Methods for processing biomass to produce oligosaccharides and related compositions

Abstract

A method for forming an ingredient for human consumption is provided herein. The method can include isolating one or more soluble polysaccharides from biomass, producing one or more oligosaccharides from the biomass, and combining the isolated soluble polysaccharides with the produced oligosaccharides to form an ingredient. A method for pre-treating biomass is also provided. The method can include subjecting biomass to physical pre-treatment, subjecting the physically pre-treated biomass to mild pre-treatment, and subjecting the mildly pre-treated biomass to vigorous pre-treatment. An ingredient for human consumption is also provided.

Description

[Technical field]

[0001] cross reference This application claims the benefit of UK Patent Application No. 1911762.1, filed August 16, 2019, UK Patent Application No. 1911764.7, filed August 16, 2019, and UK Patent Application No. 2002315.6, filed February 19, 2020, each of which is incorporated by reference in its entirety. [Background technology]

[0002] background Sugar-containing foods and beverages are an important part of cultural habits and lifestyles around the world, but the sugar they contain has been linked to obesity, diabetes, poor dental health and destructive behavior in humans. Because of this, consumer preferences are shifting away from sugar-containing foods and governments are increasingly enacting regulations to encourage less sugar consumption.

[0003] Therefore, the industry has been searching for suitable low-calorie sweeteners for decades to replace sugar in food and beverages.Unfortunately, many sugar substitutes are produced from non-natural sources and often bring a potential bitterness or other unpleasant taste along with their sweetness, neither of which is appealing to consumers.In addition, although many sweeteners can mimic the sweetness of sugar in food and beverages, few can comprehensively mimic the role that sugar plays in food, such as adding bulk, adjusting texture, providing structure, acting as a preservative, and adjusting color and flavor through caramelization and Maillard reaction.In addition, many bulk sweeteners that can mimic such physical properties of sugar have tolerance problems in the gastrointestinal tract that limit the use of bulk sweeteners to levels well below the amount required to replace sugar in a standard Western diet.

[0004] Dietary fiber is an important part of a positive diet and helps maintain digestive health and a well-regulated intestinal flora. Such fiber includes sugars of various chain lengths and types. In addition to being found naturally in a wide range of foods, fiber can also be produced separately or added to other foods during their manufacture. Summary of the Invention [Means for solving the problem]

[0005] Abstract Additional aspects and advantages of the present disclosure will become readily apparent to those skilled in the art from the following detailed description, in which only illustrative embodiments of the present disclosure have been 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 present disclosure. Accordingly, the drawings and description are to be regarded as illustrative in nature, and not as restrictive.

[0006] In some embodiments, a method for producing an ingredient for human consumption is described herein.The method for producing an ingredient for human consumption can include the steps of (a) isolating one or more soluble polysaccharides from biomass; (b) contacting the remaining biomass with one or more enzymes to form one or more oligosaccharides; (c) isolating one or more oligosaccharides; and (d) combining a portion of the one or more soluble polysaccharides from step (a) with a portion of the one or more oligosaccharides from step (c) to form an ingredient.

[0007] In some cases, the methods may include a step of purifying the isolated soluble polysaccharide or polysaccharides.

[0008] In some cases, the methods can include a step of purifying the isolated oligosaccharide or oligosaccharides.

[0009] In some cases, the methods may include treating the biomass to dissolve one or more soluble polysaccharides.

[0010] In some cases, the methods may include a step of purifying the isolated soluble polysaccharide or polysaccharides.

[0011] In some cases, the treating step includes a thermochemical treatment.

[0012] In some cases, the thermochemical treatment includes at least one of a hot water treatment or a high temperature alkaline treatment.

[0013] In some cases, the hot alkaline treatment uses an alkali having a pH of 10-14.

[0014] In some cases, the hot alkaline treatment uses at least one of sodium hydroxide, potassium hydroxide, sodium carbonate, calcium carbonate, calcium hydroxide, ammonium sulfate, ammonium hydroxide, or aqueous ammonia.

[0015] In some cases, the treating can be carried out at a temperature between 30°C and 180°C.

[0016] In some cases, the treating can occur for between 10 minutes and 24 hours.

[0017] In some cases, the one or more soluble polysaccharides and / or one or more oligosaccharides are dried prior to step (d).

[0018] In some cases, the one or more soluble polysaccharides and / or one or more oligosaccharides are dried after step (d).

[0019] In some cases, the ingredients may be water soluble.

[0020] In some cases, the solubility of an ingredient in water may be at least 80 g of ingredient per 100 g of water at 50°C.

[0021] In some cases, the method includes combining the component with a liquid to form a liquid component.

[0022] In some cases, the viscosity of the liquid component may be similar to that of corn syrup.

[0023] In some cases, the viscosity of the liquid component may be similar to that of high fructose corn syrup.

[0024] In some cases, the liquid ingredient has fewer calories per gram than corn syrup or high fructose corn syrup.

[0025] In some cases, the liquid component has a lower glycemic index than corn syrup or high fructose corn syrup.

[0026] In some cases, the liquid comprises water.

[0027] In some cases, the liquid component comprises at least 20% by dry weight of at least one oligosaccharide, and at least 2% by dry weight of at least one polysaccharide.

[0028] In some cases, the liquid component has a viscosity of 5 cps to 100,000 cps, 8,000 cps to 100,000 cps, 10,000 cps to 50,000 cps, or 15,000 cps to 25,000 cps.

[0029] In some cases, the liquid component comprises at least 2% xylan by dry weight.

[0030] In some cases, the liquid component comprises at least 2% mannan, by dry weight.

[0031] In some cases, the liquid component comprises at least 2% cellulose derivative by dry weight.

[0032] In some cases, the liquid component has a concentration of polysaccharide between 0.1% and 50% w / v.

[0033] In some cases, the liquid component includes polysaccharides and oligosaccharides in an amount ranging from a ratio of 1:100 to 1:1.

[0034] In some cases, the one or more soluble polysaccharides include at least one of mannan, xylan, mixed linkage glucan, lignocellulose, hemicellulose, cellulose derivatives, chitosan, or xyloglucan.

[0035] In some cases, the cellulose derivative includes at least one of cellulose acetate, hydroxyethyl cellulose, or hydroxymethyl cellulose.

[0036] In some cases, the biomass includes at least one of sugarcane biomass, corn biomass, wheat biomass, hardwood biomass, or softwood biomass.

[0037] In some cases, the one or more oligosaccharides include at least one of: i) cello-oligosaccharides having a degree of polymerization (DP) of 2-6; ii) xylo-oligosaccharides having a DP of 2-12; iii) arabinoxylo-oligosaccharides having a DP of 3-15; iv) manno-oligosaccharides having a DP of 2-12; v) mixed linkage glucan oligosaccharides having a DP of 2-5; vi) xyloglucan oligosaccharides having a DP of 4-12; or vii) chitooligosaccharides having a DP of 2-12.

[0038] In some cases, the component includes at least two of the oligosaccharides listed in (i)-(vii).

[0039] In some cases, the components include at least two oligosaccharides in a ratio of 1:9 to 1:1 with respect to each other.

[0040] In some embodiments, a composition for human consumption is described herein.The composition for human consumption may include soluble polysaccharides, and the oligosaccharides may include at least one of the following: (i) cello-oligosaccharides with a degree of polymerization (DP) of 2-6; (ii) xylo-oligosaccharides with a DP of 2-12; (iii) manno-oligosaccharides with a DP of 2-12; (iv) arabinoxylo-oligosaccharides with a DP of 3-15; (v) mixed-linked glucan oligosaccharides with a DP of 2-5; or (vi) chitooligosaccharides with a DP of 2-12, and the composition includes less than 5% insoluble polysaccharides by dry weight.

[0041] In some cases, the composition may be substantially free of insoluble polysaccharides.

[0042] In some cases, the compositions may be water-soluble.

[0043] In some cases, the solubility of the composition in water may be at least 80 g of the composition per 100 g of water at 50°C.

[0044] In some cases, the composition further comprises a liquid, thereby forming a liquid component.

[0045] In some cases, the liquid may be water.

[0046] In some cases, the liquid component comprises at least 20% by dry weight of at least one oligosaccharide, and at least 2% by dry weight of at least one polysaccharide.

[0047] In some cases, the liquid component has a viscosity of 5 cps to 100,000 cps, 8,000 cps to 100,000 cps, 10,000 cps to 50,000 cps, or 15,000 cps to 25,000 cps.

[0048] In some cases, the liquid component comprises at least 2% xylan by dry weight.

[0049] In some cases, the liquid component comprises at least 2% mannan, by dry weight.

[0050] In some cases, the liquid component comprises at least 2% cellulose derivative by dry weight.

[0051] In some cases, the liquid component has a concentration of polysaccharide between 0.1% and 50% w / v.

[0052] In some cases, the liquid component includes polysaccharides and oligosaccharides in an amount ranging from a ratio of 1:100 to 1:1.

[0053] In some cases, the one or more soluble polysaccharides include at least one of mannan, xylan, mixed linkage glucan, lignocellulose, hemicellulose, cellulose derivatives, chitosan, or xyloglucan.

[0054] In some cases, the cellulose derivative includes at least one of cellulose acetate, hydroxyethyl cellulose, or hydroxymethyl cellulose.

[0055] In some cases, the biomass includes at least one of sugarcane biomass, corn biomass, wheat biomass, hardwood biomass, or softwood biomass.

[0056] In some cases, the one or more oligosaccharides include at least one of: i) cello-oligosaccharides having a degree of polymerization (DP) of 2-6; ii) xylo-oligosaccharides having a DP of 2-12; iii) arabinoxylo-oligosaccharides having a DP of 3-15; iv) manno-oligosaccharides having a DP of 2-12; v) mixed linkage glucan oligosaccharides having a DP of 2-5; vi) xyloglucan oligosaccharides having a DP of 4-12; or vii) chitooligosaccharides having a DP of 2-12.

[0057] In some cases, the composition comprises at least two of the oligosaccharides listed in (i)-(vii).

[0058] In some cases, the components include at least two oligosaccharides in a ratio of 1:9 to 1:1 with respect to each other.

[0059] In some embodiments, a method for producing ingredients for human consumption is described herein.The method for producing ingredients for human consumption can include: (a) subjecting biomass to physical pretreatment to reduce the average size of biomass; (b) subjecting the physically pretreated biomass to mild pretreatment, where mild pretreatment can include (i) incubating the physically pretreated biomass in an aqueous solution to dissolve monosaccharides and / or disaccharides derived from the physically pretreated biomass; and (ii) removing some of the dissolved monosaccharides and / or disaccharides from the aqueous solution; (c) subjecting the mild pretreated biomass to strong pretreatment to increase the digestibility of the biomass; (d) contacting one or more polysaccharide-cutting enzymes and the strong pretreated biomass in a solution or suspension to form one or more oligosaccharides; and (e) concentrating the solution or suspension to increase the concentration of one or more oligosaccharides to form ingredients.

[0060] In some cases, the mild pretreatment may be an incubation cycle.

[0061] In some cases, the aggressive pre-treatment may be a thermochemical treatment and may include incubating the mildly pre-treated biomass in one of an acidic or alkaline solution.

[0062] In some cases, the method may further comprise removing at least 25% or 50% of the dissolved monosaccharides and / or disaccharides from the incubation solution in step (b)(ii).

[0063] In some cases, the robust pre-treated biomass composition after step (c) comprises less than 10% w / w monosaccharides.

[0064] In some cases, the method can further include purifying one or more oligosaccharides from the solution or suspension.

[0065] In some cases, the strong pre-treated biomass composition after step (c) comprises less than 20% w / w monosaccharides.

[0066] In some cases, the method may further include repeating step (b).

[0067] In some cases, step (b) may be performed two, three, four or five times.

[0068] In some cases, the method may further include repeating step (c).

[0069] In some cases, step (c) may be performed two, three, four or five times.

[0070] In some cases, the method may further comprise the step of concentrating a portion of the dissolved monosaccharides and / or disaccharides removed in step (b).

[0071] In some cases, the method may further comprise the step of discarding a portion of the dissolved monosaccharides and / or disaccharides removed in step (b).

[0072] In some cases, a portion of the dissolved monosaccharides and / or disaccharides removed in step (b) may not be combined with a portion of the one or more oligosaccharides in step (e) to form a component.

[0073] In some cases, the ingredients contain less than 15% monosaccharides by dry weight.

[0074] In some cases, the component contains less than 50% disaccharides, by dry weight.

[0075] In some cases, the ingredients may be substantially free of simple sugars.

[0076] In some cases, the component may be substantially free of disaccharides.

[0077] In some cases, the one or more oligosaccharides include at least one of: i) cello-oligosaccharides having a degree of polymerization (DP) of 2-6; ii) xylo-oligosaccharides having a DP of 2-12; iii) arabinoxylo-oligosaccharides having a DP of 3-15; iv) manno-oligosaccharides having a DP of 2-12; v) mixed linkage glucan oligosaccharides having a DP of 2-5; vi) xyloglucan oligosaccharides having a DP of 4-12; or vii) chitooligosaccharides having a DP of 2-12.

[0078] In some cases, the component includes at least two of the oligosaccharides listed in (i)-(vii).

[0079] In some cases, the components include at least two oligosaccharides in a ratio of 1:9 to 1:1 with respect to each other.

[0080] In some cases, the ingredients include at least one of sucrose, maltose, lactose, glucose, fructose or galactose in a dry sum of less than 50% w / w of the total dry w / w of all oligosaccharides i-vii.

[0081] In some cases, the monosaccharides and / or disaccharides include at least one of sucrose, maltose, lactose, glucose, fructose, or galactose.

[0082] In some cases, step (b) dissolves one or more organic acids in addition to the monosaccharides and / or disaccharides.

[0083] In some cases, the one or more organic acids include at least one of oxalate, tartrate, succinate, formate, citrate, maleate, lactate, or acetate.

[0084] In some cases, the total weight of oxalate, tartrate, succinate, formate, citrate, maleate, lactate and acetate may be present in the portion dissolved and removed in step b) at more than 10% of the total weight of sucrose, maltose, lactose, glucose, fructose and galactose.

[0085] In some cases, the physical pre-treatment of step (a) includes at least one of chipping, chopping, milling, ball milling, grinding, shaping, or blending the biomass.

[0086] In some cases, the mild pre-treatment of step (b) is carried out in an aqueous solution, which may include water.

[0087] In some cases, the mild pre-treatment of step (b) is carried out at a temperature between 5°C and 150°C.

[0088] In some cases, the mild pre-treatment of step (b) may be carried out for 15 minutes to 1 hour.

[0089] In some cases, the aggressive pre-treatment of step (c) involves heating the mildly pre-treated biomass in an acidic or alkaline solution.

[0090] In some cases, the heating may be to a temperature of 50°C to 150°C.

[0091] In some cases, heating may occur for 30 minutes to 4 hours.

[0092] In some cases, step (c) includes treating the mild pre-treated biomass in an alkaline solution having a pH of 8-11.

[0093] In some cases, the alkaline solution includes at least one of sodium hydroxide, potassium hydroxide, sodium carbonate, calcium carbonate, aqueous ammonia, ammonium sulfate, or ammonium hydroxide.

[0094] In some cases, step (c) includes treating the mild pre-treated biomass in an acidic solution having a pH of 4-6.

[0095] In some cases, the acidic solution includes at least one of sulfuric acid, hydrochloric acid, nitric acid, phosphoric acid, acetic acid, maleic acid, fumaric acid, or oxalic acid.

[0096] In some cases, the biomass includes at least one of sugar cane, corn stover, corn cob, bran, wheat straw, hardwood, or softwood.

[0097] In some cases, the biomass comprises at least one of cellulose, chitin, chitosan, xylan, xyloglucan, mixed linkage glucan, mannan, or lignocellulose.

[0098] In some cases, the one or more polysaccharide cleaving enzymes include at least one of cellulase, xylanase, xyloglucanase, endo-glucanase, cellobiohydrolase, mannanase, lichenase, or lytic polysaccharide monooxygenase (LPMO).

[0099] In some cases, the one or more polysaccharide cleaving enzymes include at least one of AA9, AA10, AA11, AA13, AA14, or AA15.

[0100] In some cases, the one or more polysaccharide cleaving enzymes can be prepared from a filamentous fungus, such as Trichoderma reesei.

[0101] In some cases, the one or more polysaccharide-cleaving enzymes may be operably linked to a catalytic module.

[0102] In some cases, one or more polysaccharide-cleaving enzymes may be operably linked to a non-catalytic module.

[0103] In some cases, the non-catalytic module may be a carbohydrate-binding module.

[0104] In some cases, the water soluble composition for human consumption can include at least one of the following oligosaccharides: i) cellooligosaccharides having a degree of polymerization (DP) of 2-6; ii) xylooligosaccharides having a DP of 2-12; iii) arabinoxylooligosaccharides having a DP of 3-15; iv) mannooligosaccharides having a DP of 2-12; v) mixed linkage glucan oligosaccharides having a DP of 2-5; vi) xyloglucan oligosaccharides having a DP of 4-12; or vii) chitooligosaccharides having a DP of 2-12; and at least one of the following monosaccharides or disaccharides: sucrose, maltose, lactose, fructose, or galactose, wherein the total dry weight of the monosaccharides or disaccharides comprises less than 10% of the total dry weight of the oligosaccharides having a DP of 2-12.

[0105] In some cases, the components can include at least two of the oligosaccharides listed in (i)-(vii). In some cases, the components can further include at least one of the following organic acids: oxalate, tartrate, succinate, formate, citrate, maleate, lactate, or acetate. In some cases, the components can include at least two of the organic acids. In some cases, the components can include at least two of the oligosaccharides listed in (i)-(vii) in a ratio of 1:9 to 1:1 with respect to each other.

[0106] In some cases, the composition may comprise at least one monosaccharide or disaccharide selected from the group consisting of glucose, fructose, or sucrose; at least one organic acid selected from the group consisting of oxalate, tartrate, succinate, formate, citrate, maleate, lactate, or acetate, wherein the total weight of the organic acids is greater than 10% of the total weight of the monosaccharides or disaccharides.

[0107] In some cases, a method of producing an ingredient for human consumption can include the steps of: (a) subjecting a biomass to a physical pre-treatment to reduce the average size of the biomass; (b) subjecting the physically pre-treated biomass to a mild pre-treatment, the mild pre-treatment including: (i) incubating the physically pre-treated biomass in an aqueous solution to dissolve monosaccharides and / or disaccharides from the physically pre-treated biomass; and (ii) removing a portion of the dissolved monosaccharides and / or disaccharides from the aqueous solution; (c) subjecting the mildly pre-treated biomass to a strong pre-treatment to dissolve polysaccharides and increase digestibility of the plant biomass; (d) isolating one or more dissolved polysaccharides from the biomass; (e) contacting the remaining biomass with one or more enzymes to form one or more oligosaccharides; (f) isolating the one or more oligosaccharides; and (g) combining a portion of the one or more soluble polysaccharides from step (d) with a portion of the one or more oligosaccharides from step (f) to form the ingredient.

[0108] The novel features of the invention are set forth with particularity in the appended claims. A better understanding of the features and advantages of the present invention will be obtained by reference to the following detailed description that sets forth illustrative embodiments, in which the principles of the invention are utilized, and the accompanying drawings (also referred to herein as "Figure" and "FIG."). [Brief description of the drawings]

[0109] [Figure 1] FIG. 1 shows the results of thin layer chromatography (TLC) analysis showing the mono- and disaccharide content of the biomass shown after 1 to 5 washing cycles (pre-incubation treatment).

[0110] [Diagram 2] FIG. 2 shows the results of a TLC analysis comparing the levels of mono- and disaccharides in the mixture of enzymatic hydrolysis products when washing (pre-incubation treatment) was performed (+) or not (-).

[0111] [Diagram 3] FIG. 3 shows the results of making a glaze with the composition of the present disclosure and the comparative composition.

[0112] [Figure 4] FIG. 4 shows a baked product using the liquid components of the present disclosure.

[0113] [Diagram 5] FIG. 5 shows a high performance anion exchange chromatography (HPAEC) analysis of sample 4.

[0114] [Figure 6] FIG. 6 shows a cereal bar produced using the liquid ingredients of the present disclosure.

[0115] [Figure 7] FIG. 7 shows an HPAEC chromatogram of sugars in water after washing of corn cobs.

[0116] [Figure 8] FIG. 8 is a simplified flow diagram illustrating a method of processing biomass according to some embodiments of the present disclosure.

[0117] [Figure 9] FIG. 9 is a simplified flow diagram illustrating another method of processing biomass according to some embodiments of the present disclosure.

[0118] [Figure 10-1] FIG. 10A is a diagram illustrating a method for processing biomass.

[0119] [Figure 10-2] FIG. 10B shows the measurement of sugars in a number of samples.

[0120] [Figure 10-3] FIG. 10C shows the measured organic acids in a number of samples.

[0121] [Figure 10-4] FIG. 10D shows a visual observation of a sample processed by the method of FIG. 10A.

[0122] [Figure 11-1] FIG. 11A shows a comparison between cereal bars made using soluble and insoluble polysaccharides.

[0123] [Figure 11-2] FIG. 11B shows the hardness and stickiness of the cereal bars of FIG. 11A using a penetration test.

[0124] [Figure 11-3] FIG. 11C shows the hardness of the cereal bars of FIG. 11A using a cut test. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0125] Detailed Description overview A method for forming one or more components is provided herein. The method can include isolating one or more soluble polysaccharides from a biomass or raw material (e.g., a plant biomass). The remaining biomass can then be contacted or treated with one or more enzymes to form one or more oligosaccharides, which can be enriched or isolated. Furthermore, at least a portion of the one or more soluble polysaccharides isolated from the biomass can be combined with a portion of the one or more enriched or isolated oligosaccharides to form a component.

[0126] The texture of the ingredient is consistent and smooth. Furthermore, the ingredient can have certain properties so that the ingredient can be used as a sweetener and / or sugar substitute. The ingredient properties can include sweetness, smooth texture, desirable mouthfeel, binding ability, glaze or shine-forming ability, moistness, viscosity, bulking ability, and / or caramelization ability. Compared to corn syrup or high fructose corn syrup, the ingredient can also have lower calories, lower glycemic index, lower glycemic load, rich in fiber, and / or less sugar.

[0127] Similarly, methods of producing ingredients for incorporation into food products, dietary supplements and / or cosmetics are provided herein that can include one or more pre-processing steps performed on the biomass. For example, the method can include a first pre-processing step, a second pre-processing step, a third pre-processing step or an additional pre-processing step performed on the biomass. The pre-processing steps can be performed in a specified order.

[0128] The biomass used to produce one or more oligosaccharides can be plant biomass. Examples of plant biomass include, but are not limited to, sugarcane, corn stover, corn cob, bran, wheat straw, hardwood or softwood. In some cases, the biomass can include cellulose, chitin, chitosan, xylan, xyloglucan, mixed-linked glucan, mannan, or lignocellulose.

[0129] The biomass can be digested into one or more oligosaccharides. The biomass digestion can be enzymatic. The enzymatic digestion can be performed after one or more pre-treatment steps. The one or more pre-treatment steps can be performed to reduce the size of the biomass and / or to increase the surface area of ​​the biomass available for digestion. The one or more pre-treatment steps can include one or more washing steps, dissolution steps, or pre-digestion. In some cases, the one or more pre-treatment steps can be performed to reduce the monosaccharides and / or disaccharides present in the biomass. In some cases, the one or more pre-treatment steps can be performed to recover a soluble polysaccharide fraction from the biomass.

[0130] During one or more pre-treatment steps, monosaccharides and / or disaccharides may be removed from the starting material (e.g., biomass). In other words, monosaccharides and / or disaccharides may be removed from the biomass during one or more pre-treatment steps. Thus, no or substantially no monosaccharides and / or disaccharides may be generated upon completion of one or more pre-treatment steps. That is, the pre-treated biomass may not contain any or substantially no monosaccharides and / or disaccharides. This can improve the efficiency of the method of producing ingredients for incorporation into food products, dietary supplements and / or cosmetics provided herein. For example, less purification of the ingredients (e.g., to remove monosaccharides and / or disaccharides) is required because some of the monosaccharides and / or disaccharides have already been removed from the biomass during one or more pre-treatment steps. In particular, fewer or less rigorous filtration steps may be required during purification to produce the ingredients disclosed herein.

[0131] The first pretreatment step (pretreatment step 1) can include physically treating the biomass (e.g., pulverizing the biomass). The second pretreatment step (pretreatment step 2 or mild pretreatment) can include subjecting the physically treated biomass to an incubation cycle or a washing cycle. The incubation cycle can include incubating the physically treated biomass (from pretreatment step 1) in an aqueous solution to dissolve monosaccharides and / or disaccharides from the physically treated biomass. The incubation cycle can also include removing a portion of the dissolved monosaccharides and / or disaccharides from the aqueous solution. In some cases, the aqueous solution can include water. In some other cases, the incubation cycle can be performed at about 25° C. for a period of about 30 minutes to about 1.5 hours. Pretreatment step 2 can be a mild pretreatment step. For example, the conditions (e.g., solution, temperature, time, etc.) of pretreatment step 2 can be milder than the conditions of pretreatment step 3, as described in more detail below.

[0132] The third pretreatment step (pretreatment step 3 or strong pretreatment) can include treating the incubated biomass from pretreatment step 2 in one of an acidic or alkaline solution. Pretreatment step 3 can improve the digestibility of the biomass (e.g., by enzymes). Pretreatment step 3 can also improve the access of enzymes to the biomass. In various examples, pretreatment step 3 can be performed in an alkaline solution (e.g., 1% w / v NaOH solution) at a temperature higher than room temperature (e.g., about 90°C to about 110°C). Furthermore, pretreatment step 3 can be performed by holding at the desired temperature for about 30 minutes to 1 hour. For example, the effective temperature, in this example 90°C, is held for 1 hour (this can be varied depending on the desired characteristics). The solution in the third pretreatment step can be held. In other words, the solution does not have to be discarded when the treated biomass is moved from the pretreatment step to a step after the pretreatment step, which is described in more detail below. In some cases, pretreatment step 3 can include a thermochemical treatment. That is, the pre-treatment step 3 may be carried out in an acidic or alkaline solution and / or the pre-treatment step 3 may be carried out at a temperature above room temperature.

[0133] In various examples, after one or more pre-processing steps, the method of producing an ingredient for human consumption can include contacting, in a solution or suspension, one or more polysaccharide cleaving enzymes and the biomass from pre-processing step 3 to form one or more oligosaccharides. The method can further include concentrating the solution or suspension to increase the concentration of the one or more oligosaccharides to form an ingredient (e.g., an ingredient for human consumption).

[0134] The pre-treatment step may improve the efficiency of the method compared to some other methods. Efficiency may be improved insofar as less expensive downstream processing may be required. In some cases, downstream processing may include a step of extracting monosaccharides and / or disaccharides from the pre-treated biomass. In some embodiments, it may be difficult to extract only disaccharides from, for example, intermediate solutions or fractions produced by enzymatic digestion. Therefore, it may be more effective to extract disaccharides during the pre-treatment step.

[0135] Downstream processing steps can include ion exchange chromatography, ultrafiltration, microfiltration, nanofiltration, etc. Part of the role of the nanofiltration step can be to remove excess monosaccharides from the oligosaccharide mixture. This nanofiltration can be performed multiple times to reach the desired monosaccharide level. If few monosaccharides are present in the pre-treated biomass (e.g., monosaccharides have been removed by washing or incubation), the number of such nanofiltration steps can be reduced. Furthermore, ultrafiltration generally cannot distinguish between desired disaccharides (e.g., cellobiose) and undesired disaccharides. Thus, the number of steps involved or required during downstream processing can also be reduced by removing undesired disaccharides during pre-treatment and generating desired disaccharides during the enzymatic treatment step.

[0136] While various embodiments of the present invention have been shown and described herein, it will be apparent to those skilled in the art that such embodiments are presented 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 present invention described herein may be used.

[0137] As used in this specification and claims, the singular forms "a," "an," and "the" include plural references unless the context clearly dictates otherwise. For example, the term "sample" includes multiple samples, including mixtures thereof.

[0138] The term "about" as used herein can mean within one standard deviation or greater than one standard deviation. Alternatively, about can mean a range of up to 10%, up to 5%, or up to 1% of a given value. For example, about can mean up to ±10%, ±9%, ±8%, ±7%, ±6%, ±5%, ±4%, ±3%, ±2%, or ±1% of a given value.

[0139] As used herein, "food" and "foodstuff" generally refer to any item that can be or is intended to be eaten by humans or any other animal. It can be a food, feed, beverage, or an ingredient used in the production of any of the above.

[0140] As used herein, "dietary supplement" generally refers to any composition that is introduced into a human or other animal, whether by ingestion, injection, absorption, or any other method, to provide nutrition to the human or other animal. Such dietary supplement applications may take the form of a beverage with added dietary fiber, a prebiotic additive, a pill or other capsule, or any other suitable application.

[0141] As used herein, "cosmetics" generally refers to any composition intended for use on humans or other animals to enhance its aesthetic appeal or prevent future loss of aesthetic appeal, and any other composition known in the general industry as cosmetics. Aesthetic appeal is not limited to appearance aesthetics, but also applies to texture appeal or any other appeal. Cosmetics can be mascara, foundation, lip gloss, eye shadow, eye liner, primer, lipstick, blusher, nail polish, bronzer, or any other makeup product; shampoo, conditioner, styling mousse, styling gel, hairspray, hair dye, hair wax, or any other hair product; moisturizer, exfoliant, sunscreen, cleanser, toothpaste, cream, lotion, ointment, or any other composition that is effective in repairing teeth, skin, hair, or other parts of the body in some aesthetic way. Additionally, the cosmetic product may be a composition used as a component of a face mask, a brush, a hair roller, other styling device, other solid structure, or any other suitable composition.

[0142] As used herein, "ingredients" generally refers to any composition suitable for incorporation into foodstuff, cosmetic or dietary supplement products, and may include any composition that can be used directly as a product itself.An ingredient can be a dry ingredient or a liquid ingredient, unless it is specifically referred to as "dry" or "liquid".This includes compositions that can be considered intermediates during the method of the present disclosure, such as the composition formed after combining one or more oligosaccharides with one or more soluble polysaccharides, before any further purification, optimization, drying, dissolving or any other such step, and includes the final composition obtained from the method.

[0143] As used herein, "polysaccharide" generally refers to any length of sugar polymer longer than about 20 residues. Polysaccharides may be hyperbranched, lightly branched, or unbranched. Polysaccharides may include any combination of glycosidic linkages in any manner; any number of α-linked or β-linked groups; and any combination of monomer types, such as glucose, glucosamine, mannose, xylose, galactose, fucose, fructose, glucuronic acid, arabinose, or derivatives thereof, such as any combination of the above monomers modified by acetyl or other groups. Polysaccharides may be cellulose or hemicellulose polymers. Contemplated hemicellulose polymers include xylan, glucuronoxylan, arabinoxylan, glucomannan, and xyloglucan. In some embodiments, the cellulose polymer may be cellulose.

[0144] As used herein, "lignocellulose" generally refers to aggregates containing polysaccharides that are or are derived from plant cell wall material. For example, lignocellulose can contain one or more of the following polysaccharides bound together: cellulose, xylan, mannan, and mixed-linked glucans.

[0145] As used herein, "highly branched," "lightly branched," and "unbranched" generally refer to the number of side chains per backbone stretch in a sugar. Highly branched sugars have an average of 4-10 side chains per 10 backbone residues, lightly branched sugars have an average of 1-3 side chains per 10 backbone residues, and unbranched sugars have only one backbone and no side chains. The average is calculated by dividing the number of side chains in the sugar by the number of backbone residues.

[0146] As used herein, "sugar" generally refers to either polysaccharides and / or oligosaccharides, such as monosaccharides and / or disaccharides.

[0147] As used herein, "oligosaccharide" generally refers to a sugar polymer having a chain length of less than or equal to about 20 sugar residues. Oligosaccharides may be highly branched, lightly branched or unbranched and may contain any combination of glycosidic linkages, any number of α or β linking groups, and any combination of monomer types such as glucose, glucosamine, mannose, xylose, galactose, fucose, fructose, glucuronic acid, arabinose or derivatives thereof. Suitable derivatives include the above monomers, including acetyl or other groups.

[0148] As used herein, "monosaccharide" and "disaccharide" generally refer to sugar compounds consisting of one or two residues, respectively. Monosaccharides are compounds such as glucose, glucosamine, xylose, galactose, fucose, fructose, glucuronic acid, arabinose, galacturonic acid, or epimers or other derivatives thereof. Suitable derivatives include acetyl or other groups. Disaccharides are compounds consisting of two monosaccharides linked via any glycosidic bond.

[0149] As used herein, "cellooligosaccharide" refers generally to an oligosaccharide consisting of one or more glucose residues linked by β-1,4-glycosidic bonds and may be chemically related to the oligosaccharide by oxidation, reduction, esterification, epimerization or another chemical modification.

[0150] As used herein, "xylooligosaccharide" refers generally to an oligosaccharide composed primarily of xylose residues (usually linked by β-1,4-glycosidic bonds) and may also contain glucuronic acid residues and / or arabinose residues and / or acetyl groups and / or any other modification, which may be chemically related to the oligosaccharide by oxidation, reduction, esterification, epimerization, further glycosylation or another chemical modification.

[0151] As used herein, "arabinoxylooligosaccharide" refers generally to an oligosaccharide composed of xylose residues (usually linked by β-(1→4) linkages substituted with arabinose side chains) that are typically linked by (1→2) or (1→3) linkages, and may be chemically related to the oligosaccharide by oxidation, reduction, esterification, epimerization, further glycosylation, or another chemical modification.

[0152] As used herein, "mixed linkage glucan oligosaccharide" refers generally to an oligosaccharide composed of one or more glucose residues linked by at least one β-1,3-glycosidic bond and at least one β-1,4-glycosidic bond, and may be chemically related to the oligosaccharide by oxidation, reduction, esterification, epimerization, or another chemical modification.

[0153] As used herein, "mannooligosaccharides" refers generally to oligosaccharides composed of one or more mannose residues and optionally containing one or more glucose and / or galactose residues, and may be chemically related to the oligosaccharides by oxidation, reduction, esterification, epimerization or another chemical modification.

[0154] As used herein, "chitooligosaccharide" refers generally to an oligosaccharide composed of one or more glucosamine and / or N-acetyl-glucosamine residues, and may be chemically related to the oligosaccharide by oxidation, reduction, esterification, epimerization, or another chemical modification.

[0155] As used herein, "cellulose" generally refers to polysaccharides composed of glucose residues linked by β-1,4-glycosidic bonds, and derivatives thereof. As used herein, "xylan" generally refers to polysaccharides composed of a backbone of xylose residues, and may also contain glucuronic acid residues and / or arabinose residues and / or acetyl groups and / or any other modifications. As used herein, "mixed linkage glucan" generally refers to polysaccharides composed of glucose residues linked by β-1,3-glycosidic and β-1,4-glycosidic bonds. As used herein, "mannan" generally refers to polysaccharides composed of more than 40% mannose residues, and optionally containing glucose and / or galactose residues. As used herein, "chitin" or "chitosan" generally refers to polysaccharides composed of glucosamine and / or N-acetyl-glucosamine residues. The polysaccharides of cellulose, xylan, mixed linkage glucan, mannan, chitin or chitosan can include chemical variants modified by oxidation, reduction, esterification, epimerization or another chemical modification.

[0156] As used herein, "soluble," "solubility," and grammatical variations thereof generally refer to solubility in an aqueous solution (e.g., water). As used herein, "dissolve" generally refers to the state in which a solid becomes incorporated into an aqueous solution or liquid to form a solution.

[0157] As used herein, a "suspension" generally refers to a composition comprising at least two immiscible phases, e.g., a solid phase and a liquid phase, where the weight of the solid phase, as a percentage of the weight of the composition, can range from about 0.5% to about 30%, about 1% to about 20%, about 2% to about 15%, or about 3% to about 10%. The suspension may include a suitable solvent, which may be water.

[0158] As used herein, "viscosity" generally refers to a quantity that expresses the amount of internal friction in a fluid, measured by the force per unit area that resists uniform flow. Viscosity can be measured by a variety of methods, but values ​​given herein refer to those obtained using a Brookfield HDB VE Roto Viscometer, unless otherwise indicated, using standard test procedures operated according to the manufacturer's instructions for ranges and with 400 mL samples taken in tall beakers to ensure that no container effects occur.

[0159] As used herein, "dissolved" generally refers to a solid being combined with a liquid to form a solution. I. Pre-processing Physical Pretreatment

[0160] Mechanical or physical pretreatment can be performed on the biomass to digest it into one or more oligosaccharides. Mechanical and / or physical pretreatment can be the first pretreatment step in the process of digesting the biomass. Alternatively, the mechanical or physical pretreatment step can be performed after another pretreatment step. For example, the mechanical or physical pretreatment step can be performed after another pretreatment step, such as a washing pretreatment step.

[0161] Biomass can be mechanically or physically pretreated to reduce the size of the biomass. Examples of mechanical or physical pretreatment steps include, but are not limited to, chipping, chopping, milling, ball milling, grinding, shaping, blending, and / or steam exposure of the biomass. More than one physical pretreatment may be performed on the biomass. Lysis step

[0162] The biomass may be subjected to a lysis step. The lysis step may be or may be part of a mild pretreatment step that dissolves the polysaccharide fraction or the monosaccharide and / or disaccharide fractions derived from the biomass. The lysis pretreatment step may be a washing step, an incubation step, a thermochemical step or a chemical treatment step. The lysis pretreatment may be performed before the physical or mechanical pretreatment step. The lysis pretreatment may be performed after the physical or mechanical pretreatment step.

[0163] In some embodiments, the dissolution step may be performed to remove a fraction of soluble polysaccharides. The soluble polysaccharides may be added to one or more oligosaccharides, for example, during purification of the soluble polysaccharides. The soluble polysaccharide fraction may be used to produce food types of desired taste, texture, quality, adhesiveness and odor. The presence of dissolved polysaccharides may also help produce a good quality product that does not produce sediment (or graininess) in the food product.

[0164] The dissolution step can be a chemical or thermochemical treatment of the biomass. The chemical or thermochemical treatment can include one or more aqueous solutions. The aqueous solutions can include one or more salts, acids, alkalis or ions. The aqueous solutions can include one or more of sodium hydroxide, potassium hydroxide, sodium carbonate, calcium carbonate, calcium hydroxide, ammonium sulfate, ammonium hydroxide, aqueous ammonia, dilute sulfuric acid, dilute acetic acid, dilute hydrochloric acid or dilute phosphoric acid.

[0165] The aqueous solution may be an alkaline solution having a pH of 10 to 14. The aqueous solution may be an alkaline solution having a pH of 10 to 11, 10 to 12, 10 to 13, 10 to 14, 11 to 12, 11 to 13, 11 to 14, 12 to 13, 12 to 14, or 13 to 14. The aqueous solution may be an acid solution having a pH of 2 to 6. The aqueous solution may be an acid solution having a pH of 2 to 3, 2 to 4, 2 to 5, 2 to 6, 3 to 4, 3 to 5, 3 to 6, 4 to 5, 4 to 6, or 5 to 6.

[0166] The solubilization step may be carried out at a temperature between 30°C and 180°C. The solubilization step may be carried out at a temperature of at least 30°C. The solubilization step may be carried out at a temperature of at most 180°C. The solubilization step may be carried out at a temperature between 30°C and 60°C, between 30°C and 90°C, between 30°C and 120°C, between 30°C and 150°C, between 30°C and 180°C, between 60°C and 90°C, between 60°C and 120°C, between 60°C and 150°C, between 60°C and 180°C, between 90°C and 120°C, between 90°C and 150°C, between 90°C and 180°C, between 120°C and 150°C, between 120°C and 180°C, or between 150°C and 180°C. The solubilization step may be carried out at a temperature of at least 30°C, 60°C, 90°C, 120°C, 150°C or 180°C.

[0167] The duration of the dissolution step can vary depending on the biomass used, the desired soluble polysaccharide components and / or the complexity of the desired soluble polysaccharide. The dissolution step may be carried out for about 10 minutes to about 24 hours. The solubilization step may be carried out for at least 10 minutes. The solubilization step may be carried out for at most 60 minutes. The solubilization step may be carried out for 10 minutes to 30 minutes, 10 minutes to 60 minutes or 30 minutes to 60 minutes. The solubilization step may be carried out for at least 10 minutes, 30 minutes or 60 minutes. The solubilization step may be carried out for 1 hour to 24 hours. The solubilization step may be carried out for at least 1 hour. The solubilization step may be carried out for at most 24 hours. The solubilization step may be carried out for 1 hour to 4 hours, 1 hour to 8 hours, 1 hour to 12 hours, 1 hour to 16 hours, 1 hour to 20 hours, 1 hour to 24 hours, 4 hours to 8 hours, 4 hours to 12 hours, 4 hours to 16 hours, 4 hours to 20 hours, 4 hours to 24 hours, 8 hours to 12 hours, 8 hours to 16 hours, 8 hours to 20 hours, 8 hours to 24 hours, 12 hours to 16 hours, 12 hours to 20 hours, 12 hours to 24 hours, 16 hours to 20 hours, 16 hours to 24 hours or 20 hours to 24 hours. The solubilization step may be carried out for at least 1 hour, 4 hours, 8 hours, 12 hours, 16 hours, 20 hours or 24 hours.

[0168] Soluble polysaccharide may be removed from the solution after the dissolution step. A fraction of dissolved polysaccharide may be removed from the dissolved fraction. In some cases, at least 5% of the dissolved polysaccharide may be removed and / or purified. In some cases, at most 100% of the dissolved polysaccharide may be removed and / or purified. In some cases, 5%-10%, 5%-20%, 5%-30%, 5%-40%, 5%-60%, 5%-80%, 5%-90%, 5%-100%, 10%-20%, 10%-30%, 10%-40%, 10%-60%, 10%-80%, 10%-90%, 10%-100%, 20%-30% of the dissolved polysaccharide may be removed and / or purified. , 20%-40%, 20%-60%, 20%-80%, 20%-100%, 30%-40%, 30%-60%, 30%-80%, 30%-100%, 40%-60%, 40%-80%, 40%-100%, 60%-80%, or 60%-100% may be removed and purified. In some cases, at least about 5%, 10%, 20%, 30%, 40%, 60%, 80%, or 100% of the dissolved polysaccharide may be removed and purified. Gentle pre-treatment

[0169] The biomass may be subjected to a mild pretreatment step. The mild pretreatment step may dissolve polysaccharide fractions or monosaccharide fractions and / or disaccharide fractions derived from the biomass. In some cases, the mild pretreatment step may include at least part, or all, of the dissolution step. In other words, the dissolution step may be part or a substep of the mild pretreatment step. The mild pretreatment step may be a washing step, an incubation step, a thermochemical step or a chemical treatment step. The mild pretreatment may be performed before the physical or mechanical pretreatment step. The mild pretreatment may be performed after the physical or mechanical pretreatment step. The mild pretreatment may be performed simultaneously with the physical or mechanical pretreatment step. The mild pretreatment step may be performed one or more times. The mild pretreatment may be performed 2, 3, 4, 5, 6, 7, 8, 9 or 10 times.

[0170] The mild pre-treatment step can be an incubation step or a washing step. The biomass (physically treated or not treated) may be incubated in an aqueous solution. The aqueous solution may be water or the aqueous solution may contain salts, acids, alkalis, ions, alcohols and / or other chemicals. The pH of the aqueous solution may be 6.2-8.5. The pH of the aqueous solution may be at least 6.2. The pH of the aqueous solution may be at most 8.5. The pH of the aqueous solution may be 6.2-6.5, 6.2-7, 6.2-7.2, 6.2-7.5, 6.2-7.7, 6.2-8, 6.2-8.2, 6.2-8.5, 6.5-7, 6.5-7.2, 6.5-7.5, 6.5-7.7, 6.5-8, 6.5-8.2, 6.5-8.5, 7-7.2, 7-7.5, 7-7.7, The pH of the aqueous solution may be 7 to 8, 7 to 8.2, 7 to 8.5, 7.2 to 7.5, 7.2 to 7.7, 7.2 to 8, 7.2 to 8.2, 7.2 to 8.5, 7.5 to 7.7, 7.5 to 8, 7.5 to 8.2, 7.5 to 8.5, 7.7 to 8, 7.7 to 8.2, 7.7 to 8.5, 8 to 8.2, 8 to 8.5, or 8.2 to 8.5. The pH of the aqueous solution may be 6.2, 6.5, 7, 7.2, 7.5, 7.7, 8, 8.2, or 8.5.

[0171] The pH of the aqueous solution can be 9-12. The pH of the aqueous solution can be at least 9. The pH of the aqueous solution can be at most 12. The pH of the aqueous solution can be 9-9.5, 9-10, 9-10.5, 9-11, 9-11.5, 9-12, 9.5-10, 9.5-10.5, 9.5-11, 9.5-11.5, 9.5-12, 10-10.5, 10-11, 10-11.5, 10-12, 10.5-11, 10.5-11.5, 10.5-12, 11-11.5, 11-12, or 11.5-12. The pH of the aqueous solution can be 9, 9.5, 10, 10.5, 11, 11.5, or 12.

[0172] The incubation step may be carried out for 15 minutes to 60 minutes. The incubation step may be carried out for at least 15 minutes. The incubation step may be carried out for at most 60 minutes. The incubation step may be carried out for 15 minutes to 30 minutes, 15 minutes to 45 minutes, 15 minutes to 60 minutes, 30 minutes to 45 minutes, 30 minutes to 60 minutes or 45 minutes to 60 minutes. The incubation step may be carried out for at least 15 minutes, 30 minutes, 45 minutes or 60 minutes. The incubation step may be carried out for 1 hour to 24 hours. The incubation step may be carried out for at least 1 hour. The incubation step may be carried out for at most 24 hours. The incubation step may be performed for 1 hour to 4 hours, 1 hour to 8 hours, 1 hour to 12 hours, 1 hour to 16 hours, 1 hour to 20 hours, 1 hour to 24 hours, 4 hours to 8 hours, 4 hours to 12 hours, 4 hours to 16 hours, 4 hours to 20 hours, 4 hours to 24 hours, 8 hours to 12 hours, 8 hours to 16 hours, 8 hours to 20 hours, 8 hours to 24 hours, 12 hours to 16 hours, 12 hours to 20 hours, 12 hours to 24 hours, 16 hours to 20 hours, 16 hours to 24 hours, or 20 hours to 24 hours. The incubation step may be performed for at least 1 hour, 4 hours, 8 hours, 12 hours, 16 hours, 20 hours, or 24 hours.

[0173] Dissolved polysaccharides, monosaccharides and / or disaccharides may be removed from the aqueous solution after the mild pre-treatment step. In some cases, monosaccharides and / or disaccharides may be removed from the aqueous solution after the mild pre-treatment step.

[0174] The soluble polysaccharides may be removed from the solution after the mild pre-treatment step. At least a portion of the dissolved polysaccharides may be removed from the dissolved fraction. In some cases, at least 5% of the dissolved polysaccharides may be removed and / or purified. In some cases, up to 100% of the dissolved polysaccharides may be removed and / or purified. In some cases, 5%-10%, 5%-20%, 5%-30%, 5%-40%, 5%-60%, 5%-80%, 5%-100%, 10%-20%, 10%-30%, 10%-40%, 10%-60%, 10%-80%, 10%-100%, 20%-30%, 20%-40%, 20%-60%, 20%-80%, 20%-100%, 30%-40%, 30%-60%, 30%-80%, 30%-100%, 40%-60%, 40%-80%, 40%-100%, 60%-80%, or 60%-100% of the dissolved polysaccharide may be removed and / or purified. In some cases, at least about 5%, 10%, 20%, 30%, 40%, 60%, 80% or 100% of the dissolved polysaccharides may be removed and / or purified after the mild pre-treatment step.

[0175] The monosaccharides and / or disaccharides may be removed from the solution after a mild pretreatment step. A fraction of the dissolved monosaccharides and / or disaccharides may be removed from the aqueous solution after an incubation step. In some cases, at least 5% of the dissolved monosaccharides and / or disaccharides may be removed. In some cases, at most 100% of the dissolved monosaccharides and / or disaccharides may be removed. In some cases, 5%-10%, 5%-20%, 5%-30%, 5%-40%, 5%-60%, 5%-80%, 5%-100%, 10%-20%, 10%-30%, 10%-40%, 10%-60%, 10%-80%, 10%-100%, 20%-30%, 20%-40%, 20%-60%, 20%-80%, 20%-100%, 30%-40%, 30%-60%, 30%-80%, 30%-100%, 40%-60%, 40%-80%, 40%-100%, or 60%-100% of the dissolved monosaccharides and / or disaccharides may be removed. In some cases, at least about 5%, 10%, 20%, 30%, 40%, 60%, 80% or 100% of the dissolved monosaccharides and / or disaccharides may be removed and / or purified after the mild pre-treatment step. A portion of the monosaccharides and / or disaccharides may be discarded after the incubation step. In certain instances, the portion of the dissolved monosaccharides and / or disaccharides removed in this step may not be combined with the portion of the one or more oligosaccharides produced in the biomass processing.

[0176] The aqueous solution may be removed from the biomass after the mild pre-treatment step. A portion of the aqueous solution may be removed from the biomass after the mild pre-treatment step. Between 5% and 100% of the aqueous solution may be removed from the biomass after the mild pre-treatment step. At least 5% of the aqueous solution may be removed from the biomass after the mild pre-treatment step. At most 98% of the aqueous solution may be removed from the biomass after the mild pre-treatment step. Aqueous solution of 5%-10%, 5%-20%, 5%-40%, 5%-50%, 5%-60%, 5%-80%, 5%-90%, 5%-98%, 10%-20%, 10%-40%, 10%-50%, 10%-60%, 10%-80%, 10%-90%, 10%-98%, 20%-40%, 20%-50%, 20%-60%, 20%-80%, 20%-90%, 20% ~98%, 40%-50%, 40%-60%, 40%-80%, 40%-90%, 40%-98%, 50%-60%, 50%-80%, 50%-90%, 50%-98%, 60%-80%, 60%-90%, 60%-98%, 80%-90%, 80%-98%, or 90%-98% may be removed from the biomass after the mild pre-treatment step. At least 5%, 10%, 20%, 40%, 50%, 60%, 80%, 90% or 98% of the aqueous solution may be removed from the biomass after the mild pre-treatment step. A portion of the aqueous solution may be removed using pressure filtration, centrifugation, sedimentation, filtration and / or any other suitable method. Powerful pre-processing

[0177] The biomass may be subjected to a strong pretreatment step. The strong pretreatment step may dissolve polysaccharide fractions or monosaccharide fractions and / or disaccharide fractions derived from the biomass. The strong pretreatment step may be performed to digest the biomass more by enzymes. The strong pretreatment step may help to break hydrogen bonds in the biomass. The strong pretreatment step may be a washing step, a thermochemical step or a chemical treatment step. The strong pretreatment step may be performed before a physical or mechanical pretreatment step. The strong pretreatment step may be performed after a physical or mechanical pretreatment step. The strong pretreatment step may be performed before a mild pretreatment step. The strong pretreatment step may be performed after a mild pretreatment step. The strong pretreatment step may be performed one or more times. The strong pretreatment may be performed 2, 3, 4, 5, 6, 7, 8, 9 or 10 times.

[0178] The strong pre-treatment step may be a thermochemical treatment. The chemical or thermochemical treatment may include one or more aqueous solutions. The aqueous solutions may include one or more salts, acids, alkalis or ions. The aqueous solutions may be alkaline solutions including one or more of sodium hydroxide, potassium hydroxide, sodium carbonate, calcium carbonate, calcium hydroxide, ammonium sulfate, ammonium hydroxide or aqueous ammonia. The aqueous solutions may be acidic solutions including at least one of sulfuric acid, hydrochloric acid, nitric acid, phosphoric acid, acetic acid, maleic acid, fumaric acid or oxalic acid. In some embodiments, the aqueous solutions may be retained once the strong pre-treatment step or steps are completed. In other words, the aqueous solutions may not be discarded or excluded from acceptance.

[0179] The thermochemical pretreatment may be carried out at a pH of about 2 to 6.5. The thermochemical pretreatment may be carried out at a pH of at least 2. The thermochemical pretreatment may be carried out at a pH of at most 6.5. The thermochemical pretreatment may be carried out at a pH of 2 to 2.5, 2 to 3, 2 to 3.5, 2 to 4, 2 to 4.5, 2 to 5, 2 to 5.5, 2 to 6, 2 to 6.5, 2.5 to 3, 2.5 to 3.5, 2.5 to 4, 2.5 to 4.5, 2.5 to 5, 2.5 to 5.5, 2.5 to 6, 2.5 to 6.5, 3 to 3.5, 3 to 4, 3 to 4.5, 3 to 5, 3 to 5.5, 3 to 6, 3 to 6.5 The thermochemical pretreatment may be carried out at a pH of about 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, or 6.5. The thermochemical pretreatment may be carried out at a pH of about 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, or 6.5.

[0180] The thermochemical pretreatment may be carried out at a pH of 7.5 to 12. The thermochemical pretreatment may be carried out at a pH of at least 7.5. The thermochemical pretreatment may be carried out at a pH of at most 12. The thermochemical pretreatment may be carried out at a pH of 7.5 to 8, 7.5 to 8.5, 7.5 to 9, 7.5 to 9.5, 7.5 to 10, 7.5 to 10.5, 7.5 to 11, 7.5 to 11.5, 7.5 to 12, 8 to 8.5, 8 to 9, 8 to 9.5, 8 to 10, 8 to 10.5, 8 to 11, 8 to 11.5, 8 to 12, 8.5 to 9, 8.5 to 9.5, 8.5 to 10, 8.5 to 10.5, 8.5 to 11, 8.5 to 11.5, 8 The thermochemical pretreatment may be carried out at a pH of about 7.5, 8, 8.5, 9, 9.5, 10, 10.5, 11, 11.5, or 12. The thermochemical pretreatment may be carried out at a pH of about 7.5, 8, 8.5, 9, 9.5, 10, 10.5, 11, 11.5, or 12.

[0181] The thermochemical pretreatment may be carried out at a temperature of 50°C to 150°C. The thermochemical pretreatment may be carried out at a temperature of at least 50°C. The thermochemical pretreatment may be carried out at a temperature of at most 150°C. The thermochemical pretreatment may be carried out at a temperature of 50°C to 75°C, 50°C to 80°C, 50°C to 90°C, 50°C to 100°C, 50°C to 120°C, 50°C to 130°C, 50°C to 150°C, 75°C to 80°C, 75°C to 90°C, 75°C to 100°C, 75°C to 120°C, 75°C to 130°C, 75°C to 150°C, 80°C to 90°C, 80°C to 100°C. , 80°C to 120°C, 80°C to 130°C, 80°C to 150°C, 90°C to 100°C, 90°C to 120°C, 90°C to 130°C, 90°C to 150°C, 100°C to 120°C, 100°C to 130°C, 100°C to 150°C, 120°C to 130°C, 120°C to 150°C, or 130°C to 150°C. Thermochemical pretreatment may be carried out at a temperature of at least 50°C, 75°C, 80°C, 90°C, 100°C, 120°C, 130°C or 150°C.

[0182] The thermochemical treatment may be carried out for 0.5 hours to 4 hours. The thermochemical treatment may be carried out for at least 0.5 hours. The thermochemical treatment may be carried out for at most 4 hours. The thermochemical treatment may be carried out for 0.5 hours to 0.75 hours, 0.5 hours to 1 hour, 0.5 hours to 1.5 hours, 0.5 hours to 2 hours, 0.5 hours to 2.5 hours, 0.5 hours to 3 hours, 0.5 hours to 3.5 hours, 0.5 hours to 4 hours, 0.75 hours to 1 hour, 0.75 hours to 1.5 hours, 0.75 hours to 2 hours, 0.75 hours to 2.5 hours, 0.75 hours to 3 hours, 0.75 hours to 3.5 hours, 0.75 hours to 4 hours, 1 hour to 1.5 hours, 1 hour to 2 hours. The thermochemical treatment may be carried out for at least 0.5 hours, 0.75 hours, 1 hour, 1.5 hours, 2 hours, 2.5 hours, 3 hours, 3.5 hours, or 4 hours.

[0183] Thermochemically treated biomass may contain between 1% w / w and less than 30% w / w of monosaccharides. Thermochemically treated biomass may contain between 1% w / w and 2% w / w, between 1% w / w and 5% w / w, between 1% w / w and 10% w / w, between 1% w / w and 15% w / w, between 1% w / w and 20% w / w, between 1% w / w and 25% w / w, between 1% w / w and 30% w / w, between 2% w / w and 5% w / w, between 2% w / w and 10% w / w, between 2% w / w and 15% w / w, between 2% w / w and 20% w / w, between 2% w / w and 25% w / w, between 2% w / w and 30% w / w, between 5% w / w and 10% w / w, between 5% w / w and It may contain less than 15% w / w, 5% w / w-20% w / w, 5% w / w-25% w / w, 5% w / w-30% w / w, 10% w / w-15% w / w, 10% w / w-20% w / w, 10% w / w-25% w / w, 10% w / w-30% w / w, 15% w / w-20% w / w, 15% w / w-25% w / w, 15% w / w-30% w / w, 20% w / w-25% w / w, 20% w / w-30% w / w, or 25% w / w-30% w / w of monosaccharides. Thermochemically treated biomass may contain less than 1% w / w, 2% w / w, 5% w / w, 10% w / w, 15% w / w, 20% w / w, 25% w / w or 30% w / w of monosaccharides.

[0184] Thermochemically treated biomass can contain 5% w / w to 50% w / w of disaccharides. Thermochemically treated biomass can contain 5% w / w to 10% w / w, 5% w / w to 15% w / w, 5% w / w to 20% w / w, 5% w / w to 25% w / w, 5% w / w to 30% w / w, 5% w / w to 35% w / w, 5% w / w to 40% w / w, 5% w / w to 50% w / w, 10% w / w to 15% w / w / w, 10%w / w~20%w / w, 10%w / w~25%w / w, 10%w / w~30%w / w, 10%w / w~35%w / w, 10%w / w~ 40%w / w, 10%w / w~50%w / w, 15%w / w~20%w / w, 15%w / w~25%w / w, 15%w / w~30%w / w, 15% w / w~35%w / w, 15%w / w~40%w / w, 15%w / w~50%w / w, 20%w / w~25%w / w, 20%w / w~30%w / w , 20%w / w~35%w / w, 20%w / w~40%w / w, 20%w / w~50%w / w, 25%w / w~30%w / w, 25%w / w~35 % w / w, 25% w / w-40% w / w, 25% w / w-50% w / w, 30% w / w-35% w / w, 30% w / w-40% w / w, 30% w / w-50% w / w, 35% w / w-40% w / w, 35% w / w-50% w / w, or 40% w / w-50% w / w of disaccharides. Thermochemically treated biomass may contain less than 5% w / w, 10% w / w, 15% w / w, 20% w / w, 25% w / w, 30% w / w, 35% w / w, 40% w / w or 50% w / w of disaccharides. II. Enzymatic Treatment and Downstream Processing

[0185] The method of the present disclosure may also include contacting one or more polysaccharide-cleaving enzymes and the thermochemically treated biomass in a solution or suspension to form one or more oligosaccharides. Additionally, the method may include concentrating the solution or suspension to increase the concentration of one or more oligosaccharides to form the components. One or more oligosaccharides may be purified from the solution or suspension provided herein.

[0186] One or more steps of the method of forming or producing the composition can be an enzymatic reaction, in which one or more enzymes are placed in a suitable reaction vessel together with one or more feedstocks or biomasses (e.g., plant biomass), which may be soluble or insoluble in water and a suitable solvent. As used herein, the term "plant biomass" may be replaced by the terms "feedstock" or "biomass" (e.g., biomass not derived from a plant), unless otherwise indicated.

[0187] A variety of enzymes may be suitable for use in the enzymatic reaction. Any enzyme that produces oligosaccharides when acting on a polysaccharide-containing feedstock may be suitable. For example, the enzymatic reaction may include cellulase, endo-glucanase, cellobiohydrolase, lytic polysaccharide monooxygenase (LPMO), lichenase, xyloglucan endoglucanase (XEG), mannanase, chitinase, xylanase, and / or one or more suitable enzymes.

[0188] In various examples, the enzymatic reaction may include cellulolytic preparations from genera such as Trichoderma reesei, which may be purified and / or pretreated and / or supplemented with one or more additional enzymes, such as beta-glucanase, beta-xylanase and cellobiohydrolase; beta-glucanase, beta-xylanase, LPMO and cellobiohydrolase; LPMO and xylanase; or LPMO, xylanase and lichenase. The enzymes may be supplied to the enzymatic reaction as purified enzymes, semi-purified mixtures, derived from a natural source or laboratory-grown cultures, in the form of microbial strains engineered to produce the enzymes, or in any other suitable manner. It is also envisioned to fuse these enzymes to either other enzymes or non-enzymatic modules, such as carbohydrate-binding modules (CBMs). For example, a LPMO fused to a CBM, a xylanase fused to a CBM, or a xylanase fused to a LPMO may be utilized.

[0189] Aerobic conditions can be used for one or more enzymatic reactions. Aerobic conditions can include the addition of oxygen, which can be provided by aeration of the substrate mixture with an oxygen-containing gas, such as air. Aeration can be achieved by introducing oxygen-containing bubbles into the aqueous substrate mixture by various systems, such as an air-injector, an aeration frit, a membrane system, or an internal loop airlift reactor. In some cases, the concentration of molecular oxygen during the enzymatic reaction can be from about 4 mg / L to about 14 mg / L.

[0190] Another exemplary enzyme is lichenase, which may be selected from the GH5, GH7, GH8, GH9, GH12, GH16, GH17 or GH26 family. For example, a GH16 enzyme, such as a GH16 enzyme from Bacillus subtilis, may be utilized. This enzyme may be able to act on mixed linkage glucans, for example, glucans that contain a mixture of β-1,3 and β-1,4 linkages, and cleave these linkages at the β-1,4 glycosidic bond. When lichenase acts on mixed linkage glucans, the resulting β-glucans may largely fall within the size range of 3 to about 7 residues, and thus, this β-glucan may be particularly useful in the food, cosmetic and nutraceutical industries. Mixed linkage glucans are abundant in grasses and members of the Equisetaceae family, and thus, grass-based feedstocks such as straw generally have high levels of mixed linkage glucans and may be usefully acted upon with lichenase. The lichenase may include a GH5 lichenase derived from Bacillus subtilis.

[0191] Another enzyme option is xylanase, which can act on raw materials that contain a xylan backbone. Xylanases can be, for example, glucuronoxylanase, arabinoxylanase, or glucuronoarabinoxylanase. The enzymes can be active on various polymers with a xylan backbone, such as glucuronoxylan, arabinoxylan, and glucuronoarabinoxylan. These polymers are generally abundant in raw materials from various plants, for example, both hardwoods and softwoods can contain suitable polysaccharides, hardwoods often contain glucuronoxylan, and softwoods often contain arabinoglucuronoxylan. Xylanases can include GH5 xylanases from Ruminiclostridium thermocellum and Gonapodya prolifera, and GH30 xylanases from Dickeya chrysanthemi, Bacillus subtilis, Bacteroides ovatus, and Trichoderma reesei.

[0192] Another alternative enzyme is mannanase, which can act on raw materials that contain mannan backbones. Mannanase can be, for example, mannanase, glucomannanase, galactomannanase or galactoglucomannanase. This enzyme can be active on various polymers with mannan backbones, such as mannan, glucomannan, galactomannan or galactoglucomannan. These polymers are generally abundant in raw materials from various plants, for example, both broadleaf and softwoods can contain suitable polysaccharides. Suitable mannanases include GH5 mannanase from Trichoderma reesei and Aspergillus niger, and GH26 mannanase from Aspergillus niger.

[0193] Other enzymes can include xyloglucanases and xyloglucan endoglucanases (XEGs), which are produced by various organisms, including plant pathogenic microorganisms. Xyloglucanases and XEGs can act on xyloglucan, a hemicellulosic β-1,4 glucan chain abundant in the primary cell walls of higher plants, which is modified with xylose, and some of the xylose residues are further modified with other residues, such as galactose. When the appropriate xyloglucanases or XEGs act on xyloglucan, the products can include xyloglucan oligosaccharides with backbone lengths useful in the food, cosmetic and nutraceutical industries. Suitable xyloglucanases can include GH5 xyloglucanases from Bacteroides ovatus and GH74 xyloglucanases from Trichoderma reesei.

[0194] The enzyme reaction can be carried out in a solution and / or suspension. The enzyme reaction can be carried out in a suitable reaction vessel. In some cases, the enzyme reaction can be carried out at a temperature or temperature protocol suitable for a particular combination of enzyme and raw materials, and the reaction can be allowed to proceed for a certain amount of time (e.g., a predetermined amount of time) until the product reaches a desired concentration or meets some other requirement.

[0195] To ensure optimal contact between the enzyme and the ingredients, the reaction mixture may be stirred, either constantly or at intervals. Agitation can take the form of (i) regular movement of the entire reaction vessel, (ii) a fan or other stirring device, (iii) sparging with air bubbles, or (iv) any other suitable method of agitation.

[0196] The enzymatic reaction can be a microbial fermentation. The temperature and reaction time can be suitable for the growth of the microorganism used. The microorganism can be genetically modified to produce enzymes suitable for the production of the oligosaccharide composition. The microorganism can be a bacterium, for example Escherichia coli, or a fungus, such as Saccharomyces cerevisiae or Trichoderma reesei.

[0197] In some embodiments, an expression vector suitable for modifying the microorganism of interest may be used to produce an enzyme or mixture of enzymes as described elsewhere herein. If desired, the expression vector may be a plasmid or any other nucleic acid capable of inducing the production of an enzyme. In some examples, the expression vector may include one or more of the following regulatory sequences to control the expression of the exogenous enzyme: a heat shock gene regulatory sequence, a virulence gene regulatory sequence, a sporulation gene regulatory sequence, or any other suitable regulatory sequence.

[0198] The enzymatic reaction may be carried out at a temperature or temperature protocol suitable for the enzyme and substrate used. For example, the enzymatic reaction may be carried out at a constant temperature ranging from 10°C to 100°C, 20°C to 80°C or 40°C to 60°C. In some cases, if the enzymatic reaction takes the form of a microbial fermentation, the temperature may be appropriate for this. For example, the enzymatic reaction may include growth of E. coli and / or the temperature may be substantially constant, about 37°C.

[0199] The pH of the solution or suspension can affect the activity of the enzyme. Control of the pH can help ensure that the enzymatic reaction proceeds at a suitable rate. Enzymatic reactions can be carried out at pH ranges from 2 to 10, 3 to 8, or 4 to 6.

[0200] The enzymatic reaction can be allowed to continue for a certain period of time before being quenched and the product isolated or otherwise collected. This period can be from 1 minute to 6 days, from 0.5 days to 5 days, or from 16 hours to 96 hours. The reaction can alternatively be allowed to proceed until no further catalysis occurs.

[0201] The one or more raw materials added to the enzymatic reaction can include polysaccharides. Such polysaccharides can be produced by separate reactions proceeding simultaneously or substantially simultaneously in a reaction vessel. The polysaccharides present in the enzymatic reaction can be partially cleaved by the enzyme into useful oligosaccharides, leaving partially cleaved or uncleaved polysaccharides, including, but not limited to, cellulose, xylan (such as glucuronoxylan, arabinoxylan or glucuronoarabinoxylan), mannan (such as glucomannan, galactomannan or galactoglucomannan), mixed linkage glucan, xyloglucan chitin, chitosan or lignocellulose.

[0202] The enzymatic reaction may be continued until there is 5%-75%, 5%-70%, 5%-65%, 5%-55% or 10%-50% of the polysaccharide-containing material remaining undigested, which can be monitored or confirmed by a reducing endo assay, such as an anthrone assay, and / or by chromatographic methods, such as thin layer chromatography and / or high performance anion exchange chromatography.

[0203] Any material that contains suitable polysaccharides may form part of the raw material. The food, cosmetic and nutraceutical industries generally use a wide variety of oligosaccharides, so suitable polysaccharides that participate in the enzymatic reaction are not particularly limited. Suitable raw materials for generating oligosaccharide profiles can include, for example, cellulose, lignocellulose, chitin, chitosan, xylan (such as glucuronoxylan, arabinoxylan and glucuronoarabinoxylan) and / or mannan (such as glucomannan, galactomannan or galactoglucomannan), but any raw material that can be suitably acted upon is envisioned. The raw material can include sugar cane, corn stover, corn cob, bran, wheat straw, hardwood, softwood or any other suitable biomass or plant biomass.

[0204] The raw material containing such polysaccharides is also not particularly limited, since most plant materials are abundant in such polymers. Thus, the raw material can include plant biomass such as grain, grain husk, bean pod, seed coat and / or other seed material; seaweed; corn stover, straw, bagasse, Japanese silver grass, sorghum residue, switchgrass, bamboo and / or other monocotyledonous plant tissue; water hyacinth, leaf tissue, root and / or other plant material; hardwood, hardwood chips, hardwood pulp, softwood, softwood chips, softwood pulp, paper, paper pulp, cardboard and / or other wood-based raw material; crab shell, squid biomass, shrimp shell and / or other marine biomass, and / or any combination of suitable raw materials. The raw material may include wheat straw or timber. Since any given natural raw material is likely to contain a mixture of various polysaccharides, sometimes a mixture of various enzymes may be beneficial. Such a mixture can include one or more of any suitable enzymes discussed herein. For example, such a mixture would include an LPMO and an endo-glucanase, a xylanase and a lichenase, a cellobiohydrolase and a mannanase, or an endo-glucanase and a cellobiohydrolase. In some embodiments, the enzyme partners can be present in a molar ratio of, for example, 1:100 to 100:1. Additionally, many of the suitable feedstocks are difficult to handle, so pre-treatment of the feedstock is envisioned.

[0205] After the enzymatic reaction has proceeded to a desired point, one or more oligosaccharides and one or more polysaccharides from the enzymatic reaction mixture may be separated. This process can be carried out in a variety of ways, depending on the composition of the biomass used and the specificity of the enzymes used. Since the reaction mixture often contains a mixture of soluble oligosaccharides and insoluble polysaccharides, the reaction mixture can be filtered to remove the insoluble material and the resulting soluble oligosaccharides prepared for further processing.

[0206] Oligosaccharides can also be separated from polysaccharides in several ways. Oligosaccharides may be isolated based on solubility, so that the composition of soluble sugars is only extracted for further processing and / or isolated by chromatography to produce a composition with a narrower range of oligosaccharide chain lengths. Isolation can be based on, for example, precipitation, size exclusion chromatography, ion exchange chromatography, filtration, ultrafiltration, microfiltration or nanofiltration. When solubility-based isolation is performed, the sugar profile present in the isolated composition will generally depend on the original enzymatic reaction, since different polysaccharides generally decrease in solubility with length at different rates.

[0207] Similarly, it is envisaged that all or part of the oligosaccharides produced may be further processed to produce further products before being incorporated into foodstuffs, cosmetics or dietary supplements.This further processing may include any chemical, physical or enzymatic steps, such as reduction, for example reductive amination, if necessary, oxidation, caramelization, modification with Schiff base, etc., or Maillard reaction, or any combination of such steps, to obtain various products with properties that are realized or improved for the desired purpose.For example, caramelization properties, caloric value, flavour and colour may be modified.The oligosaccharides may also be purified, for example, by precipitation, size exclusion chromatography, ion exchange chromatography, filtration, ultrafiltration, microfiltration or nanofiltration.

[0208] Similarly, it is envisaged that all or a portion of the resulting polysaccharide fraction may be further processed to produce a product with improved properties prior to incorporation into a foodstuff, cosmetic or dietary supplement. This further processing may include either chemical, physical or enzymatic steps, such as alkylation or acid treatment. The polysaccharide may also be purified, for example, by precipitation, size exclusion chromatography, ion exchange chromatography, filtration, ultrafiltration, microfiltration or nanofiltration.

[0209] In certain instances, after modification and / or purification of the oligosaccharide and polysaccharide fractions, all or a portion of the fractions can then be recombined in a polysaccharide:oligosaccharide ratio of 1:100 to 1:1, for example 1:10 to 1:1, 1:90 to 1:2, 1:80 to 1:3, 1:70 to 1:4 or 1:60 to 1:5. The particular ratio may depend on the desired properties of the final component, as well as the modifications and purification applied to the fractions. It may not be necessary to recombine all of the oligosaccharides and polysaccharides isolated from the enzymatic reaction.

[0210] The fractions can be recombined in various ways, for example by mixing a solution containing all or a portion of the oligosaccharide fraction with a solution and / or suspension containing all or a portion of the polysaccharide fraction, which may be further dried, lyophilized or concentrated in some other way. The fractions may also be recombined by mixing a dry form containing all or a portion of the oligosaccharide fraction produced by drying, lyophilization or concentration in some other way with a dry form containing all or a portion of the polysaccharide fraction produced by drying, lyophilization or concentration in some other way.

[0211] The oligosaccharide component of the final composition can include one or more of any type of oligosaccharides. For example, the oligosaccharide component can include cellooligosaccharides, xylooligosaccharides, mixed linkage glucan oligosaccharides, mannooligosaccharides, xyloglucan oligosaccharides, chitooligosaccharides, arabinoxylooligosaccharides, or derivatives of any of the above-mentioned oligosaccharides.

[0212] Any such dry or liquid composition may be considered at any stage of the process as an ingredient suitable for incorporation into a foodstuff, cosmetic or dietary supplement, including compositions that may be considered intermediates during the process, such as the composition formed after recombining the oligosaccharide and polysaccharide fractions, prior to any further purification, optimization, drying, dissolving or any other such steps, and including the final composition obtained from the process.

[0213] As described herein, the dry composition may be formed by drying and / or freeze-drying. The dry composition can be dissolved in various liquid solutions, including water, syrup, paste, solvent, alcohol, etc., to form a component of a liquid composition suitable for incorporation into food products, cosmetics, or dietary supplements. The liquid composition may be particularly useful in foods that require a smooth texture, such as candy, chocolate, and yogurt.

[0214] In some embodiments, after modification and / or purification of the oligosaccharide and polysaccharide fractions, all or a portion of the fractions can then be recombined in a polysaccharide:oligosaccharide ratio of 1:100 to 1:1, for example 1:10 to 1:1, 1:90 to 1:2, 1:80 to 1:3, 1:70 to 1:4 or 1:60 to 1:5. The particular ratio may depend on the desired properties of the final component, as well as the modifications and purification applied to the fractions.

[0215] Once the composition of oligosaccharide products suitable for the intended use is obtained, further processing and / or isolation can be performed. The derivation of foodstuffs, cosmetics or dietary supplements from the composition can provide a wide range of potential uses. The components described herein can be useful in applications where oligosaccharides, sugars, bulk sweeteners, low-intensity sweeteners or other related food ingredients are conventionally used.

[0216] The polysaccharide cleaving enzyme can be one of cellulase, xylanase, xyloglucanase, endo-glucanase, cellobiohydrolase, mannanase, lichenase, or a lytic polysaccharide monooxygenase (LPMO) selected from the group consisting of, for example, AA9, AA10, AA11, AA13, AA14, and AA15. The polysaccharide cleaving enzyme can be prepared from T. reesei fungus and / or enzymatic reaction until 5-75%, 5-65% or 5-50% of the undigested polysaccharide-containing feedstock remains.

[0217] The polysaccharide-cleaving enzyme may be operably linked to a catalytic module or a non-catalytic module, for example, the polysaccharide-cleaving enzyme may be operably linked to a non-catalytic module, where the non-catalytic module is a carbohydrate-binding module.

[0218] In various embodiments, after separation of the oligosaccharide(s) and polysaccharide(s), the oligosaccharide(s) and polysaccharide(s) may be purified and / or may be subjected to chemical, physical or enzymatic treatments such as reduction, oxidation, caramelization or the Maillard reaction, and / or may be recombined by combining dry powders of the oligosaccharides and dry polysaccharide powders.

[0219] In some embodiments, the components may include three or more oligosaccharides of different molecular weights, and the method may include forming the three or more oligosaccharides by an enzymatic reaction, where the enzymatic reaction includes contacting, in solution or suspension, one or more polysaccharide-cleaving enzymes and one or more feedstocks. III. Food, cosmetic or dietary supplement ingredients composition

[0220] The polysaccharide component of the composition can include one or more of any type of polysaccharide, for example, the polysaccharide can include cellulose, lignocellulose, xylan, mixed linkage glucan, mannan, xyloglucan, chitin, chitosan, or derivatives of any of the above-mentioned polysaccharides.

[0221] The composition or ingredients may include various oligosaccharides. The composition may include various amounts of oligosaccharides, depending, for example, on the desired properties of the composition. In some examples, the composition may include at least 20% by dry weight, e.g., at least 30% by dry weight, of cellooligosaccharides having a degree of polymerization of 2-6, and / or the composition may include at least 20% by dry weight, e.g., at least 30% by dry weight, of xylooligosaccharides having a degree of polymerization of 2-12, and / or the composition may include at least 20% by dry weight, e.g., at least 30% by dry weight, of mixed linkage glucan oligosaccharides having a degree of polymerization of 2-5, and / or the composition may include at least a small amount by dry weight of mannooligosaccharides having a degree of polymerization of 2-12. and / or the composition may comprise at least 20% by dry weight, e.g., at least 30% by dry weight, of xyloglucan oligosaccharides having a degree of polymerization of 4 to 12, and / or the composition may comprise at least 20% by dry weight, e.g., at least 30% by dry weight, of chitooligosaccharides having a degree of polymerization of 2 to 12, and / or the composition may comprise at least 20% by dry weight, e.g., at least 30% by dry weight, of arabinoxylooligosaccharides having a degree of polymerization of 3 to 15. In certain embodiments, the composition may comprise up to 100% by dry weight of the above oligosaccharides, and thus it may be understood that the above embodiments in which oligosaccharides are present at at least 20% by dry weight do not include all seven types of oligosaccharides.

[0222] In various embodiments, the composition or ingredient can comprise about 5% to about 50% w / w of cellooligosaccharides having a degree of polymerization of 2 to 6. In certain embodiments, the composition or ingredient can comprise about 5% to about 50%, about 10% to about 40%, about 15% to about 35% w / w of cellooligosaccharides having a degree of polymerization of 2 to 6. The composition or ingredient can comprise at least 5%, 8%, 10%, 15%, 20% or 25% w / w of cellooligosaccharides having a degree of polymerization of 2 to 6. In some embodiments, the composition or ingredient can comprise about 20% to about 90% w / w of cellooligosaccharides having a degree of polymerization of 2 to 6. In certain embodiments, the compositions or components may comprise about 5% to about 95%, about 10% to about 92.5%, about 30% to about 80%, about 40% to about 70%, or about 50% to about 60% w / w cellooligosaccharides having a degree of polymerization of 2-6.

[0223] In various embodiments, the composition or ingredients can comprise about 20% to about 90% w / w xylooligosaccharides having a degree of polymerization of 2 to 5. In certain embodiments, the composition or ingredients can comprise about 5% to about 95%, about 10% to about 92.5%, about 30% to about 80%, about 40% to about 70%, or about 50% to about 60% w / w xylooligosaccharides having a degree of polymerization of 2 to 5. For example, the composition can comprise at least 30% w / w xylooligosaccharides having a degree of polymerization of 2 to 5. The composition or ingredients can comprise at least 5%, 8%, 10%, 15%, 20%, or 25% w / w xylooligosaccharides having a degree of polymerization of 2 to 5.

[0224] In certain embodiments, the composition or ingredients can comprise from about 0.1% to about 15% w / w arabinoxylooligosaccharides having a degree of polymerization of 3 to 12. The composition or ingredients can comprise at least 0.1%, 0.3%, 0.5%, 0.8%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w arabinoxylooligosaccharides having a degree of polymerization of 3 to 12. In various embodiments, the composition or ingredients can comprise from about 0.5% to about 25% w / w arabinoxylooligosaccharides having a degree of polymerization of 3 to 15. The composition or ingredient may contain at least 0.1%, 0.3%, 0.5%, 0.8%, 1%, 2%, 3%, 4%, 5%, 10%, 15%, 20%, 25%, 30% or 35% w / w arabinoxylooligosaccharides having a degree of polymerization of 3-15.

[0225] In some embodiments, provided herein is the use of an oligosaccharide mixture in the formation of a foodstuff, cosmetic or dietary supplement, the oligosaccharide mixture comprising: i) cellooligosaccharides having a degree of polymerization of 2 to 6; ii) xylooligosaccharides having a degree of polymerization between 2 and 12; iii) mixed linkage glucan oligosaccharides having a degree of polymerization of 2-5; iv) mannooligosaccharides having a degree of polymerization of 2 to 12; v) xyloglucan oligosaccharides having a degree of polymerization of 4 to 10; vi) chitooligosaccharides having a degree of polymerization of 2 to 12; and / or vii) arabinoxylooligosaccharides having a degree of polymerization of 3 to 15 wherein the two oligosaccharides may be present in a ratio of 1:9 to 9:1, 1:4 to 4:1 or 2:3 to 3:2 with respect to each other.

[0226] In certain cases, the arabinoxylooligosaccharides may contain at least 0.1% arabinosyl residues. The arabinoxylooligosaccharides may contain at least 0.1%, 0.2%, 0.5%, 1%, 5% or 10% arabinosyl residues.

[0227] The amount of each of the oligosaccharides can vary depending on the desired properties of the resulting foodstuff, cosmetic or dietary supplement. For example, the two oligosaccharides can be present in a ratio of 1:9 to 1:1, 1:2 to 1:1 or 2:3 to 1:1 with respect to each other.

[0228] The oligosaccharide mixture may further comprise a third oligosaccharide. The oligosaccharide mixture may comprise a third oligosaccharide and a fourth oligosaccharide. The oligosaccharide mixture may comprise a third oligosaccharide, a fourth oligosaccharide and a fifth oligosaccharide. The oligosaccharide mixture may further comprise a third oligosaccharide, a fourth oligosaccharide, a fifth oligosaccharide and a sixth oligosaccharide. The oligosaccharide mixture may further comprise a third oligosaccharide, a fourth oligosaccharide, a fifth oligosaccharide, a sixth oligosaccharide and a seventh oligosaccharide. These oligosaccharides may be selected from the same list of at least two oligosaccharides presented above.

[0229] The oligosaccharide mixture of at least two oligosaccharides may comprise cellooligosaccharides, such as cellooligosaccharides in combination with xylooligosaccharides. Alternative compositions may comprise cellooligosaccharides in combination with mannooligosaccharides. In some embodiments, the oligosaccharide mixture may comprise cellooligosaccharides, xylooligosaccharides and arabinoxylooligosaccharides in combination with each other.

[0230] The oligosaccharide mixture of at least two oligosaccharides can further include a polysaccharide, for example a cellulosic polysaccharide such as cellulose, or a polysaccharide derivative, for example a cellulose derivative such as carboxymethylcellulose, or a polysaccharide aggregate, for example a portion of a lignocellulosic biomass. In some examples, the ratio in the combination can be 1:100 to 1:1, for example, 1:90 to 1:2, 1:80 to 1:3, 1:70 to 1:4, or 1:60 to 1:5 polysaccharide / polysaccharide derivative / polysaccharide aggregate:oligosaccharide. Thus, the ratio between the first oligosaccharide, the second oligosaccharide, and the polysaccharide can be 2:2:1 to 30:30:1, for example, about 3:3:1. Oligosaccharide Combinations

[0231] The composition may comprise a mixture of one or more oligosaccharides. The mixture of oligosaccharides may comprise two forms or types of oligosaccharides, such as cellooligosaccharides and xylooligosaccharides. The mixture of oligosaccharides may comprise three forms of oligosaccharides, such as cellooligosaccharides, mannooligosaccharides and xylooligosaccharides. The mixture of oligosaccharides may comprise four forms of oligosaccharides, such as cellooligosaccharides, mannooligosaccharides, mixed-linked glucan oligosaccharides and chitooligosaccharides.

[0232] The oligosaccharide mixture may comprise two forms of oligosaccharides, for example a first oligosaccharide and a second oligosaccharide. The oligosaccharide mixture may comprise about 5% w / w of the first oligosaccharide and about 95% w / w of the second oligosaccharide. The oligosaccharide mixture may comprise about 10% w / w of the first oligosaccharide and about 90% w / w of the second oligosaccharide. The oligosaccharide mixture may comprise about 15% w / w of the first oligosaccharide and about 85% w / w of the second oligosaccharide. The oligosaccharide mixture may comprise about 20% w / w of the first oligosaccharide and about 80% w / w of the second oligosaccharide. The oligosaccharide mixture may comprise about 25% w / w of the first oligosaccharide and about 75% w / w of the second oligosaccharide. The oligosaccharide mixture may comprise about 30% w / w of the first oligosaccharide and about 70% w / w of the second oligosaccharide. The oligosaccharide mixture may comprise about 35% w / w of the first oligosaccharide and about 65% w / w of the second oligosaccharide. The oligosaccharide mixture may comprise about 40% w / w of the first oligosaccharide and about 50% w / w of the second oligosaccharide. The oligosaccharide mixture may comprise 45% w / w of the first oligosaccharide and 55% w / w of the second oligosaccharide. The oligosaccharide mixture may comprise 50% w / w of the first oligosaccharide and 50% w / w of the second oligosaccharide. The oligosaccharide mixture may comprise 55% w / w of the first oligosaccharide and 45% w / w of the second oligosaccharide. The oligosaccharide mixture may comprise 60% w / w of the first oligosaccharide and 30% w / w of the second oligosaccharide. The oligosaccharide mixture may comprise 65% w / w of the first oligosaccharide and 35% w / w of the second oligosaccharide. The oligosaccharide mixture may comprise 70% w / w of the first oligosaccharide and 30% w / w of the second oligosaccharide. The oligosaccharide mixture may comprise 75% w / w of the first oligosaccharide and 25% w / w of the second oligosaccharide. The oligosaccharide mixture may comprise 80% w / w of the first oligosaccharide and 20% w / w of the second oligosaccharide. The oligosaccharide mixture may comprise 85% w / w of the first oligosaccharide and 15% w / w of the second oligosaccharide. The oligosaccharide mixture may comprise 90% w / w of the first oligosaccharide and 10% w / w of the second oligosaccharide. The oligosaccharide mixture may comprise 95% w / w of a first oligosaccharide and 5% w / w of a second oligosaccharide.In some cases, the first oligosaccharide may be a cello-oligosaccharide and the second oligosaccharide may be a xylo-oligosaccharide. In some examples, the first oligosaccharide may be a cello-oligosaccharide and the second oligosaccharide may be a manno-oligosaccharide. In some embodiments, the first oligosaccharide may be a xylo-oligosaccharide and the second oligosaccharide may be a manno-oligosaccharide. Other combinations of the first and second oligosaccharides are also within the scope of the present disclosure.

[0233] The oligosaccharide mixture may comprise three forms of oligosaccharides, for example a first oligosaccharide, a second oligosaccharide and a third oligosaccharide. The oligosaccharide mixture may comprise about 20% w / w of the first oligosaccharide, 40% w / w of the second oligosaccharide and 40% w / w of the third oligosaccharide. The oligosaccharide mixture may comprise about 30% w / w of the first oligosaccharide, 30% w / w of the second oligosaccharide and 40% w / w of the third oligosaccharide. The oligosaccharide mixture may comprise about 10% w / w of the first oligosaccharide, 10% w / w of the second oligosaccharide and 80% w / w of the third oligosaccharide. The oligosaccharide mixture may comprise about 20% w / w of the first oligosaccharide, 20% w / w of the second oligosaccharide and 60% w / w of the third oligosaccharide. The oligosaccharide mixture may comprise about 20% w / w of the first oligosaccharide, 30% w / w of the second oligosaccharide, and 50% w / w of the third oligosaccharide. In some examples, the first oligosaccharide may be a mannooligosaccharide, the second oligosaccharide may be a xylooligosaccharide, and the third oligosaccharide may be a cellooligosaccharide. In some examples, the first oligosaccharide may be a xyloglucan oligosaccharide, the second oligosaccharide may be a xylooligosaccharide, and the third oligosaccharide may be a cellooligosaccharide. Other combinations of the first oligosaccharide, the second oligosaccharide, and the third oligosaccharide are also within the scope of the present disclosure.

[0234] The oligosaccharide mixture may contain two or more oligosaccharides, i.e., a first oligosaccharide and a second oligosaccharide different from the first oligosaccharide. For example, the first oligosaccharide may be a xylooligosaccharide, or a cellooligosaccharide, or a mannooligosaccharide, or other oligosaccharides presented herein, while the second oligosaccharide can be a xylooligosaccharide, or a cellooligosaccharide, or a mannooligosaccharide, or other oligosaccharides not used as the first oligosaccharide. In other words, the first oligosaccharide can be different from the second oligosaccharide (e.g., the first oligosaccharide can be a different type of oligosaccharide from the second oligosaccharide). The ratio of the first oligosaccharide to the second oligosaccharide in the mixture can be about 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8 or 1:9.

[0235] The ratio of the first oligosaccharide to the second oligosaccharide in the mixture can be about 2:1, 2:3, 2:5, 2:7 or 2:9. The oligosaccharides can be cello-oligosaccharides, manno-oligosaccharides, xylo-oligosaccharides, xyloglucan-oligosaccharides, mixed linkage oligosaccharides, chitooligosaccharides, arabinoxylo-oligosaccharides, or other oligosaccharides provided herein, and the first oligosaccharide is selected to be a different oligosaccharide from the second oligosaccharide. In other words, the first oligosaccharide can be a different type of oligosaccharide from the second oligosaccharide.

[0236] The ratio of the first oligosaccharide to the second oligosaccharide in the mixture can be about 3:1, 3:2, 3:4, 3:5, 3:7 or 3:8. The oligosaccharides can be cello-oligosaccharides, manno-oligosaccharides, xylo-oligosaccharides, xyloglucan oligosaccharides, mixed linkage oligosaccharides, chitooligosaccharides, arabinoxylo-oligosaccharides, or other oligosaccharides provided herein, and the first oligosaccharide is selected to be a different oligosaccharide than the second oligosaccharide.

[0237] The ratio of the first oligosaccharide to the second oligosaccharide in the oligosaccharide mixture containing two or more oligosaccharides can be 1:9 to 9:1, 1:4 to 4:1, 1:3 to 3:1, or 2:3 to 3:2. The oligosaccharides can be cellooligosaccharides, mannooligosaccharides, xylooligosaccharides, xyloglucan oligosaccharides, mixed linkage oligosaccharides, chitooligosaccharides, arabinoxylooligosaccharides, or other oligosaccharides provided herein, where the first oligosaccharide is selected to be a different oligosaccharide than the second oligosaccharide.

[0238] In some cases, the compositions or ingredients may include at least 1% w / w, 2% w / w, 3% w / w, 4% w / w, 5% w / w, 10% w / w, 15% w / w, 20% w / w, 25% w / w, 30% w / w, or more of cellobiose, xylobiose, mannobiose (e.g., Man-β-1,4-Man), Glc-β-1,4-Man, Man-β-1,4-Glc, laminaribiose, gentiobiose, sophorose, maltose, lactose, or sucrose. In certain cases, the composition or ingredient may contain at least 1% w / w, 2% w / w, 3% w / w, 4% w / w, 5% w / w, 10% w / w, 15% w / w, 20% w / w, 25% w / w, 30% w / w or more of cellotriose, xylotriose, monoarabinosylated xylobiose, monoglucuronosylated xylobiose, maltotriose, mannotriose (e.g., Man-β-1,4-Man-β-1,4-Man), Glycotriose, glycerol ... c-β-1,4-Man-β-1,4-Man, Man-β-1,4-Glc-β-1,4-Man, Man-β-1,4-Man-β-1,4-Glc, Man-β-1,4-Glc-β-1,4-Glc, Glc-β -1,4-Man-β-1,4-Glc, Glc-β-1,4-Glc-β-1,4-Man, Glc-β-1,3-Glc-β-1,4-Glc or Glc-β-1,4-Glc-β-1,3-Glc.In certain examples, the composition or ingredient comprises at least 1% w / w, 2% w / w, 3% w / w, 4% w / w, 5% w / w, 10% w / w, 15% w / w, 20% w / w, 25% w / w, 30% w / w or more of xylotetraose, cellotetraose, monoarabinosylated xylotriose, monoglucuronosylated xylotriose, diarabinosylated xylobiose, diglucuronosylated xylose, glycerol ... Cyllobiose, maltotetraose, mannotetraose (e.g. Man-β-1,4-Man-β-1,4-Man-β-1,4-Man), Glc-β-1,4-Man-β-1,4-Man- β-1,4-Man, Man-β-1,4-Glc-β-1,4-Man-β-1,4-Man, Man-β-1,4-Man-β-1,4-Glc-β-1,4-Man, Man-β-1,4-Man -β-1,4-Man-β-1,4-Glc, Glc-β-1,4-Glc-β-1,4-Man-β-1,4-Man, Man-β-1,4-Glc-β-1,4-Glc-β-1,4-Man, M an-β-1,4-Man-β-1,4-Glc-β-1,4-Glc, Glc-β-1,4-Man-β-1,4-Glc-β-1,4-Man, Glc-β-1,4-Man-β-1,4-Man- β-1,4-Glc, Man-β-1,4-Glc-β-1,4-Man-β-1,4-Glc, Glc-β-1,3-Glc-β-1,4-Glc-1,4-Glc, Glc-β-1,4-Glc- It may contain β-1,3-Glc-1,4-Glc, Glc-β-1,4-Glc-β-1,4-Glc-1,3-Glc or Glc-β-1,3-Glc-β-1,4-Glc-1,3-Glc.In certain cases, the composition or ingredient may comprise at least 0.01% w / w, 0.05% w / w, 0.1% w / w, 0.5% w / w, 1% w / w, 2% w / w, 5% w / w, 10% w / w, 15% w / w, 20% w / w or more of xylopentaose, cellopentaose, monoarabinosylated xylotetraose, monoglucuronosylated xylotetraose, diarabinosylated xylotriose, diglucuronosylated xylotriose, maltopentaose, mannopentaose (e.g., Man-β-1,4-Man-β-1,4-Man-β-1,4-Man-β-1,4-Man), a mixed linkage glucan-derived pentasaccharide or a mannan-derived pentasaccharide.

[0239] The composition or ingredients may contain 1%-50%, 5%-40%, 10%-30% or 15%-25% w / w cellobiose. The composition or ingredients may contain 2.5%-90%, 5%-80%, 10%-70% or 20%-60% w / w xylobiose. The composition or ingredients may contain 2.5%-75%, 5%-50%, 10%-40% or 20%-30% w / w xylotriose. Oligosaccharide compositions with various degrees of polymerization

[0240] The average degree of polymerization of the oligosaccharides in the composition can be 1-50, 1.5-25, 2-15, 2.1-10, 2.1-7, or 2.2-5.

[0241] The concentration of xylooligosaccharides having a degree of polymerization 2 in the xylooligosaccharide mixture can be from about 2% to about 80% w / w. The concentration of xylooligosaccharides having a degree of polymerization 2 can be at least 2%, 4%, 6%, 8%, 10%, 12%, 15%, 18%, 20%, 25% or 30% w / w. The concentration of xylooligosaccharides having a degree of polymerization 2 can be higher, in some cases, for example, up to 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75% or 80% w / w.

[0242] The concentration of xylooligosaccharides having a degree of polymerization 3 in the xylooligosaccharide mixture can be from about 2% to about 20% w / w. The concentration of xylooligosaccharides having a degree of polymerization 3 can be at least 2%, 4%, 6%, 8%, 10%, 12%, 15%, 18% or 20% w / w.

[0243] The concentration of xylooligosaccharides having a degree of polymerization 4 in the xylooligosaccharide mixture can be from about 5% to about 20% w / w. The concentration of xylooligosaccharides having a degree of polymerization 4 can be at least 5%, 8%, 10%, 12%, 15%, 18% or 20% w / w.

[0244] The concentration of xylooligosaccharides having a degree of polymerization of 5 in the xylooligosaccharide mixture can be from about 5% to about 20% w / w. The concentration of xylooligosaccharides having a degree of polymerization of 5 can be at least 5%, 7%, 8%, 10%, 12%, 15%, 18% or 20% w / w.

[0245] The concentration of xylooligosaccharides having a degree of polymerization 6 in the xylooligosaccharide mixture can be from about 5% to about 25% w / w. The concentration of xylooligosaccharides having a degree of polymerization 6 can be at least 5%, 8%, 10%, 12%, 15%, 18%, 20% or 25% w / w.

[0246] The concentration of xylooligosaccharides having a degree of polymerization 7 in the xylooligosaccharide mixture can be from about 2% to about 20% w / w. The concentration of xylooligosaccharides having a degree of polymerization 7 can be at least 2%, 4%, 6%, 8%, 10%, 12%, 15%, 17% or 20% w / w.

[0247] The concentration of xylooligosaccharides having a degree of polymerization 8 in the xylooligosaccharide mixture can be from about 1% to about 15% w / w. The concentration of xylooligosaccharides having a degree of polymerization 8 can be at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w.

[0248] The concentration of xylooligosaccharides having a degree of polymerization of 9 in the xylooligosaccharide mixture can be from about 2% to about 15% w / w. The concentration of xylooligosaccharides having a degree of polymerization of 9 can be at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w.

[0249] The concentration of xylooligosaccharides having a degree of polymerization of 10 in the xylooligosaccharide mixture can be from about 2% to about 15% w / w. The concentration of xylooligosaccharides having a degree of polymerization of 10 can be at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w.

[0250] The concentration of xylooligosaccharides having a degree of polymerization of 11 in the xylooligosaccharide mixture can be from about 2% to about 15% w / w. The concentration of xylooligosaccharides having a degree of polymerization of 11 can be at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w.

[0251] The concentration of xylooligosaccharides having a degree of polymerization 12 in the xylooligosaccharide mixture can be from about 2% to about 15% w / w. The concentration of xylooligosaccharides having a degree of polymerization 12 can be at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w.

[0252] The concentration of cellooligosaccharides having a degree of polymerization 2 in the cellooligosaccharide mixture can be about 2% to about 80% w / w. The concentration of cellooligosaccharides having a degree of polymerization 2 can be at least 2%, 4%, 6%, 8%, 10%, 12%, 15%, 18%, 20%, 25% or 30% w / w. The concentration of cellooligosaccharides having a degree of polymerization 2 can be higher, in some cases, for example, at least 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75% or 80% w / w.

[0253] The concentration of cellooligosaccharides having a degree of polymerization 3 in the cellooligosaccharide mixture can be about 2% to about 20% w / w. The concentration of cellooligosaccharides having a degree of polymerization 3 can be at least 2%, 4%, 6%, 8%, 10%, 12%, 15%, 18% or 20% w / w.

[0254] The concentration of cellooligosaccharides having a degree of polymerization 4 in the cellooligosaccharide mixture can be about 5% to about 20% w / w. The concentration of cellooligosaccharides having a degree of polymerization 4 can be at least 5%, 8%, 10%, 12%, 15%, 18% or 20% w / w.

[0255] The concentration of cellooligosaccharides having a degree of polymerization of 5 in the cellooligosaccharide mixture can be about 5% to about 20% w / w. The concentration of cellooligosaccharides having a degree of polymerization of 5 can be at least 5%, 7%, 8%, 10%, 12%, 15%, 18% or 20% w / w.

[0256] The concentration of cellooligosaccharides having a degree of polymerization 6 in the cellooligosaccharide mixture can be about 5% to about 25% w / w. The concentration of cellooligosaccharides having a degree of polymerization 6 can be at least 5%, 8%, 10%, 12%, 15%, 18%, 20% or 25% w / w.

[0257] The concentration of mannooligosaccharides having a degree of polymerization 2 in the mannooligosaccharide mixture can be about 2% to about 30% w / w. The concentration of mannooligosaccharides having a degree of polymerization 2 can be at least 2%, 4%, 6%, 8%, 10%, 12%, 15%, 18%, 20%, 25% or 30% w / w.

[0258] The concentration of mannooligosaccharides having a degree of polymerization 3 in the mannooligosaccharide mixture can be about 2% to about 20% w / w. The concentration of mannooligosaccharides having a degree of polymerization 3 can be at least 2%, 4%, 6%, 8%, 10%, 12%, 15%, 18% or 20% w / w.

[0259] The concentration of mannooligosaccharides having a degree of polymerization 4 in the mannooligosaccharide mixture can be about 5% to about 20% w / w. The concentration of mannooligosaccharides having a degree of polymerization 4 can be at least 5%, 8%, 10%, 12%, 15%, 18% or 20% w / w.

[0260] The concentration of mannooligosaccharides having a degree of polymerization of 5 in the mannooligosaccharide mixture can be about 5% to about 20% w / w. The concentration of mannooligosaccharides having a degree of polymerization of 5 can be at least 5%, 7%, 8%, 10%, 12%, 15%, 18% or 20% w / w.

[0261] The concentration of mannooligosaccharides having a degree of polymerization 6 in the mannooligosaccharide mixture can be about 5% to about 25% w / w. The concentration of mannooligosaccharides having a degree of polymerization 6 can be at least 5%, 8%, 10%, 12%, 15%, 18%, 20% or 25% w / w.

[0262] The concentration of mannooligosaccharides having a degree of polymerization 7 in the mannooligosaccharide mixture can be about 2% to about 20% w / w. The concentration of mannooligosaccharides having a degree of polymerization 7 can be at least 2%, 4%, 6%, 8%, 10%, 12%, 15%, 17% or 20% w / w.

[0263] The concentration of mannooligosaccharides having a degree of polymerization of 8 in the mannooligosaccharide mixture can be about 1% to about 15% w / w. The concentration of mannooligosaccharides having a degree of polymerization of 8 can be at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w.

[0264] The concentration of mannooligosaccharides having a degree of polymerization of 9 in the mannooligosaccharide mixture can be about 2% to about 15% w / w. The concentration of mannooligosaccharides having a degree of polymerization of 9 can be at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w.

[0265] The concentration of mannooligosaccharides having a degree of polymerization of 10 in the mannooligosaccharide mixture can be about 2% to about 15% w / w. The concentration of mannooligosaccharides having a degree of polymerization of 10 can be at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w.

[0266] The concentration of mannooligosaccharides having a degree of polymerization of 11 in the mannooligosaccharide mixture can be about 2% to about 15% w / w. The concentration of mannooligosaccharides having a degree of polymerization of 11 can be at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w.

[0267] The concentration of mannooligosaccharides having a degree of polymerization of 12 in the mannooligosaccharide mixture can be about 2% to about 15% w / w. The concentration of mannooligosaccharides having a degree of polymerization of 12 can be at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w.

[0268] The concentration of xyloglucan oligosaccharides having a degree of polymerization 4 in the xyloglucan oligosaccharide mixture can be about 5% to about 20% w / w. The concentration of xyloglucan oligosaccharides having a degree of polymerization 4 can be at least 5%, 8%, 10%, 12%, 15%, 18% or 20% w / w.

[0269] The concentration of xyloglucan oligosaccharides having a degree of polymerization of 5 in the xyloglucan oligosaccharide mixture can be about 5% to about 20% w / w. The concentration of xyloglucan oligosaccharides having a degree of polymerization of 5 can be at least 5%, 7%, 8%, 10%, 12%, 15%, 18% or 20% w / w.

[0270] The concentration of xyloglucan oligosaccharides having a degree of polymerization 6 in the xyloglucan oligosaccharide mixture can be about 5% to about 25% w / w. The concentration of xyloglucan oligosaccharides having a degree of polymerization 6 can be at least 5%, 8%, 10%, 12%, 15%, 18%, 20% or 25% w / w.

[0271] The concentration of xyloglucan oligosaccharides having a degree of polymerization 7 in the xyloglucan oligosaccharide mixture can be about 2% to about 20% w / w. The concentration of xyloglucan oligosaccharides having a degree of polymerization 7 can be at least 2%, 4%, 6%, 8%, 10%, 12%, 15%, 17% or 20% w / w.

[0272] The concentration of xyloglucan oligosaccharides having a degree of polymerization 8 in the xyloglucan oligosaccharide mixture can be about 1% to about 15% w / w. The concentration of xyloglucan oligosaccharides having a degree of polymerization 8 can be at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w.

[0273] The concentration of xyloglucan oligosaccharides having a degree of polymerization of 9 in the xyloglucan oligosaccharide mixture can be about 2% to about 15% w / w. The concentration of xyloglucan oligosaccharides having a degree of polymerization of 9 can be at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w.

[0274] The concentration of xyloglucan oligosaccharides having a degree of polymerization of 10 in the xyloglucan oligosaccharide mixture can be about 2% to about 15% w / w. The concentration of xyloglucan oligosaccharides having a degree of polymerization of 10 can be at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w.

[0275] The concentration of mixed linkage glucan oligosaccharides having a degree of polymerization 2 in the mixed linkage glucan oligosaccharide mixture can be from about 2% to about 30% w / w. The concentration of mixed linkage glucan oligosaccharides having a degree of polymerization 2 can be at least 2%, 4%, 6%, 8%, 10%, 12%, 15%, 18%, 20%, 25% or 30% w / w.

[0276] The concentration of mixed linkage glucan oligosaccharides having a degree of polymerization 3 in the mixed linkage glucan oligosaccharide mixture can be about 2% to about 20% w / w. The concentration of mixed linkage glucan oligosaccharides having a degree of polymerization 3 can be at least 2%, 4%, 6%, 8%, 10%, 12%, 15%, 18% or 20% w / w.

[0277] The concentration of mixed linkage glucan oligosaccharides having a degree of polymerization 4 in the mixed linkage glucan oligosaccharide mixture can be about 5% to about 20% w / w. The concentration of mixed linkage glucan oligosaccharides having a degree of polymerization 4 can be at least 5%, 8%, 10%, 12%, 15%, 18% or 20% w / w.

[0278] The concentration of mixed linkage glucan oligosaccharides having a degree of polymerization of 5 in the mixed linkage glucan oligosaccharide mixture can be about 5% to about 20% w / w. The concentration of mixed linkage glucan oligosaccharides having a degree of polymerization of 5 can be at least 5%, 7%, 8%, 10%, 12%, 15%, 18% or 20% w / w.

[0279] The concentration of the chitooligosaccharides having a degree of polymerization 2 in the chitooligosaccharide mixture can be about 2% to about 30% w / w. The concentration of the chitooligosaccharides having a degree of polymerization 2 can be at least 2%, 4%, 6%, 8%, 10%, 12%, 15%, 18%, 20%, 25% or 30% w / w.

[0280] The concentration of the chitooligosaccharides having a degree of polymerization 3 in the chitooligosaccharide mixture can be from about 2% to about 20% w / w. The concentration of the chitooligosaccharides having a degree of polymerization 3 can be at least 2%, 4%, 6%, 8%, 10%, 12%, 15%, 18% or 20% w / w.

[0281] The concentration of the chitooligosaccharides having a degree of polymerization 4 in the chitooligosaccharide mixture can be about 5% to about 20% w / w. The concentration of the chitooligosaccharides having a degree of polymerization 4 can be at least 5%, 8%, 10%, 12%, 15%, 18% or 20% w / w.

[0282] The concentration of the chitooligosaccharides having a degree of polymerization of 5 in the chitooligosaccharide mixture can be about 5% to about 20% w / w. The concentration of the chitooligosaccharides having a degree of polymerization of 5 can be at least 5%, 7%, 8%, 10%, 12%, 15%, 18% or 20% w / w.

[0283] The concentration of the chitooligosaccharides having a degree of polymerization of 6 in the chitooligosaccharide mixture can be about 5% to about 25% w / w. The concentration of the chitooligosaccharides having a degree of polymerization of 6 can be at least 5%, 8%, 10%, 12%, 15%, 18%, 20% or 25% w / w.

[0284] The concentration of the chitooligosaccharides having a degree of polymerization 7 in the chitooligosaccharide mixture can be from about 2% to about 20% w / w. The concentration of the chitooligosaccharides having a degree of polymerization 7 can be at least 2%, 4%, 6%, 8%, 10%, 12%, 15%, 17% or 20% w / w.

[0285] The concentration of the chitooligosaccharides having a degree of polymerization of 8 in the chitooligosaccharide mixture can be from about 1% to about 15% w / w. The concentration of the chitooligosaccharides having a degree of polymerization of 8 can be at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w.

[0286] The concentration of the chitooligosaccharides having a degree of polymerization of 9 in the chitooligosaccharide mixture can be from about 2% to about 15% w / w. The concentration of the chitooligosaccharides having a degree of polymerization of 9 can be at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w.

[0287] The concentration of the chitooligosaccharides having a degree of polymerization of 10 in the chitooligosaccharide mixture can be from about 2% to about 15% w / w. The concentration of the chitooligosaccharides having a degree of polymerization of 10 can be at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w.

[0288] The concentration of the chitooligosaccharides having a degree of polymerization of 11 in the chitooligosaccharide mixture can be from about 2% to about 15% w / w. The concentration of the chitooligosaccharides having a degree of polymerization of 11 can be at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w.

[0289] The concentration of the chitooligosaccharides having a degree of polymerization 12 in the chitooligosaccharide mixture can be from about 2% to about 15% w / w. The concentration of the chitooligosaccharides having a degree of polymerization 12 can be at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w.

[0290] The concentration of arabinoxylooligosaccharides having a degree of polymerization 3 in the arabinoxylooligosaccharide mixture can be about 2% to about 20% w / w. The concentration of arabinoxylooligosaccharides having a degree of polymerization 3 can be at least 2%, 4%, 6%, 8%, 10%, 12%, 15%, 18% or 20% w / w.

[0291] The concentration of arabinoxylooligosaccharides having a degree of polymerization 4 in the arabinoxylooligosaccharide mixture can be about 5% to about 20% w / w. The concentration of arabinoxylooligosaccharides having a degree of polymerization 4 can be at least 5%, 8%, 10%, 12%, 15%, 18% or 20% w / w.

[0292] The concentration of arabinoxylooligosaccharides having a degree of polymerization of 5 in the arabinoxylooligosaccharide mixture can be about 5% to about 20% w / w. The concentration of arabinoxylooligosaccharides having a degree of polymerization of 5 can be at least 5%, 7%, 8%, 10%, 12%, 15%, 18% or 20% w / w.

[0293] The concentration of arabinoxylooligosaccharides having a degree of polymerization of 6 in the arabinoxylooligosaccharide mixture can be about 5% to about 25% w / w. The concentration of arabinoxylooligosaccharides having a degree of polymerization of 6 can be at least 5%, 8%, 10%, 12%, 15%, 18%, 20% or 25% w / w.

[0294] The concentration of arabinoxylooligosaccharides having a degree of polymerization 7 in the arabinoxylooligosaccharide mixture can be from about 2% to about 20% w / w. The concentration of arabinoxylooligosaccharides having a degree of polymerization 7 can be at least 2%, 4%, 6%, 8%, 10%, 12%, 15%, 17% or 20% w / w.

[0295] The concentration of arabinoxylooligosaccharides having a degree of polymerization 8 in the arabinoxylooligosaccharide mixture can be from about 1% to about 15% w / w. The concentration of arabinoxylooligosaccharides having a degree of polymerization 8 can be at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w.

[0296] The concentration of arabinoxylooligosaccharides having a degree of polymerization of 9 in the arabinoxylooligosaccharide mixture can be from about 2% to about 15% w / w. The concentration of arabinoxylooligosaccharides having a degree of polymerization of 9 can be at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w.

[0297] The concentration of arabinoxylooligosaccharides having a degree of polymerization of 10 in the arabinoxylooligosaccharide mixture can be from about 2% to about 15% w / w. The concentration of arabinoxylooligosaccharides having a degree of polymerization of 10 can be at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w.

[0298] The concentration of arabinoxylooligosaccharides having a degree of polymerization of 11 in the arabinoxylooligosaccharide mixture can be from about 2% to about 15% w / w. The concentration of arabinoxylooligosaccharides having a degree of polymerization of 11 can be at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w.

[0299] The concentration of arabinoxylooligosaccharides having a degree of polymerization 12 in the arabinoxylooligosaccharide mixture can be from about 2% to about 15% w / w. The concentration of arabinoxylooligosaccharides having a degree of polymerization 12 can be at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w.

[0300] The concentration of arabinoxylooligosaccharides having a degree of polymerization of 13 in the arabinoxylooligosaccharide mixture can be from about 2% to about 15% w / w. The concentration of arabinoxylooligosaccharides having a degree of polymerization of 13 can be at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w.

[0301] The concentration of arabinoxylooligosaccharides having a degree of polymerization of 14 in the arabinoxylooligosaccharide mixture can be from about 2% to about 15% w / w. The concentration of arabinoxylooligosaccharides having a degree of polymerization of 14 can be at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w.

[0302] The concentration of arabinoxylooligosaccharides having a degree of polymerization of 15 in the arabinoxylooligosaccharide mixture can be from about 2% to about 15% w / w. The concentration of arabinoxylooligosaccharides having a degree of polymerization of 15 can be at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w.

[0303] The concentration of arabinoxylooligosaccharides having a degree of polymerization of 3 to 12 in the arabinoxylooligosaccharide mixture can be about 0.1% to about 15% w / w. The concentration of arabinoxylooligosaccharides having a degree of polymerization of 3 to 12 can be at least 0.1%, 0.3%, 0.5%, 0.8%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or 15% w / w. Compositions Comprising Combinations of Monosaccharides, Polysaccharides and / or Oligosaccharides

[0304] In some embodiments, the composition or ingredients (e.g., ingredients for human consumption) may be water-soluble. The solubility of the ingredients in water may be at least 80 g of the ingredients per 100 g of water at 50° C.

[0305] The ingredients can be combined with a liquid to form a liquid ingredient. In some cases, the viscosity of the liquid ingredient can be comparable to or similar to the viscosity of corn syrup. In some other cases, the viscosity of the liquid ingredient can be comparable to or similar to the viscosity of high fructose corn syrup. For example, the liquid ingredient can have a viscosity of 5 cps to 100,000 cps, 8,000 cps to 100,000 cps, 10,000 cps to 50,000 cps, or 15,000 cps to 25,000 cps. Additionally, the liquid ingredient can have fewer calories per gram than corn syrup or high fructose corn syrup. The liquid ingredient can have a lower glycemic index than corn syrup or high fructose corn syrup.

[0306] In some embodiments, the liquid can comprise water or any other suitable liquid. The liquid component can comprise at least 5%, 10%, 20%, 30%, 40% or 50% by dry weight of at least one oligosaccharide. Furthermore, the liquid component can comprise at least 0.2%, 0.5%, 1%, 2%, 3%, 5% or 10% by dry weight of at least one polysaccharide. For example, the liquid component can comprise at least 20% by dry weight of at least one oligosaccharide and at least 2% by dry weight of at least one polysaccharide. Other combinations of at least one oligosaccharide and at least one polysaccharide are also within the scope of the present disclosure.

[0307] The liquid component comprises at least 0.2%, 0.5%, 1%, 2%, 3%, 5% or 10% xylan by dry weight. The liquid component may comprise at least 0.2%, 0.5%, 1%, 2%, 3%, 5% or 10% mannan by dry weight. The liquid component may comprise at least 0.2%, 0.5%, 1%, 2%, 3%, 5% or 10% cellulose derivatives by dry weight.

[0308] In various examples, the liquid component can have a polysaccharide concentration of 0.1% to 50%, 0.1% to 40%, 0.1% to 30%, 0.1% to 20%, 0.1% to 10%, 0.5% to 50%, or 1% to 50% w / v. For example, the liquid component can have a polysaccharide concentration of 0.1% to 50% w / v. The liquid component can include polysaccharides and oligosaccharides in amounts of 1:200 to 1:1, 1:150 to 1:1, 1:125 to 1:1, 1:100 to 1:1, 1:90 to 1:1, 1:80 to 1:1, 1:70 to 1:1, 1:60 and 1:1, 1:50 and 1:1, 1:25 and 1:1, or 1:10 and 1:1. For example, the liquid component can include polysaccharides and oligosaccharides in amounts ranging from a ratio of 1:100 to 1:1.

[0309] The one or more soluble polysaccharides can include at least one of mannan, xylan, mixed linkage glucan, lignocellulose, hemicellulose, cellulose derivatives, chitosan, xyloglucan, or any other suitable soluble polysaccharides. The cellulose derivatives can include at least one of cellulose acetate, hydroxyethyl cellulose, hydroxymethyl cellulose, or any other suitable cellulose derivatives.

[0310] The biomass can include at least one of sugarcane biomass, corn biomass, wheat biomass, hardwood biomass, softwood biomass, or any other suitable biomass.

[0311] In certain examples, the composition for human consumption can include soluble polysaccharides and oligosaccharides including at least one of: (i) cellooligosaccharides having a degree of polymerization (DP) of 2-6; (ii) xylooligosaccharides having a DP of 2-12; (iii) mannooligosaccharides having a DP of 2-12; (iv) arabinoxylooligosaccharides having a DP of 3-15; (v) mixed linkage glucan oligosaccharides having a DP of 2-5; or (vi) chitooligosaccharides having a DP of 2-12. The composition can include less than 5% soluble polysaccharides by dry weight. In some cases, the composition can include less than 1%, 2%, 5%, 7.5%, 10% or 20% soluble polysaccharides by dry weight. In some embodiments, the composition can be free or substantially free of insoluble polysaccharides.

[0312] The composition may include a combination of polysaccharides and oligosaccharides. In some embodiments, the composition may include a combination of oligosaccharides and soluble polysaccharides. The polysaccharide (or soluble polysaccharide) source in such compositions may contain cellulose, such as biomass, e.g., undigested components of partially digested biomass, such as undigested biomass from the same reaction that produced the oligosaccharides. The polysaccharides in the undigested biomass may include lignin, polyphenols, cellulose, lignocellulose, or any other suitable polysaccharides described herein. The addition of polysaccharides (e.g., soluble polysaccharides) to the oligosaccharide mixture may be performed to improve the gastrointestinal tolerance of the oligosaccharide mixture. Eating or drinking oligosaccharides may cause gastrointestinal distress, including diarrhea, discomfort, and bloating. The compositions described herein may have improved gastrointestinal tolerance, such as little or no discomfort, bloating, diarrhea, or gastrointestinal distress, compared to commercially available sugar compositions or sugar compositions that contain primarily monosaccharides and / or disaccharides. For example, a subject ingesting one or more of the compositions provided herein can have improved gastrointestinal tolerance, such as little or no discomfort, bloating, diarrhea, or gastrointestinal distress, as compared to if the subject ingested or ingested a commercially available sugar composition, or a sugar composition that includes primarily monosaccharides and / or disaccharides.

[0313] The concentration of undigested biomass in the composition can be 1% to 50% w / w. The concentration of undigested biomass in the composition can be 1% to 5%, 1% to 10%, 1% to 15%, 1% to 20%, 1% to 25%, 1% to 30%, 1% to 35%, 1% to 40%, 1% to 45%, 1% to 50%, 5% to 10%, 5% to 15%, 5% to 20%, 5% to 25%, 5% to 30%, 5% to 35%, 5% to 40%, 5% to 45%, 5% to 50%, 10% to 15%, 10% to 20%, 10% to 25%, 10% to 30%, 10% to 35%, 10% to 40%, 10% to 45%, 10% to 50%, 15% to 20%, 15% to It can be 25%, 15%-30%, 15%-35%, 15%-40%, 15%-45%, 15%-50%, 20%-25%, 20%-30%, 20%-35%, 20%-40%, 20%-45%, 20%-50%, 25%-30%, 25%-35%, 25%-40%, 25%-45%, 25%-50%, 30%-35%, 30%-40%, 30%-45%, 30%-50%, 35%-40%, 35%-45%, 35%-50%, 40%-45%, 40%-50%, or 45%-50% w / w. The concentration of undigested biomass in the composition can be about 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45% or 50% w / w. The concentration of undigested biomass in the composition can be at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40% or 45% w / w. The concentration of undigested biomass in the composition can be at most 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45% or 50% w / w.

[0314] The concentration of the soluble polysaccharide in the composition can be 1% to 50% w / w. The concentration of the soluble polysaccharide in the composition can be 1% to 5%, 1% to 10%, 1% to 15%, 1% to 20%, 1% to 25%, 1% to 30%, 1% to 35%, 1% to 40%, 1% to 45%, 1% to 50%, 5% to 10%, 5% to 15%, 5% to 20%, 5% to 25%, 5% to 30%, 5% to 35%, 5% to 40%, 5% to 45%, 5% to 50%, 10% to 15%, 10% to 20%, 10% to 25%, 10% to 30%, 10% to 35%, 10% to 40%, 10% to 45%, 10% to 50%, 15% to 20%, 15% to 25%. %, 15%-30%, 15%-35%, 15%-40%, 15%-45%, 15%-50%, 20%-25%, 20%-30%, 20%-35%, 20%-40%, 20%-45%, 20%-50%, 25%-30%, 25%-35%, 25%-40%, 25%-45%, 25%-50%, 30%-35%, 30%-40%, 30%-45%, 30%-50%, 35%-40%, 35%-45%, 35%-50%, 40%-45%, 40%-50%, or 45%-50% w / w. The concentration of the soluble polysaccharide in the composition may be about 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45% or 50% w / w. The concentration of the soluble polysaccharide in the composition may be at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40% or 45% w / w. The concentration of the soluble polysaccharide in the composition may be at most 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45% or 50% w / w.

[0315] The concentration of xylooligosaccharides in the composition can be 1% to 50% w / w. The concentration of xylooligosaccharides in the composition can be 1% to 5%, 1% to 10%, 1% to 15%, 1% to 20%, 1% to 25%, 1% to 30%, 1% to 35%, 1% to 40%, 1% to 45%, 1% to 50%, 5% to 10%, 5% to 15%, 5% to 20%, 5% to 25%, 5% to 30%, 5% to 35%, 5% to 40%, 5% to 45%, 5% to 50%, 10% to 15%, 10% to 20%, 10% to 25%, 10% to 30%, 10% to 35%, 10% to 40%, 10% to 45%, 10% to 50%, 15% to 2 ... It can be 5%, 15%-30%, 15%-35%, 15%-40%, 15%-45%, 15%-50%, 20%-25%, 20%-30%, 20%-35%, 20%-40%, 20%-45%, 20%-50%, 25%-30%, 25%-35%, 25%-40%, 25%-45%, 25%-50%, 30%-35%, 30%-40%, 30%-45%, 30%-50%, 35%-40%, 35%-45%, 35%-50%, 40%-45%, 40%-50%, or 45%-50% w / w. The concentration of xylooligosaccharides in the composition can be about 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45% or 50% w / w. The concentration of xylooligosaccharides in the composition can be at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40% or 45% w / w. The concentration of xylooligosaccharides in the composition can be at most 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45% or 50% w / w.

[0316] The concentration of cellooligosaccharide in the composition can be 1% to 50% w / w. The concentration of cellooligosaccharide in the composition can be 1% to 5%, 1% to 10%, 1% to 15%, 1% to 20%, 1% to 25%, 1% to 30%, 1% to 35%, 1% to 40%, 1% to 45%, 1% to 50%, 5% to 10%, 5% to 15%, 5% to 20%, 5% to 25%, 5% to 30%, 5% to 35%, 5% to 40%, 5% to 45%, 5% to 50%, 10% to 15%, 10% to 20%, 10% to 25%, 10% to 30%, 10% to 35%, 10% to 40%, 10% to 45%, 10% to 50%, 15% to 2 ... It can be 5%, 15%-30%, 15%-35%, 15%-40%, 15%-45%, 15%-50%, 20%-25%, 20%-30%, 20%-35%, 20%-40%, 20%-45%, 20%-50%, 25%-30%, 25%-35%, 25%-40%, 25%-45%, 25%-50%, 30%-35%, 30%-40%, 30%-45%, 30%-50%, 35%-40%, 35%-45%, 35%-50%, 40%-45%, 40%-50%, or 45%-50% w / w. The concentration of cellooligosaccharides in the composition can be about 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45% or 50% w / w. The concentration of cellooligosaccharides in the composition can be at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40% or 45% w / w. The concentration of cellooligosaccharides in the composition can be at most 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45% or 50% w / w.

[0317] In some embodiments, the composition can comprise at least 5% w / w cellooligosaccharides and at least 5% w / w of a second oligosaccharide (e.g., at least 5% w / w of a xylooligosaccharide, mannooligosaccharide, mixed linkage glucan oligosaccharide, xyloglucan oligosaccharide, chitooligosaccharide, arabinoxylooligosaccharide, or any other suitable oligosaccharide).

[0318] The concentration of mannooligosaccharides in the composition can be 1% to 50% w / w. The concentration of mannooligosaccharides in the composition can be 1% to 5%, 1% to 10%, 1% to 15%, 1% to 20%, 1% to 25%, 1% to 30%, 1% to 35%, 1% to 40%, 1% to 45%, 1% to 50%, 5% to 10%, 5% to 15%, 5% to 20%, 5% to 25%, 5% to 30%, 5% to 35%, 5% to 40%, 5% to 45%, 5% to 50%, 10% to 15%, 10% to 20%, 10% to 25%, 10% to 30%, 10% to 35%, 10% to 40%, 10% to 45%, 10% to 50%, 15% to 2 ... It can be 5%, 15%-30%, 15%-35%, 15%-40%, 15%-45%, 15%-50%, 20%-25%, 20%-30%, 20%-35%, 20%-40%, 20%-45%, 20%-50%, 25%-30%, 25%-35%, 25%-40%, 25%-45%, 25%-50%, 30%-35%, 30%-40%, 30%-45%, 30%-50%, 35%-40%, 35%-45%, 35%-50%, 40%-45%, 40%-50%, or 45%-50% w / w. The concentration of mannooligosaccharides in the composition can be about 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45% or 50% w / w. The concentration of mannooligosaccharides in the composition can be at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40% or 45% w / w. The concentration of mannooligosaccharides in the composition can be at most 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45% or 50% w / w.

[0319] The concentration of chitooligosaccharide in the composition can be 1% to 50% w / w. The concentration of chitooligosaccharide in the composition can be 1% to 5%, 1% to 10%, 1% to 15%, 1% to 20%, 1% to 25%, 1% to 30%, 1% to 35%, 1% to 40%, 1% to 45%, 1% to 50%, 5% to 10%, 5% to 15%, 5% to 20%, 5% to 25%, 5% to 30%, 5% to 35%, 5% to 40%, 5% to 45%, 5% to 50%, 10% to 15%, 10% to 20%, 10% to 25%, 10% to 30%, 10% to 35%, 10% to 40%, 10% to 45%, 10% to 50%, 15% to 2 ... It can be 5%, 15%-30%, 15%-35%, 15%-40%, 15%-45%, 15%-50%, 20%-25%, 20%-30%, 20%-35%, 20%-40%, 20%-45%, 20%-50%, 25%-30%, 25%-35%, 25%-40%, 25%-45%, 25%-50%, 30%-35%, 30%-40%, 30%-45%, 30%-50%, 35%-40%, 35%-45%, 35%-50%, 40%-45%, 40%-50%, or 45%-50% w / w. The concentration of chitooligosaccharides in the composition can be about 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45% or 50% w / w. The concentration of chitooligosaccharides in the composition can be at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40% or 45% w / w. The concentration of chitooligosaccharides in the composition can be at most 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45% or 50% w / w.

[0320] The concentration of the xyloglucan oligosaccharide in the composition can be from 1% to 50% w / w. The concentration of xyloglucan oligosaccharides in the composition is 1%-5%, 1%-10%, 1%-15%, 1%-20%, 1%-25%, 1%-30%, 1%-35%, 1%-40%, 1%-45%, 1%-50%, 5%-10%, 5%-15%, 5%-20%, 5%-25%, 5%-30%, 5%-35%, 5%-40%, 5%-45%, 5%-50%, 10%-15%, 10%-20%, 10%-25%, 10%-30%, 10%-35%, 10%-40%, 10%-45%, 10%-50%, 15%-20%, 15% It can be ~25%, 15%-30%, 15%-35%, 15%-40%, 15%-45%, 15%-50%, 20%-25%, 20%-30%, 20%-35%, 20%-40%, 20%-45%, 20%-50%, 25%-30%, 25%-35%, 25%-40%, 25%-45%, 25%-50%, 30%-35%, 30%-40%, 30%-45%, 30%-50%, 35%-40%, 35%-45%, 35%-50%, 40%-45%, 40%-50%, or 45%-50% w / w. The concentration of xyloglucan oligosaccharides in the composition can be about 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45% or 50% w / w. The concentration of xyloglucan oligosaccharides in the composition can be at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40% or 45% w / w. The concentration of xyloglucan oligosaccharides in the composition can be at most 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45% or 50% w / w.

[0321] The concentration of the mixed linkage glucan oligosaccharide in the composition can be 1% to 50% w / w. The concentration of the mixed linkage glucan oligosaccharide in the composition can be 1% to 5%, 1% to 10%, 1% to 15%, 1% to 20%, 1% to 25%, 1% to 30%, 1% to 35%, 1% to 40%, 1% to 45%, 1% to 50%, 5% to 10%, 5% to 15%, 5% to 20%, 5% to 25%, 5% to 30%, 5% to 35%, 5% to 40%, 5% to 45%, 5% to 50%, 10% to 15%, 10% to 20%, 10% to 25%, 10% to 30%, 10% to 35%, 10% to 40%, 10% to 45%, 10% to 50%, 15% to 20%, 15% to 20%, 15% to 30%, 15% to 35%, 15% to 40%, 15% to 45%, 15% to 50%, 15% to 20%, 15% to 3 ... % to 25%, 15% to 30%, 15% to 35%, 15% to 40%, 15% to 45%, 15% to 50%, 20% to 25%, 20% to 30%, 20% to 35%, 20% to 40%, 20% to 45%, 20% to 50%, 25% to 30%, 25% to 35%, 25% to 40%, 25% to 45%, 25% to 50%, 30% to 35%, 30% to 40%, 30% to 45%, 30% to 50%, 35% to 40%, 35% to 45%, 35% to 50%, 40% to 45%, 40% to 50%, or 45% to 50% w / w. The concentration of the mixed linkage glucan oligosaccharide in the composition can be about 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45% or 50% w / w. The concentration of the mixed linkage glucan oligosaccharide in the composition can be at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40% or 45% w / w. The concentration of the mixed linkage glucan oligosaccharide in the composition can be at most 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45% or 50% w / w.

[0322] The concentration of the arabinoxylooligosaccharides in the composition can be from 1% to 50% w / w. The concentration of arabinoxylooligosaccharides in the composition is 1%-5%, 1%-10%, 1%-15%, 1%-20%, 1%-25%, 1%-30%, 1%-35%, 1%-40%, 1%-45%, 1%-50%, 5%-10%, 5%-15%, 5%-20%, 5%-25%, 5%-30%, 5%-35%, 5%-40%, 5%-45%, 5%-50%, 10%-15%, 10%-20%, 10%-25%, 10%-30%, 10%-35%, 10%-40%, 10%-45%, 10%-50%, 15%-20%, 15% It can be ~25%, 15%-30%, 15%-35%, 15%-40%, 15%-45%, 15%-50%, 20%-25%, 20%-30%, 20%-35%, 20%-40%, 20%-45%, 20%-50%, 25%-30%, 25%-35%, 25%-40%, 25%-45%, 25%-50%, 30%-35%, 30%-40%, 30%-45%, 30%-50%, 35%-40%, 35%-45%, 35%-50%, 40%-45%, 40%-50%, or 45%-50% w / w. The concentration of arabinoxylooligosaccharides in the composition can be about 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45% or 50% w / w. The concentration of arabinoxylooligosaccharides in the composition can be at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40% or 45% w / w. The concentration of arabinoxylooligosaccharides in the composition can be at most 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45% or 50% w / w.

[0323] The composition may include one or more polysaccharides (e.g., one or more soluble polysaccharides) and one or more oligosaccharides. The composition may include a polysaccharide and one type of oligosaccharide. The composition may include one polysaccharide or multiple polysaccharides and two forms of oligosaccharides. The composition may include one polysaccharide or multiple polysaccharides and three forms of oligosaccharides. The composition may include one polysaccharide or multiple polysaccharides and four forms of oligosaccharides. The composition may include one polysaccharide or multiple polysaccharides and five forms of oligosaccharides. The oligosaccharides can be xylooligosaccharides, cellooligosaccharides, mannooligosaccharides, mixed linkage glucan oligosaccharides, xyloglucan oligosaccharides, chitooligosaccharides, arabinoxylooligosaccharides, or any other suitable oligosaccharides described herein.

[0324] The composition may comprise about 1% to 50% w / w polysaccharides, such as undigested biomass or extracted soluble polysaccharide types, and about 5% to about 95% w / w oligosaccharides. The polysaccharide composition may be at least about 1%, 2%, 2.5%, 3%, 4%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45% or 50% w / w. The oligosaccharides in such mixtures may be present in greater than 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90% or 95% w / w. The oligosaccharides may be a mixture of one or more oligosaccharides. For example, the composition may comprise 5% w / w undigested biomass and 50% w / w of an oligosaccharide mixture described elsewhere herein. In another example, the composition may comprise 2.5% w / w of soluble polysaccharides and 50% w / w of an oligosaccharide mixture described elsewhere herein.

[0325] The composition may comprise about 5% w / w of polysaccharides, such as a type of undigested biomass, and about 5% to about 95% w / w of oligosaccharides. The oligosaccharides in such a mixture may be present at greater than 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90% or 95% w / w. The oligosaccharides may be a mixture of one or more oligosaccharides. For example, the composition may comprise 5% w / w of undigested biomass, and 50% w / w of an oligosaccharide mixture described elsewhere herein. In another example, the composition may comprise 5% w / w of soluble polysaccharides, and 50% w / w of an oligosaccharide mixture described elsewhere herein.

[0326] The composition may comprise about 7% w / w polysaccharides, such as a type of undigested biomass, and about 5% to about 93% w / w oligosaccharides. The oligosaccharides may form at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90% or 93% w / w of such mixtures. The oligosaccharides may be a mixture of one or more oligosaccharides. For example, the composition may comprise 7% w / w undigested biomass, and 50% w / w of an oligosaccharide mixture described elsewhere herein. In another example, the composition may comprise 7% w / w soluble polysaccharides, and 50% w / w of an oligosaccharide mixture described elsewhere herein.

[0327] The composition may comprise about 10% w / w polysaccharides, such as a type of undigested biomass, and about 5% to about 90% w / w oligosaccharides. The oligosaccharides may form at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85% or 90% w / w of such mixtures. The oligosaccharides may be a mixture of one or more oligosaccharides. For example, the composition may comprise 10% w / w undigested biomass, and 50% w / w of an oligosaccharide mixture described elsewhere herein. In another example, the composition may comprise 10% polysaccharides, and 50% w / w of an oligosaccharide mixture described elsewhere herein.

[0328] The composition may comprise about 12% w / w polysaccharides, such as a type of undigested biomass, and about 5% to about 95% w / w oligosaccharides. The oligosaccharides may form at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85% or 88% w / w of such mixtures. The oligosaccharides may be a mixture of one or more oligosaccharides. For example, the composition may comprise 12% w / w undigested biomass, and 50% w / w of an oligosaccharide mixture described elsewhere herein. In another example, the composition may comprise 12% w / w soluble polysaccharides, and 50% w / w of an oligosaccharide mixture described elsewhere herein.

[0329] The composition may comprise about 15% w / w polysaccharides, such as a type of undigested biomass, and about 5% to about 85% w / w oligosaccharides. The oligosaccharides may form at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80% or 85% w / w of such mixtures. The oligosaccharides may be a mixture of one or more oligosaccharides. For example, the composition may comprise 15% w / w undigested biomass, and 50% w / w of an oligosaccharide mixture described elsewhere herein. In another example, the composition may comprise 15% w / w soluble polysaccharides, and 50% w / w of an oligosaccharide mixture described elsewhere herein.

[0330] The composition may comprise about 20% w / w polysaccharides, such as a type of undigested biomass, and about 5% to about 80% w / w oligosaccharides. The oligosaccharides may form at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75% or 80% w / w of such mixtures. The oligosaccharides may be a mixture of one or more oligosaccharides. For example, the composition may comprise 20% w / w undigested biomass, and 50% w / w of an oligosaccharide mixture described elsewhere herein. In another example, the composition may comprise 20% w / w soluble polysaccharides, and 50% w / w of an oligosaccharide mixture described elsewhere herein.

[0331] The composition may comprise about 25% w / w polysaccharides, such as a type of undigested biomass, and about 5% to about 75% w / w oligosaccharides. The oligosaccharides may form at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70% or 75% w / w of such mixtures. The oligosaccharides may be a mixture of one or more oligosaccharides. For example, the composition may comprise 25% w / w undigested biomass, and 50% w / w of an oligosaccharide mixture described elsewhere herein. In another example, the composition may comprise 25% w / w soluble polysaccharides, and 50% w / w of an oligosaccharide mixture described elsewhere herein.

[0332] The composition may comprise about 30% w / w polysaccharides, such as a type of undigested biomass, and about 5% to about 70% w / w oligosaccharides. The oligosaccharides may form at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65% or 70% w / w of such mixtures. The oligosaccharides may be a mixture of one or more oligosaccharides. For example, the composition may comprise 30% w / w undigested biomass, and 50% w / w of an oligosaccharide mixture described elsewhere herein. In another example, the composition may comprise 30% w / w soluble polysaccharides, and 50% w / w of an oligosaccharide mixture described elsewhere herein.

[0333] The composition may comprise about 40% w / w polysaccharides, such as a type of undigested biomass, and about 5% to about 60% w / w oligosaccharides. The oligosaccharides may form at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55% or 60% w / w of such mixtures. The oligosaccharides may be a mixture of one or more oligosaccharides. For example, the composition may comprise 40% w / w undigested biomass, and 50% w / w of an oligosaccharide mixture described elsewhere herein. In another example, the composition may comprise 40% w / w soluble polysaccharides, and 50% w / w of an oligosaccharide mixture described elsewhere herein.

[0334] The composition may comprise about 50% w / w polysaccharides, such as a type of undigested biomass, and about 5% to about 50% w / w oligosaccharides. The oligosaccharides may form at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45% or 50% w / w of such mixtures. The oligosaccharides may be a mixture of one or more oligosaccharides. For example, the composition may comprise 50% w / w undigested biomass, and 50% w / w of an oligosaccharide mixture described elsewhere herein. In another example, the composition may comprise 50% w / w soluble polysaccharides, and 50% w / w of an oligosaccharide mixture described elsewhere herein.

[0335] In some embodiments, the composition or ingredients may contain less than 1%, 5%, 10%, 15%, 20%, 25%, 30% or 40% w / w of monosaccharides. For example, the composition may contain less than 20% w / w of monosaccharides. The composition may contain 10%-40%, 15%-30%, 18%-25% or about 20% w / w of monosaccharides. In some embodiments, the composition or ingredients may contain less than 1%, 5%, 10%, 15%, 20%, 25%, 30% or 40% w / w of glucose. For example, the composition may contain less than 10% w / w of glucose. The composition may contain 10%-40%, 15%-30%, 18%-25% or about 20% w / w of glucose. In some embodiments, the compositions or ingredients may contain less than 1%, 5%, 10%, 15%, 20%, 25%, 30% or 40% w / w xylose. For example, the compositions may contain less than 10% w / w xylose. The compositions may contain 10%-40%, 15%-30%, 18%-25% or about 20% w / w xylose.

[0336] In certain cases, the ratio of glucose residues to xylose residues (e.g., glucose:xylose) in the composition or ingredient can be 1:1 to 1:9, 1:1 to 1:7, 1:1 to 1:5, 1:1 to 1:3, or 1:1 to 1:2.

[0337] In certain embodiments, the composition may comprise less than 30%, 40%, 50%, 60%, 65%, 70%, 75% or 80% w / w of disaccharides. For example, the composition may comprise less than 70% w / w of disaccharides. The composition may comprise 10%-95%, 15%-90%, 20%-80%, 30%-70% or 40%-60% w / w of disaccharides. The composition may comprise 5%-95%, 10%-92.5%, 15%-90%, 20%-70%, 30%-60% or 40%-50% of disaccharides. In various embodiments, the composition may comprise at least 0.5%, 1%, 2.5%, 5%, 7.5%, 10%, 15% or 20% w / w of trisaccharides. For example, the composition may comprise at least 5% w / w of trisaccharides. In various embodiments, the composition may comprise at least 0.5%, 1%, 2.5%, 5%, 7.5%, 10%, 15%, or 20% w / w of trisaccharides. For example, the composition may comprise at least 5% w / w of trisaccharides. The composition may comprise 1%-75%, 2.5%-60%, 5%-50%, 10%-40%, or 20%-30% of trisaccharides. In some cases, the composition may comprise at least 0.1%, 0.5%, 1%, 2.5%, 5%, 7.5%, 10%, 15%, or 20% w / w of tetrasaccharides. For example, the composition may comprise at least 1% w / w of tetrasaccharides. In various cases, the composition may comprise at least 0.01%, 0.05%, 0.1%, 0.2%, 0.3%, 0.5%, 0.5%, 1%, 2.5%, 5%, 7.5% w / w of the pentasaccharide. For example, the composition may comprise at least 0.1% w / w of the pentasaccharide. Use of the composition as an ingredient

[0338] In some embodiments, the composition is an ingredient (e.g., in a food product). In certain embodiments, the ingredient comprises at least 50%, 60%, 70%, 80%, 90%, 95%, 99% or 99.5% sugar by dry weight. The ingredient can consist essentially of sugar. For example, the ingredient may comprise less than 0.5%, 0.3% or 0.1% of other substances by dry weight.

[0339] The component may comprise an oligosaccharide mixture as described elsewhere herein. The component may comprise at least two types of oligosaccharides. For example, the component may comprise three types of oligosaccharides. The component may comprise four types of oligosaccharides. The component may comprise five types of oligosaccharides. The component may comprise six types of oligosaccharides. The component may comprise seven types of oligosaccharides.

[0340] In some embodiments, the composition includes cellooligosaccharides, e.g., cellooligosaccharides in combination with xylooligosaccharides. Alternative compositions can include cellooligosaccharides in combination with mannooligosaccharides.

[0341] The ingredients can be used to prepare a final product. The ingredients can also be processed in some physical or chemical way before or during incorporation into a foodstuff, cosmetic or dietary supplement. The ingredients can be directly incorporated into the product, or can be incorporated into, for example, dough, cake mixture, chocolate mixture or other foodstuff precursor; cosmetic base composition; or dietary supplement, and can be cooked or otherwise processed in a way that can cause, for example, chemical modification, texture change, color change or other modification.

[0342] Foodstuffs, cosmetics or dietary supplements may be produced from the ingredients described herein. For example, in the food industry, the sugar blends produced by the current method may be used as sweeteners, bulking agents, added dietary fiber or water retention agents. The ingredients may be used as sugar substitutes. The ingredients may be incorporated into cakes, breads or other baked goods, or chocolates or other confectioneries such as toffees, fudges, meringues, jams, jellies or caramels; or beverages, for example, to provide a desired taste or color characteristic or to increase the fiber content of the diet. In certain examples, the ingredients may be incorporated into animal feed, for example, either as isolated ingredients or by utilizing the enzyme reaction mixture directly as feed.

[0343] In the cosmetics industry, sugars can be useful as ingredients because they can improve texture and water retention, act as UV absorbing molecules, maintain the structure of gels or creams, and / or act as bulking agents.The dietary fiber provided by the compositions described herein can promote digestive health, well-regulated intestinal flora, and other benefits to healthy living, so they can be incorporated into dietary supplement compositions.In this context, the compositions described herein can also function as ingredients in probiotic drinks, or other prebiotic or probiotic formulations.

[0344] The compositions or ingredients described herein can be used to modify one or more properties of the final product, including, but not limited to, sweetness, texture, mouthfeel, binding, gloss, smoothness, moistness, viscosity, color, hygroscopicity, flavor, bulking, water retention, caramelization, surface texture, crystallization, structural properties, and dissolution.

[0345] In some cases, the compositions and / or ingredients described herein can provide the final product with properties equivalent to or superior to the same properties achieved by sugar mixtures that mainly contain monosaccharides and / or disaccharides. The control composition can be a sugar commonly used in food and beverages, for example, a monosaccharide composition such as glucose, fructose, a disaccharide composition such as sucrose, or an artificial sugar composition. The control composition can be table sugar, corn syrup, high fructose corn syrup, or any other suitable composition. The term "equivalent" as used herein generally means that the two compositions can be up to 100%, up to 95%, up to 90%, or up to 80% identical. For example, equivalent can mean that the composition is up to 90% identical to the control composition.

[0346] In some cases, the compositions described herein may be used as sweetener compositions. The sweetener compositions may be used by themselves or as an ingredient in a final product. The compositions described herein may provide approximately the same level of sweetness as, or greater than, the same amount of a control composition, where the control composition mainly comprises monosaccharides and / or disaccharides. The compositions described herein may be used to replace the control composition as a sweetener in a final product. In some cases, the sweetness in the composition may be 5%, 10%, 15%, 20%, 30%, 40%, 50%, 70%, 80%, 90% or 100% higher than the same amount of the control composition.

[0347] The compositions described herein can provide a flavor profile that is comparable to or better than the flavor profile of an equivalent amount of a control composition, where the control composition mainly comprises monosaccharides and / or disaccharides.The compositions described herein can be used to replace the control composition as a flavor enhancer in the final product.In some cases, the flavor in the composition can be 5%, 10%, 15%, 20%, 30%, 40%, 50%, 70%, 80%, 90% or 100% higher than the equivalent amount of the control composition.

[0348] The compositions described herein can provide a texture profile that is equivalent to or better than an equivalent amount of a control composition, where the control composition comprises primarily monosaccharides and / or disaccharides. The compositions described herein may be used to replace the control composition as a texture enhancer in the final product.

[0349] The compositions described herein can provide binding profiles that are comparable to or better than an equivalent amount of a control composition, where the control composition comprises primarily monosaccharides and / or disaccharides. The compositions described herein may be used to replace the control composition as a binding enhancer in the final product.

[0350] The compositions described herein can provide a polish profile that is equivalent to or better than an equivalent amount of a control composition, where the control composition comprises primarily monosaccharides and / or disaccharides. The compositions described herein may be used to replace the control composition as a polish enhancer in the final product.

[0351] The compositions described herein can provide moisturization that is comparable to or better than an equivalent amount of a control composition, where the control composition comprises primarily monosaccharides and / or disaccharides. The compositions described herein may be used to replace the control composition to provide moisturization in a final product.

[0352] The compositions described herein can provide a similar or better shade profile to an equivalent amount of a control composition, where the control composition comprises primarily monosaccharides and / or disaccharides. The compositions described herein may be used to replace the control composition as a shade enhancer in the final product.

[0353] The compositions described herein can provide a dissolution profile that is comparable to or better than that of an equivalent amount of a control composition, where the control composition mainly comprises monosaccharides and / or disaccharides.The compositions described herein can be used to replace the control composition as a dissolution enhancer in the final product.In some cases, the dissolution of the composition can be 5%, 10%, 15%, 20%, 30%, 40%, 50%, 70%, 80%, 90% or 100% higher than that of the equivalent amount of the control composition.

[0354] The compositions described herein can provide a mouthfeel that is comparable to or better than an equivalent amount of a control composition, where the control composition comprises primarily monosaccharides and / or disaccharides.

[0355] The compositions described herein can provide a viscosity that is comparable to or better than an equivalent amount of a control composition, where the control composition comprises primarily monosaccharides and / or disaccharides.

[0356] The compositions described herein can provide hygroscopicity that is comparable to or greater than an equivalent amount of a control composition, where the control composition comprises primarily monosaccharides and / or disaccharides. In some cases, the hygroscopicity of the composition can be 5%, 10%, 15%, 20%, 30%, 40%, 50%, 70%, 80%, 90% or 100% greater than an equivalent amount of the control composition.

[0357] The compositions described herein can provide moisture retention that is comparable to or better than an equivalent amount of a control composition, where the control composition comprises primarily monosaccharides and / or disaccharides. In some cases, the moisture retention of the composition can be 5%, 10%, 15%, 20%, 30%, 40%, 50%, 70%, 80%, 90% or 100% higher than an equivalent amount of the control composition.

[0358] The compositions described herein can provide compositions that are lower in calories than an equivalent amount of a control composition, where the control composition comprises primarily monosaccharides and / or disaccharides. In some cases, the calorie count of the composition can be 5%, 10%, 15%, 20%, 30%, 40%, 50%, 70%, 80%, 90% or 100% lower than an equivalent amount of the control composition.

[0359] The compositions described herein can provide a lower glycemic index than an equivalent amount of a control composition, where the control composition comprises primarily monosaccharides and / or disaccharides. In some cases, the glycemic index of the composition can be 5%, 10%, 15%, 20%, 30%, 40%, 50%, 70%, 80%, 90% or 100% lower than an equivalent amount of the control composition.

[0360] The compositions described herein can provide bulking that is comparable to or better than an equivalent amount of a control composition, where the control composition comprises primarily monosaccharides and / or disaccharides.

[0361] The compositions described herein can provide caramelization that is comparable to or better than an equivalent amount of a control composition, where the control composition contains primarily monosaccharides and / or disaccharides.

[0362] The compositions described herein can provide surface texture that is equivalent to or better than an equivalent amount of a control composition, where the control composition comprises primarily monosaccharides and / or disaccharides.

[0363] The compositions described herein can provide crystallization that is comparable to or better than an equivalent amount of a control composition, where the control composition comprises primarily monosaccharides and / or disaccharides.

[0364] The compositions described herein can achieve structural characteristics comparable to an equivalent amount of a control composition, where the control composition comprises primarily monosaccharides and / or disaccharides.

[0365] The compositions described herein can provide a reduced aftertaste compared to an equivalent amount of a control composition, where the control composition comprises primarily monosaccharides and / or disaccharides.

[0366] Different compositions of oligosaccharides can have improved dissolution profiles, hygroscopicity profiles and taste profiles compared to the oligosaccharides used alone.

[0367] The compositions or ingredients described herein may be used to increase the fiber content of a final product, such as a food product or dietary supplement. The compositions may provide a higher level of fiber in the final product compared to an equivalent amount of a control composition, where the control composition mainly comprises monosaccharides and / or disaccharides. In some cases, the compositions may improve the fiber content of the final product without negatively affecting or substantially negatively affecting any other properties, such as taste, sweetness, mouthfeel, texture, binding, or any other property described herein. In some cases, the fiber content of the composition may be 5%, 10%, 15%, 20%, 30%, 40%, 50%, 70%, 80%, 90% or 100% more than an equivalent amount of a control composition.

[0368] The ingredients can be used to modify the properties of the final product, such as a food product or a dietary supplement or a cosmetic product. To modify the properties of the final product, the final product can further include a polysaccharide, such as a cellulosic polysaccharide, such as cellulose, or a polysaccharide derivative, such as a cellulose derivative, such as carboxymethylcellulose, or a polysaccharide aggregate, such as a portion of a lignocellulosic biomass. In some examples, the final product can include from 0% to 40% polysaccharide, polysaccharide derivative, or polysaccharide aggregate by dry weight, such as from 1% to 30% polysaccharide, polysaccharide derivative, or polysaccharide aggregate by dry weight, such as from 5% to 25% polysaccharide, polysaccharide derivative, or polysaccharide aggregate by dry weight, such as from 10% to 20% polysaccharide, polysaccharide derivative, or polysaccharide aggregate by dry weight.

[0369] The concentration of the composition containing the mixture of polysaccharides and oligosaccharides in the final product can be any of 0.1% to 40% w / w. The concentration of the composition containing the mixture of polysaccharides and oligosaccharides in the final product can be about 0.1% to about 0.5%, about 0.1% to about 1%, about 0.1% to about 5%, about 0.1% to about 10%, about 0.1% to about 15%, about 0.1% to about 20%, about 0.1% to about 25%, about 0.1% to about 30%, about 0.1% to about 35%, about 0.1% to about 40%, about 0.5% to about 1%. , about 0.5% to about 5%, about 0.5% to about 10%, about 0.5% to about 15%, about 0.5% to about 20%, about 0.5% to about 25%, about 0.5% to about 30%, about 0.5% to about 35%, about 0.5% to about 40%, about 1% to about 5%, about 1% to about 10%, about 1% to about 15%, about 1% to about 20%, about 1% to about 25%, about 1% to about 30%, about 1% to about 35% , about 1% to about 40%, about 5% to about 10%, about 5% to about 15%, about 5% to about 20%, about 5% to about 25%, about 5% to about 30%, about 5% to about 35%, about 5% to about 40%, about 10% to about 15%, about 10% to about 20%, about 10% to about 25%, about 10% to about 30%, about 10% to about 35%, about 10% to about 40%, about 15% to about 20%, about 15% to about The concentration of the composition containing the mixture of polysaccharides and oligosaccharides in the final product can be about 0.1%, about 0.5%, about 1%, about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 20% to about 25%, about 20% to about 30%, about 20% to about 35%, about 20% to about 40%, about 25% to about 30%, about 25% to about 35%, about 25% to about 40%, about 30% to about 35%, about 30% to about 40%, or about 35% to about 40% w / w. The concentration of the composition comprising the mixture of polysaccharides and oligosaccharides in the final product can be at least 0.1%, 0.5%, 1%, 5%, 10%, 15%, 20%, 25%, 30%, or 35% w / w. The concentration of the composition comprising the mixture of polysaccharides and oligosaccharides in the final product can be at most 0.5%, 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, or 40% w / w.

[0370] In some cases, the oligosaccharide mixture (e.g., cellooligosaccharides and xylooligosaccharides) can form at least 20%, 30%, 40%, 50%, 60% or 70% w / w of the edible composition or ingredient. For example, a combination of 50% w / w cellooligosaccharides and xylooligosaccharides can form the edible composition or ingredient.

[0371] In some examples, the ingredients can include corn cob extract (MCE). The MCE can be a mixture of oligosaccharides consisting primarily of arabinoxylooligosaccharides, xylooligosaccharides, cellooligosaccharides, and cellulose. In certain examples, the oligosaccharides can be non-digestible or substantially non-digestible. The arabinoxylooligosaccharides can be oligomers having a xylose backbone linked by β-(1→4) linkages substituted by arabinose side chains. The arabinoxylooligosaccharides can be non-digestible. The arabinoxylooligosaccharides can be produced by hydrolysis of arabinoxylan (a polysaccharide consisting of β-(1→4) linked xylose units substituted by arabinose side chains). Additionally, the arabinoxylooligosaccharides can have a degree of polymerization (DP) of 3 to 15.

[0372] In various examples, xylooligosaccharides can be oligomers having a xylose backbone linked by β-(1→4) bonds. Xylooligosaccharides can be non-digestible. Xylooligosaccharides can be produced by hydrolysis of arabinoxylan. Furthermore, xylooligosaccharides can have a DP of 2-8.

[0373] In certain instances, cellooligosaccharides can be oligomers with glucose backbones linked by β-(1→4) bonds. Cellooligosaccharides can be non-digestible. Cellooligosaccharides can be produced by hydrolysis of cellulose (a polysaccharide consisting of β-(1→4) linked glucose units). Furthermore, cellooligosaccharides can have a DP of 2-4, with the majority having a DP of 2. IV. Illustrative Embodiments Exemplary Methods for Extracting Soluble Polysaccharides for Subsequent Combination with Produced Oligosaccharides

[0374] FIG. 8 is a simplified flow diagram illustrating an embodiment of a method for extracting soluble polysaccharides for subsequent combination with the resulting oligosaccharides to form components.

[0375] In the illustrated method, extraction or removal of at least a portion of the soluble polysaccharides 140 prior to enzymatic treatment 150 may ensure that at least a portion of the soluble polysaccharides is retained or reserved for combination 144 with the resulting oligosaccharides to form component 105. In some other methods (not shown), the soluble polysaccharides are not extracted prior to enzymatic treatment so that the soluble polysaccharides may be digested by one or more enzymes. Component 105 of the illustrated method may be a sweetener or sugar substitute, which may be in a substantially soluble or fully soluble state. Thus, component 105 may be delivered as a syrup-like product (e.g., a viscous liquid). In some cases, ingredient 105 can be a replacement or partial replacement for corn syrup, high fructose corn syrup, maple syrup, honey, treacle, golden syrup, molasses, dextrose syrup, fructose syrup, agave syrup, date syrup, brown rice malt syrup, coconut syrup, corn syrup, or other suitable liquid sweeteners in food products.

[0376] As shown, biomass 107 (e.g., corn cobs or any other suitable biomass) can be physically pre-treated 110 (e.g., by comminuted or any other suitable method of physically pre-treating biomass). The physically treated biomass 112 can then be or can be subjected to thermochemical pre-treatment 130 (e.g., 15% w / v comminuted corn cobs can be heated in 1% w / v NaOH for 1 hour). In some embodiments, the thermochemical pre-treatment 130 is followed by a neutralization step (not shown) followed by extraction of soluble compounds or substances 140 from the physically pre-treated biomass 112. Extraction of the soluble compounds or substances 140 can include removing a liquid portion (e.g., supernatant) of the physically pre-treated biomass 112. The liquid portion can include soluble compounds 146 from the physically pre-treated biomass 112. The soluble compounds 146 can include soluble polysaccharides. In certain embodiments, 15% of the liquid portion (including soluble compounds or substances 146) can then be extracted or removed from the thermochemically pre-treated biomass 112. The extracted portion containing the soluble compounds or substances 146 (e.g., soluble polysaccharides) can then be subjected to one or more purification steps 142b (e.g., ultrafiltration) to concentrate the soluble polysaccharides 147.

[0377] Additionally, the liquid portion not extracted in step 140 can include soluble compounds or substances 146 and insoluble compounds or substances 148. For example, if 15% of the liquid portion is extracted as described above, the remaining portion of the physically pre-treated biomass 112 (including 85% of the liquid portion) can include soluble and insoluble polysaccharides. The solution containing the soluble compounds or substances 146 and the insoluble compounds or substances 148 can then be subjected to or can undergo the enzyme treatment 150 disclosed herein. For example, the solution containing the soluble compounds or substances 146 and the insoluble compounds or substances 148 can be added with one or more polysaccharide cutting enzymes to 0.5% w / v and incubated at 50° C. for 24 hours. The enzyme-treated biomass 151 can then become treated (e.g., filtered) 152, and at least a portion of the undigested biomass can be removed. In certain cases, the removed undigested biomass can be discarded or excluded from acceptance. The digested biomass 141 can then be subjected to purification 142a (e.g., ion exchange chromatography, nanofiltration, microfiltration, ultrafiltration or any other suitable method of purification) or the biomass can be subjected to purification as described herein to enrich for oligosaccharides 154. The extracted, isolated and / or purified oligosaccharides 154 and the extracted, isolated and / or purified soluble polysaccharides 147 can be combined, mixed and / or spray dried to form component 105. Exemplary Methods for Pretreating Biomass to Remove Monosaccharides and / or Disaccharides

[0378] FIG. 9 is a simplified flow diagram illustrating an embodiment of a method for pretreating biomass to remove mono- and / or disaccharides prior to enzymatic treatment.

[0379] The embodiment of FIG. 9 may include components or steps similar in some respects to those of the embodiment of FIG. 8. For example, the embodiment of FIG. 9 includes a physical pretreatment step 110, which may be similar to the physical pretreatment 210 of FIG. 8. It will be understood that the illustrated embodiments may have similar features. Thus, similar features are indicated with similar reference numbers, with the leading digits increasing by 100 for each reference number. For example, a physical pretreatment is designated as "110" in FIG. 8, and a similar physical pretreatment is designated as "210" in FIG. 9. As such, the relevant disclosure above regarding similarly identified features may not be repeated hereafter. Moreover, certain features of the method and related components or steps illustrated in FIG. 9 may not be indicated or identified by reference numbers in the drawings or specifically discussed in the written description that follows. However, such features may be clearly the same or substantially the same as features shown in and / or described with respect to other embodiments. Thus, the relevant descriptions of such features apply equally to the features of the method and relevant components or steps of Figure 9. Any suitable combination of features and variations thereof described with respect to the method illustrated in Figure 8 may be used in the method and components or steps of Figure 9, and vice versa. This pattern of disclosure applies equally to further embodiments shown in subsequent figures and / or in the description below.

[0380] As illustrated, mild pretreatment 220 (e.g., washing or incubation cycles as provided herein) of physically pretreated biomass 212 can include removing 224 soluble compounds 246. In certain cases, the soluble compounds 246 can include monosaccharides and / or disaccharides. The removed soluble monosaccharides and / or disaccharides can then be discarded and / or rejected. Thus, mild pretreatment 220 can be performed or carried out to remove soluble monosaccharides and / or disaccharides from biomass 207.

[0381] The biomass 207 can be physically pretreated 210 (e.g., shredded), and then the physically pretreated biomass 212 can be subjected to or undergo a mild pretreatment 220, such as washing or incubation (e.g., in water at 25° C. for 30 minutes). Soluble sugars 246 (e.g., soluble monosaccharides and / or disaccharides) can then be removed from the solution containing the mildly pretreated biomass 226. The mildly pretreated biomass 226 can then be subjected to or undergo a strong pretreatment 230. In certain cases, the strong pretreatment 230 can be a thermochemical pretreatment. For example, the mildly pretreated biomass 226 can be treated in 1% w / v NaOH at 100° C. for 60 minutes. In some embodiments, the strong pretreatment 230 can be followed by a neutralization step (not shown), followed by an enzyme treatment 250. The strong pretreated biomass 232 can then be treated with enzymes 250 as discussed herein. Further downstream processing 260 can then be performed on the enzyme-treated biomass 251 to generate components 205. Additional Exemplary Embodiments for Extracting Soluble Polysaccharides for Combination with Oligosaccharides

[0382] In some cases, the present disclosure relates to novel methods of processing plant biomass material to produce foodstuff, cosmetic, or nutritional supplement ingredients.

[0383] Sugary foods and beverages are an important part of cultures and lifestyles around the world, but the sugar they contain has been linked to obesity, diabetes, poor dental health, and destructive behaviors in people. As a result, consumer preferences are shifting away from sugar-containing foods, and governments are increasingly enacting regulations to encourage less sugar consumption.

[0384] Therefore, the industry has been searching for suitable low-calorie sweeteners for decades to replace sugar in food and beverages.Unfortunately, many sugar substitutes are produced from non-natural sources, and often bring along their sweetness with a potential bitterness or other unpleasant taste, neither of which is appealing to consumers.In addition, although many sweeteners can mimic the sweetness of sugar in food and beverages, few can comprehensively mimic the role that sugar plays in food, such as adding bulk, adjusting texture, providing structure, acting as a preservative, and adjusting color and flavor through caramelization and Maillard reaction.

[0385] Dietary fiber is an important part of a positive diet and helps maintain digestive health and a well-regulated intestinal flora. Such fiber includes sugars of various chain lengths and types. In addition to being found naturally in a wide range of foods, fiber can also be produced separately or added to other foods during their manufacture.

[0386] Biomass is a good source of sugars that can be used to replace sugar and add fiber to foods. However, there is still a need to optimize the processes by which these sugars are obtained from biomass and processed into compositions useful as foodstuffs, cosmetics, or nutritional supplement ingredients.

[0387] First, sugars generally need to be produced by controlled degradation. Different amounts of different sizes of sugars in an ingredient can affect its nutritional value and other properties such as hygroscopicity, which affect properties such as texture of the product the ingredient is used to make. It can also be desirable for an ingredient to contain polysaccharides, since polysaccharides can improve gastrointestinal tolerance. However, due to the rate at which some particularly desirable polysaccharides are destroyed during the enzymatic reactions of already known methods, it is difficult to isolate and then incorporate these desirable polysaccharides into the ingredient.

[0388] Furthermore, foods that require a smooth texture, such as candy, chocolate, and yogurt, generally require ingredients to be soluble to achieve a smooth texture. Ingredients that contain insoluble polymeric materials can result in a grainy texture. However, due to the insolubility of certain polysaccharides, it can be difficult to make compositions that contain polysaccharides that are entirely soluble, especially in a one-pot process from a single piece of biomass. Typically, soluble polysaccharides degrade faster than insoluble polysaccharides, so that after exposing plant biomass to enzymes, the soluble polysaccharides are entirely degraded and only insoluble polysaccharides remain, as in previously known methods.

[0389] Surprisingly, it has been discovered herein that polysaccharides can be isolated and incorporated into soluble foodstuffs, cosmetics, or nutraceutical ingredients that contain oligosaccharides, thus maintaining the benefits of increasing the ingredient's gastrointestinal tolerance and enabling it to be used in smooth textured foods. Polysaccharides can be isolated from the same plant biomass from which other desired sugars are obtained providing an efficient and streamlined production process.

[0390] Thus, in a first aspect of the present disclosure, there is provided a method for producing a foodstuff, cosmetic, or nutraceutical ingredient, comprising: a) providing a plant biomass comprising one or more soluble polysaccharides and one or more insoluble polysaccharides; b) treating the plant biomass to solubilize one or more soluble polysaccharides; c) removing a portion of the dissolved soluble polysaccharide(s); d) reacting the remaining plant biomass with one or more enzymes to form one or more oligosaccharides; e) removing one or more oligosaccharides; and f) combining a portion of the dissolved soluble polysaccharide(s) from step (c) with one or more oligosaccharides from step (e) to form the component. A method is provided that includes:

[0391] Thus, also provided are foodstuffs, cosmetic or nutraceutical ingredients obtainable by the methods of the present disclosure.

[0392] In another aspect of the present disclosure: i) cellooligosaccharides having a degree of polymerization from 2 to 6; ii) xylooligosaccharides with a degree of polymerization between 2 and 12; iii) mannooligosaccharides having a degree of polymerization between 2 and 12; iv) mixed linkage glucan oligosaccharides having a degree of polymerization from 2 to 5; v) xyloglucan oligosaccharides having a degree of polymerization of 4 to 12; and vi) chitooligosaccharides having a degree of polymerization of 2 to 12 At least one oligosaccharide selected from the list consisting of: i) xylan; ii) Mannan; iii) cellulose derivatives; iv) mixed linkage glucans; v) xyloglucan; and vi) Chitosan At least one polysaccharide selected from the list consisting of wherein the liquid component comprises at least 20% by dry weight of at least one oligosaccharide and at least 2% by dry weight of at least one polysaccharide, and the liquid component has a viscosity of 5 to 100,000 cps. A foodstuff, cosmetic, or nutritional liquid ingredient is provided.

[0393] The preparation of foodstuff, cosmetic, or nutritional supplement ingredients with the methods provided herein can allow for efficient use of biomass by incorporating oligomeric and polymeric materials from the same biomass source as the soluble ingredients are made. Furthermore, the methods can allow for purification, derivatization, or other modifications, as well as control of the proportions of oligomers and polymers, which can improve the functional properties, nutritional properties, and tolerance of the ingredients.

[0394] Any material containing suitable polysaccharides may be plant biomass. The foodstuff, cosmetic, and nutraceutical industries use a wide variety of oligosaccharides, so the polysaccharides suitable for the method are not particularly limited. Plant biomass suitable for generating the oligosaccharide profile of the present disclosure may include, for example, cellulose, lignocellulose, chitin, chitosan, xylan (such as glucuronoxylan, arabinoxylan, and glucuronoarabinoxylan), xyloglucan, mixed linkage glucan, and / or mannan (such as glucomannan, galactomannan, or galactoglucomannan), however, any plant biomass that can be appropriately worked with is contemplated. The plant biomass may include one or more soluble polysaccharides, such as any one of the following: mannan, mixed linkage glucan, lignocellulose, hemicellulose, certain cellulose derivatives, such as cellulose acetate, hydroxyethylcellulose, and hydroxymethylcellulose, and chitosan. In some embodiments, the plant biomass includes hemicellulose. In certain embodiments, the hemicellulose includes xylan and / or mannan.

[0395] Thus, the plant biomass may be grain, grain husks, bean husks, seed coats, and / or other seed material; seaweed; corn stover, straw, bagasse, miscanthus, sorghum residue, switchgrass, bamboo, and / or other monocotyledonous tissue; water hyacinth, leaf tissue, roots, and / or other vegetable matter; and / or any combination of suitable plant biomass. In some cases, the plant biomass comprises or suitably consists of sugarcane biomass (such as sugarcane bagasse), corn biomass (such as corn cobs or corn stover), wheat biomass (such as wheat straw, bran, etc.), hardwood or softwood. In some cases, the plant biomass comprises corn cobs, sugarcane bagasse, wheat straw, or rice straw.

[0396] In various cases, in step (b), "treating" is a thermochemical treatment of the plant biomass. As used herein, "thermochemical" generally refers to heating the plant biomass in a chemical above room temperature (room temperature can be about 20°C to 22°C), such as heating in a solution containing water, alkali, or an ionic solvent. The thermochemical step can physically and chemically modify the chemical components of the plant biomass. For example, free hydroxide ions from water or alkali can break hydrogen bonds between sugars, such as hemicellulose, that allow solubilization of some types of sugars, making them more readily available for further enzyme splitting in subsequent steps. These broken hydrogen bonds may be between monomers of the same sugar chain that contribute to the tertiary structure of the chain. The broken hydrogen bonds may be between monomers of different sugar chains that contribute to the quaternary structure of one or more chains. The treatment can then result in one or more polysaccharides that are soluble (i.e., polysaccharides that are particularly susceptible to hydroxide ion destruction, such as hemicellulose), being dissolved in the chemicals used. One or more polysaccharides that are insoluble, such as cellulose, do not dissolve in the chemicals.

[0397] The heating in processing step (b) may suitably be at a temperature range of from 30° C. to 180° C., from 50° C. to 150° C., or from 70° C. to 120° C. Higher temperatures can help soluble polysaccharides to dissolve quickly, but temperatures that are too high can be difficult to achieve efficiently and cost-effectively, and can chemically modify biomass components including sugars (e.g., in undesirable ways).

[0398] Heating may occur over a range of time scales, with particularly large amounts of biomass being exposed to heating for longer periods that can be adjusted accordingly, for example, heating of plant biomass can be for 1 minute to 72 hours, 10 minutes to 24 hours, 20 minutes to 12 hours, or 25 minutes to 8 hours.

[0399] In some cases, the thermochemical treatment may involve heating the plant biomass in water, i.e., at a neutral pH of about pH 7.

[0400] In certain cases, the thermochemical treatment may include heating the plant biomass in an alkaline solution having a pH of 8 to 14, 9 to 14, or 10 to 14. The solution may include or be suitable from any one of the alkalis selected from: sodium hydroxide, potassium hydroxide, sodium carbonate, calcium carbonate, calcium hydroxide, ammonium sulfate, ammonium hydroxide, and aqueous ammonia. In various cases, the alkali may be sodium hydroxide. Combinations of the listed alkalis are also contemplated.

[0401] A number of different sequential thermochemical treatment steps are also contemplated. For example, there may be two sequential thermochemical treatments, three sequential thermochemical treatments, or four or more sequential thermochemical treatments. In some cases, the biomass may be thermochemically treated in a neutral aqueous solution and then in an alkaline aqueous solution.

[0402] After the treatment step, step (c) may include removing a portion of the dissolved soluble polysaccharide or polysaccharides. The purpose of this step may be to isolate and remove the soluble polysaccharides from the plant biomass so that they are not degraded and therefore not lost in subsequent enzymatic reactions. This may allow the use of these polysaccharides in forming the components in a later step. All or a portion of the dissolved polysaccharides may be removed depending on the desired amount in the final component. The soluble polysaccharides may be removed using a simple step such as filtration of the chemical in which the soluble polysaccharides are dissolved.

[0403] Step d) involves reacting the remaining plant biomass, which may be in the form of a solution and / or suspension, with one or more enzymes to form one or more oligosaccharides. The soluble and insoluble polysaccharides present in the remaining plant biomass solution and / or suspension can be partially or fully cleaved by the one or more enzymes into oligosaccharides (e.g., useful oligosaccharides), possibly including partially cleaved or uncleaved polysaccharides, which may include cellulose, xylan (such as glucuronoxylan, arabinoxylan, or glucuronoarabinoxylan), mannan (such as glucomannan, galactomannan, or galactoglucomannan), mixed linkage glucan, xyloglucan, chitin, chitosan, or lignocellulose.

[0404] The enzymatic reaction may occur in solution and / or suspension in a suitable reaction vessel. The enzymatic reaction may occur at a temperature or temperature protocol suitable for the particular combination of enzyme and plant biomass, and the reaction may be allowed to proceed for a certain length of time until the product reaches a desired concentration or meets some other requirement.

[0405] The reaction mixture may be stirred constantly or at intervals to ensure optimal contact between the enzyme and the plant biomass. Agitation may take the form of rhythmic movement throughout the reaction vessel, a fan or other agitation device, sparging with air bubbles, or any other method of agitation.

[0406] The enzymatic reaction may be a microbial fermentation. The temperature and reaction time may be suitable for the growth of the microbial organism used. The microbial organism may be genetically modified to produce enzymes suitable for the production of the oligosaccharides of the present disclosure. The microorganism may be, for example, a bacterium, such as Escherichia coli, or a fungus, such as Saccharomyces cerevisiae, Aspergillus niger, or Trichoderma reesei.

[0407] The present disclosure further embodies an expression vector suitable for modifying a target microorganism to produce an enzyme or mixture of enzymes of the present disclosure. If desired, the expression vector, which may be a plasmid or any other nucleic acid capable of inducing the production of the enzyme, may contain one or more of the following regulatory sequences to control the expression of the exogenous enzyme: a heat shock gene regulatory sequence, a virulence gene regulatory sequence, and a sporulation gene regulatory sequence.

[0408] The enzymatic reaction can be carried out at a temperature or temperature protocol appropriate for the enzyme and substrate used, and may be carried out at a constant temperature ranging, for example, from about 10° C. to about 100° C., from about 20° C. to about 70° C., or from about 30° C. to about 60° C. If the enzymatic reaction takes the form of a microbial fermentation, the temperature may be appropriate, for example, such that the enzymatic reaction may include growth of E. coli, and / or the temperature may be constant and be about 37° C.

[0409] The pH of the solution or suspension can affect the activity of the enzyme. Control of the pH may ensure that the enzymatic reaction proceeds at an appropriate rate. The enzymatic reaction of the present disclosure may be carried out at a pH ranging from about 2 to about 10, from about 3 to about 8, or from about 4 to about 6.

[0410] The enzymatic reaction can be allowed to continue for a certain period of time before being quenched and the product isolated or otherwise collected. This period can be from about 1 minute to about 6 days, from about 0.5 days to about 5 days, or from about 16 hours to about 96 hours. The reaction can alternatively be allowed to proceed until no further catalysis occurs.

[0411] The enzymatic reaction can be allowed to continue until less than 75%, 70%, 65%, 55%, or 50% of the plant biomass containing polysaccharides is left undigested. This can be monitored or confirmed by a reducing end assay, such as anthrone assay, and / or by chromatographic methods, such as thin layer chromatography and high performance anion exchange chromatography. The reaction may be allowed to continue until all polysaccharides are converted to oligosaccharides.

[0412] There are many enzymes suitable for use in the enzymatic reaction of the method of the present invention. For example, "lytic polysaccharide monooxygenase" and "LPMO" are a class of enzymes that can oxidatively cleave polysaccharides using a copper-containing moiety and an oxygen source, such as a molecule of dioxygen, peroxide, or any other oxygen source, and a suitable reducing agent. Thus, when a LPMO is used, the enzymatic reaction may be carried out under aerobic conditions. Suitable reducing agents are not particularly limited, and examples include ascorbic acid, gallic acid, cysteine, NADH, NADPH, pyrogallol, dithiothreitol, cyanoborohydride, borohydride, photosynthetic pigments, lignin, lignols, and combinations of cellobiose and cellobiose dehydrogenase. A wide variety of photosynthetic pigments may be used. In some embodiments, thylakoids and purified fractions or chlorophyllin may be used, and light may be provided. The LPMO can be selected from the following families: AA9, AA10, AA11, AA13, AA14, and AA15. In various cases, the LPMO can be PaLPMO9E (SEQ ID NO: 1), an AA9 LPMO originally isolated from the ascomycete Podospora anserina, or the LPMO can be an AA9 LPMO from Trichoderma reesei (SEQ ID NO: 23).

[0413] Aerobic conditions may include the addition of oxygen, which may be provided by aeration of the substrate mixture with an oxygen-containing gas, such as air. Aeration may be performed by the introduction of oxygen-containing gas bubbles into the aqueous substrate mixture by various systems, such as an air injector, an aeration frit, a membrane system, or an internal loop airlift reactor. The concentration of molecular oxygen in the enzyme reaction may be from about 4 mg / L to about 14 mg / L.

[0414] Another type of enzyme that can be used in the method is a "cellulase" that has hydrolytic activity on cellulose, such as endo-1,4-beta-glucanase, cellobiohydrolase, and / or beta-glucosidase activity. Such enzymes can cleave glycosidic bonds in one or more forms of cellulose, including cellulose found in plant biomass. In doing so, they produce products that include glucose and cellooligosaccharides. In certain cases, the beta-glucanase may include enzymes from the GH5, GH7, and GH12 enzymes, such as those derived from Aspergillus niger (SEQ ID NOs: 12, 13, and 14) and Trichoderma reesei (SEQ ID NOs: 24 and 25).

[0415] Another type of enzyme is the "cellobiohydrolase" which has hydrolytic activity against cellulose and produces mainly cellobiose as a product. Cellobiose is a disaccharide and a cellooligosaccharide. Such enzymes can cleave glycosidic bonds in one or more forms of cellulose, including cellulose found in plant biomass. In various cases, the cellobiohydrolase may be from the GH6 and GH7 enzyme families, the Cel6A or Cel7A enzymes from Trichoderma reesei (SEQ ID NOs: 10 and 11, respectively).

[0416] Another type of enzyme is a "beta-glucosidase" that has hydrolytic activity on cellulose and produces primarily glucose as a product. Such enzymes can cleave the glycosidic bonds of one or more forms of cellulose, including cellulose found in plant biomass. In some embodiments, the beta-glucosidase may include a GH3 beta-glucosidase, such as that from Trichoderma reesei (SEQ ID NO: 22).

[0417] Another type of enzyme is lichenase, which may be selected from the: GH5, GH7, GH8, GH9, GH12, GH16, GH17, or GH26 family. In some embodiments, the lichenase may be a GH16 enzyme, such as the GH16 enzyme from Bacillus subtilis (SEQ ID NO: 2). The enzyme may act on mixed linkage glucans, for example, glucans that contain a mixture of β-1,3 and β-1,4 linkages, and cleave them at the β-1,4 glycosidic linkage. When lichenase acts on mixed linkage glucans, the β-glucans produced may be predominantly contained within a size range of about 3 to about 7 residues, and thus are particularly useful in foods, cosmetics, and nutritional supplements. Mixed linkage glucans are abundant in members of the grass and horsetail families, and thus grass-based biomass, such as straw, may have high levels of mixed linkage glucans and act usefully with lichenase.

[0418] Another type of enzyme is xylanase, which can act on plant biomass, for example, containing a xylan backbone. The xylanase can be, for example, glucuronoxylanase, arabinoxylanase, or glucuronoarabinoxylanase. The enzyme can be active on various polymers with a xylan backbone, such as glucuronoxylan, arabinoxylan, and glucuronoarabinoxylan. These polymers are abundant in various plant biomass, for example, both hardwoods and softwoods can contain suitable polysaccharides, with hardwoods often containing glucuronoxylan and softwoods often containing arabinoglucuronoxylan. In some cases, the xylanases may include GH5 xylanases from Ruminiclostridium thermocellum (SEQ ID NO:3) and Gonapodya prolifera (SEQ ID NO:4), as well as GH30 xylanases from Dickeya chrysanthemi (SEQ ID NO:5), Bacillus subtilis (SEQ ID NO:6), Bacteroides ovatus (SEQ ID NO:7), and Trichoderma reesei (SEQ ID NO:15).

[0419] Other enzymes useful in the present disclosure may include xyloglucanases and xyloglucan endoglucanases (XEGs), which are produced by numerous organisms, including plant pathogenic microorganisms. They can act on xyloglucan, a hemicellulose β-1,4 glucan chain abundant in the primary cell walls of higher plants, decorated with xylose, some of the xylose residues being further decorated with other residues, such as galactose. When the appropriate xyloglucanases or XEGs act on xyloglucan, the products include xyloglucan oligosaccharides with backbone lengths useful in the foodstuff, cosmetic, and nutraceutical industries. In some cases, the xyloglucanases may include GH5 xyloglucanase (SEQ ID NO: 8) from Bacteroides ovatus and GH74 xyloglucanase from Trichoderma reesei.

[0420] Any given natural plant biomass is likely to contain a mixture of various polysaccharides, so it is likely that a mixture of various enzymes may be beneficial in some cases. Such a mixture may also contain one or more of any other enzymes. For example, such a mixture may contain LPMO with endo-glucanase, xylanase with lichenase, cellobiohydrolase with mannase, or endo-glucanase with cellobiohydrolase, where the enzyme partners are present in a molar ratio of, for example, 1:100 to 100:1.

[0421] In certain cases, the one or more enzymes may be a cocktail of various enzymes, for example, a crude or semi-crude enzyme preparation. The term "crude enzyme preparation" as used herein generally refers to a soluble preparation extracted from microbial fermentation that has undergone minimal processing after extraction, for example, typically, the preparation may only undergo filtration to remove insoluble components. The term "semi-crude enzyme preparation" as used herein generally refers to a soluble preparation extracted from microbial fermentation that has undergone some processing after extraction, for example, the preparation may undergo filtration with increasing enzyme concentration to remove insoluble components and / or nanofiltration to remove small molecular weight compounds.

[0422] In certain cases, the crude or semi-crude enzyme preparation may be from bacteria or fungi. In some embodiments, the crude or semi-crude enzyme preparation may be from fungi, such as filamentous cellulolytic fungi, such as from Trichoderma or Aspergillus species. In certain embodiments, the enzyme may be a crude or semi-crude enzyme preparation from a Trichoderma reesei strain.

[0423] In step (e), one or more oligosaccharides formed in step (d) are removed, which may be done in several ways. They may be isolated based on solubility, so that a composition of only soluble sugars is extracted for further processing and / or isolated by chromatography to generate a composition with a narrower band of oligosaccharide chain lengths. Isolation may be based, for example, on precipitation, size exclusion chromatography, ion exchange chromatography, filtration, ultrafiltration, microfiltration, or nanofiltration. When solubility-based isolation is performed, the profile of sugars present in the isolated composition may depend on the initial enzymatic reaction, since the solubility of different sugars decreases with length at different rates.

[0424] Within the scope of the present disclosure, further processing of all or a portion of the removed oligosaccharide(s) to generate further products prior to combining with one or more dissolved polysaccharides to form the component is also contemplated. This further processing may include any chemical, physical, or enzymatic step, such as reduction, e.g. reductive amination, where appropriate; oxidation, caramelization, modification with Schiff base, or via Maillard reaction, or any combination of such steps, to provide various products with improved properties for the desired purpose. For example, caramelization characteristics, heating value, flavor, and color may be modified. The oligosaccharides may be purified, for example, through precipitation, size exclusion chromatography, ion exchange chromatography, filtration, ultrafiltration, microfiltration, or nanofiltration.

[0425] Also contemplated within the scope of the present disclosure is further processing of all or a portion of the dissolved soluble polysaccharide(s) to produce a product with improved properties prior to combining with the removed oligosaccharide(s) to form a component. This further processing may include any chemical, physical, or enzymatic step, such as alkylation or acid treatment. The polysaccharide may be purified, for example, through precipitation, size exclusion chromatography, ion exchange chromatography, filtration, ultrafiltration, microfiltration, or nanofiltration.

[0426] In some cases, after modification and / or purification of the oligosaccharides and polysaccharides, all or portions thereof are then combined as in step (f), which may be in a polysaccharide:oligosaccharide ratio of 1:100 to 1:1, 1:10 to 1:1, 1:90 to 1:2, 1:80 to 1:3, 1:70 to 1:4, or 1:60 to 1:5. The particular ratio may depend on the desired properties of the final component and the modifications and purifications that have been applied to the saccharides. In certain embodiments, it is not necessary to recombine all of the removed oligosaccharides and polysaccharides.

[0427] In step (f), the combination may be carried out in various ways, for example by mixing a solution containing all or a portion of the soluble polysaccharide(s) with a solution and / or suspension containing all or a portion of the removed oligosaccharide(s), which may be further condensed by spray drying, freeze drying or some other method. The soluble polysaccharides and the removed oligosaccharides may be combined by mixing a dry form containing all or a portion of the removed oligosaccharide(s) produced by spray drying, freeze drying or some other method of condensation after their removal in step (e), which dry form contains all or a portion of the soluble polysaccharide(s) produced by spray drying, freeze drying or some other method of condensation after their removal in step (c). Alternatively, (i) the soluble polysaccharide(s) or (ii) one of the removed oligosaccharides may be in a dry form, and the other may be in a solution when combined.

[0428] When the component is in a dry form, the method may further include the step (g) of mixing and dissolving the component in a liquid to form a liquid component. In various cases, the liquid may be an aqueous solution, such as water.

[0429] The component formed in step (f) and the liquid component formed in step (g) and of the second aspect of the present disclosure may contain different oligosaccharides in different amounts depending on the desired properties. In some cases, the ingredients and liquid ingredients may comprise at least 20% or at least 30% by dry weight of cello-oligosaccharides having a degree of polymerization of 2 to 6, the ingredients may comprise at least 20% or at least 30% by dry weight of xylo-oligosaccharides having a degree of polymerization of 2 to 12, the ingredients may comprise at least 20% or at least 30% by dry weight of mixed linkage glucan oligosaccharides having a degree of polymerization of 2 to 5, the ingredients may comprise at least 20% or at least 30% by dry weight of manno-oligosaccharides having a degree of polymerization of 2 to 12, the ingredients may comprise at least 20% or at least 30% by dry weight of xyloglucan oligosaccharides having a degree of polymerization of 4 to 12, and / or the ingredients may comprise at least 20% or at least 30% by dry weight of chitooligosaccharides having a degree of polymerization of 2 to 12. In some cases, the ingredients can contain up to 100% by dry weight of the oligosaccharides and polysaccharides described herein, and thus the above embodiments in which oligosaccharides are present at at least 20% by dry weight do not contain type 5 or 6 oligosaccharides.

[0430] The ingredients and liquid components may contain at least 50%, 60%, 70%, 80%, 90%, 95%, 99%, or 99.5% by dry weight of sugar present. The ingredients and liquid components may consist essentially of sugar. For example, the ingredients may have less than 0.5% by dry weight or less than 0.3% by dry weight, such as 0.1% by dry weight, of other substances.

[0431] In certain cases, the liquid ingredient may have a viscosity of 5 to 100,000 cps, 10 to 80,000 cps, 20 to 60,000 cps, 30 to 40,000 cps, 40 to 20,000 cps, or 50 to 10,000 cps. If the liquid ingredient is to be included in a beverage, a low syrup viscosity, such as about 20 to 300 cps, 50 to 200 cps, or 100 to 150 cps, may be desirable. Higher viscosity values ​​may be desirable in applications such as making chocolate, and thus if the liquid ingredient is desired to be in a syrup, it may have a viscosity in the range of about 8,000 to 100,000, about 10,000 to 50,000 cps, or about 15,000 to 25,000 cps. Viscosity values ​​are according to testing using a Brookfield HDB VE Roto Viscometer using standard test procedures, with 400 mL samples taken in tall beakers to ensure no container effects occur. The instrument is operated according to the manufacturer's instructions for range (Roto Viscometer using spindle code 61, spindle speed 100 rpm, and 22°C).

[0432] In some cases, the liquid component may have a flow rate of 100 to 350 seconds, 150 to 300 seconds, or 200 to 250 seconds. As used herein, "flow rate" generally refers to the volume of fluid passing per unit time. Flow rate values ​​specified herein are determined by timing the flow rate of 5 mL of liquid component from a vertically standing syringe (BD Plastipak 300613) filled with 20 mL of test liquid under gravity at room temperature, unless otherwise indicated.

[0433] In various cases, the liquid component may have a concentration of oligosaccharides from 1 to 200% w / v, 10 to 150% w / v, 20 to 140% w / v, 30 to 130% w / v, 40 to 120% w / v, 50 to 115% w / v, or 60 to 110% w / v.

[0434] In some cases, the liquid component may have a polysaccharide concentration of 0.1 to 50% w / v, 0.2 to 40% w / v, 0.3 to 30% w / v, 0.5 to 20% w / v, or 1 to 20% w / v.

[0435] In certain cases, the liquid component may have a total concentration of oligosaccharides and polysaccharides of 1 to 200% w / v, 10 to 160% w / v, 20 to 150% w / v, 30 to 140% w / v, 40 to 130% w / v, 50 to 120% w / v, or 60 to 110% w / v. In various cases, the higher the concentration of oligosaccharides and polysaccharides in the liquid, the thicker and more viscous the viscosity of the liquid may be. The liquid component may be a homogenous solution.

[0436] In another embodiment, the ingredients and liquid ingredients may contain at least two types of oligosaccharides. The amount of each of the oligosaccharides may vary depending on the desired properties of the resulting foodstuff, cosmetic, or nutritional supplement. The two types of oligosaccharides may be present in a ratio of 1:9 to 9:1 or 1:2 to 2:1. Furthermore, the ingredients and liquid ingredients may contain three types of oligosaccharides, they may contain four types of oligosaccharides, they may contain five types of oligosaccharides, or they may contain six types of oligosaccharides.

[0437] The ingredients and liquid components may include cellooligosaccharides, for example cellooligosaccharides in combination with xylooligosaccharides. Alternatively, the ingredients and liquid components may include cellooligosaccharides in combination with mannooligosaccharides.

[0438] The one or more soluble polysaccharides in the component may be particularly soluble in water or alkali.For example, the soluble polysaccharides used in the present disclosure can include hemicellulose, such as xylan, mannan, mixed linkage glucan, and certain cellulose derivatives, such as cellulose acetate, hydroxyethyl cellulose, and hydroxymethyl cellulose, and chitosan.In some embodiments, the one or more soluble polysaccharides can include hemicellulose.In certain embodiments, the hemicellulose can include xylan and / or mannan.

[0439] In some cases, the ingredients and liquid components can include at least 2% or at least 3% by dry weight of xylan. In various cases, the ingredients and liquid components can include at least 2% or at least 3% by dry weight of mannan. In certain cases, the ingredients and liquid components can include at least 2% or at least 3% by dry weight of cellulose derivatives. In some cases, the ingredients and liquid components can include at least 2% or at least 3% by dry weight of mixed-linked glucan. In various cases, the ingredients and liquid components can include at least 2% or at least 3% by dry weight of xyloglucan. In certain cases, the ingredients and liquid components can include at least 2% or at least 3% by dry weight of chitosan.

[0440] In certain cases, the ingredients and liquid components may comprise 2 to 40% by dry weight of one or more soluble polysaccharides, including polysaccharide derivatives, 3 to 30% by dry weight of one or more soluble polysaccharides, 5 to 25% by dry weight of one or more soluble polysaccharides, or 8 to 20% by dry weight of one or more soluble polysaccharides.

[0441] In some cases, the ingredients and liquid components have a polysaccharide:oligosaccharide ratio of 1:100 to 1:1, 1:10 to 1:1, 1:90 to 1:2, 1:80 to 1:3, 1:70 to 1:4, or 1:60 to 1:5.

[0442] The resulting ingredients and liquid ingredients may be useful in applications where oligosaccharides, sugars, bulk sweeteners, low intensity sweeteners, or other related food ingredients are conveniently used. For example, bulking agents as sweeteners, dietary fiber or humectants are added. The use of reduced cane sugar in food products is prominent. For example, it may be incorporated into cakes, breads, or other baked goods; chocolates or other confectioneries such as toffees, fudges, meringues, jams, jellies, or caramels; or beverages to impart desirable taste or color characteristics or increase dietary fiber content. Alternatively, the ingredients may be incorporated into animal feed, for example as an isolated ingredient or by utilizing the enzyme reaction mixture directly in the feed.

[0443] The compositions or ingredients described herein may be used to modify one or more properties of the finished product, including, but not limited to, sweetness, texture, mouthfeel, binding, gloss, smoothness, moistness, viscosity, color, hygroscopicity, flavor, bulk, moisture retention, caramelization, surface texture, crystallization, structural properties, low calorie, low glycemic index, low glycemic load, increased fiber, reduced sugar, and dissolution. These may be improvements over what is currently possible with various types of sugars, sugar substitutes, and / or other such compounds.

[0444] In the cosmetic industry, the ingredient can improve texture and water retention, act as UV absorbing molecules, maintain gel or cream structure, and / or act as a bulking agent.Furthermore, the ingredient and liquid ingredient can be useful in nutritional supplement compositions, since the dietary fiber provided has been shown to support digestive health, regulate gut flora well, and provide other healthy living benefits.In this context, the ingredient provided herein can function as an ingredient in probiotic beverages or other prebiotic or probiotic formulations.

[0445] The detailed description is further supplemented by reference to the following numbered embodiments: 1) A method for producing a foodstuff, cosmetic, or nutraceutical ingredient comprising one or more oligosaccharides and one or more soluble polysaccharides, comprising: (a) providing a plant biomass comprising one or more soluble polysaccharides and one or more insoluble polysaccharides; (b) treating the plant biomass to dissolve the one or more soluble polysaccharides; (c) removing a portion of the dissolved one or more soluble polysaccharides; (d) reacting the remaining plant biomass with one or more enzymes to form one or more oligosaccharides; (e) removing the one or more oligosaccharides; and (f) combining a portion of the dissolved one or more soluble polysaccharides from step (c) with the one or more oligosaccharides from step (e) to form the ingredient. 2) A method according to the numbered embodiments, wherein the treatment in step (b) is a thermochemical treatment. 3) The method according to numbered embodiment 2, wherein the thermochemical treatment is a hot water treatment or a high temperature alkaline treatment. 4) The method according to numbered embodiment 3, wherein the alkaline treatment uses an alkali having a pH of 10 to 14. 5) The method according to any of numbered embodiments 3 or 4, wherein the alkaline treatment uses sodium hydroxide, potassium hydroxide, sodium carbonate, calcium carbonate, calcium hydroxide, ammonium sulfate, ammonium hydroxide, and aqueous ammonia. 6) The method according to any preceding numbered embodiment, wherein the treatment in step (b) occurs at a temperature of 30 to 180° C. 7) The method according to any preceding numbered embodiment, wherein the treatment in step (b) occurs for a period of 10 minutes to 24 hours. 8) The method of any preceding numbered embodiment, wherein after removing the one or more oligosaccharides, the one or more oligosaccharides and / or the dissolved one or more soluble polysaccharides are subjected to a chemical, physical, or enzymatic treatment, such as reduction, oxidation, caramelization, or Maillard reaction. 9) The method of any preceding numbered embodiment, wherein the dissolved soluble polysaccharide(s) and / or oligosaccharide(s) are dried before being combined together in step (f).10) The method according to numbered embodiment 9, wherein the method further comprises: (g) mixing and dissolving the components in a liquid to form a liquid component, the liquid component having a viscosity of 5 to 100,000 cps. 11) The method according to numbered embodiment 10, wherein the concentration of oligosaccharides and polysaccharides in the liquid component is from 1 to 200% w / v. 12) The method according to any preceding numbered embodiment, comprising one or more soluble polysaccharides: at least one selected from the group consisting of mannan, mixed linkage glucan, lignocellulose, hemicellulose, certain cellulose derivatives such as cellulose acetate, hydroxyethylcellulose, and hydroxymethylcellulose, and chitosan. 13) The method according to numbered embodiment 12, wherein the hemicellulose comprises xylan and / or mannan. 14) The method according to any preceding numbered embodiment, wherein the plant biomass comprises sugarcane biomass, corn biomass, wheat biomass, hardwood, or softwood. 15) A foodstuff, cosmetic or nutraceutical ingredient obtainable by the method of any preceding numbered embodiment. 16) A foodstuff, cosmetic or nutraceutical ingredient obtainable by the method of any preceding numbered embodiment. 17) A foodstuff, cosmetic or nutraceutical ingredient obtainable by the method of any preceding numbered embodiment. 18) A foodstuff, cosmetic or nutraceutical ingredient obtainable by the method of any preceding numbered embodiment. 19) A foodstuff, cosmetic or nutraceutical ingredient obtainable by the method of any preceding numbered embodiment. 20) A foodstuff, cosmetic or nutraceutical ingredient obtainable by the method of any preceding numbered embodiment. 21) A foodstuff, cosmetic or nutraceutical ingredient obtainable by the method of any preceding numbered embodiment. 22) A foodstuff, cosmetic or nutraceutical ingredient obtainable by the method of any preceding numbered embodiment. 17) The liquid ingredient of numbered embodiment 16, wherein the liquid ingredient comprises at least two of the oligosaccharides listed in (i) through (vi).18) The liquid component of any of numbered embodiments 16 or numbered embodiment 17, wherein the liquid component comprises at least 20% by dry weight of cellooligosaccharides having a degree of polymerization of 2 to 6. 19) The liquid component of any of numbered embodiments 16 to 18, wherein the liquid component comprises at least 20% by dry weight of xylooligosaccharides having a degree of polymerization of 2 to 12. 20) The liquid component of any of numbered embodiments 16 to 19, wherein the liquid component comprises at least 20% by dry weight of mannooligosaccharides having a degree of polymerization of 2 to 12. 21) The liquid component of any of numbered embodiments 16 to 20, wherein the liquid component comprises at least 2% by dry weight of xylan. 22) The liquid component of any of numbered embodiments 16 to 21, wherein the liquid component comprises at least 2% by dry weight of mannan. 23) The liquid component of any of numbered embodiments 16 to 22, wherein the liquid component comprises at least 2% by dry weight of a cellulose derivative. 24) The liquid component of any one of numbered embodiments 16 to 23, wherein the liquid component has a polysaccharide concentration of 0.1 to 50% (w / v). 25) The liquid component of any one of numbered embodiments 16 to 24, wherein the liquid component has an oligosaccharide concentration of 1 to 200% (w / v). 26) The liquid component of any one of numbered embodiments 16 to 25, wherein the liquid component has a polysaccharide and oligosaccharide concentration of 1 to 200% (w / v). 27) The liquid component of any one of numbered embodiments 16 to 26, wherein the liquid component comprises polysaccharide and oligosaccharide in an amount of 1:100 to 1:1 ratio. 28) The liquid component of any one of numbered embodiments 16 to 27, wherein the liquid component comprises two oligosaccharides relative to each other in a ratio of 1:9 to 9:1. 29) Use of a liquid ingredient of any of numbered embodiments 16 to 28 in a foodstuff, cosmetic, or nutritional supplement product. Additional Exemplary Embodiments in Which Biomass is Pretreated to Remove Monosaccharides and / or Disaccharides

[0446] In some cases, the present disclosure relates to novel methods of physically and thermochemically processing plant biomass materials to produce foodstuff, cosmetic, or nutritional supplement ingredients.

[0447] Sugary foods and beverages are an important part of cultures and lifestyles around the world, but the sugar they contain has been linked to obesity, diabetes, poor dental health, and destructive behaviors in people. As a result, consumer preferences are shifting away from sugar-containing foods, and governments are increasingly enacting regulations to encourage less sugar consumption.

[0448] Therefore, the industry has been searching for suitable low-calorie sweeteners for decades to replace sugar in food and beverages.Unfortunately, many sugar substitutes are produced from non-natural sources, and often bring along their sweetness with a potential bitterness or other unpleasant taste, neither of which is appealing to consumers.In addition, although many sweeteners can mimic the sweetness of sugar in food and beverages, few can comprehensively mimic the role that sugar plays in food, such as adding bulk, adjusting texture, providing structure, acting as a preservative, and adjusting color and flavor through caramelization and Maillard reaction.

[0449] Dietary fiber is an important part of a positive diet and helps maintain digestive health and a well-regulated intestinal flora. Such fiber includes sugars of various chain lengths and types. In addition to being found naturally in a wide range of foods, fiber can also be produced separately or added to other foods during their manufacture.

[0450] Biomass is a good source of sugars that can be used to replace sugar and add fiber to food. Compositions made from feedstocks have been provided. However, there is still a need to optimize the process by which these sugars are obtained from biomass on a large and commercial scale and processed into compositions useful as foodstuffs, cosmetics, or nutritional supplement ingredients. Enzymatic destruction of large amounts of plant biomass can take a significant amount of time. Furthermore, sugars generally need to be produced by controlled destruction. Enzymatic destruction can be desirable because it allows for exquisite control of product size, ensuring that no and / or little monosaccharides are obtained.

[0451] The methods provided herein can economically and efficiently enable the production of foodstuff, cosmetic, or nutraceutical ingredients from plant biomass starting materials faster than previously used methods, resulting in a purer end product and can be used on a large and commercial scale. The methods can do so by performing a pre-washing step to remove endogenous monosaccharides and / or disaccharides from the biomass and using a thermochemical pretreatment step to ensure the controlled destruction and release of sugars that may be required for the production of the ingredient. Together, these steps can ensure that no and / or no monosaccharides are introduced during pre-enzymatic processing. This can maximize the efficiency and limit the amount of post-reaction purification required.

[0452] Thus, in another aspect of the present disclosure, there is provided a method for producing a foodstuff, cosmetic, or nutraceutical ingredient comprising one or more oligosaccharides, the method comprising: a) a step of physical treatment of plant biomass containing mono- and / or disaccharides; b) a washing cycle (also referred to herein as an incubation cycle) step comprising: (i) washing (e.g., incubating) the plant biomass to solubilize at least a portion of the monosaccharides and / or disaccharides, and (ii) removing at least a portion of the monosaccharides and / or polysaccharides; c) a step of thermochemical pretreatment of plant biomass; d) forming one or more oligosaccharides by an enzymatic reaction, the enzymatic reaction comprising contacting the plant biomass in solution or suspension with one or more polysaccharide cleaving enzymes; e) separating (also referred to herein as enriching or isolating) one or more oligosaccharides from the enzymatic reaction mixture and forming a component using the one or more oligosaccharides. A method is provided that includes:

[0453] Preparation of foodstuff, cosmetic, or nutraceutical ingredients in the manner provided herein can allow for efficient use of biomass by: incorporating, purifying, derivatizing, or otherwise modifying oligomeric and polymeric materials from the same biomass source, and controlling the proportions of oligomers and polymers that can improve the functional properties, nutritional properties, and tolerance of the ingredients.

[0454] Steps (a), (b), and (c) of the disclosed method are all "pretreatment" steps performed on the plant biomass starting material. As used herein, "pretreatment" generally refers to what is done to the plant biomass before the polysaccharide-cleaving enzyme comes into contact with the plant biomass.

[0455] Step (a) is a physical treatment of the plant biomass that may have the purpose of physically breaking down the plant biomass in preparation for subsequent steps. The physical step may help speed up the overall process by increasing the available surface area of ​​the plant biomass, for example, allowing chemicals used in subsequent steps to be active on more of the plant biomass at once. The physical pretreatment step may include chipping, chopping, milling, ball milling, grinding, shaping, blending, or combinations thereof, of the plant biomass.

[0456] Any material that contains suitable polysaccharides may be plant biomass. The foodstuff, cosmetic, and nutraceutical industries use a wide variety of oligosaccharides, and the suitable polysaccharides in the method are not particularly limited. Suitable plant biomass for generating the oligosaccharide profile of the present disclosure may include, for example, cellulose, lignocellulose, chitin, chitosan, xylan (such as glucuronoxylan, arabinoxylan, and glucuronoarabinoxylan), xyloglucan, and mixed-linkage glucan, and / or mannan (such as glucomannan, galactomannan, or galactoglucomannan). However, any plant biomass that can be appropriately worked with is contemplated.

[0457] Thus, the plant biomass may be grain, grain husk, bean husk, seed coat, and / or other seed material; seaweed; corn stover, straw, bagasse, miscanthus, sorghum residue, switchgrass, bamboo, and / or other monocotyledonous tissue; water hyacinth, leaf tissue, roots, and / or other vegetable matter; and / or any combination of suitable plant biomass. In some embodiments, the plant biomass may include sugarcane, corn stover, corn cob, bran, wheat straw, hardwood or softwood. In certain embodiments, the plant biomass may include corn cob.

[0458] Step (b) is a washing cycle (or incubation cycle) pretreatment of the plant biomass that may occur after a physical pretreatment step. The purpose of step (b) may be to solubilize and remove monosaccharides and / or disaccharides from the biomass. For example, the monosaccharides and / or disaccharides may include, but are not limited to, free sucrose, maltose, lactose, glucose, fructose, or galactose. Removal of free disaccharides such as sucrose may be of interest since disaccharides cannot be easily removed subsequently from the oligosaccharide fraction, for example filtration methods can be used, but may generally cause an equal loss of other disaccharides.

[0459] Step (i) of the wash cycle may occur at a temperature range of, for example, 5 to 150° C., 10 to 100° C., or 15 to 50° C. In some cases, the wash cycle can occur at room temperature, for example, at about 15 to 25° C. or about 20 to 22° C. Higher temperatures may allow for more rapid solubilization of monosaccharides and / or disaccharides, but too high a temperature may be more difficult to achieve in an efficient and cost-effective manner and may damage biomass compounds or solubilize compounds that are not desired to be solubilized during this step.

[0460] Step (i) of the wash cycle may occur over a range of time scales, e.g., larger amounts of biomass may be exposed to this step for longer periods and can be adjusted accordingly. For example, time scales may be from 0.5 minutes to 72 hours, 1 minute to 12 hours, 5 minutes to 24 hours, or 10 minutes to 3 hours. In certain embodiments, this step may occur as a batch or continuous.

[0461] In some embodiments, step (i) of the wash cycle may comprise washing the plant biomass at room temperature in water, i.e. in water provided at a neutral pH of about pH 7. In another aspect, step (i) of the wash cycle may comprise heating the plant biomass in water, i.e. in water provided at a neutral pH of about pH 7. During the wash cycle, the neutral water provided may become slightly acidic as mono- and / or disaccharides solubilize.

[0462] In certain embodiments, step (i) of the wash cycle may include heating the plant biomass in an alkaline solution having a pH of 7.1 to 14, 7.5 to 12, or 8 to 11. The solution may include or be suitable from any one of the alkalis selected from: sodium hydroxide, potassium hydroxide, sodium carbonate, calcium carbonate, calcium hydroxide, ammonium sulfate, ammonium hydroxide, and aqueous ammonia. In various embodiments, the alkali may be sodium hydroxide. Combinations of the listed alkalis are also contemplated.

[0463] In various cases, step (i) of the wash cycle may include heating the plant biomass in an acidic solution having a pH of 1 to 6.9, 2 to 6.5, or 4 to 6. The solution may include or be suitable from any organic or inorganic acid, such as one of the acids selected from: sulfuric acid, hydrochloric acid, nitric acid, phosphoric acid, acetic acid, maleic acid, fumaric acid, and oxalic acid. In certain cases, the acid may be sulfuric acid. Combinations of the listed acids are also contemplated.

[0464] In certain cases, some or all of the monosaccharides and / or disaccharides in the plant biomass are solubilized and removed during step (b). Other contaminants, such as other soluble sugars and minerals, may also be removed during this step.

[0465] Step (b) may be repeated to remove monosaccharides and / or disaccharides that were not removed from the plant biomass during the washing cycle. In various cases, step (b) may be performed at least two times, at least three times, at least four times, or at least five times. Step (c) is a thermochemical pretreatment of the plant biomass that may occur after the physical and washing pretreatment steps. As used herein, "thermochemical" generally refers to heating the plant biomass in a chemical above room temperature (room temperature being, for example, about 15 to 25°C or about 20 to 22°C), such as heating in a solution of water, acid, or alkali. The purpose of the thermochemical step may be to help speed up the overall process, as it may chemically modify the chemical components of the plant biomass, for example by breaking hydrogen bonds between sugars, making them more easily broken down by enzymes in a subsequent step.

[0466] Heating may be to a range of temperatures, for example, 50 to 150° C., 60 to 130° C., 65 to 120° C., or 70 to 110° C. Higher temperatures can allow for more rapid chemical and / or physical modifications, but too high a temperature can be more difficult to achieve in an efficient and cost-effective manner.

[0467] Heating may occur over a range of time scales, and particularly large amounts of biomass may be exposed to heating for longer periods of time and can be adjusted accordingly, for example, heating of plant biomass can be for 5 minutes to 72 hours, 15 minutes to 24 hours, 30 minutes to 12 hours, or 1 hour to 4 hours.

[0468] In some embodiments, the thermochemical pretreatment may involve heating the plant biomass in water, i.e., at a neutral pH of about pH 7.

[0469] In certain embodiments, the thermochemical treatment may include heating the plant biomass in an alkaline solution having a pH of 7.1 to 14, 9 to 13, or 10 to 13. The solution may include or be suitable from any one of the alkalis selected from: sodium hydroxide, potassium hydroxide, sodium carbonate, calcium carbonate, calcium hydroxide, ammonium sulfate, ammonium hydroxide, and aqueous ammonia. In various embodiments, the alkali may be sodium hydroxide. Combinations of the listed alkalis are also contemplated.

[0470] In some cases, the thermochemical treatment may include heating the plant biomass in an acidic solution having a pH of 1 to 6.9, 2 to 6.5, or 4 to 6. The solution may include or be suitable from any organic or inorganic acid, such as one of the acids selected from: sulfuric acid, hydrochloric acid, nitric acid, phosphoric acid, acetic acid, maleic acid, fumaric acid, and oxalic acid. In certain cases, the acid may be sulfuric acid. Combinations of the listed acids are also contemplated.

[0471] A number of different sequential thermochemical treatment steps are also contemplated for step (c), for example step (c) may be carried out at least two times, at least three times, at least four times, or at least five times.

[0472] In certain cases, the washing in step (b) may be carried out in water and the pretreatment in step (c) may be carried out in alkali.

[0473] After the pretreatment step, step (d) may include an enzymatic reaction to form one or more oligosaccharides from the plant biomass. The polysaccharides present in the plant biomass may be partially cleaved by the enzyme into oligosaccharides (e.g., useful oligosaccharides) or may remain partially cleaved or uncleaved polysaccharides, which may include cellulose, xylan (such as glucuronoxylan, arabinoxylan, or glucuronoarabinoxylan), mannan (such as glucomannan, galactomannan, or galactoglucomannan), mixed linkage glucan, xyloglucan chitin, chitosan, or lignocellulose.

[0474] The reaction may occur in solution and / or suspension. The reaction may occur in a suitable reaction vessel. In some cases, the reaction may occur at a temperature or temperature protocol appropriate for the particular combination of enzyme and plant biomass, and the reaction may proceed for a certain length of time until the product reaches a desired concentration or until some other requirement is met and the product is isolated or otherwise collected. This period may be from about 1 minute to about 6 days, from about 0.5 days to about 5 days, or from about 16 hours to about 96 hours. The reaction may alternatively proceed until no further catalysis occurs.

[0475] To ensure optimal contact between the enzymes and the plant biomass, the reaction mixture may be stirred constantly or at intervals. Agitation may take the form of rhythmic movement of the entire reaction vessel, a fan or other agitation device, sparging of air bubbles, or any other method of agitation.

[0476] The enzymatic reaction may be a microbial fermentation. The temperature and reaction time may be suitable for the growth of the microbial organism used. The microbial organism may be genetically modified to produce enzymes suitable for the production of the oligosaccharides of the present disclosure. The microorganism may be, for example, a bacterium, such as Escherichia coli, or a fungus, such as Saccharomyces cerevisiae or Trichoderma reesei.

[0477] The present disclosure further embodies an expression vector suitable for modifying a subject microorganism to produce an enzyme or mixture of enzymes of the present disclosure. If desired, the expression vector, which may be a plasmid or any other nucleic acid capable of inducing the production of the enzyme, may contain one or more of the following regulatory sequences to control the expression of the exogenous enzyme: a heat shock gene regulatory sequence, a virulence gene regulatory sequence, and a sporulation gene regulatory sequence.

[0478] The enzymatic reaction can be carried out at a temperature or temperature protocol appropriate for the enzyme and substrate used. For example, it may be carried out at a constant temperature ranging from about 10° C. to about 100° C., from about 20° C. to about 70° C., or from about 30° C. to about 40° C. If the enzymatic reaction takes the form of microbial fermentation, the temperature may be appropriate, for example, such that the enzymatic reaction may include growth of E. coli and / or the temperature may be constant and be about 37° C.

[0479] The pH of the solution or suspension can affect the activity of the enzyme. Control of the pH may ensure that the enzymatic reaction proceeds at an appropriate rate. The enzymatic reaction of the present disclosure may occur at a pH ranging from about 2 to about 10, from about 3 to about 8, or from about 4 to about 6.

[0480] The enzymatic reaction may be allowed to continue to run until 5-75%, 5-70%, 5-65%, 5-55%, or more, or 10-50% of the plant biomass containing polysaccharides remains undigested, which can be monitored or confirmed by a reducing end assay, such as the anthrone assay, and / or by chromatographic methods, such as thin layer chromatography and high performance anion exchange chromatography.

[0481] There are many enzymes that may be suitable for use in the enzymatic reaction of the method of the present invention. For example, "lytic polysaccharide monooxygenases" and "LPMOs" are a class of enzymes that can oxidatively cleave polysaccharides using a copper-containing moiety and an oxygen source, such as molecules of dioxygen, peroxide, or any other oxygen source; and a suitable reducing agent. Thus, when LPMOs are used, the enzymatic reaction can be carried out under aerobic conditions. Suitable reducing agents are not particularly limited, and examples include ascorbic acid, gallic acid, cysteine, NADH, NADPH, pyrogallol, dithiothreitol, cyanoborohydride, borohydride, photosynthetic pigments, lignin, lignols, and combinations of cellobiose and cellobiose dehydrogenase. A wide variety of photosynthetic pigments, such as thylakoids and purified fractions or chlorophyllins, may be used, and light may be provided. LPMOs can be selected from the following families: AA9, AA10, AA11, AA13, AA14, and AA15. The LPMO may be PaLPMO9E (SEQ ID NO: 1), an AA9 LPMO originally isolated from the ascomycete Podospora anserina. The LPMO may be an AA9 LPMO from Trichoderma reesei (SEQ ID NO: 23).

[0482] Aerobic conditions may include the addition of oxygen, which may be provided by aeration of the substrate mixture with an oxygen-containing gas, such as air. Aeration may be performed by the introduction of oxygen-containing gas bubbles into the aqueous substrate mixture by various systems, such as an air injector, an aeration frit, a membrane system, or an internal loop airlift reactor. The concentration of molecular oxygen in the enzyme reaction may be from about 4 mg / L to about 14 mg / L.

[0483] Another type of enzyme that can be used in the method is a "cellulase" that has hydrolytic activity on cellulose, such as endo-1,4-beta-glucanase, cellobiohydrolase, and / or beta-glucosidase activity. Such enzymes can cleave glycosidic bonds in one or more forms of cellulose, including cellulose found in plant biomass. In doing so, they produce products that include glucose and cellooligosaccharides. Beta-glucanases include enzymes from the GH5, GH7, and GH12 enzymes, such as those derived from Aspergillus niger (SEQ ID NOs: 12, 13, and 14) and Trichoderma reesei (SEQ ID NOs: 24 and 25).

[0484] Another type of enzyme is the "cellobiohydrolase" which has hydrolytic activity against cellulose and produces mainly cellobiose as a product. Cellobiose is a disaccharide and a cellooligosaccharide. Such enzymes can cleave glycosidic bonds in one or more forms of cellulose, including cellulose found in plant biomass. The cellobiohydrolase may be from the GH6 and GH7 enzyme families, the Cel6A or Cel7A enzymes from Trichoderma reesei (SEQ ID NOs: 10 and 11, respectively).

[0485] Another type of enzyme is a "beta-glucosidase" that has hydrolytic activity on cellulose and produces primarily glucose as a product. Such enzymes can cleave the glycosidic bonds of one or more forms of cellulose, including the cellulose found in plant biomass. Beta-glucosidases can include GH3 beta-glucosidases, such as those from Trichoderma reesei (SEQ ID NO: 22).

[0486] Another type of enzyme is lichenase, which may be selected from the: GH5, GH7, GH8, GH9, GH12, GH16, GH17, or GH26 family. In some cases, the lichenase may be a GH16 enzyme. The GH16 enzyme may be derived from Bacillus subtilis (SEQ ID NO: 2). The enzyme may act on mixed linkage glucans, which are glucans that contain a mixture of β-1,3 and β-1,4 linkages, and may cleave them at the β-1,4 glycosidic linkage. When lichenase acts on mixed linkage glucans, the β-glucans produced may be predominantly contained within a size range of about 3 to about 7 residues, and thus may be particularly useful in the food, cosmetic, and nutraceutical industries. Mixed linkage glucans are abundant in members of the grass and horsetail families, and thus grass-based biomass, such as straw, has high levels of mixed linkage glucans and may usually act usefully with lichenase.

[0487] Another type of enzyme is xylanase, which can act on plant biomass containing xylan backbones. Xylanases can be, for example, glucuronoxylanase, arabinoxylanase, or glucuronoarabinoxylanase. The enzymes can be active on various polymers with xylan backbones, such as glucuronoxylan, arabinoxylan, and glucuronoarabinoxylan. These polymers are abundant in various plant biomass, for example, both broadleaf and softwoods can contain suitable polysaccharides, with broadleaf often containing glucuronoxylan and softwood often containing arabinoglucuronoxylan. In some embodiments, xylanases may include GH5 xylanases from Ruminiclostridium thermocellum (SEQ ID NO:3) and Gonapodya prolifera (SEQ ID NO:4), as well as GH30 xylanases from Dickeya chrysanthemi (SEQ ID NO:5), Bacillus subtilis (SEQ ID NO:6), Bacteroides ovatus (SEQ ID NO:7), and Trichoderma reesei (SEQ ID NO:15).

[0488] Other enzymes useful in the present disclosure may include xyloglucanases and xyloglucan endoglucanases (XEGs), which are produced by numerous organisms, including plant pathogenic microorganisms. They can act on xyloglucan, a hemicellulose β-1,4 glucan chain that is abundant in the primary cell walls of higher plants and modified with xylose, some of the xylose residues being further modified with other residues, such as galactose. When the appropriate xyloglucanases or XEGs act on xyloglucan, the products include xyloglucan oligosaccharides with a backbone length that can be useful in the foodstuff, cosmetic, and nutraceutical industries. Xyloglucanases can include GH5 xyloglucanase (SEQ ID NO: 8) from Bacteroides ovatus and GH74 xyloglucanase from Trichoderma reesei.

[0489] Any given natural plant biomass is likely to contain a mixture of various polysaccharides, so it is likely that a mixture of various enzymes may be beneficial in some cases. Such a mixture may also contain one or more of any other enzymes. For example, such a mixture may contain LPMO with endo-glucanase, xylanase with lichenase, cellobiohydrolase with mannase, or endo-glucanase with cellobiohydrolase, with the enzyme partners being present in a molar ratio of 1:100 to 100:1.

[0490] In some cases, the enzyme or enzymes may be a cocktail of various enzymes, for example, a crude or semi-crude enzyme preparation. The term "crude enzyme preparation" as used herein generally refers to a soluble preparation extracted from microbial fermentation that has undergone minimal processing after extraction. For example, typically, the preparation may only undergo filtration to remove insoluble components. The term "semi-crude enzyme preparation" as used herein generally refers to a soluble preparation extracted from microbial fermentation that has undergone some processing after extraction, for example, the preparation may undergo filtration with increasing enzyme concentration to remove insoluble components and / or nanofiltration to remove small molecular weight compounds.

[0491] In some cases, the crude or semi-crude enzyme preparation may be from a bacterium or a fungus. For example, the preparation may be from a fungus, such as a filamentous cellulolytic fungus from a Trichoderma or Aspergillus species. The enzyme may be a crude or semi-crude enzyme preparation from a Trichoderma reesei strain.

[0492] In step (e), the oligosaccharides may be separated from the enzymatic reaction mixture in several ways. They may be isolated based on solubility, so that a soluble sugar-only composition is extracted for further processing and / or isolated by chromatography to generate a composition with a narrower band of oligosaccharide chain lengths. Isolation may be based, for example, on precipitation, size-exclusion chromatography, ion-exchange chromatography, filtration, ultrafiltration, microfiltration, or nanofiltration. When solubility-based isolation is performed, the profile of sugars present in the isolated composition may depend on the initial enzymatic reaction, since the solubility of different sugars decreases with length at different rates.

[0493] Within the scope of the present disclosure, further processing of all or part of the generated oligosaccharides is also contemplated to generate further products before incorporation into foodstuffs, cosmetics, or nutritional supplements. This further processing may include any chemical, physical, or enzymatic step, such as reduction, e.g. reductive amination, where appropriate; oxidation, caramelization, modification with Schiff base, or via Maillard reaction, or any combination of such steps, to provide various products with improved properties for the desired purpose. For example, caramelization characteristics, heating value, flavor, and color may be modified. The oligosaccharides may be purified, for example, through precipitation, size exclusion chromatography, ion exchange chromatography, filtration, ultrafiltration, microfiltration, or nanofiltration.

[0494] The component formed in step (e) may contain various oligosaccharides in various amounts depending on the desired properties. In various cases, the component may contain at least 20% or at least 30% by dry weight cello-oligosaccharides having a degree of polymerization of 2 to 6, the component may contain at least 20% or at least 30% by dry weight xylo-oligosaccharides having a degree of polymerization of 2 to 12, the component may contain at least 20% or at least 30% by dry weight mixed linkage glucan oligosaccharides having a degree of polymerization of 2 to 5, the component may contain at least 20% or at least 30% by dry weight manno-oligosaccharides having a degree of polymerization of 2 to 12, the component may contain at least 20% or at least 30% by dry weight xyloglucan oligosaccharides having a degree of polymerization of 4 to 12, and / or the component may contain at least 20% or at least 30% by dry weight chitooligosaccharides having a degree of polymerization of 2 to 12. In some embodiments, the ingredients can contain up to 100% by dry weight of the oligosaccharides and polysaccharides described herein, and thus the above embodiments in which oligosaccharides are present at at least 20% by dry weight do not contain all six types of oligosaccharides.

[0495] In some cases, the ingredients may contain at least 50%, 60%, 70%, 80%, 90%, 95%, 99%, or 99.5% by dry weight of sugars present. The ingredients may consist essentially of sugars. For example, the ingredients may have less than 0.5% by dry weight or less than 0.3% by dry weight, e.g., 0.1% by dry weight, of other substances.

[0496] In various cases, the composition may contain at least two types of oligosaccharides. The amount of each of the oligosaccharides may vary depending on the desired properties of the resulting foodstuff, cosmetic, or nutritional supplement. The two types of oligosaccharides may be present in a ratio of 1:9 to 9:1 or 1:2 to 2:1. Additionally, the composition may contain three types of oligosaccharides, four types of oligosaccharides, five types of oligosaccharides, or six types of oligosaccharides.

[0497] In some embodiments, the ingredients may include cellooligosaccharides, for example cellooligosaccharides in combination with xylooligosaccharides. In certain embodiments, the ingredients may include cellooligosaccharides in combination with mannooligosaccharides.

[0498] The resulting ingredients can be useful in applications where oligosaccharides, sugars, bulk sweeteners, low intensity sweeteners, or other related food ingredients are advantageously used. For example, bulking agents as sweeteners, dietary fiber or humectants are added. The use of reduced cane sugar in foods can be prominent. The ingredients may be incorporated into cakes, breads, or other baked goods; into chocolates or other confectioneries, such as toffees, fudges, meringues, jams, jellies, or caramels; or into beverages, for example to impart desirable taste or color characteristics or increase dietary fiber content. In some cases, the ingredients may be incorporated into animal feed, for example, as an isolated ingredient or by utilizing the enzyme reaction mixture directly in the feed.

[0499] The compositions or ingredients described herein may be used to modify one or more properties of the finished product, including, but not limited to, sweetness, texture, mouthfeel, binding, gloss, smoothness, moistness, viscosity, color, hygroscopicity, flavor, bulk, water retention, caramelization, surface texture, crystallization, structural properties, reduced calories, low glycemic index, low glycemic load, increased fiber, reduced sugar, and dissolution. These may be improvements over what is currently possible with various types of sugars, sugar substitutes, and / or other such compounds.

[0500] In the cosmetic industry, the ingredient can improve texture and moisturization, act as UV absorbing molecules, maintain gel or cream structure, and / or serve as a bulking agent.Furthermore, the ingredient can be useful in nutritional supplement compositions, since the dietary fiber provided has been shown to support digestive health, regulate gut flora well, and provide other healthy living benefits.In this context, the ingredient provided herein can function as an ingredient in probiotic beverages or other prebiotic or probiotic formulations.

[0501] The detailed description is further supplemented by reference to the following numbered embodiments: 1) A method for producing a foodstuff, cosmetic, or nutraceutical ingredient comprising one or more oligosaccharides, comprising: a) a step of physical treatment of a plant biomass comprising monosaccharides and / or disaccharides; b) a step of a washing cycle comprising (i) washing the plant biomass to solubilize at least a portion of the monosaccharides and / or disaccharides and (ii) removing at least a portion of the monosaccharides and / or disaccharides; c) a step of thermochemical pretreatment of the plant biomass; d) a step of forming one or more oligosaccharides by an enzymatic reaction comprising contacting one or more polysaccharide-cleaving enzymes with the plant biomass in a solution or suspension; e) a step of separating the one or more oligosaccharides from the enzymatic reaction mixture and forming an ingredient using the one or more oligosaccharides. 2) The method of numbered embodiment 1, wherein the physical pretreatment step comprises chipping, chopping, milling, ball milling, grinding, shaping, or blending the plant biomass. 3) The method of any of numbered embodiment 1 or numbered embodiment 2, wherein step (i) of the wash cycle occurs in water, acid, or alkali. 4) The method of any preceding numbered embodiment, wherein step (i) of the wash cycle occurs at a temperature of 5 to 150°C, 10 to 100°C, or 15 to 50°C. 5) The method of any preceding numbered embodiment, wherein step (i) of the wash cycle occurs on a time scale of 0.5 minutes to 72 hours, 1 minute to 12 hours, 5 minutes to 24 hours, or 10 minutes to 3 hours. 6) The method of any preceding numbered embodiment, wherein the thermochemical pretreatment comprises heating the plant biomass in water, acid, or alkali solution. 7) The method of numbered embodiment 6, wherein the heating of the plant biomass is at a temperature of 50 to 150°C, 60 to 130°C, 65 to 120°C, or 70 to 110°C. 8) The method of any of numbered embodiments 6 or 7, wherein heating the plant biomass is for 5 minutes to 72 hours, 15 minutes to 24 hours, 30 minutes to 12 hours, or 1 hour to 4 hours. 9) The method of any one of numbered embodiments 6 to 8, wherein the solution has a pH of 7.1 to 14, 7.5 to 12, or 8 to 11.10) The method of numbered embodiment 9, wherein the solution comprises sodium hydroxide, potassium hydroxide, sodium carbonate, calcium carbonate, aqueous ammonia, ammonium sulfate, or ammonium hydroxide. 11) The method of any one of numbered embodiments 6 to 8, wherein the solution has a pH of 1 to 6.9, 2 to 6.5, or 4 to 6. 12) The method of numbered embodiment 11, wherein the solution comprises sulfuric acid, hydrochloric acid, nitric acid, phosphoric acid, acetic acid, maleic acid, fumaric acid, or oxalic acid. 13) The method of any one of numbered embodiments 1 to 12, wherein the plant biomass is sugarcane, corn stover, corn cob, bran, wheat straw, hardwood or softwood. 14) The method of any one of numbered embodiments 1 to 13, wherein the plant biomass comprises cellulose, chitin, chitosan, xylan, xyloglucan, mixed linkage glucan, mannan, or lignocellulose. 15) The method of any one of numbered embodiments 1 to 14, wherein one or more of the polysaccharide cleaving enzymes is a cellulase, xylanase, xyloglucanase, endo-glucanase, cellobiohydrolase, mannase, lichenase, or lytic polysaccharide monooxygenase (LPMO), such as one selected from the group consisting of: AA9, AA10, AA11, AA13, AA14, and AA15. 16) The method of any one of numbered embodiments 1 to 15, wherein one or more of the polysaccharide cleaving enzymes is prepared from a T. reesei fungus. 17) The method of any one of numbered embodiments 1 to 16, wherein the one or more oligosaccharides comprises one of β-glucan, cello, MLG-, mannan-, or xylooligosaccharides. 18) The method of any one of numbered embodiments 1 to 17, wherein the polysaccharide cleaving enzyme is operably linked to a catalytic or non-catalytic module, e.g., the polysaccharide cleaving enzyme is operably linked to a non-catalytic module, the non-catalytic module being a carbohydrate-binding module. 19) The method of any one of numbered embodiments 1 to 18, wherein after separation of the one or more oligosaccharides, the one or more oligosaccharides are subjected to a chemical, physical, or enzymatic treatment, such as reduction, oxidation, caramelization, or Maillard reaction.

[0502] The detailed description is further supplemented by reference to the following numbered embodiments. 1) A method for producing an ingredient for human consumption, comprising: (a) physically treating a plant biomass; (b) subjecting the physically treated plant biomass to an incubation cycle comprising: (i) incubating the physically treated plant biomass in an incubation solution having a pH of 6.6 to 7.4 to solubilize monosaccharides and / or disaccharides from the physically treated plant biomass; and (ii) removing a portion of the solubilized monosaccharides and / or disaccharides from the incubation solution; (c) thermochemically treating the incubated plant biomass with one of (i) an acidic solution having a pH of 2 to 6.5 or (ii) an alkaline solution having a pH of 7.5 to 12; (d) contacting the thermochemically treated plant biomass in a solution or suspension with one or more polysaccharide cleaving enzymes to form one or more oligosaccharides; and (e) concentrating the solution or suspension to increase the concentration of the one or more oligosaccharides to form an ingredient. 2) The method of numbered embodiment 1, further comprising removing at least a portion of the monosaccharides and / or disaccharides from the incubation solution in step (b)(ii). 3) The method of any preceding numbered embodiment, wherein the thermochemically treated plant biomass is free of any or substantially no monosaccharides. 4) The method of any preceding numbered embodiment, further comprising purifying one or more oligosaccharides from the solution or suspension. 5) The method of any preceding numbered embodiment, further comprising repeating step (b). 6) The method of numbered embodiment 5, wherein step (b) is performed 2, 3, 4, or 5 times. 7) The method of any preceding numbered embodiment, further comprising repeating step (c). 8) The method of numbered embodiment 7, wherein step (c) is performed 2, 3, 4, or 5 times. 9) The method of any preceding numbered embodiment, further comprising discarding the portion of the solubilized monosaccharides and / or disaccharides removed in step (b).10) The method of any preceding numbered embodiment, wherein the portion of solubilized monosaccharides and / or disaccharides removed in step (b) is not combined with the portion of one or more oligosaccharides of step (e) to form the component. 11) The method of any preceding numbered embodiment, wherein the component is substantially free of monosaccharides. 12) The method of any preceding numbered embodiment, wherein the component is substantially free of disaccharides. 13) The method of any preceding numbered embodiment, wherein the one or more oligosaccharides comprise at least one of: i) cello-oligosaccharides having a degree of polymerization (DP) of 2 to 6; ii) xylo-oligosaccharides having a DP of 2 to 12; iii) arabinoxylo-oligosaccharides having a DP of 3 to 15; iv) manno-oligosaccharides having a DP of 2 to 12; v) mixed linkage glucan oligosaccharides having a DP of 2 to 5; vi) xyloglucan oligosaccharides having a DP of 4 to 12; or vii) chitooligosaccharides having a DP of 2 to 12. 14) The method of numbered embodiment 13, wherein the component comprises at least two of the oligosaccharides listed in (i) to (vii). 15) The method of numbered embodiment 14, wherein the component comprises at least two oligosaccharides in a ratio of 1:9 to 1:1 relative to each other. 16) The method of any preceding numbered embodiment, wherein the monosaccharides and / or disaccharides comprise at least one of sucrose, glucose, maltose, lactose, glucose, fructose, or galactose. 17) The method of any preceding numbered embodiment, wherein the physical treatment of step (a) comprises at least one of chipping, chopping, milling, ball milling, grinding, shaping, or blending the plant biomass. 18) The method of any preceding numbered embodiment, wherein the incubation of step (b) occurs in an incubation solution comprising water. 19) The method of any preceding numbered embodiment, wherein the incubation of step (b) occurs at a temperature between 15° C. and 95° C. 20) The method of any preceding numbered embodiment, wherein the incubation of step (b) occurs for a period between 15 minutes and 1 hour. 21) The method of any preceding numbered embodiment, wherein the thermochemical treatment of step (c) comprises heating the physically treated plant biomass in an acidic or alkaline solution.22) The method of numbered embodiment 21, wherein the heating is at a temperature of 50° C. to 150° C. 23) The method of numbered embodiment 21 or 22, wherein the heating is carried out for 30 minutes to 4 hours. 24) The method of any preceding numbered embodiment, wherein the incubated plant biomass is thermochemically treated in an alkaline solution having a pH of 8 to 11. 25) The method of numbered embodiment 24, wherein the alkaline solution comprises at least one of sodium hydroxide, potassium hydroxide, sodium carbonate, calcium carbonate, aqueous ammonia, ammonium sulfate, or ammonium hydroxide. 26) The method of any preceding numbered embodiment, wherein the incubated plant biomass is thermochemically treated in an acidic solution having a pH of 4 to 6. 27) The method of numbered embodiment 26, wherein the acidic solution comprises at least one of sulfuric acid, hydrochloric acid, nitric acid, phosphoric acid, acetic acid, maleic acid, fumaric acid, or oxalic acid. 28) The method of any preceding numbered embodiment, wherein the plant biomass comprises at least one of sugarcane, corn stover, corn cob, bran, wheat straw, hardwood or softwood. 29) The method of any preceding numbered embodiment, wherein the plant biomass comprises at least one of cellulose, chitin, chitosan, xylan, xyloglucan, mixed linkage glucan, mannan, or lignocellulose. 30) The method of any preceding numbered embodiment, wherein the one or more polysaccharide cleaving enzymes comprise at least one of cellulase, xylanase, xyloglucanase, endo-glucanase, cellobiohydrolase, mannanase, lichenase, or lytic polysaccharide monooxygenase (LPMO). 31) The method of any preceding numbered embodiment, wherein the one or more polysaccharide cleaving enzymes comprise at least one of AA9, AA10, AA11, AA13, AA14, or AA15. 32) The method of any preceding numbered embodiment, wherein one or more of the polysaccharide cutting enzymes are prepared from Trichoderma reesei fungus. 33) The method of any preceding numbered embodiment, wherein one or more polysaccharide cutting enzymes are operably linked to a catalytic module. 34) The method of any preceding numbered embodiment, wherein one or more polysaccharide cutting enzymes are operably linked to a non-catalytic module.35) The method of numbered embodiment 34, wherein the non-catalytic module is a carbohydrate-binding module. 36) A method for producing an ingredient for human consumption, comprising: (a) (i) physically treating a plant biomass; (ii) incubating the plant biomass in an incubation solution having a pH of 6.6 to 7.4 to solubilize a portion of the monosaccharides and / or disaccharides and removing a portion of the solubilized monosaccharides and / or disaccharides; and (iii) pretreating the plant biomass with a pretreatment comprising thermochemically treating the plant biomass with one of: (i) an acidic solution having a pH of 2 to 6.5 or (ii) an alkaline solution having a pH of 7.5 to 12; (b) contacting the pretreated plant biomass with one or more polysaccharide-cleaving enzymes in a solution or suspension to form one or more oligosaccharides; and (c) isolating a portion of the one or more oligosaccharides to form an ingredient. EXAMPLES

[0503] The following illustrative examples are representative of embodiments of the compositions and methods described herein, and are not intended to be limiting in any way. Example 1 Example Process

[0504] The following steps can be performed to generate the components as provided herein: 1. Physical pretreatment of plant biomass: 100 g of milled corn cob is mixed with water to a solids concentration of 10% (w / w) and mixed for 60 minutes at room temperature, at the end of which mixing is stopped and the liquid is filtered off, retaining the solids. 2. Resuspend the solids in water at a solids concentration of 10% (w / w) in the suspension. Begin mixing, heat to 95°C, and mix at 95°C for 60 minutes. 3. At the end of 60 minutes of heating, add 6 g of sodium hydroxide (0.2-3% by weight of the corn cob) and continue to stir. Heat to a temperature of 95°C and mix for 60 minutes to break down the hemicellulose present in the corn cob. Stop heating at the end of 60 minutes and cool to a temperature of 50°C. 4. Hydrolysis: Add cellulolytic enzymes (eg, from Trichoderma reesei) and incubate at 50° C., pH 5.5 (adjusted with 1 mol / L sulfuric acid and / or 1 mol / L sodium hydroxide) for 72 hours. 5. Biomass Separation: At the end of hydrolysis, the liquid is separated from the products via a solid-liquid separator. 6. Enzymatic separation following enzymatic hydrolysis: The liquid fraction from the slurry contains enzymes, oligosaccharides, water, and salts that need to be separated. A 10 kDa hollow fiber membrane is used to separate the enzyme proteins and other macromolecules. 7. The salts are removed using an ion exchange column at ≦45° C. a. Cation column: Strongly acidic cation exchange resin, cross-linked polystyrene matrix, sulfonate functional group, Na + Counter ion. b. Anion column: macroporous, weakly basic anion exchange resin, cross-linked polystyrene matrix, dimethyl-tertiary amine functional groups, OH - Counter ion. 8. Oligosaccharide concentration: The desired oligosaccharides are selectively concentrated via nanofiltration at room temperature. 9. Concentration: The liquid is concentrated to 40-75% at 60-80°C. 10. Spray drying: Spray drying at an inlet temperature of 130-160°C and an outlet temperature of 65-85°C. Example 2 Removal of soluble sugars by a wash cycle (i.e., incubation cycle)

[0505] The following steps were performed to remove soluble sugars from the plant biomass: 1. Water was added to 100 mg of the four plant biomass types (i.e., sugarcane, wheat, cob, and willow) to a concentration of 10% (w / v) and incubated at 45° C. for 30 min, after which the suspensions were centrifuged and the supernatant removed. 2. Step 1 was repeated five times. 3. 2.5 μl of each supernatant fraction was analyzed by thin layer chromatography (TLC).

[0506] The thin layer chromatograms in FIG. 1 show the presence of washed soluble sugars from four types of plant biomass in five successive washing or incubation cycles (1, 2, 3, 4, and 5). Undesirable monosaccharides and disaccharides, such as glucose, sucrose, and maltose, are indicated by arrows. The TLC results showed that for all plant biomasses, the supernatant removed after the first washing cycle had abundant monosaccharides and disaccharides present therein. Thus, the washing cycle successfully removed monosaccharides and disaccharides from the plant biomass. The supernatants from the subsequent washing cycles had significantly fewer monosaccharides and disaccharides present therein, indicating that the plant biomass has minimal, if any, monosaccharides and disaccharides in the supernatant after washing cycles as provided herein. Example 3 Absence of washed soluble sugars in enzymatic hydrolysis

[0507] The following steps are carried out to demonstrate the absence of soluble sugars in the enzymatic hydrolysis: 1. Washed cob and willow plant biomass from Example 2 were each incubated in 1% (w / v) NaOH at 99° C. for 30 minutes and then cooled. 100 mg of unwashed cob and willow biomass were also each incubated in 1% (w / v) NaOH at 99° C. for 30 minutes and then cooled. 2. 150 μl of the resulting suspension was mixed with 150 μl each of 1 M ammonium acetate (pH 5.5) and 150 μl of an enzyme composition containing beta-xylanase and cellobiohydrolase. The suspension was then incubated at 50° C. for 16 hours to allow the enzyme reaction with the plant biomass to occur. 3. 2.5 μl of each supernatant fraction was analyzed by TLC.

[0508] The thin layer chromatograms in Figure 2 show the products of enzymatic digestion of the four types of biomass washed (+) or unwashed (-) in Example 2. The results show that after enzymatic digestion, the final products contained glucose, sucrose, and maltose for the unwashed cob and willow plant biomass. However, the final products after enzymatic digestion did not contain glucose, sucrose, or maltose for the washed cob and willow plant biomass. Example 4 The addition of polymers to the oligosaccharide solution allows the solution to dry into a hard glaze.

[0509] The following steps were performed to demonstrate that the addition of polymer to an oligosaccharide solution allows the solution to be dried into a hard glaze: 1. 100 μl of 10-320 mM (10, 20, 40, 80, 160, and 320 mM) cellobiose ± 1% w / v birchwood xylan was pipetted onto a glass plate. 2. The samples were dried at 37°C. 3. Samples were scored with a knife to test whether the ingredients would settle into a solid glaze. With cellobiose alone, no strong glaze was formed and the dried powder easily cracked when pressure was applied with a knife. In contrast, with 1% w / v xylan added and with cellobiose added at or below 80 mM, the compositions dried to form a solid, off-white, translucent surface that was strong enough to be scored with a knife leaving a dent but no cracks. At cellobiose concentrations of 160 mM or higher (5.5% w / v or 550% w / w compared to xylan), the glaze morphology reverted to that in the absence of xylan; i.e., no strong glaze was formed and the dried powder easily cracked when pressure was applied with a knife (see Figure 3). Example 5 Demonstration of the composition of two types of oligosaccharides and polysaccharides in foods

[0510] The following steps were performed to demonstrate a composition containing two oligosaccharides and a polysaccharide in a food product: 1. 4 g of birchwood xylan was dissolved in 75 ml of boiling water. 2. 12 g of cellobiose and 24 g of xylooligosaccharides (mainly with a degree of polymerization (DP) of 2 to 6) were added in 3 g portions and dissolved by boiling. 3. The mixture was reduced to 50 ml with heating to form a thick solution with a cloudy honey-like consistency and appearance, but not very sweet. 4. 10 mL of the mixture was mixed with 12 g of oats to form a flapjack / cereal bar mixture and another 10 mL of the mixture was also mixed with 6 g of fruit and 6 g of nuts to make a cereal bar mixture. 5. The samples were baked at 100°C for 10 minutes and then allowed to cool and dry overnight.

[0511] As shown in FIG. 4, Panel A (flapjack / cereal bar) and Panel B (fruit and nut bar), the resulting flapjacks and fruit and nut bars were of desirable texture and consistency that matched flapjack / cereal bars and fruit and nut bars made with known syrups typically used in baking.

[0512] For the flapjacks / cereal bars, the thickened solution from step 3 helped bind the mixture from step 4 together. The combined effect of the ingredients and the properties of the oats resulted in a grainy surface texture consistent with flapjacks / cereal bars produced using traditional sugar. The ingredients created a firm, chewy, moist, viscous texture resulting in a mouthfeel consistent with that expected for these types of foods but not present with oats alone. The product was mild in its sweetness and did not contain any bitterness or off-flavors that can be characteristic of high intensity sweeteners.

[0513] In the case of the fruit and nut bars, the thickened solution from step 3 helped bind the mixture from step 4 together. It also added a smooth, shiny glazed surface that is a core part of the aesthetic quality of such foods and that could not be created if the ingredients contained only oligosaccharides (i.e., oligosaccharides without polysaccharides). The ingredients created a firm, chewy, moist, viscous texture resulting in a mouthfeel consistent with that expected from these types of foods but not present with either nuts or fruits alone. The product was mild in its sweetness and did not contain bitterness or off-flavors that can be characteristic of high intensity sweeteners. Example 6 Process for making the components

[0514] The following steps can be performed to make the compositions provided herein: 1.15% w / v corncob (ground through a 1 mm pore size filter) in 1% w / v NaOH is heated at 90° C. for 1 hour, thereby solubilizing some of the polymeric components of the biomass. 2. Adjust the pH to 5.5 with sulfuric acid. 3. Extract a volume that contains only the liquid components of the reaction, which is 15% of the total volume of the reaction ("soluble polymer"); retain the remaining 85% volume of the reaction, which contains all of the insoluble biomass fraction ("remaining biomass"). 4. The remaining biomass was supplemented with a cellulolytic enzyme cocktail (e.g., an enzyme cocktail from Trichoderma reesei containing cellulase, xylanase, arabinofuranosidase, LPMO, etc.) to 0.5% w / v and incubated at 50°C for 24 hours. 5. Separate the soluble oligomeric reaction products from the undigested, insoluble polymeric compounds by filtration. 6. Purify the oligomeric reaction product by sequential microfiltration, ultrafiltration, and ion exchange chromatography (e.g., cross-flow filtration on ceramic membranes; filtration can be performed using 110 0.45 μm cutoff Inside Ceram candle filters (TiO2, Φ25 mm x L 1178 mm HD 6 mm, 8 channels per membrane) supplied by TAMI Industries with a maximum feed pressure of 3 bar). 7. Purify the soluble polymer by using ultrafiltration (e.g., 10 kDa spiral wound membrane (Snyder ST-2B-6338, PES, feed spacer thickness 31 mm) run on an Alfa Laval ultrafiltration unit). 8. The solutions formed in steps 6 and 7 are recombined and further purified and concentrated using nanofiltration to form the component. Example 7 Viscosity measurements of various solutions

[0515] The following steps were performed to measure the viscosity of the various solutions: 1. A solution of three sugars, including cellobiose (Cell2), xylooligosaccharides (XOS) mainly with DP 2-6, and polymeric beechwood xylan (BWX), was created by boiling the sugars in water. The final concentrations were: a. Sample 1: 0.33 g / ml Cell2, 0.66 g / ml XOS, 0.13 g / ml BWX b. Sample 2: 0.17 g / ml Cell2, 0.33 g / ml XOS, 0.07 g / ml BWX; c. Sample 3: 0.54g / ml Cell2, 0.52g / ml XOS, 0.07g / ml BWX. 2. Samples were tested using a Brookfield HDB VE Roto Viscometer using standard test procedures. 400 mL samples were collected in tall beakers to ensure no container effects occurred. The instrument was operated according to the manufacturer's instructions for the range: Roto Viscometer using spindle code 61, spindle speed 100 rpm, and 22°C. [Table A]

[0516] Sample 1 had a consistency similar to that of thick honey, requiring mixing to dilute into aqueous solutions. In contrast, Samples 2 and 3 were very flowable and easily mixed into aqueous solutions. The results showed that the viscosity of the compositions was more affected by the concentration of polysaccharides than the total concentration of oligosaccharides and polysaccharides. Samples 2 and 3 have the same polysaccharide concentration, but Sample 3 has twice the total concentration of oligosaccharides and polysaccharides than Sample 2. The viscosity of Sample 3 is twice that of Sample 2, following a linear relationship between total concentration and viscosity. However, there is an exponential increase in viscosity values ​​as the concentration of polysaccharides increases. The polysaccharide concentration of Sample 1 is twice that of Sample 3, and the concentrations of total oligosaccharides and polysaccharides are the same, yet the viscosity of Sample 1 is 15 times greater than Sample 3. Example 8 Preparation of Water-Soluble Liquid Products / Components

[0517] The following steps were performed to generate a water soluble liquid product / component (Sample 4): 1. 100 g of milled corn cob was heated in 1 L of deionized water containing 2.5 g of sodium chlorite for 1.5 hours at 80° C. with constant stirring. The remaining volume was reconstituted to 900 mL by adding 200 mL of deionized water containing an additional 5 g of sodium chlorite and heated at 80° C. for 1 hour with constant stirring. 2. The solution was filtered through a 2 mm pore size ceramic filter funnel under vacuum until the filtrate was clear. 3. The retained solids were incubated in 1 L of 0.5 M sodium hydroxide 0.1% (w / v) sodium borohydride for 17 hours at 50° C. and 115 rpm with shaking. 4. The pH was then adjusted to 7 with concentrated sulfuric acid and dialyzed against tap water for 24 hours in 12,000 dalton cut-off dialysis tubing. 5. The contents of the dialysis tube were transferred to a 2 L beaker and the insoluble fraction was allowed to settle by gravity. 6. The supernatant was decanted twice and concentrated by evaporation at 80°C to a volume of 120 mL. 7. The water-soluble polymer was precipitated by centrifugation after the addition of 3 volumes of ethanol. The resulting supernatant was discarded and the precipitate was air-dried to constant weight at room temperature. 8. The oligosaccharides were added to a final w / w of 10% cellobiose, 75% xylooligosaccharides, 15% extracted water soluble polymers and mixed until homogenous in a Waring Xtreme blender at the lowest power setting. 94 g of solids were recovered from the blender. 9. Surprisingly, all 94 g of solids dissolved in 60 mL of water at 50° C. with constant gentle stirring (approximately 100 rpm), indicating a solubility of greater than 150 g / 100 g.

[0518] The generated sample (eg, steps 8 and 9 above) is referred to as Sample 4. Example 9 Physicochemical properties of the water-soluble liquid product of Example 8

[0519] Flow properties: The flow properties of the water-soluble liquid product according to the present disclosure described in Example 8 (Sample 4) are detailed in Table 1 along with comparative compositions of water, 20% w / v glucose, 40% w / v glucose, 60% w / v glucose, 80% w / v glucose, and ≧99% glycerol (Fisher G / 0650 / 17 as supplied). Glucose solutions were made by weighing out 6, 12, 18, and 24 g each of D-glucose and making up to 30 mL with water at 90° C. Flow properties were measured by flow rates of 5 mL over time and, where appropriate, 20 mL of liquid from an upright syringe (BD Plastipak 300613) filled with 20 mL of test liquid under gravity at room temperature. Table 1: Flow characteristics [Table 1] The longer it takes for the sample to flow out of the bottom of the syringe (i.e., it has a lower flow rate) correlates to a higher viscosity of the sample. The lower the flow rate, the more syrupy / sticky and viscous the liquid sample becomes. The flow rate measured for Sample 4 is similar to glycerol and lower than all glucose solutions tested. This property of Sample 4 makes it more useful than glucose solutions, water as a binder in foodstuffs such as cereal bars, as well as providing sweetness to the product.

[0520] Color: The color of sample 4 corresponds to Standard Reference Method (SRM) No. 30, a method for assessing the color of wort or beer published in the Recommended Methods of the American Society of Brewing Chemists (ASBC Methods of Analysis, Beer 10. Spectrophotometric Color Method Approved 1958, rev. 2015. American Society of Brewing Chemists, St. Paul, MN, USA). Briefly, the absorbance of the sample is measured at a wavelength of 430 nm in a cell with a path length of 1 cm. The obtained absorbance value is multiplied by 12.7 to obtain the value. Due to the turbidity of sample 4, the absorbance could not be measured and the evaluation was made by optical comparison with SRM No. 30 (dark red / brown).

[0521] Anion exchange chromatography: Analysis of sample 4 by high performance anion exchange chromatography (HPAEC) was performed using a Thermo Fisher Scientific DIONEX ICS-6000 system equipped with a CarboPac PA200 analytical column (3 x 250 mm) and a CarboPac PA200G guard column (3 x 50 mm) and a Dionex ED electrochemical detector. Data was acquired with Chromeleon 7 software.

[0522] Eluents A (milli Q water), B (250 mM NaOH), and C (250 mM NaOH + 1 M sodium acetate) were used to generate a mobile phase with the gradient profile shown in Table 2. Table 2: Gradient profile [Table 2]

[0523] Samples for analysis were prepared by diluting sample 4 100-fold and passing it through a 0.45 μm syringe filter, the injection volume of the analyte was 10 μl.

[0524] HPAEC analysis (see FIG. 5) confirmed that Sample 4 was a mixture of monosaccharides, disaccharides, and other oligosaccharides composed of glucose and xylose. This is in contrast to syrups typically used in the food industry, such as corn syrup and high fructose corn syrup, which contain primarily glucose and fructose monosaccharides. As a result, the product of Sample 4, when used as an ingredient, is expected to have fewer calories, a lower glycemic index, and contain fiber, as opposed to corn syrup and / or high fructose corn syrup. Example 10 Cold-pressed fruit cereal bars

[0525] A cold pressed fruit cereal bar was prepared as follows: 1. 120g of sample 4 from Example 8 was heated with 30g of coconut oil and 1 / 4 teaspoon of cinnamon was added. 2. Once frothy, remove the mixture from the heat and add 40g oats, 40g dried dates, 10g crisp rice and 10g seeds. 3. The ingredients were mixed thoroughly until all components were coated, the mixture was transferred to a freezer bag and placed in the freezer. The contents of the bag were rolled to a thickness of 7-10 mm, chilled at 4°C overnight, and then cut into rectangles.

[0526] The resulting product (see Figure 6) was a chewy, sticky cereal bar that was loosely set and contained oats and crisp rice with perceptible sweetness imparted by Sample 4 and chopped dates. Example 11 Generating ingredients in large-scale manufacturing processes

[0527] The following steps were taken to produce the components in a large scale manufacturing process: 1. Physical pretreatment of plant biomass: Mix 100 kg of milled corn cob with water at 15% (w / w) solids concentration in suspension. Start mixing, heat to 95°C and mix at 95°C for 60 minutes. Add 6 kg of sodium hydroxide (0.2-3% by weight of corn cob) and continue stirring. Heat to a temperature of 95°C and mix for 60 minutes to release the hemicellulose present in the corn cob. At the end of 60 minutes, cool to 50°C and adjust the pH to 5.5 with sulfuric acid. 2. Removal of the soluble polysaccharide portion: A portion of the soluble phase, representing 5-30% of the total xylan, is removed. It is neutralized with sulfuric acid, concentrated and purified by ultrafiltration. Any precipitated polymer is removed. 3. Hydrolysis: Cellulolytic enzymes (e.g., from Trichoderma reesei) are added to the milled corn cob mixture and incubated at about 50°C for 12-72 hours. 4. Biomass Separation: At the end of hydrolysis, the liquid is separated from the products by passing it through a solid-liquid separator such as a filter press or a decantation centrifuge. 5. Enzymatic separation after enzymatic hydrolysis: The liquid fraction from the slurry contains enzymes, oligosaccharides, water, and salts that need to be separated. A 3 kDa or 10 kDa hollow fiber membrane is used to separate the enzyme proteins and other macromolecules. 6. Salts are removed using an ion exchange column at ≦45° C. a. Cation column: Strongly acidic cation exchange resin, cross-linked polystyrene matrix, sulfonate functional groups, and Na + Counter ion. b. Anion column: macroporous, weakly basic anion exchange resin, cross-linked polystyrene matrix, dimethyl-tertiary amine functional groups, and OH - Counter ion. 7. Oligosaccharide concentration: The desired oligosaccharides are selectively concentrated through nanofiltration at room temperature. 8. Concentration: Concentrate the liquid to 40-75% at 60-80°C as required. 9. Reconjugation: Combine the purified soluble polymer with the enzymatically generated oligomer in a dry weight ratio of 5:95 to 20:80. 10. Spray drying: The resulting solution is spray dried at an inlet temperature of 130-160°C and an outlet temperature of 65-85°C. Example 12 Use of liquid ingredients to produce extruded cereal bars

[0528] Extruded cereal bars can be produced using the liquid ingredients as provided herein by the following steps: 1. 120 kg of solution containing 10.5 kg xylan, 57 kg xylooligosaccharides, and 7.5 kg cellobiose is heated with 130 kg coconut oil and transferred into a high speed mixer. 200 kg rolled oats and 25 kg chopped dates / raisin mixture are added and mixed thoroughly. This is pulsed into the dough mixer through the dough feeding system and set at 20 psi. 2. The mixture is transferred through belt and ramshorn and baked for 20 minutes at 180°C. The product is then transferred through oven traveler onto biscuit cutting line, then made into various shapes of fills and cooled to <5°C before being sealed and packaged. 3. The resulting product is a soft, sticky cereal bar, loosely set and filled with oats. Sweetness is provided by a liquid solution containing 10.5 kg xylan, 57 kg xylooligosaccharides, 7.5 kg cellobiose, and chopped dates in the bar. The liquid solution containing 10.5 kg xylan, 57 kg xylooligosaccharides, 7.5 kg cellobiose, and coconut oil acts as a binder that holds the other ingredients together inside the bar and gives the bar its structure. Example 13 Use of liquid ingredients to produce extruded breakfast cereals

[0529] The following steps can be performed to use the liquid ingredients provided herein to produce an extruded breakfast cereal: 1. A dough is formed by combining cereal flour (approximately 85-75% w / v) with a solution containing 22.5 g xylan, 30 g cellobiose, and 97.5 g xylooligosaccharides per 100 g water (approximately 15-25% v / v) and optional additives such as preservatives and vitamins and minerals for nutritional value. The dough is extruded using a twin-screw extruder, where the product is cooked using the addition of heat and moisture, and / or a combination of steam and mechanical shear, and the product shape is formed by forcing through a nozzle. The product is then allowed to rise until light in texture and golden in color, and cooled. 2. The result is a light, crisp, formed breakfast cereal product. A liquid solution containing 22.5g xylan, 30g cellobiose, and 97.5g xylooligosaccharides gives the product a sweet taste and helps form the dough structure before extrusion. Example 14 Use of liquid ingredients for producing tomato ketchup

[0530] To use the liquid ingredients provided herein to produce tomato ketchup, the following steps can be performed: 1. Blend 4 onions and 250g celery in a food processor until finely chopped. Fry it in 5 tablespoons vegetable oil over low heat for 5 minutes. Add 4 cloves garlic and cook for another 5 minutes. Add 1 teaspoon crushed coriander, 1 short cinnamon stick, 1 teaspoon allspice, 1 / 2 teaspoon ground black pepper, and 2 teaspoons celery salt and cook for another minute. 2. Add 2 kg ripe chopped tomatoes, 3 tablespoons tomato puree, one-half (1 / 2) teaspoon chili sauce, 200 mL white wine vinegar, and 285 mL of a solution containing 22.5 g xylan, 30 g cellobiose, and 97.5 g xylooligosaccharides per 100 g water. Return the mixture to the heat, bring to a boil, and simmer, uncovered, for 1 hour until the tomatoes are tender. Discard the cinnamon stick and blend the sauce mixture until smooth, then strain. 3. The resulting product is a smooth and tangy tomato ketchup. The liquid solution containing 22.5g xylan, 30g cellobiose, and 97.5g xylooligosaccharides sweetens the product, gives body to the sauce, and helps thicken and bulk the sauce. Example 15 HPAEC chromatography of sugars in water after washing of corncobs.

[0531] Corncobs were incubated at 100 g / L in room temperature water ("Wash" in Table 3), then the water was decanted. Water was added to the original total volume and heated to 90° C. for 60 minutes ("Wet" in Table 3), and then heated in dilute NaOH at 90° C. for 60 minutes ("Pretreatment" in Table 3).

[0532] HPAEC was performed on the washed, wet, and pretreated samples to identify sugar peaks (see FIG. 7, e.g., chromatogram from "Wash"). As indicated in Table 3, approximately 2% of the corncob at the start of the process is glucose that can be washed out, and presumably more glucose could be washed out. Note also that the pH after the wash step drops to 4.5. Table 3 [Table 3] (Example 16) Sugar and organic acid determination

[0533] To quantify the impact of the pre-wash step on the process, three separate batches of corncobs were treated according to the procedure outlined in FIG. 10A. Samples were analyzed for sugars by HPLC (FIG. 10B) and for organic acids by HPLC (FIG. 10C). The differences in sugar and organic acid composition isolated from the various samples (samples A-E and from 5 minutes (1 / 12 hours) to 4 hours) show the impact of the wash. The impact of the pre-wash was likely greater than 150 mL out of 600 mL was extracted per wash. These data therefore indicate a course of action where washing can be performed but is not limiting.

[0534] As shown, glucose, fructose, and sucrose were all reduced by the pre-wash. Fructose and glucose are largely destroyed during NaOH treatment, while sucrose is resistant to NaOH treatment. Sucrose generally cannot be removed from other disaccharides through filtration, so washing can help remove it. Table 4 shows a comparison of the sugars in Sample D ("No Wash" and "Double Wash"), and Table 5 shows a comparison of the sugars in the 4-hour samples ("No Wash" and "Double Wash"). Table 4 [Table 4] Table 5 [Table 5]

[0535] Before the start of the "caustic digestion" step (e.g., a thermochemical step), pre-washing results in about a 50% reduction of glucose and fructose, and about an 80% reduction of sucrose. At the end of hydrolysis, pre-washing results in a small difference in the amount of sugars. Xylose-based sugars appear to increase in concentration (as evidenced by a negative change), while glucose and cellobiose decrease in concentration.

[0536] The large amounts of organic acids detected are not a product of washing, but rather of pretreatment. However, the unwashed biomass appears to have a higher total load of organic acids than the twice-washed material.

[0537] The acid concentration for the washed biomass also appears to be lower for the steps leading up to the pretreatment. Table 6 shows a comparison of Sample D and Table 7 shows a comparison of the 4 hour samples. Table 6 [Table 6] Table 7 [Table 7]

[0538] The effect of washing on the organic acids content, as shown by the "change" (calculated by "no washing" minus "double washing"), is evident both at the end of the wetting stage (sample D) and at the end of the hydrolysis reaction (4 h sample) (see, for example, Tables 6 and 7). At the end of the wetting stage (sample D), all other acids (n / d) except lactate and formate were detected and were at lower concentrations for the material that had been washed twice.

[0539] At the end of hydrolysis, the acetate content is higher (hydrolysis releases acetate), while the other acids remain lower for the washed biomass than for the unwashed one (except for lactate, which is present in low concentrations).

[0540] Visual observation of the samples is shown in Figure 10D. After two washes, the corncob released fewer color compounds and the liquor was lighter. Without being bound to any particular theory, the color compounds appear to be phenols and organic acids released during washing. (Example 17) Comparison of cold-pressed cereal bars

[0541] Cold pressed cereal bars were prepared following the recipe as before (see Example 10).

[0542] Soluble and insoluble polysaccharides were used in the cereal bars for comparison. The soluble and insoluble polysaccharides were as follows: Soluble polysaccharides: 60 mL water containing 94 g dry components with the composition 10% dry w / w cellobiose, 75% xylooligosaccharides, and 15% extracted water soluble polymers (Sample 4 as described above in Example 8). Insoluble polysaccharides: 60 mL of water containing 94 g of dry ingredients with the composition 10% dry w / w cellobiose, 75% xylooligosaccharides, and 15% microcrystalline cellulose.

[0543] Referring to Figure 11A, the cereal bars made with the insoluble polysaccharides looked like solid bars when placed on a table, but began to crumble as soon as they were lifted off the table and picked up due to their soft texture and ingredients that were not well bound together. In contrast, the cereal bars made with the soluble polysaccharides could be easily handled and maintained their shape.

[0544] The hardness and stickiness of the cereal bars were measured using a TA-XTPlusC texture analyzer (Stable Microsystems, UK) using "ExponentC" software. A sample with dimensions (L x W x H) of 9.6 cm x 3.8 cm x 1 cm was placed centred under the probe. A 6 mm diameter aluminium cylindrical probe was used in the penetration test in "return to start" mode and a 30 kg load cell. When the probe was induced above the surface it penetrated a distance of 2 mm into the sample at a speed of 2 mm / s. At this point (2 mm depth) the force value was recorded and taken as a measure of the "hardness" of the sample. The probe was then withdrawn from the sample and the maximum force to withdraw or "stickiness" at that point was recorded. The pre-test speed was 1 mm / s and the post-test speed was 10 mm / s.

[0545] The results showed that the stiffest bars were those made with the soluble components, while lower values ​​were obtained for the insoluble components (Figure 11B). Similarly, the soluble polysaccharide-containing bars had higher stickiness, and the insoluble polysaccharide-containing bars had reduced stickiness. These results confirmed the visual and tactile observations. Table 8: Results [Table 8] Table 9: Texture analyzer settings [Table 9]

[0546] The hardness of the cereal bars was further measured using a TA-XTPlusC texture analyzer (Stable Microsystems, UK) using "ExponentC" software. Samples with dimensions of 9.6 cm x 3.8 cm x 1 cm (L x W x H) were placed centred under the knife. The edge of the knife was used in the cut test in "return to start" mode and a 30 kg load cell. When the knife was induced on the surface, it penetrated 5 mm into the sample at a test speed of 2 mm / s. The maximum force measured during the cut test was recorded as the hardness of the sample. The pre-test speed was 1.5 mm / s and the post-test speed was 10 mm / s.

[0547] The results obtained using the cutting method confirm those obtained using the penetration method described above: the results show that the hardest bars were those made with soluble polysaccharide, while lower values ​​were obtained for the insoluble polysaccharide-containing bars, indicating that the latter were softer (Figure 11C). Table 10: Results [Table 10] Table 11: Texture analyzer settings [Table 11] (Example 18) Viscosity of sugar compositions containing insoluble versus soluble polysaccharides

[0548] The following samples were prepared: Sample 1: 60 mL of water containing 94 g of dry components with a composition of 10% dry w / w cellobiose, 75% xylooligosaccharides, and 15% extracted water-soluble polymers. Sample 2: 60 mL of water containing 94 g of dry ingredients with the following composition: 10% dry w / w cellobiose, 75% xylooligosaccharides, and 15% microcrystalline cellulose.

[0549] "Sample 4" in Example 8, "soluble polysaccharide" in Example 17, and "Sample 1" in Example 18 are substantially identical and / or synonymous. Furthermore, "Sample 2" in Example 18 and "insoluble polysaccharide" in Example 17 are substantially identical and / or synonymous.

[0550] The samples were analyzed for flow properties as described in Example 9. The data confirm that soluble polysaccharides modulate the viscosity of the oligosaccharide composition (Table 12), allowing fine tuning of the viscometric properties of the solution in a way that is not generally possible with oligosaccharides alone. Table 12 [Table 12] Sequence Listing

[0551] LPMO AA9 LPMO, from Podospora anserina (SEQ ID NO:1). Genbank ID CAP67740 [ka]

[0552] Lichenase GH16 lichenase, from Bacillus subtilis subsp. subtilis str. 168 (SEQ ID NO:2). GenBank ID CAA86922.1 [ka]

[0553] Xylanase GH5 arabinoxylanase, from Ruminiclostridium thermocellum (SEQ ID NO:3). GenBank ID ABN53395.1 [ka]

[0554] GH5 xylanase, from Gonapodya prolifera (SEQ ID NO:4). GenBank ID KXS18720.1 [ka]

[0555] GH30 xylanase, from Dickeya chrysanthemi (SEQ ID NO:5). GenBank ID AAB53151.1 [ka]

[0556] GH30 xylanase, from Bacillus subtilis subsp. subtilis str. 168 (SEQ ID NO:6). GenBank ID CAA97612.1 [ka]

[0557] GH30 xylanase, from Bacteroides ovatus (SEQ ID NO:7). GenBank ID SDY64378.1 [ka]

[0558] Xyloglucanase GH5 xyloglucanase, from Bacteroides ovatus (SEQ ID NO:8). GenBank ID ALJ47680.1 [ka]

[0559] GH74 xyloglucanase, from Trichoderma reesei (SEQ ID NO:9) GenBank ID AAP57752.1 [ka]

[0560] Cellobiohydrolase GH7 Cel7A cellobiohydrolase, from Trichoderma reesei (SEQ ID NO: 10) GenBank ID CAH10320.1 [ka]

[0561] GH6 Cel6A cellobiohydrolase, from Trichoderma reesei (SEQ ID NO:11) GenBank ID AAA34210.1 [ka]

[0562] Endoglucanase A eglA-Aspergillus niger GH12 (SEQ ID NO: 12) [ka]

[0563] Aspergillus niger endo-β-1,4-glucanase GH5, CBM1 (SEQ ID NO: 13) [ka]

[0564] Aspergillus niger endo-β-1,4-glucanase B GH5 (SEQ ID NO: 14) [ka]

[0565] GH30 xylanase, from Trichoderma reesei (SEQ ID NO: 15) [ka]

[0566] Aspergillus niger endo-β-1,4-xylanase 1 GH11 (SEQ ID NO: 16) [ka]

[0567] GH5 Mannase, from Trichoderma reesei (SEQ ID NO: 17) [ka]

[0568] Aspergillus niger endo-β-1,4-mannanase GH26 (SEQ ID NO: 18) [ka]

[0569] Aspergillus niger β-mannanase GH5 (SEQ ID NO: 19) [ka]

[0570] Aspergillus niger cellobiohydrolase A GH7 (SEQ ID NO: 20) [ka]

[0571] Aspergillus niger cellobiohydrolase B GH7, CBM1 (SEQ ID NO:21) [ka]

[0572] GH3 beta-glucosidase from Trichoderma reesei (SEQ ID NO:22) [ka]

[0573] AA9 LPMO, from Trichoderma reesei (SEQ ID NO:23) [ka]

[0574] GH7 beta-gluanase (EGI), from Trichoderma reesei (SEQ ID NO:24) GenBank: AAA34212.1 [ka]

[0575] GH5 beta-glucanase (EGII), from Trichoderma reesei (SEQ ID NO:25) GenBank: ABA64553.1 [ka]

[0576] While preferred embodiments of the present invention have been shown and described herein, it will be apparent to those skilled in the art that such embodiments are provided by way of example only. Numerous variations, modifications, and substitutions will now 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 in practicing the invention.

[0577] All publications, patent applications, issued patents, and other documents mentioned herein are incorporated by reference herein as if each individual publication, patent application, issued patent, or other document was specifically and individually indicated to be incorporated by reference in its entirety. Definitions contained in the texts incorporated by reference are excluded to the extent that they conflict with definitions in the present disclosure. In certain embodiments, for example, the following are provided: (Item 1) 1. A method of forming an ingredient for human consumption, comprising the steps of: (a) isolating one or more soluble polysaccharides from a biomass; (b) contacting the remaining biomass with one or more enzymes to form one or more oligosaccharides; (c) isolating the one or more oligosaccharides; and (d) combining a portion of the one or more soluble polysaccharides from step (a) with a portion of the one or more oligosaccharides from step (c) to form the component. A method comprising: (Item 2) 2. The method of claim 1, further comprising purifying the isolated one or more soluble polysaccharides. (Item 3) 3. The method of claim 1 or 2, further comprising purifying the isolated one or more oligosaccharides. (Item 4) 4. The method according to any one of the preceding claims, further comprising treating the biomass to dissolve the one or more soluble polysaccharides. (Item 5) 5. The method according to any one of items 1 to 4, further comprising a step of purifying the isolated one or more soluble polysaccharides. (Item 6) 5. The method of claim 4, wherein the treating step comprises a thermochemical treatment. (Item 7) 7. The method according to claim 6, where...

Claims

1. A method for producing an ingredient for human consumption, said method comprising: (a) subjecting a biomass to a first pre-treatment to reduce the average size of the biomass to produce a pre-treated biomass; (b) subjecting the pretreated biomass to a second pretreatment, the second pretreatment comprising: (i) incubating the pre-treated biomass in an aqueous solution to dissolve monosaccharides and / or disaccharides from the pre-treated biomass; and (ii) removing a portion of the dissolved monosaccharides and / or disaccharides from the aqueous solution to produce a second pre-treated biomass; (c) subjecting the second pretreated biomass to a third pretreatment to solubilize polysaccharides and increase digestibility of the second pretreated biomass to produce a third pretreated biomass; (d) contacting the third pre-treated biomass with one or more enzymes in solution or suspension to form one or more oligosaccharides; (e) concentrating the solution or suspension to increase the concentration of the one or more oligosaccharides; and (f) combining a portion of the one or more soluble polysaccharides with a portion of the one or more oligosaccharides from step (f) to form the component. Including, The one or more soluble polysaccharides are isolated from the third pre-treated biomass from step c).

2. The method of claim 1, wherein the component comprises 1% to 50% w / w polysaccharide.

3. 3. The method of claim 1 or claim 2, further comprising treating the biomass to dissolve the one or more soluble polysaccharides, the treating comprising a thermochemical treatment including hot water treatment or high temperature alkaline treatment.

4. 2. The method of claim 1, wherein the one or more soluble polysaccharides and / or the one or more oligosaccharides are dried prior to step f) or the one or more soluble polysaccharides and / or the one or more oligosaccharides are dried after step (f).

5. 2. The method of claim 1, wherein the solubility of said component in water is at least 80 g of said component per 100 g of water at 50°C.

6. The method of claim 1 further comprising combining the component and a liquid to form a liquid component.

7. The method of claim 6, wherein the liquid component has a viscosity of from 5 cps to 100,000 cps.

8. 10. The method of claim 1, wherein the third pretreatment is a thermochemical treatment comprising incubating the second pretreated biomass in one of an acidic or alkaline solution.

9. 9. The method of claim 1 or 8, further comprising removing at least 25% or 50% of the dissolved mono- and / or disaccharides from the aqueous solution in step (b)(ii).

10. 10. The method of any one of claims 1, 8, and 9, wherein the third pre-treated biomass composition after step (c) comprises less than 10% w / w monosaccharides.

11. The method of any one of claims 1 and 8 to 10, wherein step (b) and / or step (c) is performed two or more times.

12. 12. The method of any one of claims 1 and 8-11, wherein the component comprises less than 15% by dry weight monosaccharides or less than 60% by dry weight disaccharides.

13. The one or more oligosaccharides are selected from the group consisting of: i) cellooligosaccharides having a degree of polymerization (DP) of 2 to 6; ii) xylooligosaccharides with a DP of 2-12; iii) arabinoxylooligosaccharides having a DP of 3 to 15; iv) mannooligosaccharides with a DP of 2-12; v) mixed linkage glucan oligosaccharides having a DP of 2-5; vi) xyloglucan oligosaccharides having a DP of 4 to 12; or The method according to any one of claims 1 to 12, comprising at least one of the following: vii) chitooligosaccharides having a DP of 2 to 12.

14. 14. The method of claim 13, wherein the component comprises at least two of the oligosaccharides listed in (i) to (vii) in a weight ratio of 1:9 to 1:

1.

15. 14. The method of claim 13, wherein the component comprises at least one of sucrose, maltose, lactose, glucose, fructose or galactose in a total dry w / w of less than 50% of the total dry w / w of the one or more oligosaccharides of i) to vii).

16. 16. The method of any one of claims 1 to 15, wherein step (b) further comprises dissolving one or more organic acids.

17. 17. The method of claim 16, wherein the one or more organic acids comprise at least one of oxalate, tartrate, succinate, formate, citrate, maleate, lactate or acetate, and the total weight of the oxalate, tartrate, succinate, formate, citrate, maleate, lactate and acetate is more than 10% of the total weight of sucrose, maltose, lactose, glucose, fructose and galactose in the portion dissolved and removed in step (b).

18. 2. The method of claim 1, wherein the first pre-treatment of step (a) comprises at least one of chipping, chopping, milling, ball milling, grinding, shaping, or blending the biomass.

19. 10. The method of claim 1, wherein the biomass comprises at least one of sugarcane biomass, corn biomass, wheat biomass, hardwood biomass, or softwood biomass.

20. 20. The method of any one of claims 1-7 or 19, wherein the one or more enzymes comprise at least one of cellulase, xylanase, xyloglucanase, endo-glucanase, cellobiohydrolase, mannanase, lichenase, or lytic polysaccharide monooxygenase (LPMO).

21. 21. The method of any one of claims 1 to 7 or 20, wherein the one or more enzymes include at least one of AA9, AA10, AA11, AA13, AA14 or AA15.

22. 22. The method of any one of claims 1 to 7, 20 or 21, wherein the one or more of the enzymes are prepared from a filamentous fungus.

23. 10. The method of claim 1, further comprising incorporating the ingredient into at least one of a baked good, a chocolate, a confectionery, or a beverage.

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