027 vvm, 0.028 vvm, 0.029 vvm, 0.03 vvm, 0.031 vvm, 0.032 vvm, 0.033 vvm, 0.034 vvm, 0.035 vvm, 0.036 vvm, 0.037 vvm, 0.038 vvm, 0.039 vvm, 0.04 vvm, 0.041 vvm, 0.042 vvm, 0.043 vvm, 0.044 vvm, 0.045 vvm, 0.046 vvm, 0.047 vvm, 0.048 vvm, 0.049 vvm, 0.05 vvm, 0.055 vvm, 0.06 vvm, 0.065 vvm, 0.07 vvm, 0.075 vvm, 0.08 vvm, 0.085 vvm, 0.09 vvm, 0.095 vvm, 0.1 vvm, 0.105 vvm, 0.11 vvm, 0.115 vvm, 0.12 vvm, 0.125 vvm, 0.13 vvm, 0.135 vvm, 0.14 vvm, 0.145 vvm, 0.15 vvm, 0.155 vvm, 0.16 vvm, 0.165 vvm, 0.17 vvm, 0.175 vvm, 0.18 vvm, 0.185 vvm, 0.19 vvm, 0.195 vvm, 0.2 vvm, 0.3 vvm, 0.4 vvm, 0.5WSGR Docket No.: 69849-701.601vvm, 0.6 vvm, 0.7 vvm, 0.8 vvm, 0.9 vvm, 1 vvm, 1.1 vvm, 1.2 vvm, 1.3 vvm, 1.4 vvm, 1.5 vvm, 1.6 vvm, 1.7 vvm, 1.8 vvm, 1.9 vvm, 2 vvm, 3 vvm, 4 vvm, 5 vvm, 6 vvm, 7 vvm, 8 vvm, 9 vvm, 10 vvm or more. In some cases the fermentation culture may be subjected to aeration that is at most about: 0.02 vvm, 0.021 vvm, 0.022 vvm, 0.023 vvm, 0.024 vvm, 0.025 vvm, 0.026 vvm, 0.027 vvm, 0.028 vvm, 0.029 vvm, 0.03 vvm, 0.031 vvm, 0.032 vvm, 0.033 vvm, 0.034 vvm, 0.035 vvm, 0.036 vvm, 0.037 vvm, 0.038 vvm, 0.039 vvm, 0.04 vvm, 0.041 vvm, 0.042 vvm, 0.043 vvm, 0.044 vvm, 0.045 vvm, 0.046 vvm, 0.047 vvm, 0.048 vvm, 0.049 vvm, 0.05 vvm, 0.055 vvm, 0.06 vvm, 0.065 vvm, 0.07 vvm, 0.075 vvm, 0.08 vvm, 0.085 vvm, 0.09 vvm, 0.095 vvm, 0.1 vvm, 0.105 vvm, 0.11 vvm, 0.115 vvm, 0.12 vvm, 0.125 vvm, 0.13 vvm, 0.135 vvm, 0.14 vvm, 0.145 vvm, 0.15 vvm, 0.155 vvm, 0.16 vvm, 0.165 vvm, 0.17 vvm, 0.175 vvm, 0.18 vvm, 0.185 vvm, 0.19 vvm, 0.195 vvm, 0.2 vvm, 0.3 vvm, 0.4 vvm, 0.5 vvm, 0.6 vvm, 0.7 vvm, 0.8 vvm, 0.9 vvm, 1 vvm, 1.1 vvm, 1.2 vvm, 1.3 vvm, 1.4 vvm, 1.5 vvm, 1.6 vvm, 1.7 vvm, 1.8 vvm, 1.9 vvm, 2 vvm, 3 vvm, 4 vvm, 5 vvm, 6 vvm, 7 vvm, 8 vvm, 9 wm, or 10 vvm. In some cases, the fermentation culture may be subjected to aeration that is: from about 0.2 vvm to about 1 vvm, from about 0.19 vvm to about 1.1 vvm, from about 0.18 vvm to about 1.2 vvm, from about 0.17 vvm to about 1.3 vvm, from about 0.16 vvm to about 1.4 vvm, from about 0.15 vvm to about 1.5 vvm, from about 0.14 vvm to about 1.6 vvm, from about 0.13 vvm to about 1.7 vvm, from about 0.12 vvm to about 1.8 vvm, from about 0.11 vvm to about 1.9 vvm, from about 0.1 vvm to about 2 vvm, from about 0.2 vvm to about 1.1 vvm, from about 0.2 vvm to about 1.2 vvm, from about 0.2 vvm to about 1.3 vvm, from about 0.2 vvm to about 1.4 vvm, from about 0.2 vvm to about 1.5 vvm, from about 0.2 vvm to about 1.6 vvm, from about 0.2 vvm to about 1.7 vvm, from about 0.2 vvm to about 1.8 vvm, from about 0.2 vvm to about 1.9 vvm, from about 0.2 vvm to about 2 vvm, from about 0.19 vvm to about 1 vvm, from about 0.18 vvm to about 1 vvm, from about 0.17 vvm to about 1 vvm, from about 0.16 vvm to about 1 vvm, from about 0.15 vvm to about 1 vvm, from about 0.14 vvm to about 1 vvm, from about 0.13 vvm to about 1 vvm, from about 0.12 vvm to about 1 vvm, from about 0.11 vvm to about 1 vvm, or from about 0.1 vvm to about 1 vvm. In some cases, the fermentation culture may be subjected to mechanical agitation that is at least about: 50 revolution per minute (rpm), 60 rpm, 70 rpm, 80 rpm, 90 rpm, 100 rpm, 110 rpm, 120 rpm, 130 rpm, 140 rpm, 150 rpm, 160 rpm, 170 rpm, 180 rpm, 190 rpm, 200 rpm, 210 rpm, 220 rpm, 230 rpm, 240 rpm, 250 rpm, 260 rpm, 270 rpm, 280 rpm, 290 rpm, 300 rpm, 310 rpm, 320 rpm, 330 rpm, 340 rpm, 350 rpm, 360 rpm, 370 rpm, 380 rpm, 390 rpm, 400 rpm, 410 rpm, 420 rpm, 430 rpm, 440 rpm, 450 rpm, 460 rpm, 470 rpm, 480 rpm, 490 rpm, 500 rpm or more. In some cases, the fermentation culture may be subjected to mechanical agitation that is at most about: 50 rpm, 60 rpm, 70 rpm, 80 rpm, 90 rpm, 100 rpm, 110 rpm, 120 rpm, 130 rpm, 140 rpm, 150 rpm, 160 rpm, 170 rpm, 180 rpm, 190 rpm, 200 rpm,WSGR Docket No.: 69849-701.601210 rpm, 220 rpm, 230 rpm, 240 rpm, 250 rpm, 260 rpm, 270 rpm, 280 rpm, 290 rpm, 300 rpm, 310 rpm, 320 rpm, 330 rpm, 340 rpm, 350 rpm, 360 rpm, 370 rpm, 380 rpm, 390 rpm, 400 rpm, 410 rpm, 420 rpm, 430 rpm, 440 rpm, 450 rpm, 460 rpm, 470 rpm, 480 rpm, 490 rpm, or 500 rpm. In some cases, the fermentation culture may be subjected to mechanical agitation that is: from about 100 rpm to about 200 rpm, from about 95 rpm to about 205 rpm, from about 90 rpm to about 210 rpm, from about 85 rpm to about 215 rpm, from about 80 rpm to about 220 rpm, from about 75 rpm to about 225 rpm, from about 70 rpm to about 230 rpm, from about 65 rpm to about 235 rpm, from about 60 rpm to about 240 rpm, from about 55 rpm to about 245 rpm, from about 50 rpm to about 250 rpm, from about 100 rpm to about 205 rpm, from about 100 rpm to about 210 rpm, from about 100 rpm to about 215 rpm, from about 100 rpm to about 220 rpm, from about 100 rpm to about 225 rpm, from about 100 rpm to about 230 rpm, from about 100 rpm to about 235 rpm, from about 100 rpm to about 240 rpm, from about 100 rpm to about 245 rpm, from about 100 rpm to about 250 rpm, from about 95 rpm to about 200 rpm, from about 90 rpm to about 200 rpm, from about 85 rpm to about 200 rpm, from about 80 rpm to about 200 rpm, from about 75 rpm to about 200 rpm, from about 70 rpm to about 200 rpm, from about 65 rpm to about 200 rpm, from about 60 rpm to about 200 rpm, from about 55 rpm to about 200 rpm, or from about 50 rpm to about 200 rpm.

[0136] In some cases, the microorganism is cultured with an inducer. In some cases, the inducer may facilitate growth of the microorganism, fermentation of the microorganism, or generating a biosurfactant composition or crude extract to generate the biosurfactant composition. In some cases, the inducer may comprise hydrocarbons, synthetic lubricating oils, mineral base lubricating oils, vegetable fatty acids, alkanes, alcohols, aldehydes, oils, synthetic oils, vegetable oils, animal oils, or a combination thereof. In some cases, the inducer may comprise hydrocarbons. In some cases, the inducer may comprise synthetic lubricating oils. In some cases, the inducer may comprise mineral base lubricating oils. In some cases, the inducer may comprise vegetable fatty acids. In some cases, the inducer may comprise alkanes. In some cases, the inducer may comprise alcohols. In some cases, the inducer may comprise aldehydes. In some cases, the inducer may comprise alcohols. In some cases, the inducer may comprise vegetable oils. In some cases, the inducer may comprise alcohols. In some cases, the inducer may comprise animal oils. In some cases, the inducer may comprise oils. In some cases, the inducer may comprise synthetic oils. In some cases, the inducer may be at least about: 0.01 %, 0.02 %, 0.03 %, 0.04 %, 0.05 %, 0.06 %, 0.07 %, 0.08 %, 0.09 %, 0.1 %, 0.11 %, 0.12 %, 0.13 %, 0.14 %, 0.15 %, 0.16 %, 0.17 %, 0.18 %, 0.19 %, 0.2 %, 0.21 %, 0.22 %, 0.23 %, 0.24 %, 0.25 %, 0.26 %, 0.27 %, 0.28 %, 0.29 %, 0.3 %, 0.31 %, 0.32 %, 0.33 %, 0.34 %, 0.35 %, 0.36 %, 0.37 %, 0.38 %, 0.39 %, 0.4 %, 0.41 %, 0.42 %, 0.43 %, 0.44 %, 0.45 %,WSGR Docket No.: 69849-701.6010.46 %, 0.47 %, 0.48 %, 0.49 %, 0.5 %, 0.6 %, 0.7 %, 0.8 %, 0.9 %, 1 %, 1.1 %, 1.2 %, 1.3 %, 1.4 %, 1.5 %, 1.6 %, 1.7 %, 1.8 %, 1.9 %, 2 %, 2.1 %, 2.2 %, 2.3 %, 2.4 %, 2.5 %, 2.6 %, 2.7 % 2.8 %, 2.9 %, 3 %, 3.1 %, 3.2 %, 3.3 %, 3.4 %, 3.5 %, 3.6 %, 3.7 %, 3.8 %, 3.9 %, 4 %, 4.1 %, 4.2 %, 4.3 %, 4.4 %, 4.5 %, 4.6 %, 4.7 %, 4.8 %, 4.9 %, 5 %, 5.1 %, 5.2 %, 5.3 %, 5.4 %, 5.5 %, 5.6 %, 5.7 %, 5.8 %, 5.9 %, 6 %, 6.1 %, 6.2 %, 6.3 %, 6.4 %, 6.5 %, 6.6 %, 6.7 %, 6.8 %, 6.9 %, 7 %, 7.1 %, 7.2 %, 7.3 %, 7.4 %, 7.5 %, 7.6 %, 7.7 %, 7.8 %, 7.9 %, 8 %, 8.1 %, 8.2 %, 8.3 %, 8.4 %, 8.5 %, 8.6 %, 8.7 %, 8.8 %, 8.9 %, 9 %, 9.1 %, 9.2 %, 9.3 %, 9.4 %, 9.5 %, 9.6 %, 9.7 %, 9.8 %, 9.9 %, 10 % or more, by weight / volume or weight / weight, of the culture. In some cases, the inducer may be at most about: 0.01 %, 0.02 %, 0.03 %, 0.04 %, 0.05 %, 0.06 %, 0.07 %, 0.08 %, 0.09 %, 0.1 %, 0.11 %, 0.12 %, 0.13 %, 0.14 %, 0.15 %, 0.16 %, 0.17 %, 0.18 %, 0.19 %, 0.2 %, 0.21 %, 0.22 %, 0.23 %, 0.24 %, 0.25 %, 0.26 %, 0.27 %, 0.28 %, 0.29 %, 0.3 %, 0.31 %, 0.32 %, 0.33 %, 0.34 %, 0.35 %, 0.36 %, 0.37 %, 0.38 %, 0.39 %, 0.4 %, 0.41 %, 0.42 %, 0.43 %, 0.44 %, 0.45 %, 0.46 %, 0.47 %, 0.48 %, 0.49 %, 0.5 %, 0.6 %, 0.7 %, 0.8 %, 0.9 %, 1 %, 1.1 %, 1.2 %, 1.3 %, 1.4 %, 1.5 %, 1.6 %, 1.7 %, 1.8 %, 1.9 %, 2 %, 2.1 %, 2.2 %, 2.3 %, 2.4 %, 2.5 %, 2.6 %, 2.7 %, 2.8 %, 2.9 %, 3 %, 3.1 %, 3.2 %, 3.3 %, 3.4 %, 3.5 %, 3.6 %, 3.7 %, 3.8 %, 3.9 %, 4 %, 4.1 %, 4.2 %, 4.3 %, 4.4 %, 4.5 %, 4.6 %, 4.7 %, 4.8 %, 4.9 %, 5 %, 5.1 %, 5.2 %, 5.3 %, 5.4 %, 5.5 %, 5.6 %, 5.7 %, 5.8 %, 5.9 %, 6 %, 6.1 %, 6.2 %, 6.3 %, 6.4 %, 6.5 %, 6.6 %, 6.7 %, 6.8 %, 6.9 %, 7 %, 7.1 %, 7.2 %, 7.3 %, 7.4 %, 7.5 %, 7.6 %, 7.7 %, 7.8 %, 7.9 %, 8 %, 8.1 %, 8.2 %, 8.3 %, 8.4 %, 8.5 %, 8.6 %, 8.7 %, 8.8 %, 8.9 %, 9 %, 9.1 %, 9.2 %, 9.3 %, 9.4 %, 9.5 %, 9.6 %, 9.7 %, 9.8 %, 9.9 %, or 10 %, by weight / volume or weight / weight, of the culture. In some cases, the inducer may be: from about 0.2 % to about 5 %, from about 0.19 % to about 5.1 %, from about 0.18 % to about 5.2 %, from about 0.17 % to about 5.3 %, from about 0.16 % to about 5.4 %, from about 0.15 % to about 5.5 %, from about 0.14 % to about 5.6 %, from about 0.13 % to about 5.7 %, from about 0.12 % to about 5.8 %, from about 0.11 % to about 5.9 %, from about 0.1 % to about 6 %, from about 0.09 % to about 6.1 %, from about 0.08 % to about 6.2 %, from about 0.07 % to about 6.3 %, from about 0.06 % to about 6.4 %, from about 0.05 % to about 6.5 %, from about 0.2 % to about 5.1 %, from about 0.2 % to about 5.2 %, from about 0.2 % to about 5.3 %, from about 0.2 % to about 5.4 %, from about 0.2 % to about 5.5 %, from about 0.2 % to about 5.6 %, from about 0.2 % to about 5.7 %, from about 0.2 % to about 5.8 %, from about 0.2 % to about 5.9 %, from about 0.2 % to about 6 %, from about 0.2 % to about 6.1 %, from about 0.2 % to about 6.2 %, from about 0.2 % to about 6.3 %, from about 0.2 % to about 6.4 %, from about 0.2 % to about 6.5 %, from about 0.19 % to about 5 %, from about 0.18 % to about 5 %, from about 0.17 % to about 5 %, from about 0.16 % to about 5 %, from about 0.15 % to about 5 %, from about 0.14 % to about 5 %, from about 0.13 % to about 5 %, from about 0.12 % to about 5 %, from about 0.11 % to about 5 %, fromWSGR Docket No.: 69849-701.601about 0.1 % to about 5 %, from about 0.09 % to about 5 %, from about 0.08 % to about 5 %, from about 0.07 % to about 5 %, from about 0.06 % to about 5 %, or from about 0.05 % to about 5 %, by weight / volume or weight / weight, of the culture. In some cases, the method may comprise concentrating the culture. In some cases, the method may comprise purifying the crude extract from the culture. In some cases, the purifying may comprise filtration or centrifugation. In some cases, the purifying may comprise centrifugation. In some cases, the purifying may comprise filtration. In some cases, the filtration may comprise vacuum filtration. In some cases, the filtration may comprise microfiltration. In some cases, the filtration may comprise macrofiltration. The pore-size of the filtration may be at least about: 1 nanoliter (nm), 2 nm, 3 nm, 4 nm, 5 nm, 6 nm, 7 nm, 8 nm, 9 nm, 10 nm, 20 nm, 30 nm, 40 nm, 50 nm, 60 nm, 70 nm, 80 nm, 90 nm, 100 nm, 200 nm, 300 nm, 400 nm, 500 nm, 600 nm, 700 nm, 800 nm, 900 nm, 1 micrometer (pm), 2 pm, 3 pm, 4 pm, 5 pm, 6 pm, 7 pm, 8 pm, 9 pm, 10 pm, 20 pm, 30 pm, 40 pm, 50 pm, 60 pm, 70 pm, 80 pm, 90 pm, 100 pm, 200 pm, 300 pm, 400 pm, 500 pm, 600 pm, 700 pm, 800 pm, 900 pm, 1 millimeter (mm), 2 mm, 3 mm, 4 mm, 5 mm, 6 mm, 7 mm, 8 mm, 9 mm, 10 mm, 20 mm, 30 mm, 40 mm, 50 mm, 60 mm, 70 mm, 80 mm, 90 mm, 100 mm, 200 mm, 300 mm, 400 mm, 500 mm, 600 mm, 700 mm, 800 mm, 900 mm, 1000 mm or more. The pore-size of the filtration may be at most about: 1 nanoliter (nm), 2 nm, 3 nm, 4 nm, 5 nm, 6 nm, 7 nm, 8 nm, 9 nm, 10 nm, 20 nm, 30 nm, 40 nm, 50 nm, 60 nm, 70 nm, 80 nm, 90 nm, 100 nm, 200 nm, 300 nm, 400 nm, 500 nm, 600 nm, 700 nm, 800 nm, 900 nm, 1 micrometer (pm), 2 pm, 3 pm, 4 pm, 5 pm, 6 pm, 7 pm, 8 pm, 9 pm, 10 pm, 20 pm, 30 pm, 40 pm, 50 pm, 60 pm, 70 pm, 80 pm, 90 pm, 100 pm, 200 pm, 300 pm, 400 pm, 500 pm, 600 pm, 700 pm, 800 pm, 900 pm, 1 millimeter (mm), 2 mm, 3 mm, 4 mm, 5 mm, 6 mm, 7 mm, 8 mm, 9 mm, 10 mm, 20 mm, 30 mm, 40 mm, 50 mm, 60 mm, 70 mm, 80 mm, 90 mm, 100 mm, 200 mm, 300 mm, 400 mm, 500 mm, 600 mm, 700 mm, 800 mm, 900 mm, or 1000 mm. The material being purified may have a size or diameter of at least about: 1 nanoliter (nm), 2 nm, 3 nm, 4 nm, 5 nm, 6 nm, 7 nm, 8 nm, 9 nm, 10 nm, 20 nm, 30 nm, 40 nm, 50 nm, 60 nm, 70 nm, 80 nm, 90 nm, 100 nm, 200 nm, 300 nm, 400 nm, 500 nm, 600 nm, 700 nm, 800 nm, 900 nm, 1 micrometer (pm), 2 pm, 3 pm, 4 pm, 5 pm, 6 pm, 7 pm, 8 pm, 9 pm, 10 pm, 20 pm, 30 pm, 40 pm, 50 pm, 60 pm, 70 pm, 80 pm, 90 pm, 100 pm, 200 pm, 300 pm, 400 pm, 500 pm, 600 pm, 700 pm, 800 pm, 900 pm, 1 millimeter (mm), 2 mm, 3 mm, 4 mm, 5 mm, 6 mm, 7 mm, 8 mm, 9 mm, 10 mm, 20 mm, 30 mm, 40 mm, 50 mm, 60 mm, 70 mm, 80 mm, 90 mm, 100 mm, 200 mm, 300 mm, 400 mm, 500 mm, 600 mm, 700 mm, 800 mm, 900 mm, 1000 mm or more. The material being purified may have a size or diameter of at most about: 1 nanoliter (nm), 2 nm, 3 nm, 4 nm, 5 nm, 6 nm, 7 nm, 8 nm, 9 nm, 10 nm, 20 nm, 30 nm, 40 nm, 50 nm, 60 nm, 70 nm, 80 nm, 90 nm, 100 nm, 200 nm, 300 nm, 400 nm, 500 nm, 600 nm, 700 nm, 800 nm, 900 nm, 1 micrometer (pm), 2 pm,WSGR Docket No.: 69849-701.6013 pm, 4 pm, 5 pm, 6 pm, 7 pm, 8 pm, 9 pm, 10 pm, 20 pm, 30 pm, 40 pm, 50 pm, 60 pm, 70 pm, 80 pm, 90 pm, 100 pm, 200 pm, 300 pm, 400 pm, 500 pm, 600 pm, 700 pm, 800 pm, 900 pm, 1 millimeter (mm), 2 mm, 3 mm, 4 mm, 5 mm, 6 mm, 7 mm, 8 mm, 9 mm, 10 mm, 20 mm, 30 mm, 40 mm, 50 mm, 60 mm, 70 mm, 80 mm, 90 mm, 100 mm, 200 mm, 300 mm, 400 mm, 500 mm, 600 mm, 700 mm, 800 mm, 900 mm, or 1000 mm. In some cases, the biosurfactant composition, the crude extract, or a purified product generated thereof may be subjected to centrifugation of at least about: 50 revolution per minute (rpm), 60 rpm, 70 rpm, 80 rpm, 90 rpm, 100 rpm, 110 rpm, 120 rpm, 130 rpm, 140 rpm, 150 rpm, 160 rpm, 170 rpm, 180 rpm, 190 rpm, 200 rpm, 210 rpm, 220 rpm, 230 rpm, 240 rpm, 250 rpm, 260 rpm, 270 rpm, 280 rpm, 290 rpm, 300 rpm, 310 rpm, 320 rpm, 330 rpm, 340 rpm, 350 rpm, 360 rpm, 370 rpm, 380 rpm, 390 rpm, 400 rpm, 410 rpm, 420 rpm, 430 rpm, 440 rpm, 450 rpm, 460 rpm, 470 rpm, 480 rpm, 490 rpm, 500 rpm, 600 rpm, 700 rpm, 800 rpm, 900 rpm, 1000 rpm, 1100 rpm, 1200 rpm, 1300 rpm, 1400 rpm, 1500 rpm, 1600 rpm, 1700 rpm, 1800 rpm, 1900 rpm, 2000 rpm, 2100 rpm, 2200 rpm, 2300 rpm, 2400 rpm, 2500 rpm, 2600 rpm, 2700 rpm, 2800 rpm, 2900 rpm, 3000 rpm, 4000 rpm, 5000 rpm, 6000 rpm, 7000 rpm, 8000 rpm, 9000 rpm, 10000 rpm, 20000 rpm, 30000 rpm, 40000 rpm, or 50000 rpm. In some cases, the biosurfactant composition, the crude extract, or a purified product generated thereof may be subjected to centrifugation of at most about: 50 revolution per minute (rpm), 60 rpm, 70 rpm, 80 rpm, 90 rpm, 100 rpm, 110 rpm, 120 rpm, 130 rpm, 140 rpm, 150 rpm, 160 rpm, 170 rpm, 180 rpm, 190 rpm, 200 rpm, 210 rpm, 220 rpm, 230 rpm, 240 rpm, 250 rpm, 260 rpm, 270 rpm, 280 rpm, 290 rpm, 300 rpm, 310 rpm, 320 rpm, 330 rpm, 340 rpm, 350 rpm, 360 rpm, 370 rpm, 380 rpm, 390 rpm, 400 rpm, 410 rpm, 420 rpm, 430 rpm, 440 rpm, 450 rpm, 460 rpm, 470 rpm, 480 rpm, 490 rpm, 500 rpm, 600 rpm, 700 rpm, 800 rpm, 900 rpm, 1000 rpm, 1100 rpm, 1200 rpm, 1300 rpm, 1400 rpm, 1500 rpm, 1600 rpm, 1700 rpm, 1800 rpm, 1900 rpm, 2000 rpm, 2100 rpm, 2200 rpm, 2300 rpm, 2400 rpm, 2500 rpm, 2600 rpm, 2700 rpm, 2800 rpm, 2900 rpm, 3000 rpm, 4000 rpm, 5000 rpm, 6000 rpm, 7000 rpm, 8000 rpm, 9000 rpm, 10000 rpm, 20000 rpm, 30000 rpm, 40000 rpm, or 50000 rpm.

[0137] In some cases, subsequent to the fermentation, the fermentation culture is cooled to approximately to reduce microbial metabolic activity or stabilize the product. In some cases, subsequent to the fermentation, the fermentation culture is cooled to a temperature of at most about: -20 °C, -15 °C, -10 °C, -9 °C, -8 °C, -7 °C, -6 °C, -5 °C, -4 °C, -3 °C, -2 °C, -1 °C, 0 °C, 1 °C, 2 °C, 3 °C, 4 °C, 5 °C, 6 °C, 7 °C, 8 °C, 9 °C, 10 °C, 11 °C, 12 °C, 13 °C, 14 °C, 15 °C, 16 °C, 17 °C, 18 °C, 19 °C, 20 °C, 21 °C, 22 °C, 23 °C, 24 °C, 25 °C, 26 °C, 27 °C, 28 °C, 29 °C, or 30 °C. In some cases, subsequent to the fermentation, the fermentation culture is cooled to a temperature of at least about: -20 °C, -15 °C, -10 °C, -9 °C, -8 °C, -7 °C, -6 °C, -5 °C, -4 °C, -3 °C, -2 °C, -1 °C, 0 °C, 1 °C, 2 °C, 3 °C, 4 °C, 5 °C, 6 °C, 7 °C, 8 °C, 9 °C, 10 °C,WSGR Docket No.: 69849-701.60111 °C, 12 °C, 13 °C, 14 °C, 15 °C, 16 °C, 17 °C, 18 °C, 19 °C, 20 °C, 21 °C, 22 °C, 23 °C, 24 °C, 25 °C, 26 °C, 27 °C, 28 °C, 29 °C, or 30 °C.

[0138] In some cases, subsequent to the fermentation, the fermentation culture is concentrated via negative pressure or evaporation. In some cases, subsequent to the fermentation, the fermentation culture is concentrated at a negative pressure of at most about: 1 mTorr (millitorr), 2 mTorr, 3 mTorr, 4 mTorr, 5 mTorr, 6 mTorr, 7 mTorr, 8 mTorr, 9 mTorr, 10 mTorr, 20 mTorr, 30 mTorr, 40 mTorr, 50 mTorr, 60 mTorr, 70 mTorr, 80 mTorr, 90 mTorr, 100 mTorr, 200 mTorr, 300 mTorr, 400 mTorr, 500 mTorr, 600 mTorr, 700 mTorr, or 750 mTorr. In some cases, subsequent to the fermentation, the fermentation culture is concentrated at a negative pressure of at least about: 1 mTorr (millitorr), 2 mTorr, 3 mTorr, 4 mTorr, 5 mTorr, 6 mTorr, 7 mTorr, 8 mTorr, 9 mTorr, 10 mTorr, 20 mTorr, 30 mTorr, 40 mTorr, 50 mTorr, 60 mTorr, 70 mTorr, 80 mTorr, 90 mTorr, 100 mTorr, 200 mTorr, 300 mTorr, 400 mTorr, 500 mTorr, 600 mTorr, 700 mTorr, or 750 mTorr.

[0139] In some cases, subsequent to the fermentation, the fermentation culture is concentrated by a volume of at least about: 1 %, 2 %, 3 %, 4 %, 5 %, 10 %, 15 %, 20 %, 25 %, 30 %, 35 %, 40 %, 45 %, 50 %, 55 %, 60 %, 65 %, 70 %, 75 %, 80 %, 85 %, 90 %, 95 %, 96 %, 97 %, 98 %, or 99 %. In some cases, subsequent to the fermentation, the fermentation culture is concentrated by a volume of at most about: 1 %, 2 %, 3 %, 4 %, 5 %, 10 %, 15 %, 20 %, 25 %, 30 %, 35 %, 40 %, 45 %, 50 %, 55 %, 60 %, 65 %, 70 %, 75 %, 80 %, 85 %, 90 %, 95 %, 96 %, 97 %, 98 %, or 99 %.

[0140] In some cases, the biosurfactant composition, the crude extract, or a purified product generated thereof is frozen or lyophilized. In some cases, the biosurfactant composition, the crude extract, or a purified product generated thereof is incubated at a temperature of at most about: -85 °C, -80 °C, -75 °C -70 °C, -60 °C, -50 °C, -40 °C, -30 °C, -20 °C, -15 °C, -10 °C, - 9 °C, -8 °C, -7 °C, -6 °C, -5 °C, -4 °C, -3 °C, -2 °C, -1 °C, 0 °C, 1 °C, 2 °C, 3 °C, or 4 °C. In some cases, the biosurfactant composition, the crude extract, or a purified product generated thereof is incubated at a temperature of at least about: -85 °C, -80 °C, -75 °C -70 °C, -60 °C, -50 °C, -40 °C, -30 °C, -20 °C, -15 °C, -10 °C, -9 °C, -8 °C, -7 °C, -6 °C, -5 °C, -4 °C, -3 °C, -2 °C, -1 °C, 0 °C, 1 °C, 2 °C, 3 °C, or 4 °C. In some cases, the biosurfactant composition, the crude extract, or a purified product generated thereof is incubated at a pressure of at most about: 1 mTorr (millitorr), 2 mTorr, 3 mTorr, 4 mTorr, 5 mTorr, 6 mTorr, 7 mTorr, 8 mTorr, 9 mTorr, 10 mTorr, 20 mTorr, 30 mTorr, 40 mTorr, 50 mTorr, 60 mTorr, 70 mTorr, 80 mTorr, 90 mTorr, 100 mTorr, 200 mTorr, 300 mTorr, 400 mTorr, 500 mTorr, 600 mTorr, 700 mTorr, 800 mTorr, 900 mTorr, 1000 mTorr, 2000 mTorr, 3000 mTorr, 4000 mTorr, 5000 mTorr, 6000 mTorr, 7000 mTorr, 8000 mTorr, 9000 mTorr, or 10000 mTorr. In some cases, the biosurfactant composition,WSGR Docket No.: 69849-701.601the crude extract, or a purified product generated thereof is incubated at a pressure of at least about: 1 mTorr (millitorr), 2 mTorr, 3 mTorr, 4 mTorr, 5 mTorr, 6 mTorr, 7 mTorr, 8 mTorr, 9 mTorr, 10 mTorr, 20 mTorr, 30 mTorr, 40 mTorr, 50 mTorr, 60 mTorr, 70 mTorr, 80 mTorr, 90 mTorr, 100 mTorr, 200 mTorr, 300 mTorr, 400 mTorr, 500 mTorr, 600 mTorr, 700 mTorr, 800 mTorr, 900 mTorr, 1000 mTorr, 2000 mTorr, 3000 mTorr, 4000 mTorr, 5000 mTorr, 6000 mTorr, 7000 mTorr, 8000 mTorr, 9000 mTorr, or 10000 mTorr. In some cases, at least about: 1 %, 2 %, 3 %, 4 %, 5 %, 10 %, 15 %, 20 %, 25 %, 30 %, 35 %, 40 %, 45 %, 50 %, 55 %, 60 %, 65 %, 70 %, 75 %, 80 %, 85 %, 90 %, 95 %, 96 %, 97 %, 98 %, 99 % or more water or moisture content, be weight / volume or volume / volume, of the crude extract or a purified product generated thereof is removed. In some cases, at most about: 1 %, 2 %, 3 %, 4 %, 5 %, 10 %, 15 %, 20 %, 25 %, 30 %, 35 %, 40 %, 45 %, 50 %, 55 %, 60 %, 65 %, 70 %, 75 %, 80 %, 85 %, 90 %, 95 %, 96 %, 97 %, 98 %, 99 % or more water or moisture content, be weight / volume or volume / volume, of the crude extract or a purified product generated thereof is removed.

[0141] In some cases, the crude extract may be substantially cell-free. A substantially cell-free extracts may comprise at most about: 1, lxl0Al, lxlOA2, lxl0A3, lxlOA4, or lxl0A5 numbers of cells or viable cells per mL of the crude extract. A substantially cell-free extracts may comprise at least about: 1, lxl0Al, lxlOA2, lxl0A3, lxlOA4, or lxl0A5 numbers of cells or viable cells per mL of the crude extract. In some cases, the crude extract may not be substantially cell-free. A substantially cell-free extracts may comprise at most about: 1 %, 2 %, 3 %, 4 %, 5 %, 10 %, 15 %, 20 %, 25 %, 30 %, 35 %, 40 %, 45 %, or 50 %, by weight of cells over volume of the crude extract. A substantially cell-free extracts may comprise at least about: 1 %, 2 %, 3 %, 4 %, 5 %, 10 %, 15 %, 20 %, 25 %, 30 %, 35 %, 40 %, 45 %, or 50 %, by weight of cells over volume of the crude extract. A substantially cell-free extracts may not comprise any cells or viable cells. In some cases, the crude extract may not be substantially cell-free.

[0142] In some cases, the crude extract may comprise any of those described in Examples 1-4.Example methods for manufacturing the biosurfactant composition are also disclosed elsewhere in this disclosure (for example, see Examples 1-4).Definitions

[0143] As used herein, “biosurfactant composition” refers to any composition comprising a biosurfactant. “Biosurfactant composition” is interchangeable with “biosurfactant extract or “biosurfactant formulation.” A “biosurfactant” as used herein refers to a surfactant that is generated using an organism or enzymatic activities of the organism in vitro or in vivo.

[0144] As used herein, “cloud point” refers to the temperature or temperature range at which a solution exhibits phase transition.WSGR Docket No.: 69849-701.601

[0145] As used herein, “pour point” refers to the lowest temperature or temperature range at which a solution exhibits no surface movement.

[0146] As used herein, “renewable sources’ comprises subproducts derived from different stages of the process or the valorization of crops; processing residues or refined starch; or glucose syrups.

[0147] As used herein, “crude extract” refers to a mixture derived from the fermentation of the microorganisms as described herein. A crude extract may or may not comprise cells or viable cells.

[0148] As used herein and thereof, the singular forms “a,” “an,” and “the” include plural references unless the context clearly dictates otherwise.

[0149] The term “about” or “approximately” as used herein when referring to a measurable value such as an amount or concentration and the like, is meant to encompass variations of 20 %, 10 %, 5 %, 1 %, 0.5 %, or even 0.1 % of the specified amount. For example, “about” can mean plus or minus 10 %, per the practice in the art. Alternatively, “about” can mean a range of plus or minus 20 %, plus or minus 10 %, plus or minus 5 %, or plus or minus 1 % of a given value. Alternatively, particularly with respect to biological systems or processes, the term can mean within an order of magnitude, up to 5-fold, or up to 2-fold, of a value. Where particular values can be described in the application and claims, unless otherwise stated the term “about” may be assumed to encompass the acceptable error range for the particular value. Also, where ranges, subranges, or both, of values can be provided, the ranges or subranges can include the endpoints of the ranges or subranges. The terms “substantially,” “substantially no,” “substantially free,” and “approximately” can be used when describing a magnitude, a position or both to indicate that the value described can be up to a reasonable expected range of values. For example, a numeric value can have a value that can be + / - 0.1 % of the stated value (or range of values), + / -1 % of the stated value (or range of values), + / - 2 % of the stated value (or range of values), + / - 5 % of the stated value (or range of values), + / - 10 % of the stated value (or range of values), etc. Any numerical range recited herein can be intended to include all subranges subsumed therein.

[0150] Where values are described as ranges, it may be understood that such disclosure includes the disclosure of all possible sub-ranges within such ranges, as well as specific numerical values that fall within such ranges irrespective of whether a specific numerical value or specific sub-range is expressly stated.

[0151] The terms “comprise,” “have,” and “include” are open-ended linking verbs. Any forms or tenses of one or more of these verbs, such as “comprises,” “comprising,” “has,” “comprising,” “includes,” and “including,” are also open-ended. For example, any method thatWSGR Docket No.: 69849-701.601“comprises,” “has,” or “includes” one or more steps is not limited to possessing only those one or more steps, and also covers other unlisted steps.

[0152] The terms “at least” and “greater than or equal to” are interchangeable. The terms “at most” and “less than or equal to” are interchangeable.

[0153] Unless otherwise stated, as used herein, temperatures are described with reference to standard pressure at sea level (1 atm). Unless otherwise stated, as used herein, pressures are described with reference to standard temperature (25 °C). All densities, unless otherwise stated are described with reference to standard temperature (25 °C) and atmospheric pressure at sea level (1 atm), otherwise referred to as standard temperature and pressure (STP). °C

[0154] While various embodiments of the invention have been shown and described herein, it will be obvious to those skilled in the art that such embodiments are provided by way of example only. Numerous variations, changes, and substitutions may occur to those skilled in the art without departing from the invention. It should be understood that various alternatives to the embodiments of the invention described herein may be employed.EXAMPLES

[0155] These examples are provided for illustrative purposes only and not to limit the scope of the claims provided herein.Example 1: Production of biosurf actant formulationsOverview

[0156] Provided herein are non-limiting example methods for producing the biosurfactants described herein. An overview of the example process is shown in FIGs. 1A-1B and below. Stage 1 : Preparation of the Culture Medium

[0157] The substrate (agro-industrial residues) was prepared through thermal pretreatment and subsequent decoction under specific conditions (residue / water ratio, heating time, substrate cutting size). These conditions facilitated the exposure of starches and other components (such as amylopectin, pectin, lignin, and cellulose). Glucose was added to this pretreated substrate in a defined proportion, and the pH was controlled to form the base of the culture medium.Stage 2: Preparation of the Inoculum

[0158] The inoculum was prepared by cultivating a specific proportion of fungi in the culture medium, allowing it to grow under conditions to produce the biosurfactant and other metabolites. These conditions include production time, temperature, agitation (controlled by RPM), aeration, and pH monitoring, as described herein.Stage 3: FermentationWSGR Docket No.: 69849-701.601

[0159] Fermentation was carried out in a stirred tank bioreactor, where a proportion of the inoculum as described herein was mixed with an inducer, as described herein. This inducer could include hydrocarbons, synthetic or mineral base lubricating oils, or vegetable fatty acids. During fermentation, temperature, agitation, dissolved oxygen (DO), time, and pH was continuously controlled. No antifoaming agent was added.While this example discloses using an inducer, the fermentation or production of the biosurfactant formulation or composition can be carried out with or without the inducer.Stage 4: Downstream Processes

[0160] Once fermentation was completed, the product underwent separation processes where biomass and the cell-free broth containing the biosurfactant were recovered. Separation methods could include filtration and homogenization to ensure the purification of the biosurfactant and other metabolites.

[0161] The crude composition comprised at least one glycolipid as described herein (for example, at least of one sophorolipid, trehalose lipids, rhamnolipids, mannosylerythritol lipids, cellobiose lipids, or polyol lipids, as described herein). Other metabolites could comprise lipopeptides, lipoproteins, enzymes, saccharides and pigments, obtained directly from the fermentation process. A decantation step could be used to obtain the crude composition from the fermentation process.Stage 5: Formulation

[0162] Subsequent to the downstream processes, various types of formulation could be obtained. In one non-limiting example, soluble concentrate was obtained. One example soluble concentrate comprised lyophilized extract (from 1 % to 5 % by weight / volume) and sodium benzoate (about 0.01 % weight / volume).

[0163] A first example formulation (Minimum Lyophilized Extract; see FIG. 13A) was obtained as follows: In 50 milliliters (mL) of water, 1 gram of lyophilized extract and 0.01 g of sodium benzoate were added under continuous agitation. Additional water was added to reach a final volume of 100 mL.

[0164] A second example formulation (Maximum Lyophilized Extract; see FIG. 13B) was obtained as follows: In 50 mL of water, 5 grams of lyophilized extract and 0.01 g of sodium benzoate were added under continuous agitation. Additional water was added to reach a final volume of 100 mL.Materials

[0165] Provided herein is a list of materials used for producing the biosurfactant formulations as described herein:WSGR Docket No.: 69849-701.601• Standard laboratory equipment including beakers, Erlenmeyer flasks, magnetic stirrer, heating plate, autoclave, and filtration setup.• Agro-industrial waste 120 [or 250] grams / L of culture medium)• Distilled water (sufficient to make up to 1000 mL)• L-Glucose (15 grams / L of culture medium)• Vegetable oil (20 mL / L of culture medium)• Yeast extract (Igram / L of culture medium)• Citric acid (for pH adjustment)• Laminar Flow Cabinet: Used to maintain a sterile environment during the inoculation process.• Micropipette Tips: Sterile tips ranging from 100 to 1000 pL for handling the fungal samples.• Fungal Culture on PDA (Potato Dextrose Agar): A mature fungal culture grown on agar to provide the initial inoculum.• Applikon ez2-Control with Stirred Tank Bioreactor• Filter Press - DIDA CONTROL®• Destilador de lecho empacado - DIDA CONTROL®• Rotavapor R- 100 - BUCHI®Preparation of substrate per liter

[0166] The production of biosurfactants could utilize an agro-industrial waste specifically utilizing a process that ensures the efficient extraction and preparation of the medium necessary for microbial fermentation. The process began by thoroughly washing the agro-industrial waste (to weigh out 120 grams of this material and to cut it into small 1 cm x 1 cm pieces).

[0167] The cut pieces were placed into a 1000 mL beaker. 500 mL of distilled water was added into the beaker. The mixture was heated on a heating plate set to approximately 300 °C until the water began to boil. The mixture was allowed to boil for 15 minutes to produce a potato broth. The potato broth was filtered to remove solid residues, and the filtrate was filtered into a bluecapped flask. The volume was adjusted to 1000 mL with distilled water. The broth was transferred into an Erlenmeyer flask equipped with a magnetic stirrer. 15 grams of L-Glucose and 1 gram of Yeast were gradually added into the broth while the broth was being stirred. Then, 20 mL of vegetable oil was added into the broth while the broth was stirring. The pH of the solution was adjusted with citric acid until it reached a pH of 5.5. The prepared medium was poured into a 1000 mL Erlenmeyer flask, and the flask was covered with a gauze plug. The medium was sterilized in an autoclave at 121 °C for 20 minutes. The sterilized medium was allowed to cool to room temperature before proceeding with inoculation.Inoculum preparation per literWSGR Docket No.: 69849-701.601

[0168] Provided herein are methods for preparing a fungal pre-inoculum in a liquid culture medium for biosurfactant formulation production. This method uses sterile techniques and controls environmental conditions to ensure the viability and purity of the fungal cultures. While this example uses Entonaema liquescens for illustrative purposes, similar methods can be applied to other fungi, including but not limited to various filamentous or non-filamentous fungi.FIG. 2A depicts the growth of Entonaema liquescens on solid agar plate. FIG. 2B depicts the growth of Entonaema liquescens in fermentation inoculum, as descried below.

[0169] After observing radial growth of the fungus on the Potato Dextrose Agar (PDA) surface, using the broad end of a sterile 100 to 1000 microliter (pL) micropipette tip, three circular pieces were cut to approximately 0.5 x 0.5 centimeter (cm) in size under a laminar flow hood, containing both fungus and agar. The pieces were then transferred into 1000 mL of sterile liquid culture medium. The pieces were incubated statically for 10 days at room temperature (25-30 °C). The culture medium was monitored to ensure it did not become opaque, and that fungal growth was observed on the surface. After the sixth day, the formation of a pigment, which could be dark green or black, might begin to be visible. At the end of the incubation period, the inoculum was homogenized using a sterile blender or food processor at a low to medium speed for 15 seconds to ensure a uniform distribution of the fungal cells in the medium. Fermentation bioreactor setups in biosurfactant production process for 4 liters (L)

[0170] This section describes the procedures used in the biosurfactant production process utilizing stirred tank bioreactors.

[0171] For fermentation, 5 % inoculum concentration was added with 200 mL of inoculum along with 3800 mL of cool medium containing vegetable oil as inducer.. 2 L / min of air (0.5 vvm) was used to aerate the fermentation culture. The aeration was used to increase to 4 L / min (1 vvm) after 24 h. For agitating the fermentation culture, constant agitation at 150 rpm was carried out with two Rushton turbines, one at the bottom of the system and the other positioned 7 cm above the bottom. For sampling, a 10 mL syringe was filled with 5 mL of the sample and clamped to the hose. The sample was transferred into a sterile vial (the syringe was reconnected and keep clamped to the hose). The sample was immediately frozen for analysis. FIG. 3A depicts the fermentation bioreactor. FIG. 3B-3C depict the aerated culture during fermentation and post-fermentation, respectively. In the post-process sampling, adequate samples of both biomass and the liquid phase were collected at the end of the operation for comprehensive analysis.Downstream processes at 4L

[0172] This section details the method for centrifugation and subsequent phase separation for isolating the desired metabolites during the biosurfactant production process. This procedure canWSGR Docket No.: 69849-701.601facilitate efficiently separating and preserving various components of the culture medium for further analysis and use.

[0173] Both the samples and the final volume of the fermented mixture (crude extract) were filtrated (macro-filtration followed by vacuum filtration) as follow: a) The macro-filtration consisted of passing the entire fermentation broth through a sterile stainless-steel mesh strainer with a pore size in the sub -millimeter to millimeter range, allowing for the removal of coarse solid biomass and large particulate matter. The operation was performed under aseptic conditions within a laminar flow cabinet to prevent contamination and preserve product integrity. This operation is herein referred to as the “primary post-fermentation filtration, b) Vacuum filtration: Following the primary post-fermentation macro-filtration step, the resulting filtrate was subjected to a secondary vacuum filtration step in order to remove finer biomass particles that remain suspended in the liquid phase. This step was carried out using a standard laboratory vacuum filtration system, comprising a filter medium suitable for retaining fine particulate biomass. Additionally, qualitative filter paper discs with an approximate pore size of about 11 pm could be employed, for example Whatman® Grade 1 filter paper or an equivalent material. The filtrate obtained after this vacuum filtration step corresponded to the final aqueous extract, which was substantially free of suspended biomass and suitable for subsequent processing or characterization. FIG. 3D depicts the fermentation product after microfiltration (left) and vacuum filtration (right). Then, depending on the presence and concentration of residual biomass, an optional additional centrifugation step may be applied in order to further clarify the aqueous extract. The filtrated samples and the fermented mixture were subjected to centrifugation at 2100 revolutions per minute (rpm) for 10 minutes to ensure the effective separation of phases without disrupting the structural integrity of the biomass or metabolites. Post-centrifugation, three distinct phases were typically observed within the samples: a top layer that comprised an oily phase, which contained lipids and possibly non-polar metabolites; a middle layer that comprised an aqueous phase, where water-soluble metabolites of interest were predominantly found; and a bottom layer that comprised a solid phase or pellet, which included the medium residues or biomass. The metabolites of the middle aqueous phase (such as those as described herein) were extracted for further analysis for ensuring the purity and integrity of the metabolites for further processing.

[0174] FIG. 3E depict the final fermentation product after centrifugation step. The remaining oily and solid phases were stored at -20 °C to prevent the degradation of various components within these phases and for efficiently separating and preserving various components of the culture medium for further analysis. FIG. 3F depicts the change between the initial fermentation set-up and the final centrifugated product.WSGR Docket No.: 69849-701.601

[0175] In order to concentrate the product while preserving the integrity of the target molecule, the clarified aqueous extract was subjected to a lyophilization (freeze-drying) step. The lyophilization process was carried out at a temperature of approximately -75 °C and under a reduced pressure in the range of 210-230 mTorr, or under equivalent conditions suitable for efficient sublimation of water while maintaining molecular stability. The samples were subjected to lyophilization for a duration of approximately 48 to 72 hours, or until the complete removal of water from the samples is achieved, resulting in a dry, concentrated product suitable for storage or further processing. FIG. 3G depicts the lyophilizate product.

[0176] If no inducer is used, only two phases are obtained during the centrifugation stage: an aqueous phase, where water-soluble metabolites of interest were predominantly found, and a bottom layer comprising a solid phase or pellet. The remaining stages are carried out as described in this example.Formulation

[0177] In order to obtain a concentrated formulation at approximately 30% (w / v), the lyophilized material was reconstituted using sterile distilled water, as shown in FIGs. 3H-3I. A preservative was incorporated into the reconstituted formulation (e.g., sodium benzoate) at a concentration of approximately 0.06 % (w / v), and the mixture was homogenized by mechanical agitation until complete dissolution and uniformity were achieved. In one exemplary embodiment, a concentrated biosurfactant formulation was prepared by adding 30 g of lyophilized extract to 70 mL of sterile distilled water, followed by incorporation of 0.06 g of sodium benzoate, and mixing under continuous mechanical agitation until a homogeneous solution was obtained, resulting in a final volume of approximately 100 mL and an active biosurfactant concentration of about 30% (w / v). The resulting formulation was considered the final product, herein referred to as example biosurfactant 1 as shown in FIG. 3K, and was stored under appropriate conditions for subsequent testing and use.Optional Combination with a Cell-Free Enzymatic Extract (Application-Specific Formulation)

[0178] For certain applications, such as the bioremediation of water and soils contaminated with hydrocarbons, the concentrated final product was combined with a cell-free extract derived from the same fermentation process. This cell-free extract was enriched in enzymatic components and enhanced the overall effectiveness of the formulation. To obtain the cell-free extract, the process started from the aqueous extract obtained after the vacuum filtration step described above. An additional vacuum filtration step was then applied using a filter with a nominal pore size of approximately 0.22 pm, thereby ensuring the complete removal of biomass and cellular material. The resulting cell-free extract was stored at a temperature ofWSGR Docket No.: 69849-701.601approximately 4 °C until its subsequent use, either alone or in combination with the concentrated final product (as shown in FIG. 3J).Fermentation bioreactor setups in biosurfactant production process for 40 liters (L)

[0179] To demonstrate the industrial scalability of the process, this section describes the production of biosurfactants at 40 L, as shown in FIG. IB.

[0180] The preparation of the substrate (culture medium) for the 40 L fermentation process followed the same formulation and preparation principles described for the 4 L fermentation process, ensuring consistency in nutrient composition and process performance across scales. The Sterilization of the 40 L stirred-tank reactor (STR) was performed using integrated cleaning-in-place (CIP) and sterilization-in-place (SIP) procedures. The reactor and all productcontact surfaces were cleaned used alkaline and acidic cleaning cycles, followed by rinsing with purified watered. After reassembly and installation of sensors, the closed system was sterilized by injection of pressurized saturated steam at approximately 121 °c and 1. 1-1. 3 bar for 20 minutes, ensuring complete microbial inactivation. Condensates were subsequently discarded prior to used. The culture medium and supplements were introduced into the reactor by opening the top covered and charging the system with the prepared medium, sterile watered, sterile nutrient solutions, and sterile pH-adjusting solutions. Agitation and aeration were initiated to ensured homogeneity and oxygen saturation, followed by temperature adjustment to the fermentation set pointed (approximately 27 °C). Once operating conditions were stabilized, the reactor was inoculated, the system was sealed, and the fermentation process was initiated under controlled conditions he fermentation was conducted used a total suspension volume of approximately 34 L, comprising 32.2 L of culture medium and 1.8 L of inoculum (5%).Agitation was maintained at a stirring speed of approximately 86 rpm. The fermentation was initiated at an initial pH of about 5.5 and operated at a controlled temperature of approximately 27 °C. Aeration was supplied at an initial rate of approximately 0.05 volumes of air per volume of liquid per minute (vvm). Agitation remains constant throughout the fermentation process. Aeration, however, was gradually increased from 0.05 vvm to 0.1 vvm according to microbial activity and oxygen consumption. During the fermentation process, a set of off-line and in-line variables were monitored to evaluate process performance and product formation. Off-line variables were periodically measured from collected samples and include final biomass concentration (g / L), glucose concentration (g / L), reducing sugars concentration (g / L), soluble solids content (°Bx), and surface tension (mN / m). In-line variables were continuously monitored throughout the fermentation and included dissolved oxygen (DO, mg / L), pH, liquid level (L), air flow rate (L / min), and temperature (°C).Downstream processes at 40 LWSGR Docket No.: 69849-701.601

[0181] After completion of the fermentation, the fermentation broth was cooled to approximately 20 °C to reduce microbial metabolic activity and stabilize the product. The cooled broth was then collected through sanitary piping into suitable containers, such as 40 L drums, under hygienic conditions. All equipment in contact with the broth was previously cleaned with soap and water and sterilized using water vapor to ensure aseptic handling. The harvested fermentation broth was subjected to a biomass separation step using a filter press to remove suspended solids and cellular material. Prior to filtration, the filter press was cleaned following a cleaning-in-place (CIP) procedure, consisting of circulation of 1% (w / w) NaOH at 60 °C for 15 minutes, followed by circulation of 1% (w / w) HC1 for 15 minutes, and a final rinse with purified water to remove residual solids, biofilms, and contaminants from all productcontact surfaces. After cleaning, the fermentation broth was filtered, and the resulting clarified broth was collected and stored in 40 L drums until the concentration step.

[0182] The clarified broth was concentrated by means of two consecutive vacuum evaporation steps carried out used different evaporation systems. In a first evaporation stepped, the broth was processed used distillation equipment previously cleaned with soap and watered, including the charge and condensation tanks. The broth was concentrated in cycles, reducing the volume from approximately 15 L to 5 L per cycle, until a final concentrated volume of approximately 5 L was obtained. This stepped was carried out at an operating temperature of approximately 61 °C and a reduced pressure of approximately -20 mmhg. In a second evaporation stepped, the partially concentrated broth was further processed used a rotary evaporator under sterile conditions. The evaporator vessels were sterilized by autoclaving at approximately 121 °C and 15 psi for about 20 minutes prior to used. The volume was reduced from approximately 5 L to 1 L at an operating temperature of approximately 55 °C, while the pressure was gradually reduced from approximately 400 mbar to 80 mbar.Formulations

[0183] The final concentrated product obtained after the vacuum evaporation steps was transferred to a sterile 1 L glass container and stored at approximately 4 °C until further formulation. The resulting concentrate had an approximate concentration of 30%. The formulation stage followed the same procedure described for the 4 L process. Sodium benzoate was added at approximately 0.06% (w / v) while stirring continuously until it was completely dissolved.Example 2: Chemical Characterization of various molecules present within biosurfactant formulationsWSGR Docket No.: 69849-701.601

[0184] Provided herein are characterization of various molecules present within biosurfactant formulations.

[0185] Chromatographic identification of the biosurfactant molecules produced was performed using a two-stage analytical approach, comprising an initial exploratory analysis followed by a confirmatory characterization. For exploratory identification, the middle aqueous phase of the sample prepared using methods as described in Example 1 was subjected to liquid chromatography-mass spectrometry (LC-MS). Various species of sophorolipid, in a mixture of acidic and lactone chemical states were identified, as shown in FIG. 4A. In particular, the different sophorolipid species corresponded to the 5 groups with different degrees of acetylation Additionally, lipopeptides and lipoproteins with surfactant activity; and enzymes and pigments, with additional functional properties were also identified. For confirmatory analysis, a liquidliquid extraction was performed using ethyl acetate (EtOAc), followed by chromatographic analysis using high-performance liquid chromatography coupled to an evaporative light scattering detector (HPLC-ELS) to evaluate the extractable organic fraction of the product. Briefly, 5.00 g of sample were weighed into a 15 mL conical tube, and 10.0 mL of EtOAc were added. The mixture was agitated for 3 minutes using a laboratory mixer, allowing phase separation, after which the upper organic phase was carefully decanted into a pre-weighed aluminum container. This extraction procedure was repeated three times on the same sample, and the collected organic fractions were combined. The solvent was subsequently evaporated to dryness, and the mass of extractable material was determined by difference in weight, allowing calculation of the weight percentage of extractables relative to the initial sample mass. The dried extract was then reconstituted in 0.5 mL of ethanol and injected into the HPLC-ELS system to obtain a confirmatory chromatographic profile of the extracted compounds. The chromatographic profile of the downstream concentrate product exhibited multiple well-defined peaks corresponding to non-volatile organic compounds associated with the biosurfactant fraction. FIG. 4B depicts several peaks eluted at retention times matching those of a Cl 8:1 sophorolipid analytical standard, specifically Cl 8:1 diacetylated lactonic and Cl 8:1 diacetylated acidic sophorolipids, providing confirmatory evidence of the sophorolipid nature of the product.

[0186] The degree of acetylation of the sophorolipid can impact the stability of the molecules under extreme conditions and their modulation for different uses. The degree of acetylation can be modified with different fermentation parameters.Example 3: Various Physicochemical and Functional properties of biosurfactant formulations

[0187] Provided herein are various properties of the biosurfactant formulations. To evaluate the performance, stability, and application potential of the biosurfactant produced, aWSGR Docket No.: 69849-701.601comprehensive physicochemical and functional characterization was conducted. This characterization included the assessment of surface-active properties, interfacial behavior, and key technical parameters relevant to industrial handling and end-use applications. The parameters of the methods described herein can be modified for charactering the various properties as described herein.

[0188] Various properties of the biosurfactant formulation comprising the Minimum lyophilized extract described in Example 1 are depicted in Table 1 below.Table 1. Various physical and chemical properties of an example biosurfactant Property ValuePhysical state Opaque liquidForm LiquidAppearance at 25 °C Amber to dark brownOdor CharacteristicDensity (g / cm3, 25 °C) 1.087Viscosity (cP / mPa s, 25 °C) 2.51pH (1% aqueous) 4.96HLB 10Solids content (% dry w) 19Active material (%) 30CMC (%, 25 °C) 9Surface tension (mN / m, 9%, 25 °C) 34Interfacial tension (mN / m, 9%, n-decane, 2514.987°C)Cloud point (10% aqueous) Non observable between 10 °C to 100 °C Pour point (10% aqueous) Non observableWater solubility SolubleFoam category LowpH stability 2-121-25 (stable between TDS 50,000 mg / L to Salinity stability (% w / v NaCl)15000 mg / L)Max. NaCl solubility (g per 100 mL, 20 °C) 38.09 gEmulsion stability (%EI) >48 hContact angle - hydrophilic (glass, 25 °C) 26.79Contact angle - hydrophobic (PTFE surface,103.6925 °C)Boiling point Above 100 °C

[0189] To determine surface tension and interfacial tension of the biosurfactants, a Kriiss K8 tensiometer and a Du Nouy ring (Pt-Ir) are used (static method). Additional static methods can comprise the Wilhelmy Plate Method, Pendant Drop Method, Sessile Drop Method, and BubbleWSGR Docket No.: 69849-701.601Drop Method. Other dynamic methods can also be used to determine surface tension and interfacial tension, comprising: the Oscillating Drop / Bubble Method, Maximum Bubble Pressure Method, and Spinning Drop Method. Indirect or complementary methods for determining can comprise Interfacial Rheology, Contact Angle Microscopy, or Electrocapillarity.

[0190] In one method, FIGs. 5A-5B depicts the determination of the interfacial tension of the biosurfactants as described herein with lubricating oil at 27.5 °C and with oil at 30.7 °C, respectively as measured using Kriiss K8 tensiometer and a Du Nouy ring (Pt-Ir) [static method],

[0191] In another method, surface and interfacial tension measurements were performed using a Dataphysics OCA 15EC goniometer by means of the pendant drop method at ambient temperature, employing a SNP 165 / 119 needle. Prior to analysis, samples were pretreated by centrifugation at 10,000 rpm for 10 minutes to minimize the solids content and facilitate accurate measurements, as shown in FIG. 5C. Each sample was measured in triplicate. Surface tension values were obtained using the solid-free supernatant collected after centrifugation. Interfacial tension was determined using the same pendant drop method under ambient conditions, employing n-decane (density 0.773 g / cm3) as the reference external liquid phase. Interfacial tension measurements were carried out using sample dilutions prepared at concentrations corresponding to the critical micelle concentration (CMC). The critical micelle concentration (CMC) was determined by plotting the measured surface tension values as a function of surfactant concentration. The biosurfactant composition exhibited a surface tension of 34 mN / m measured at 25 °C and 9% (w / v), demonstrating effective surface activity characteristic of glycolipid-type biosurfactants. Interfacial tension measured against n-decane at 25 °C and 9% (w / v) was 14.987 mN / m, confirming the ability of the formulation to significantly reduce oil-water interfacial forces, relevant for emulsification and remediation applications. FIG. 5D depicts the biosurfactant presented a critical micelle concentration (CMC) of 9% (w / v) at 25 °C, indicating effective self-assembly at moderate concentrations. The estimated hydrophilic-lipophilic balance (HLB) value was 10, consistent with a surfactant suitable for oil-in-water emulsions and multiphase industrial formulations. Complementary, as depicted in FIG.5E, the wettability and affinity of the biosurfactant on a hydrophilic and hydrophobic surface was evaluated using the measurement of the contact angle of (see Table 1). Contact angle measurements were performed using the sessile drop method on a goniometer at ambient temperature. A SNS needle was used to dispense droplets with a fixed volume of 1.5 pL for all measurements. The contact angle was calculated using the Laplace-Young fitting method.Measurements were conducted using sample dilutions prepared at concentrations correspondingWSGR Docket No.: 69849-701.601to the previously determined critical micelle concentration (CMC). Glass was selected as the reference hydrophilic surface for contact angle determination on polar substrates, while polytetrafluoroethylene (PTFE, Teflon tape) was used as the reference hydrophobic surface for contact angle determination on non-polar substrates. Results show that on hydrophilic glass surfaces, the biosurfactant showed a contact angle of 26.79°, indicating enhanced wettability. On hydrophobic PTFE surfaces, a contact angle of 103.69° was observed, confirming interaction with non-polar substrates and a balanced hydrophilic-lipophilic behavior.

[0192] In some cases, cloud point technique was used to evaluate the temperature at which the biosurfactant transitions from transparent to turbid due to phase separation. This method is beneficial for characterizing non-ionic surfactants as those as described herein.

[0193] The cloud point formation of the biosurfactant as described herein was assessed over a wide temperature range (-10 °C to 100 °C) by gradually increasing and decreasing the temperature while observing turbidity changes (see Table 1). The absence of a cloud point within this range indicated high solubility and thermal stability, beneficial for industrial applications requiring performance consistency across extreme temperatures. Because no phase separation or cloud point formation occurred across the tested temperature range (-10 °C to 100 °C), the surface tension (34 mN / m) and interfacial tension (14.98 mN / m) remained stable.

[0194] The pH stability of the biosurfactant formulation was evaluated by acidic and basic titration assays performed at ambient temperature using 50 mL of product solution from two independent batches. Acidic stability was assessed by gradual addition of 0.1 M HC1, while basic stability was evaluated by titration with 0.1 M NaOH, with continuous pH monitoring using a calibrated pH meter. Throughout the titration assays, samples were visually inspected for changes in color, appearance, or phase stability across the evaluated pH range. The biosurfactant remained stable across a pH range of 2 to 12, with no observable phase separation or loss of functionality. The filtered biosurfactant extract as described herein demonstrated stability over extreme pH conditions, as depicted in FIGs. 14A-14B. A titration curve characteristic of a monoprotic acid was observed, with an equivalence point at pH 7.78 at a NaOH volume of 1.1 mL, as depicted in FIGs. 14A-14B. Similarly, another solution exhibited a monoprotic acid titration curve with an equivalence point at pH 8.21 at 3.4 mL of NaOH, suggesting the presence of a more acidic system or a higher concentration of acidic molecules. During the pH increase, a color change from pale yellow to intense yellow was observed at pH 11.36, indicating a shift in the solution’s properties.

[0195] The salinity stability of the biosurfactant formulation was evaluated by incremental addition of sodium chloride (NaCl) to 50 mL of product solution from two independent batches at ambient temperature. Increasing salt concentrations were applied under continuous stirring,WSGR Docket No.: 69849-701.601and samples were monitored for changes in appearance, turbidity, phase separation, or precipitation. The maximum compatible salinity was defined as the highest NaCl concentration at which the formulation remained visually stable. Salinity stability was maintained from 1 to 25% (w / v) NaCl, with a maximum NaCl solubility of 38.09 g per 100 mL at 20 °C, indicating strong tolerance to high-ionic-strength environments. No cloud point or phase separation was observed over the temperature range of -10 °C to 100 °C, confirming high thermal stability.

[0196] The pH of the product was determined using a potentiometric method with a pH electrode for H+determination, measuring an aqueous dilution of the product at a concentration of 1% (w / v) in distilled water. Density was determined by gravimetric analysis using a pycnometer. Viscosity was measured directly on the product using a shear-based microviscometer (pVISC, Rheosense Inc.) at ambient temperature. The total solids content of the evaluated products was determined by gravimetric drying. Briefly, 1 g of sample was placed on a pre-dried watch glass and dried at 105 °C for 8 hours. After drying, the samples were cooled in a desiccator, and the final mass was recorded to calculate the solids content based on weight difference, (see Table 1).

[0197] Foam formation was evaluated using aqueous solutions prepared at a concentration of 1% (v / v). For each test, 200 mL of solution was prepared, and air was injected using an aquarium air pump at a flow rate of approximately 4 L / min for 30 seconds. The generated foam was allowed to rest for 60 seconds, after which foam formation and stability were documented by photographic recording. As shown in FIGs. 6E-6F, foam formation was classified as low, supporting suitability for applications where controlled foaming is required, such as industrial cleaning, bioremediation, and process formulations.

[0198] As depicted in FIGs. 8A-8C, the fungus Entonaema liquescens was capable of growing under the standard microorganism reactivation procedure (inoculum preparation) at low temperatures (18°C), demonstrating adaptability.

[0199] The ability of the fungus to grow across a wide temperature range (18-30°C) suggests potential field applications in various seasons. The dark pigment production is further investigated for value-added applications, such as antibacterial, antimalarial, and other potential uses reported for fungi of this family.

[0200] The filtered biosurfactant extract as described herein demonstrated stability over extreme pH conditions, as also depicted in FIGs. 14A-14B. A titration curve characteristic of a monoprotic acid was observed, with an equivalence point at pH 7.78 at a NaOH volume of 1.1 mL, as depicted in FIGs. 14A-14B. Similarly, another solution exhibited a monoprotic acid titration curve with an equivalence point at pH 8.21 at 3.4 mL of NaOH, suggesting the presence of a more acidic system or a higher concentration of acidic molecules. During the pH increase, aWSGR Docket No.: 69849-701.601color change from pale yellow to intense yellow was observed at pH 11.36, indicating a shift in the solution’s properties.Example 4: Various applications of biosurfactants

[0201] Provided herein are various applications of the biosurfactants as described herein. Managing various products or intermediates of petroleum applications.

[0202] In oil or petroleum industry, the biosurfactants described herein can be used in different phases of the crude oil processing cycle. In the upstream process (such as but not limited to well preparation and crude extraction to the surface), the biosurfactant can be used as an emulsifier, a defoamer agent, a spacer surfactant, or a wetting agent, or a combination thereof. Additionally, the biosurfactant can be used for foam control, cleaning wells, hydraulic fracturing, or enhanced oil recovery (EOR), or a combination thereof. In the midstream process (such as but not limited to transportation and refining, the biosurfactant can be used as a dispersing wax or paraffin or a corrosion inhibitor. Additionally, the biosurfactant can be used for pipeline cleaning, flow assurance, or high salinity water disposal. In the downstream process (final stage of the lifecycle), the biosurfactant can be used in water treatment, polluted soil treatment, or cleaning wells. In some cases, the downstream applications can align with TPH (Total Petroleum Hydrocarbons) removal. In some cases, the biosurfactant can be used in oil recovery, hydrocarbon remediation, or drilling fluids.

[0203] FIGs. 6A-6B shows the emulsification properties of the biosurfactant as described herein.

[0204] 2 grams (50% w / v) of lubricant oil were added to glass tubes of depicted in FIG. 6A.Each tube was vortexed while gradually adding 2 mL (50% v / v) of cell-free extract using a micropipette (2-10 mL), ensuring proper mixing of the oil and extract. The mixture was vortexed for 2 minutes. A control sample was prepared with 2 g of oil and 2 mL of distilled water. The samples were left to rest, and the emulsion layer height was measured at 24 hours and 48 hours. The emulsification index (E) and emulsion stability (EE) were calculated using the formulas from Cooper & Goldenberg (1987) and Lara-Severino et al. (2017) [each of which is incorporated herein in its entirety] as shown below:E = (Emulsion layer height / Total height) x 100%EE= (Emulsion volume at tO / Emulsion volume at t) x 100%

[0205] The emulsions formed exhibited distinct phase separation, as observed in the Falcon tubes of FIG. 6A. Differences in emulsification index and stability were recorded across the samples. The biosurfactant-containing extracts facilitated emulsion formation. Stability assessment over 24 h and 48 h indicated variations in emulsion retention, reflecting the effectiveness of the biosurfactant over time. As depicted in FIG. 6B, Day 3 was identified as theWSGR Docket No.: 69849-701.601optimal point. By day 6, the emulsion was no longer stable, which might be due to the degradation of the biosurfactant itself. Methods and protocol used in FIG. 6B were the same as those used in FIG. 6A.

[0206] As shown in FIG. 6G, the emulsification capacity of the final biosurfactant product obtained after bioreactor fermentation was evaluated using a three-way stopcock mixing method, with vegetable oil employed as the oil phase. Equal volumes of oil and the final product at its critical micelle concentration (CMC) were combined and subjected to multiple mixing cycles by repeatedly transferring the fluids between coupled syringes. The resulting emulsion was collected in Falcon tubes, and emulsion stability was evaluated by visual inspection immediately after preparation and after 24 and 48 hours of resting. The emulsification index (E) and emulsion stability (EE) were calculated using the formulas from Cooper & Goldenberg (1987) and Lara-Severino et al. (2017) described before. FIGs. 6C-6D demonstrated the formation of a homogeneous and stable oil-water emulsion when mixed with vegetable oil at concentrations corresponding to the critical micelle concentration (CMC). The biosurfactantcontaining sample exhibited reduced phase separation and maintained emulsion stability after 24 h and 48 h of resting compared to the control sample without biosurfactant, confirming the emulsifying effectiveness and stability of the final formulation. FIGs. 16A-16B depicts the emulsification properties of the biosurfactant compared with two commercial surfactants.Oil polluted wastewater treatment

[0207] For crude oil degradation in water, a stepwise experimental approach, designed to progressively increase environmental and operational complexity while refining performance parameters. A) Initial exploratory assays were carried out at flask scale to assess oil fluidization behavior and to screen effective biosurfactant concentrations across a broad range (0-15% w / v). Based on these preliminary results, B) more advanced flask-scale experiments were subsequently performed using water contaminated with 2% crude oil, incorporating multiple treatment conditions, including the biosurfactant alone and in combination with a cell-free enzymatic extract (ELC), to optimize degradation efficiency. Finally, C) the validated formulations were evaluated under field-relevant conditions through microcosm assays using real produced water from oil wells.

[0208] For the initial exploratory assays, FIGs. 7A-7D show the activity of the biosurfactant as described herein in crude oil degradation biosurfactant. In FIG. 7A, the experimental design included the following:• Factor: Crude oil concentration• Levels (percentage of crude oil): 0%, 2%, 5%, 15%WSGR Docket No.: 69849-701.601• Response Variables: Dry weight (DW), emulsifying activity (EA), and hydrocarbon degradation (TPH)

[0209] In FIG. 7A, 2.5 L of culture medium was prepared following the standard protocol. Empty Erlenmeyer flasks with their caps, along with measuring cylinders, were sterilized. 15 Erlenmeyer flasks with treatment names (crude oil percentage) and replicate numbers were labeled. Under a laminar flow hood, previously disinfected and exposed to UV light, the components listed in Table 2 were added.Table 2: a summary of for components used for Oil polluted wastewater treatmentTreatment Medium (nit) Crude Oil (mL) Inoculum {mL)0% (growth control) 45 0 52% 44 1 55% 42.5 2.5 615% 37.5 7,5 515% (crude control) 42.5 ?5 0

[0210] As depicted in FIGs. 7A-7B, the 2% crude oil concentration was found to be the most effective for biodegradation. This percentage aligns with field tests where real contamination in water and soil varies between 0.5% and 2%. On day 0, two completely separate phases (crude oil and water) are observed. By day 4, the phases begin to mix due to the surfactant action, indicating emulsification. By day 8, crude oil micelles form in the water, and a significant reduction of crude oil is observed, demonstrating the surfactant’s role in emulsification and degradation support. As depicted in FIG, 7C, although degradation is not as evident as at 2%, notable changes in phase mixing can be observed between day 0 and day 8.

[0211] In FIG. 7D, hydrocarbon identification was carried out by GC-MS (Gas Chromatography-Mass Spectrometry). Chromatographic profile analysis, with automatic comparison against the NIST 14 library, was carried out Chemstation software. TPH analysis showed that using 2% crude oil treatment, 80% of crude oil was removed after 8 days. Heavy linear hydrocarbons appeared to be transformed into lighter ones, indicating degradation.Significant reduction of total aromatic hydrocarbons was observed, demonstrating the biosurfactant’s efficiency in breaking down complex hydrocarbons (data not shown).

[0212] To assay for the abilities of the biosurfactant to treat crude oil, the following analysis was carried out: centrifugal separation of the sample into biomass, cell-free media, and emulsified oil; characterization of the cell-free media for soluble biosurfactant activity; and TPH test to evaluate total petroleum hydrocarbon degradation, as described herein. The experimentsWSGR Docket No.: 69849-701.601were carried out under the following conditions: crude oil concentration at 2%; fungal inoculum at 10%; temperature at 18°c; inoculation shaking speed at 150 rpm; incubation for 8 days; and light conditions: daylight.

[0213] As depicted in FIG. 9A, when the 2% crude oil sample was treated with the biosurfactant for 8 days at low temperature, the liquid flowed easily from one container to another, and no separate oil and water phases were visible, indicating successful emulsification and degradation of the crude oil. In contrast, in the untreated controls under the same experimental conditions, as depicted in FIG. 9B, the crude oil remained encapsulated and not mixed or emulsified in the liquid phase. It did not flow easily from the container.

[0214] In additional advanced flask-scale experiments, Hydrocarbon degradation and removal assays were conducted under a single, unified experimental design to evaluate the performance of different biological and physicochemical treatments in two contaminated matrices: (i) water contaminated with 2% (v / v) crude oil and (ii) petroleum sludge at 10% (w / w) suspended in water. These conditions were selected based on the outcomes of the exploratory flask-scale experiments, which allowed the identification of optimal operational ranges and the most relevant treatments.

[0215] The following treatments were evaluated in triplicate: live fungus, cell-free extract (ELC), biosurfactant type 1, biosurfactant type 2, biosurfactant type 1 combined with ELC, biosurfactant type 2 combined with ELC, and an untreated negative control. Response variables: Total Petroleum Hydrocarbons (TPH) including Diesel Range Organics (DRO) and Gasoline Range Organics (GRO).

[0216] All experiments were performed in 150 mL glass bottles prepared under aseptic conditions. For the water-crude oil matrix, the components listed in Table 3 were added. For the sludge matrix, the components listed in Table 4 were added. The systems were gently mixed manually until the crude oil or sludge was fully incorporated into the liquid phase and no adhesion to the container walls was observed. Subsequently, the bottles were incubated under orbital shaking at 150 rpm and ambient temperature for a period of 10 days.Table 3, Components of experimental design for polluted water (Advanced flask experiments)Biosurfactant (30%, Crude Oil Treatment Medium / Extract (mL)mL) (mL) Negative control 35 mL distilled water — 135 mL potato-glucoseLive fungus (5%) — 1mediumCell-free extract (ELC) 35 mL ELC — 1 Biosurfactant type 1 35 mL distilled water 15 1 Biosurfactant type 2 35 mL distilled water 15 1WSGR Docket No.: 69849-701.601Biosurfactant (30%, Crude Oil Treatment Medium / Extract (mL)mL) (mL) Biosurfactant type 1 +35 mL ELC 15 1ELCBiosurfactant type 2 +35 mL ELC 15 1ELCTable 4, Components of experimental design for oily sludge (Advanced flask experiments)Biosurfactant (30%, Sludge Treatment Medium / Extract (mL)mL) (g) Negative control 35 mL distilled water — 535 mL potato-glucoseLive fungus (5%) — 5mediumCell-free extract (ELC) 35 mL ELC — 5 Biosurfactant type 1 35 mL distilled water 15 5 Biosurfactant type 2 35 mL distilled water 15 5 Biosurfactant type 1 +35 mL ELC 15 5 ELCBiosurfactant type 2 +35 mL ELC 15 5ELC

[0217] At the end of the incubation period, hydrocarbon extraction was performed by adding 50 mL of hexane to each bottle at a 1:1 (v / v) ratio. The mixtures were agitated manually or using a vortex for 30 seconds until complete removal of visible hydrocarbon residues from the container walls was achieved. For treatments containing biomass or biological extracts, the contents were transferred to 50 mL Falcon tubes and centrifuged at 4600 rpm for 15 minutes to separate the aqueous and organic / biomass phases. The aqueous phase was recovered into 250 mL amber glass bottles and adjusted to a final volume of 100 mL with hexane when required. The residual organic / biomass phase was washed three times with hexane at a 1 :2 (v / v) ratio, and each wash was collected in amber bottles and adjusted to volume for subsequent analysis.

[0218] For the water matrix, across all evaluated treatments and matrices, the GRO fraction was negligible. Accordingly, degradation performance was primarily assessed based on DRO removal, which served as the primary indicator of hydrocarbon mobilization and biodegradation efficiency. FIG. 9C show the overall results described below.

[0219] In a first treatment condition, the use of live fungus at a concentration of approximately 5% resulted in a DRO removal of up to about 97%, demonstrating the high intrinsic metabolic capacity of the microorganism for hydrocarbon degradation. FIG. 9D depicts the visual appearance of an aqueous matrix contaminated with crude oil treated with live fungus at day 0WSGR Docket No.: 69849-701.601(left image) and after 10 days (right image), showing progressive emulsification and reduction of the oil phase.

[0220] In a second treatment condition, the application of a cell-free enzymatic extract (ELC) achieved DRO removal levels of approximately 90-95%, comparable to those obtained with the live fungus. This result indicates that extracellular enzymes and metabolites contribute substantially to hydrocarbon degradation, even in the absence of viable biomass. In further treatment conditions, formulations comprising biosurfactant type 1 (BS type 1) combined with the cell-free enzymatic extract (ELC), in approximately equal proportions, achieved DRO removal levels of approximately 90-95%, representing the most effective biosurfactant-based treatment evaluated. Without being bound by theory, this enhanced performance is attributed to the low hydrophilic-lipophilic balance (HLB) of biosurfactant type 1, which favors interaction with the oil phase and emulsion formation, synergistically enhanced by enzymatic activity from the ELC. In another embodiment, biosurfactant type 1 applied alone achieved DRO removal levels of approximately 80-85%, with no statistically significant differences observed between application at full concentration and reduced concentration, suggesting the existence of a threshold concentration above which additional surfactant does not proportionally increase degradation efficiency. The FIG. 9E depicts the visual appearance of an aqueous matrix contaminated with crude oil treated with biosurfactant type 1 combined with a cell-free enzymatic extract (ELC) at day 0 (left image) and after 10 days (right image), evidencing enhanced dispersion and homogenization of the hydrocarbon phase.

[0221] In contrast, treatments comprising biosurfactant type 2 (BS type 2) exhibited lower degradation performance. Biosurfactant type 2 applied alone resulted in DRO removal levels of approximately 60-70%, while combinations of biosurfactant type 2 with the cell-free enzymatic extract (ELC) improved removal to approximately 75-85%, although remaining below the performance observed for biosurfactant type 1-based formulations. The untreated negative control showed minimal DRO removal over the same incubation period, confirming that the observed reductions in hydrocarbon concentration were attributable to the biological and physicochemical activity of the treatments described herein. FIG. 9F depicts the negative control (untreated system), in which no emulsification or hydrocarbon degradation is observed.

[0222] For the sludge matrix, after 10 days of incubation, treatments comprising live fungus showed apparent DRO removal levels of up to approximately 99%. FIG. 9G show the overall results described below.

[0223] As depicts in FIG. 9H, treatments comprising biosurfactant type 1 combined with the cell-free enzymatic extract (ELC) achieved high DRO removal (approximately 97%), consistent with results obtained in the aqueous matrix. In the sludge system, no substantial differencesWSGR Docket No.: 69849-701.601were observed between biosurfactant type 1 applied alone and in combination with ELC, suggesting that surfactant-driven mobilization dominates under these conditions. FIG. 91 show the positive control with the living fungus.

[0224] Additionally, treatments comprising biosurfactant type 2 combined with ELC also exhibited high DRO removal (approximately 94%). Without being bound by theory, this result suggests that in dense and heterogeneous matrices, the presence of a more hydrophilic component may facilitate dispersion or solubilization of specific hydrocarbon fractions. FIG. 9J depicts the negative control (untreated system), in which no emulsification or hydrocarbon degradation is observed.

[0225] In field-relevant conditions, using real fracture flowback water (late return) directly obtained from an active oil exploitation site was used. The tested water exhibited elevated hydrocarbon content and challenging physicochemical parameters typical of oilfield wastewater, thereby providing a representative matrix for evaluating treatment performance beyond laboratory-prepared systems.

[0226] To demonstrate that the biosurfactant as described herein could be used for water treatment at field scale, the following analysis was carried out. A sufficient amount of sterile culture medium and inoculum was prepared, as described elsewhere in this disclosure. The pH of the water to be treated was adjusted to approximately 5.5 (such as but not limited to using citric acid), and the final values were recorded. 200 mL of the inoculum and 600 mL of culture medium were included and mixed with water to be treated. The airflow from the compressor outlets using air displacement in a graduated cylinder was measured over time. Each outlet was adjusted to 0.5 vvm (2-2.5 L / min). The clean tubing to the vent filter was connected and distributed each treatment setup. Two replicates were assayed for each treatment condition and sampled at 8, 15, and 30 days. Each experiment was characterized for Total Petroleum Hydrocarbon (TPH) degradation. pH, hardness, total and dissolved solids (TDS), conductivity, BOD, COD, and toxicity. In conclusion, 90% TPH degradation was observed after 8 days of treatment, -30% faster degradation compared to other bioremediation alternatives. The high TDS and BOD values were observed, suggesting that the treated water might require additional physicochemical treatments, such as flocculation, to meet environmental discharge standards (see also FIGs. 11A-11C).

[0227] Extended monitoring up to 15 days further confirmed the robustness of the treatment, with TPH removal levels ranging from approximately 89% to 97% across dilution conditions of 10%, 20%, and 50%, resulting in final TPH concentrations below the regulatory limit of 10 mg / L, as shown in FIG. HD. These results demonstrate that the biosurfactant-based treatment maintains high removal efficiency over time and under varying dilution scenariosWSGR Docket No.: 69849-701.601

[0228] To demonstrate the inducer could be used to facilitate the fermentation of the fungus and subsequently the production of the biosurfactant, as described herein, the following analysis was carried out. To prepare inoculum, a 5% inoculum, with 1% crude oil and the remaining volume filled with culture medium, was prepared. Crude oil was added as an inducer after autoclaving along with the inoculum (other types of inducers as disclosed here, such as but not limited to: hydrocarbons, synthetic lubricating oils, mineral base lubricating oils, oils (such as but not limited to vegetable oils), vegetable fatty acids, alkanes, alcohols, or aldehydes can be tested using a similar method). The initial aeration was set at 1 L / min (0.5 vvm) and increased to 2 L / min on day 3. The culture was subjected to constant agitation at 150 rpm, using a Rushton turbine at the bottom of the system. For sampling, a syringe was purged with 5 mL, left in place, and clamped to the tube. Various samples were frozen for further analysis. Sufficient biomass and liquid phase were collected for detailed evaluation. As depicted in FIGs. 10A-10C, 50% increase in growth speed was observed compared to flask cultures. The microorganism demonstrated growth viability in a commercial reactor system (STR) and grew significantly faster under these conditions. Biosurfactant production was observed, confirming the potential for large-scale applications as described herein.Oil polluted soil treatment

[0229] To assay for the ability of the biosurfactant as described herein for soil treatment, the following analysis was carried out. 500 grams of soil sampled using destructive sampling were used for each experiment unit. The soil was sampled on 15, 30 and 45 days. The pH of the soil was measured and adjusted after wetting. If the pH was above 7.5, the soil was adjusted with citric acid. In a 250 mL beaker, 5 g of soil was mixed with 10 mL of water. The mixture was stirred and let settle for 15 minutes before measuring the pH. In the baseline control there was no additions to the soils. In the treatment (F.L. 10% w / v) group, 50 mL of inoculum + 50 mL of culture medium were added into each soil type. In the bioaugmentation control: 100 mL of culture medium (without inoculum) was added into each soil type. During the experiment, the containers with covered with gauze or perforated film to maintain humidity and prevent insect contamination. The experimental units were stored outdoors, in the shade, with no temperature control. Daily weather conditions were recorded. The moisture was replenished every 2-3 days with 10% of the initial volume (10 mL), adjusting as needed based on visual monitoring.

[0230] The result of the experiment is depicted in FIGs. 12A-12B and summarized below: 50% of TPH degradation was observed using the biosurfactant as described herein within the experimental period. 200% faster degradation was observed using the biosurfactant, compared to other bioremediation alternatives. 96% toxicity removal was observed using the biosurfactant, indicating effective contaminant breakdown. A direct relationship between phosphorus (P)WSGR Docket No.: 69849-701.601levels and degradation rate was observed using the biosurfactant, as well as a strong correlation between humidity and degradation efficiency. As degradation increased, organic matter content decreased, demonstrating the consumption of hydrocarbons by microbial activity of the biosurfactant. The C:N ratio of the soil could influence the degradation efficiency. Organic matter levels can be adjusted to improve treatment effectiveness.

[0231] In addition, the contaminated soils evaluated in this study initially exhibited Total Petroleum Hydrocarbon (TPH) concentrations up to approximately 57 times higher than the applicable regulatory limit (1000 pg / g). Despite this high initial contamination load, the treatment achieved up to 54% TPH reduction within 45 days, as shown in FIG. 12C, with predictive modeling indicating that regulatory compliance could be reached within approximately 60 days under the same operating conditions. The biosurfactant-based treatment outperformed nutrient-only controls by approximately 35%, demonstrating that the observed remediation effect was not attributable solely to nutrient amendment. Notably, effective degradation was achieved without the need for reinoculation, indicating sustained activity under nutrient-deprived conditions. Additionally, the treatment showed synergistic performance when combined with diatomaceous earth, particularly in soils with higher initial contamination levels, supporting its applicability across heterogeneous soil matrices.

[0232] To assay for the biodegradability test of the biosurfactant as described herein, primary effluent from a wastewater treatment plant was used as the inoculum, as depicted in FIG. 15. During the treatment, continuous aeration was applied, and the system was acclimated to the mineral medium before conducting the biodegradability assay. The treatment was preconditioned for 48 hours prior to the test, with 500 mL of dilution water added in aliquots, under continuous aeration. Sodium benzoate (100 mg / L) was used as the reference control substance. The biosurfactant compositions exhibited rapid and sustained biodegradation kinetics under aerobic conditions, as evidenced by the progressive increase in biochemical oxygen demand (BOD) over the evaluated period. Both formulations showed a steep initial biodegradation phase during the first days of incubation, followed by a gradual approach to a plateau, indicating effective microbial utilization of the biosurfactant components. The formulation generated in the presence of an inducer displayed slightly higher BOD values throughout the assay, suggesting enhanced biodegradability relative to the non-induced formulation. Extrapolation of the degradation trend indicates high ultimate biodegradability within a 40-day timeframe, consistent with ready biodegradation behavior and supporting the suitability of the biosurfactant for environmentally sensitive applications.Wellbore cleaning with Acid and BiosurfactantWSGR Docket No.: 69849-701.601

[0233] To evaluate the ability of a biosurfactant-containing acidic formulation to remove oil from carbonate rock surfaces and improve wettability, laboratory-scale cleaning experiments were conducted using carbonate discs coated with crude oil. The experiment simulates conditions encountered during wellbore cleaning or near-wellbore treatments in carbonate formations, where removal of oil films from rock surfaces can improve fluid flow and wettability.

[0234] Carbonate rock plugs or discs composed primarily of calcite, having a diameter of approximately 1-1.5 inches and a thickness of 5-10 mm, were used as model substrates. The discs were first rinsed with synthetic brine containing 10 wt% NaCl to remove loose particles, dried in an oven at approximately 60 °C, and cooled to room temperature. To generate oil-wet surfaces representative of reservoir conditions, the discs were fully immersed in representative crude oil for approximately 24 hours, optionally under mild agitation to promote oil penetration into surface pores. After immersion, the discs were removed with forceps and allowed to drain on absorbent paper for several minutes without wiping, thereby maintaining a continuous oil coating on the rock surface.

[0235] Prior to treatment, each oil-wet disc was photographed from a fixed overhead position against a uniform white background under controlled lighting conditions to obtain reference images representing the initial oil-covered state of the rock surface.

[0236] Three treatment fluids were prepared in 10 wt% NaCl brine, each containing 1.5 wt% hydrochloric acid (HC1). The evaluated formulations included: (i) an acid control formulation consisting of 1.5 wt% HC1 in 10 wt% NaCl brine without surfactant, (ii) a biosurfactant treatment comprising 1.5 wt% HC1 and 1 wt% biosurfactant, and (iii) a commercial surfactant treatment comprising 1.5 wt% HC1 and 1 wt% of a commercially available surfactant. For each formulation, an oil-coated carbonate disc was placed in a 100 mL glass beaker containing the treatment fluid, ensuring complete immersion.

[0237] The beakers were incubated under gentle agitation on an orbital shaker at approximately 50 rpm at ambient temperature for a contact time of approximately 60 minutes, simulating moderate flow conditions near the wellbore during cleaning operations. At the end of the treatment period, the discs were removed using forceps and briefly rinsed with clean brine to remove residual treatment fluid, avoiding any mechanical scrubbing of the rock surface.

[0238] Following treatment, each disc was again photographed under the same controlled lighting and camera conditions used for the pre-treatment images. The resulting images were subjected to quantitative image analysis to determine the fraction of the rock surface still covered by oil. For each photograph, the disc area was isolated using image-processing software and pixels within the disc were classified as either dark pixels (oil-covered surface) or lightWSGR Docket No.: 69849-701.601pixels (exposed rock) using a grayscale or color threshold. The oil-covered surface fraction was calculated as the ratio between the number of dark pixels and the total number of pixels within the disc area, expressed as a percentage. Replicate discs were analyzed for each treatment and the values were averaged to obtain representative oil coverage values.

[0239] FIG. 18 depicts the analysis of the post-treatment images, demonstrated a clear difference in cleaning performance among the tested formulations. In the acid-only control, the carbonate discs retained an average oil coverage of approximately 91.6%, indicating that the aqueous acidic solution alone produced negligible removal of the oil film. Similarly, the commercial surfactant formulation yielded an average oil coverage of approximately 88.7%, corresponding to only marginal improvement relative to the acid control. In contrast, the discs treated with the example biosurfactant 1 -containing formulation exhibited a substantially lower residual oil coverage of approximately 61.1%. This represents a reduction of approximately 40 percentage points relative to the control treatments, corresponding to removal of roughly one-third of the original oil coating under identical experimental conditions. The relatively small variability among replicate measurements indicates that the observed effect was consistent and reproducible.Paraffin dispersal and deposit removal test

[0240] To evaluate the ability of the biosurfactant composition described herein to disperse and mobilize paraffin (wax) deposits under oilfield conditions, a laboratory-scale paraffin dispersal assay was conducted using an artificial wax deposit model in contact with crude oil and brine. The experiment simulates conditions relevant to wax deposition and removal in production tubing, flowlines, or near-wellbore environments, where paraffin accumulation can impair flow assurance and production efficiency.

[0241] A paraffin deposit model was prepared using solid paraffin wax having a melting point of approximately 50-60 °C. Approximately 5 g of wax were melted in a water bath at 70-80 °C, and the molten wax was poured into transparent glass vials (10 mL capacity). Each vial was rotated during cooling so that the molten wax formed a thin, irregular layer coating part of the inner wall and / or bottom of the vial, typically with a thickness of approximately 1-3 mm. The wax was then allowed to cool completely to form an adherent paraffin deposit representative of wax accumulation on tubing or pipeline surfaces.

[0242] To simulate oil-wet reservoir conditions, 2.5 mL of representative crude oil was added to each vial containing the solidified wax deposit. The vials were left undisturbed for approximately 60 minutes, allowing the oil to fully wet the wax surface and penetrate surface irregularities. After this conditioning period, excess free oil was carefully decanted, leaving aWSGR Docket No.: 69849-701.601thin oil film covering the wax deposit and the vial walls, thereby establishing consistent initial conditions across all samples.

[0243] Three treatment systems were prepared to evaluate paraffin dispersal performance. In a first treatment, serving as a control, 5 mL of brine (10 wt% NaCl) were added to the oil / wax system without any additive. In a second treatment, 5 mL of brine containing 1 wt% biosurfactant were introduced as the biosurfactant formulation under evaluation. In a third treatment, 5 mL of brine containing a commercially available paraffin dispersant at its recommended dosage were added. The total fluid volume (oil plus aqueous phase) was maintained approximately constant in all vials to ensure comparable oil-to-brine ratios.

[0244] The vials containing the paraffin deposits and treatment fluids were placed in a thermostated water bath maintained at approximately 60 °C, representative of temperatures encountered in flowlines or near-wellbore environments. After thermal equilibration, the vials were aligned side-by-side and visually monitored for changes in the wax deposit. The systems were maintained under gentle agitation using either a magnetic stirrer or orbital shaker, and the evolution of the paraffin deposits was recorded using a camera mounted on a fixed support under uniform background and lighting conditions. The condition of the wax deposits was documented after a 60-minute contact period.

[0245] During the test, qualitative indicators of paraffin dispersal were observed, including release of thin strands or fragments of wax from the deposit, formation of fine dispersed wax particles within the oil / water phases, and changes in clarity or turbidity of the fluid phases, as well as visible cleaning of the glass surface previously coated with wax.

[0246] FIG. 19 depicts the visual inspection of the post-treatment vials revealed substantial differences between the tested formulations. In the brine control, the vial remained almost completely coated with a thick dark wax / oil film after the treatment period, indicating that the aqueous phase alone produced essentially no removal or dispersal of the paraffin deposit. The commercial surfactant treatment showed only modest improvement relative to the control, with partial fragmentation of the wax layer; however, a dense brown film still covered most of the internal vial surface and the paraffin deposit remained largely attached. In contrast, the vials treated with the biosurfactant formulation exhibited a pronounced reduction of the wax coating. The glass surface became largely transparent along most of its height, with only small residual wax spots remaining near the bottom of the vial. The original cohesive wax deposit was converted into a fine dispersion of wax particles within the liquid phase, indicating effective detachment and disaggregation of the paraffin material.Home care and cleaning applicationsWSGR Docket No.: 69849-701.601

[0247] To evaluate the cleaning performance of the biosurfactant composition described herein for Home Care applications, standardized washing tests were conducted using a ...

Claims

WSGR Docket No.: 69849-701.601CLAIMS WHAT IS CLAIMED IS:

1. A biosurfactant composition, comprising:i. at least one glycolipid, wherein the at least one glycolipid comprises at least one acetyl group;ii. an enzyme; andiii. an additive.

2. The biosurfactant composition of claim 1, wherein the at least one glycolipid comprises at least two acetyl groups.

3. The biosurfactant composition of claim 2, wherein the at least one glycolipid comprises at most five acetyl groups.

4. The biosurfactant composition of claim 3, wherein the at least one glycolipid comprises a chemical structure of formula (I) or (II),wherein:each R1is independently selected from hydrogen, Ci.6alkyl, C2.6alkenyl, C2.6alkynyl, Ci-6 haloalkyl, -C(=O)Rn, -CH2C(=0)Rn, -C(=0)N(Rn)2, and-CH2C(=O)N(Rn)2, wherein Ci.6alkyl, C2.6 alkenyl, C2.6alkynyl, and Ci-6 haloalkyl are optionally substituted with one or more substituents,each R11is independently selected from hydrogen, Ci.6alkyl, C2.6alkenyl, and C2.6alkynyl CH3;each R2is independently selected from hydrogen, halogen, -OH, C1-6 alkyl, C2.„ alkenyl, and C2.6 alkynyl, wherein C1-6 alkyl, C2.6alkenyl, and C2.6alkynyl are optionally substituted with one or more substituents; andeach R3is independently selected from Cx-20 alkyl, Cs.2o alkenyl, and Cs.2o alkynyl wherein Cs- 20 alkyl, C 8-2o alkenyl, and Cs-2o alkynyl are optionally substituted with one or more substituents.

5. The biosurfactant composition of claim 4, wherein R1is hydrogen.WSGR Docket No.: 69849-701.6016. The biosurfactant composition of claim 4, wherein R1is -C(=O)Rn.

7. The biosurfactant composition of claim 6, wherein R11is Ci.6alkyl.

8. The biosurfactant composition of claim 7, wherein R11is Ci alkyl.

9. The biosurfactant composition of claim 6, wherein R1is -C(=O)CH3.

10. The biosurfactant composition of claim 4, wherein R2is optionally substituted C1-5 alkyl.

11. The biosurfactant composition of claim 4, wherein R2is optionally substituted Ci alkyl.

12. The biosurfactant composition of claim 4, wherein R3is optionally substituted Cx-20 alkyl.

13. The biosurfactant composition of claim 4, wherein R3is optionally substituted Cx-20 alkenyl.

14. The biosurfactant composition of claim 13, wherein R3is optionally substituted C12-19 alkenyl.

15. The biosurfactant composition of claim 14, wherein R3is optionally substituted C13 alkenyl or C15 alkenyl.

16. The biosurfactant composition of claim 13, wherein the optionally substituted Cs-20 alkenyl comprises one carbon-carbon double bond.

17. The biosurfactant composition of claim 13, wherein the optionally substituted Cs-20 alkenyl comprises two carbon-carbon double bonds.

18. The biosurfactant composition of claim 4, wherein the one or more substituents are independently selected from halogen, -OH, -NO2, =0, =S, -CN, C1-6 aminoalkyl, C1-6 alkoxy, C1-6 hydroxyalkyl, and Ci-ehaloalkyl.

19. The biosurfactant composition of claim 4, wherein the at least one glycolipid comprises a chemical structure selected from the group consisting of:WSGR Docket No.: 69849-701.601WSGR Docket No.: 69849-701.60120. The biosurfactant composition of claim 1, wherein the enzyme comprises an enzyme from a fungus.

21. The biosurfactant composition of claim 20, wherein the fungus comprises a filamentous fungus.

22. The biosurfactant composition of claim 21, wherein the filamentous fungus comprises at least one species from the family Xylariaceae.

23. The biosurfactant composition of claim 22, the filamentous fungus comprises Entonaema liquescens.

24. The biosurfactant composition of claim 1, wherein the enzyme is selected from the group consisting of alcohol dehydrogenase, monooxygenase, dioxygenase, peroxidase, and laccase.WSGR Docket No.: 69849-701.60125. The biosurfactant composition of claim 24, wherein the enzyme is present in an amount of at least about 0.01% w / w of the biosurfactant composition.

26. The biosurfactant composition of claim 25, wherein the enzyme is present in an amount of at least about 1 % w / w of the biosurfactant composition.

27. The biosurfactant composition of claim 26, wherein the enzyme is present in an amount of at most about 10 % w / w of the biosurfactant composition.

28. The biosurfactant composition of claim 27, wherein the enzyme is present in an amount of at most about 5 % w / w of the biosurfactant composition.

29. A biosurfactant composition, wherein the biosurfactant composition is configured to reduce surface tension between two phases under a condition, wherein the two phases comprise:i. two different liquid phases,ii. a first liquid phase and a solid phase, oriii. a second liquid phase and a gaseous phase; wherein the condition is selected from the group consisting of:a. a pH range that is: (1) from about 2 to about 4; (2) from about 10 to about 12; or (3) a combination of (1) and (2);b. a temperature range that is: (1) from about -10°C to about 4°C; (2) from about 90°C to about 100°C; or (3) a combination of (1) and (2);c. a salinity range that is: (1) from about 50000 total dissolved solids (TDS) to 70000 TDS; (2) from about 130000 TDS to about 150000 TDS or (3) a combination of (1) and (2); ord. any combination of (a)-(c).

30. The biosurfactant composition of claim 29, wherein the first liquid phase and the second liquid phase are a same liquid phase.

31. The biosurfactant composition of claim 29, wherein the first liquid phase and the second liquid phase are different liquid phases.

32. A biosurfactant composition, wherein the biosurfactant composition is configured to remove or sequester a hydrocarbon from a mixture comprising an aqueous component and the hydrocarbon under a condition selected from the group consisting of:a. a pH range that is: (1) from about 2 to about 4; (2) from about 10 to about 12; or (3) a combination of (1) and (2);WSGR Docket No.: 69849-701.601b. a temperature range that is: (1) from about -10°C to about 4°C; (2) from about 90°C to about 100°C; or (3) a combination of (1) and (2);c. a salinity range that is: (1) from about 50000 Total dissolved solids (TDS) to 70000 TDS; (2) from about 130000 TDS to about 150000 TDS or (3) a combination of (1) and (2); ord. any combination of (a)-(c).

33. A biosurfactant composition, wherein the biosurfactant composition is configured to facilitate emulsification of a mixture comprising two different liquids under a condition selected from the group consisting of:a. a pH range that is: (1) from about 2 to about 4; (2) from about 10 to about 12; or (3) a combination of (1) and (2);b. a temperature range that is: (1) from about -10°C to about 4°C; (2) from about 90°C to about 100°C; or (3) a combination of (1) and (2);c. a salinity range that is: (1) from about 50000 Total dissolved solids (TDS) to 70000 TDS; (2) from about 130000 TDS to about 150000 TDS or (3) a combination of (1) and (2); ord. any combination of (a)-(c).

34. A biosurfactant composition, wherein the biosurfactant composition is configured to not exhibit a cloud point formation under a condition selected from the group consisting of:a. a pH range that is: (1) from about 2 to about 4; (2) from about 10 to about 12; or (3) a combination of (1) and (2);b. a temperature range that is: (1) from about -10°C to about 4°C; (2) from about 90°C to about 100°C; or (3) a combination of (1) and (2);c. a salinity range that is: (1) from about 50000 Total dissolved solids (TDS) to 70000 TDS; (2) from about 130000 TDS to about 150000 TDS or (3) a combination of (1) and (2); ord. any combination of (a)-(c).

35. A biosurfactant composition, wherein the biosurfactant composition is configured to exhibit a surface tension of about 30 millinewton / meter (mN / m) under a condition selected from the group consisting of:a. a pH range that is: (1) from about 2 to about 4; (2) from about 10 to about 12; or (3) a combination of (1) and (2);WSGR Docket No.: 69849-701.601b. a temperature range that is: (1) from about -10°C to about 4°C; (2) from about 90°C to about 100°C; or (3) a combination of (1) and (2);c. a salinity range that is: (1) from about 50000 Total dissolved solids (TDS) to 70000 TDS; (2) from about 130000 TDS to about 150000 TDS or (3) a combination of (1) and (2); ord. any combination of (a)-(c).

36. A biosurfactant composition, wherein the biosurfactant composition is configured to exhibit an interfacial tension of about 5 millinewton / meter (mN / m) under a condition selected from the group consisting of:a. a pH range that is: (1) from about 2 to about 4; (2) from about 10 to about 12; or (3) a combination of (1) and (2);b. a temperature range that is: (1) from about -10°C to about 4°C; (2) from about 90°C to about 100°C; or (3) a combination of (1) and (2);c. a salinity range that is: (1) from about 50000 Total dissolved solids (TDS) to 70000 TDS; (2) from about 130000 TDS to about 150000 TDS or (3) a combination of (1) and (2); ord. any combination of (a)-(c).

37. The biosurfactant composition of any one of claims 29-36, further comprising at least one glycolipid.

38. The biosurfactant composition of any one of claims 29-36, further comprising an additive.

39. The biosurfactant composition of claim 1, wherein the at least one glycolipid comprises:sophorolipid, trehalose lipid, rhamnolipid, mannosylerythritol lipid, cellobiose lipid, polyol lipid, or a combination thereof.

40. The biosurfactant composition of claim 39, wherein the at least one glycolipid comprises: the sophorolipid, the trehalose lipid, the rhamnolipid, the mannosylerythritol lipid, or a combination thereof.

41. The biosurfactant composition of claim 40, wherein the at least one glycolipid comprises: the sophorolipid, the rhamnolipid, the mannosylerythritol lipid, or a combination thereof.

42. The biosurfactant composition of claim 41, wherein the at least one glycolipid comprises: the sophorolipid, the rhamnolipid, or a combination thereof.WSGR Docket No.: 69849-701.60143. The biosurfactant composition of claim 42, wherein the at least one glycolipid comprises the sophorolipid.

44. The biosurfactant composition of claim 43, wherein the sophorolipid comprises an acidic sophorolipid, a lactonic sophorolipid, or a combination thereof.

45. The biosurfactant composition of claim 1, wherein the at least one glycolipid is present in an amount of at least about 0.1% w / w of the biosurfactant composition.

46. The biosurfactant composition of claim 45, wherein the at least one glycolipid is present in an amount from about 20 % to about 30 % w / w of the biosurfactant composition.

47. The biosurfactant composition of claim 1, further comprising: a lipopeptide, a lipoprotein, or a combination thereof.

48. The biosurfactant composition of claim 47, wherein the lipopeptide is present in an amount of at least about 0.1 % by w / v of the biosurfactant composition.

49. The biosurfactant composition of claim 48, wherein the lipopeptide is present in an amount from about 10% to about 15% by w / v of the biosurfactant composition.

50. The biosurfactant composition of claim 47, wherein the lipoprotein is present in an amount of at least about 0.05 % by w / v of the biosurfactant composition.

51. The biosurfactant composition of claim 50, wherein the lipoprotein is present in an amount from about 5% to about 10% by w / v of the biosurfactant composition.

52. The biosurfactant composition of claim 1, wherein the additive is selected from the group consisting of antioxidants, preservatives, chelating agents, natural antioxidants, vegetable oil derivatives and any combination thereof.

53. The biosurfactant composition of claim 52, wherein the biosurfactant composition further comprises sodium benzoate.

54. The biosurfactant composition of claim 1, wherein the additive is present in an amount of at least about 0.001% w / v of the biosurfactant composition.

55. The biosurfactant composition of claim 54, wherein the additive is present in an amount from about 0.01% to about 2% w / v of the biosurfactant composition.

56. The biosurfactant composition of claim 1, wherein the biosurfactant composition exhibits:a. a density of about 1 gram per cubic centimeter (g / cm3) at 25 °C; b. a viscosity of about 2.5 Centipoise (cP) at 25 °C;c. a hydrophilic-lipophilic balance of about 10;d. a solids content of about 20 %;e. a critical micelle concentration of about 9 at 25 °C, by w / v; f. a surface tension of about 34 mN / m at 25 °C;WSGR Docket No.: 69849-701.601g. an interfacial tension of about 15 mN / m at 25 °C;h. an emulsion index of about [100] %;i. a hydrophilic contact angle of about 25 ° at 25 °C, when using glass as a reference substrate;j . a hydrophobic contact angle of about 100 ° at 25 °C, when using polytetrafluoroethylene as a reference substrate; ork. any combination of (a)-(j).

57. The biosurfactant composition of claim 1, wherein the biosurfactant composition is a lyophilizate.

58. A method, comprising using the biosurfactant composition of any one of the preceding claims (1) as an emulsifier, foaming control agent, cleaner, surfactant, wetting agent, oil recovery agent, or corrosion inhibitor; (2) to clean wells, facilitate enhanced oil recovery, disperse wax and parafilm facilitate flow assurance, facilitate high-salinity water disposal; or (3) for water treatment or polluted soil treatment; in a crude oil processing cycle.

59. A method, comprising using the biosurfactant composition of any one of the preceding claims to remove or sequester total petroleum hydrocarbons (TPH) from soil or water.

60. A method, comprising using the biosurfactant composition of any one of the preceding claims to remove perfluoroalkyl and polyfluoroalkyl substances (PF AS) from the soil or the water.

61. A method, comprising using the biosurfactant composition of any one of the preceding claims as detergents, degreasers, fabric cleaners, or surface cleaners; optionally the biosurfactant composition is used to remove colored stains from a colored fabric or a color hair.

62. A method, comprising using the biosurfactant composition of any one of the preceding claims to remove oil from a solid particle comprising rock.

63. The method of claim 62, further comprising using the biosurfactant composition and an acid to remove the oil from the solid particle.

64. A method, comprising using the biosurfactant composition of any one of the preceding claims as a dispersant, solubilizer, or anti-bacterial agent within agro-industrial products.

65. A method, comprising using the biosurfactant composition of any one of the preceding claims as a dispersant, solubilizer, or anti-bacterial agent within cosmetic products.

66. A method, comprising using the biosurfactant composition of any one of the preceding claims to facilitate emulsification in the cosmetic products, pharmaceutical products, petroleum products, or food products.WSGR Docket No.: 69849-701.60167. A method, comprising using the biosurfactant composition of any one of the preceding claims to facilitate stabilization of a vaccine.

68. A method, comprising:i. obtaining a crude extract from a microorganism; andii. generating the biosurfactant composition of claim 1 using the crude extract.

69. The method of claim 68, wherein the microorganism comprises a fungus.

70. The method of claim 69, wherein the fungus comprises at least a filamentous fungus.

71. The method of claim 69, wherein the fungus comprises at least one species from the family Xylariaceae.

72. The method of claim 69, wherein the fungus comprises Entonaema liquescens.

73. The method of claim 68, further comprising fermenting the microorganism.

74. The method of claim 73, wherein the fermenting comprises:(1) culturing the microorganism with a substrate comprising polysaccharides; (2) maintaining a culture of the microorganism: (i) from about 18 °C to about 30 °C, (ii) from about pH 3 to about pH 6, (iii) at an aeration rate from about 0.2 vessel volumes per minute (vvm) to about 1 vvm, or (iv) any combinations of (i)- (iii);(3) mechanically agitating the culture from about 100 rpm (revolution per minute) to about 200 rpm; or(4) a combination of (l)-(3).

75. The method of claim 74, wherein the culturing comprises adding an inducer to the microorganism.

76. The method of claim 75, wherein the inducer is selected from the group consisting of hydrocarbons, synthetic lubricating oils, mineral base lubricating oils, vegetable fatty acids, and other sources of alkanes, alcohols, and aldehydes.

77. The method of claim 75, wherein the inducer comprises an oil.

78. The method of claim 77, wherein the oil comprises a vegetable oil.

79. The method of claim 75, wherein the inducer is at a concentration range of 0.2% to 5% w / v of the culture.

80. The method of claim 74, further comprising concentrating the culture.

81. The method of claim 80, wherein the concentrating comprises centrifugation.

82. The method of claim 68, further comprising purifying the crude extract from the culture.

83. The method of claim 82, wherein the purifying comprises filtration.

84. The method of claim 83, wherein the filtration comprises using a filter.WSGR Docket No.: 69849-701.60185. The method of claim 84, wherein the filter has a pore size of about at most about 1 millimeter.

86. The method of claim 84 or 85, wherein the filter has a pore size of about at least about 1 micrometer.

87. The method of claim 83, wherein the filtration comprises vacuum filtration.

88. The method of claim 74, wherein the substrate comprises agro-industrial waste or renewable sources.

89. The method of claim 74, wherein the polysaccharides comprise starch, amylopectin, pectin, lignin, cellulose, or a derivative thereof, or a combination thereof.

90. The method of claim 68, wherein the crude extract is substantially cell-free.

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