Fermentatively produced retinoid-containing compositions and methods of making and using same

Abstract

Disclosed herein are compositions suitable for use in food, feed, pharmaceutical, and cosmetic products, comprising a retinoid component and its fermentation residue. In one embodiment, the retinoid component is present in a weight ratio to the fermentation residue of 4:1 or greater, the retinoid contains cis- and trans-isomers, and the cis-isomer is present in an amount of less than 3% by weight of the total isomer mixture. Preferably, the composition is biobased. Biotechnology methods for producing the compositions described elsewhere are also described, as well as additional methods for processing such compositions into more stabilized, more commercially acceptable forms. Finally, food, feed, pharmaceutical, and cosmetic products incorporating the compositions described herein are also disclosed.

Description

Detailed Description of the Invention

[0001] [Technical field] The present invention relates to fermentatively produced retinoid-containing compositions, methods for producing same, methods for formulating such fermentatively produced retinoid-containing compositions, and formulations and (pre)products obtained therefrom.

[0002] [CROSS-REFERENCE TO RELATED APPLICATIONS]

[0002] This application claims priority to U.S. Provisional Patent Application No. 63 / 429802, filed December 2, 2022, the entire contents of which are incorporated herein by reference in their entirety as if fully set forth herein.

[0003] [background]

[0003] Retinoids are a class of compounds that are vitamers of or chemically related to vitamin A. Some retinoids have found use in many applications due to the various health benefits associated with them. Retinoids have many important functions in the body, including roles in vision, cell growth control and differentiation, bone tissue growth, immune function, and even the activation of genes that may suppress tumors.

[0004]

[0004] Vitamin A, one particularly preferred retinoid, is a fat-soluble vitamin and an essential nutrient for humans. Vitamins encompass several chemically related naturally occurring compounds or metabolites, or vitamers, all of which contain a β-ionone ring. Thus, it includes a group of organic compounds including retinol, retinal, retinoic acid, and retinyl acetate. Vitamin A has many uses, including embryonic development and growth, maintenance of the immune system, and vision.

[0005] Vitamin A exists in several major forms in foods, including retinol. It is also found in animal foods either as retinol or fatty acids bound to retinyl esters, or as the carotenoids alpha-carotene, beta-carotene, gamma-carotene, and the xanthophyll beta-cryptoxanthin (all containing a beta-ionone ring), which function as provitamin A in herbivorous and omnivorous animals that possess enzymes to cleave and convert the provitamin carotenoids to retinal and then to retinol.

[0006]

[0006] Vitamin A deficiency is common in developing countries. Deficiency can occur at any age, but is most common in preschool children and pregnant women. This is because pregnant women need retinol to provide the fetus. Vitamin A deficiency affects approximately one-third of children under the age of five worldwide, and it is estimated that hundreds of thousands of cases of blindness and death result from the childhood disease due to immune system failure.

[0007] Although vitamin A and its vitamers are produced biosynthetically in most animal species through the breakdown of β-carotene, their industrial production relies on chemical synthesis. The first industrialized synthesis of retinol was achieved by Hoffmann-La Roche in 1947. Over the following decades, many companies developed their own proprietary processes. β-Ionone, synthesized from acetone, is the essential starting point for all industrial syntheses. Each process involves extending an unsaturated carbon chain. Nevertheless, such chemical synthesis methods tend to be energy intensive and typically utilize raw materials from petroleum-based sources.

[0008]

[0008] Biosynthetic alternatives are beginning to emerge. Such methods that rely on fermentation tend to impart a significantly lower carbon footprint (i.e., on the order of 30-70%) than their chemical analogues, and can also incorporate biobased, and therefore more circularly sourced, starting materials.

[0009]

[0009] Some methods utilize genetically modified yeast species, such as Saccharomyces cerevisiae, to synthesize retinal and retinol using xylose as the starting substrate. This has been achieved by having the yeast first synthesize β-carotene and then produce retinal via the cleavage enzyme β-carotene 15,15'-dioxygenase.

[0010]

[0010] Further attempts to produce biosynthetically derived isoprenoids, particularly retinoids, using bio-based carbon sources during fermentation (e.g., via oleaginous yeasts including Yarrowia lipolytica) have been described by DSM IP Assets BV, for example in WO 2022090548.

[0011] Regardless of the manufacturing method used, retinoids, particularly vitamin A and its vitamers, especially retinol, are highly sensitive to oxidation and self-heat in oxygen-rich environments. Oxidation of retinoids can cause deterioration in the potency, bioavailability, and / or efficacy of their associated products, as well as pose fire and other safety concerns. Therefore, such industrially produced materials must be prepared and transported at low temperatures and in an oxygen-free atmosphere. When formulated as a dietary supplement or food additive, retinol is often stabilized as retinyl acetate or retinyl palmitate, ester derivatives that tend to be slightly more oxidatively stable. Furthermore, it is highly desirable to minimize the amount of impurities that, by themselves or in conjunction with various retinoids, accelerate the degradation and / or oxidation process.

[0012]

[0012] Notwithstanding the above, there remains a need to formulate and provide retinoid-containing compositions that are at least one of the following: contribute to greater recyclability due to a higher biobased content, impart a lower carbon footprint (especially compared to existing industrially chemically synthesized such compositions), and may provide excellent oxidative stability, preferably oxidative stability comparable to or even superior to current industrially synthesized alternatives. Additionally or alternatively, there is a need to provide fermentatively produced retinoid-containing compositions tailored for superior and / or improved performance in end-use applications. One such example is formulation for superior and / or improved antimicrobial efficacy, which may be important in various cosmetic applications, including, but not limited to, the prevention / treatment / reduction of symptoms of acne or malodor caused by sweating. The above ensures that such retinoid-containing compositions may be commercially suitable for use in a variety of products in various food and beverage (hereinafter "food"), animal feed (hereinafter "feed"), pharmaceutical, and personal care applications.

[0013] [Brief Overview]

[0013] The inventors herein have discovered that one or more of the aforementioned problems can be solved when the present invention according to various aspects and / or embodiments described herein is utilized. Accordingly, several aspects and embodiments of the present invention are described herein. A first aspect is a composition for use in a food, feed, pharmaceutical, or cosmetic product, comprising: (i) a retinoid component comprising a mixture of cis- and trans-isomers; and (ii) a fermentation residue thereof, wherein the retinoid component is present in a weight ratio of greater than 4:1 relative to the fermentation residue, and the cis-isomer is present in an amount by weight of less than 3% by weight relative to the total weight of the retinoid component; and optionally, the biobased carbon content of (i) and (ii) is greater than 50%, or greater than 60%, or greater than 70%, or greater than 90%. According to further embodiments of the first aspect, the composition may contain one or more specific retinoids, including retinyl acetate, retinol, or retinyl palmitate, and / or the fermentation residue may contain one or more fatty acid retinyl esters (FAREs), retinal, retinol, farnesol, a fermentation carbon source, and / or β-carotene in varying amounts as the fermentation residue. In another embodiment of the first aspect, the composition may contain specific amounts of FAREs and cis-isomers of the retinoid mixture. In yet further embodiments, the ratio of the retinoid component (i) to the fermentation residue (ii) may be adjusted to a range other than that specified in the first embodiment, and / or the composition may contain varying levels of biobased carbon content. In yet other embodiments, the composition may exist in a crystalline form, optionally also having a specific average crystal length, or alternatively as a "wet crystal" with a solvent or as an emulsion in oil.

[0014]

[0014] A second aspect of the present invention is a method for fermentatively producing a retinoid-containing composition, comprising: (a) culturing a microorganism under conditions that allow the production of a fermentation product; and (b) isolating the fermentation product to obtain an isolated retinoid-containing composition, wherein the isolated retinoid-containing composition comprises (i) a retinoid component comprising a mixture of cis- and trans-isomers, and (ii) a fermentation residue thereof, wherein the retinoid component is present in a weight ratio of greater than 4:1 relative to the fermentation residue, and the cis-isomer is present in an amount by weight of less than 3% relative to the weight of the total retinoid component.

[0015]

[0015] A third aspect of the present invention is a method for preparing a food, feed, pharmaceutical or personal care (pre)product, comprising the steps of: (1) providing a composition comprising, consisting of, or consisting essentially of (i) a retinoid component comprising a mixture of cis and trans isomers and (ii) a fermentation residue thereof, wherein the retinoid component is present in a weight ratio of greater than 4:1 to the fermentation residue, and the cis isomer is present in an amount by weight of less than 3% by weight relative to the weight of the total retinoid component, and optionally wherein the biobased carbon content of (i) and (ii) is greater than 50%, or greater than 60%, or greater than 70%, or greater than 90%; and (2) blending the composition to produce a blend.

[0016]

[0016] A fourth aspect of the present invention is a beadlet or extrudate produced by the method of any of the embodiments of the third aspect of the present invention and / or incorporating a composition according to any of the embodiments of the first aspect of the present invention and / or produced by any of the embodiments according to the second aspect of the present invention.

[0017]

[0017] A fifth aspect of the present invention is a food, feed, pharmaceutical or personal care product comprising any of the extrudates and / or beadlets according to the fourth aspect and / or produced according to any of the embodiments of the third aspect and / or comprising a composition according to any of the embodiments of the first aspect and / or produced by any of the methods of the second aspect. [Brief explanation of the drawings]

[0018] [Figure 1] FIG. 1 shows a photomicrograph of the crystalline morphology of a fermentatively produced retinoid-containing composition according to one or more aspects and / or embodiments of the present invention (Example 1), with selected crystal length measurements indicated thereon. [Figure 2] FIG. 1 shows a photomicrograph of the crystalline morphology of a fermentatively produced retinoid-containing composition according to one or more aspects and / or embodiments of the present invention (Example 2), with selected crystal length measurements indicated thereon. [Figure 3] FIG. 1 shows a photomicrograph of the crystalline morphology of a fermentatively produced retinoid-containing composition according to one or more aspects and / or embodiments of the present invention (Example 3), with selected crystal length measurements indicated thereon. [Figure 4] FIG. 1 shows a photomicrograph of the crystalline morphology of a fermentatively produced retinoid-containing composition according to one or more aspects and / or embodiments of the present invention (Example 4), with selected crystal length measurements indicated thereon. [Figure 5] 1 shows a photomicrograph of the crystalline morphology of a chemically synthesized retinoid-containing composition (Example 5) with selected crystal length measurements indicated thereon.

[0019] [Detailed explanation]

[0023] A first aspect of the present invention is a composition for use in a food, feed, pharmaceutical or cosmetic product, comprising: (i) a retinoid component comprising a mixture of cis and trans isomers; (ii) its fermentation residue; comprising, consisting of, or consisting essentially of The retinoid component is present in a weight ratio of more than 4:1 relative to the fermentation residue, the cis isomer is present in an amount of less than 3% by weight based on the weight of the total retinoid component; Optionally, the biobased carbon content of (i) and (ii) is greater than 50%, or greater than 60%, or greater than 70%, or greater than 90%.

[0020]

[0024] The composition according to the first aspect of the present invention contains both (i) a retinoid component and (ii) a fermentation residue thereof. As used herein, "retinoid component" refers to the specific retinoid compound that is the predominant retinoid resulting from the fermentation process. As used herein, the term "retinol" refers to a compound of formula (I) (without indication of stereochemistry): [ka] In the formula: R is -CHO, -CHOH, -COOH, -CH(R 1 )2, -CH2OR 2 , -COOR 3 , -CONHR 4 or -CO(NR 4 )2, R1 is independently lower alkoxy or R 1’ and R 1’’ taken together is a lower alkylenedioxy; R 2 is alkanoyl or aroyl, R 3 is alkyl, aryl or aralkyl, and R 4 , R 4’ and R 4’’ are independently hydrogen, alkyl, aryl, or aralkyl; The term "lower alkoxy" refers to an alkoxy group having 1 to 6 carbon atoms, such as methoxy, ethoxy, or propoxy. The term "lower alkylenedioxy" similarly refers to groups containing 1 to 6 carbon atoms, such as methylenedioxy or ethylenedioxy, where the alkyl or alkylene moiety may be linear or branched depending on the number of carbon atoms; The term "alkanoyl" refers to any straight-chain or branched alkanoyl group having from 1 to 18 carbon atoms, such as formyl, acetyl, propionyl, butyryl, stearoyl, and palmitoyl; The term "aroyl" refers to aromatic carboxylic acids having 7 and 11 carbon atoms, including benzoyl or naphthoyl, respectively; The term "alkyl" refers to a straight or branched alkyl group having 1 to 18 carbon atoms, such as methyl, ethyl, propyl, butyl, decyl, dodecyl, hexadecyl, or octadecyl; The term "aryl" by itself or as part of "aralkyl" is phenyl or naphthyl, and The term "aralkyl" includes groups having 1 to 4 carbon atoms in the aliphatic portion, such as benzyl and phenylpropyl.

[0021]

[0025] In particular, the compound of formula (I) defined herein is selected from vitamin A and / or its derivatives, including, but not limited to, one or more retinoids selected from retinol, retinal, retinoic acid, and derivatives such as esters, acetals, and / or amides, particularly esters, more particularly retinyl acetate. Particularly suitable retinyl esters according to the present invention are retinyl acetate, retinyl propionate, retinyl linoleate, retinyl palmitate, retinyl retinoate, and / or retinyl formyl aspartate. Preferably, the compound according to formula (I) is selected from retinol and / or retinyl esters, particularly retinol and / or retinyl acetate. In a preferred embodiment, the retinoid component consists of retinol. In another preferred embodiment, the retinoid component consists of retinyl acetate.

[0022]

[0026] It is understood that the retinoid component may result in multiple isoforms or isomers of retinoid. In such cases, the retinoid component includes all such isomers. When the fermentation process results in multiple retinoid species, it is meant herein that only the most predominantly present retinoid (by weight %) is considered to be the retinoid component.

[0023]

[0027] By extension, it will be understood that any other retinoids produced will not be construed as part of the "retinoid component" for purposes of the present invention. Rather, such retinoids will form part of the "fermentation residue." In addition, fermentation residue will be construed herein to include any other biologically synthesized non-retinoid substances that are nevertheless produced by the microorganisms utilized in the fermentation process, such as precursor compounds like β-carotene or derivative compounds such as various types of fatty acid retinyl esters (FAREs). Such substances may be referred to herein as "internal" fermentation residues, in the sense that they are produced by the microorganisms themselves, and not through the introduction of foreign substances into the process.

[0024]

[0028] However, in addition to other retinoids not produced by microorganisms, the fermentation residue may further contain any other substances or materials that are the result of the processes used to culture, ferment, and isolate the desired retinoid. This therefore includes any "external" impurities or by-products introduced into the composition by the more extensive fermentation and extraction process, including, but not limited to, residual amounts of the carbon source used to feed the retinoid-producing microorganism, any chemical additives used in the fermentation vessel or process, filtration media, wash fluids, solvents, biomass, or any other distillates. When interpreting a composition in the foregoing manner, the fermentatively produced and isolated active retinoid, along with all impurities associated therewith, can form a predefined composition of "active" ingredients, which can then be incorporated into one or more commercially viable (pre)products via a formulation process.

[0025]

[0029] In one embodiment, the fermentation residue of the composition comprises one or more of FARE, retinal, retinol, farnesol, a fermentation carbon source, and β-carotene.

[0026]

[0030] Fermentation residues have generally been known to be undesirable "impurities" in fermentation processes. Specifically, with regard to the fermentative production of retinoids, particularly retinyl acetate, undesirable "by-products" include, but are not limited to, retinal, retinol, FARE, or dihydro forms such as dihydro-retinol or di-dihydro-retinyl acetate, particularly the formation of FARE, which should be reduced or eliminated. The present inventors have surprisingly discovered that certain fermentation residues, such as FARE, may not be as undesirable as previously thought and, when combined with one or more other factors, can actually contribute to oxidative stability. Specifically, the present inventors have surprisingly discovered that FARE content tends to be much less detrimental (by as much as 10 times less significant) to reduced oxidative stability than the corresponding amount of cis-isomers of retinoids present in the retinoid component. Even more potentially significant, the present inventors have observed that retinoid-containing compositions with low amounts of FARE are associated with, or at least correlated with, increased oxidative stability compared to those without FARE.

[0027]

[0031] Thus, in one embodiment, the composition of the first aspect is configured to have a so-called "weighted" selected fermentation residue value (WSFRV). The WSFRV is calculated according to the following formula: WSFRV=C+0.1×F (In the formula, C = weight percent of the cis isomer present relative to the total amount of retinoid component (i) and fermentation residue (ii), and F = weight percent of FARE present relative to the total amount of retinoid component (I) and fermentation residue (II) is determined in accordance with

[0028]

[0032] In certain embodiments, the WSFRV of the composition is configured to be between 0.1 and 2.5, or between 0.1 and 2.0, or between 0.1 and 1.0, or between 0.1 and 0.9, or between 0.4 and 2.5, or between 0.4 and 2.0, or between 0.4 and 1.0, or between 0.4 and 0.9, or between 0.6 and 2.5, or between 0.6 and 2.0, or between 0.6 and 1.0, or between 0.5 and 0.9.

[0029]

[0033] As used herein, FARE is generally meant to include, but is not necessarily limited to, all fatty acid retinyl esters as commonly understood in the technical field to which the present invention applies. Three types of FARE commonly produced in the fermentative production of various retinoids include retinyl acetate, retinyl palmitate, and retinyl oleate. As used herein, when the retinoid component is a retinoid other than retinyl acetate, retinyl palmitate, or retinyl oleate (such as retinol), retinyl acetate, retinyl palmitate, and retinyl oleate are each included as FAREs in the fermentation residue. However, for example, when retinyl acetate is produced as the retinoid component (I), it is not considered a FARE either. In that case, retinyl palmitate and retinyl oleate are still considered as such.

[0030]

[0034] In other embodiments, the fermentation residue comprises one or more FAREs, wherein the one or more FAREs are present in an amount of less than 4 wt.%, or less than 3 wt.%, or less than 2 wt.%, or less than 1 wt.%, based on the total weight of the retinoid components present. In yet other alternative embodiments, the FAREs are present in an amount of 0.05-4 wt.%, or 0.1-4 wt.%, or 0.15-4 wt.%, or 0.2-4 wt.%, or 0.05-3 wt.%, or 0.1-3 wt.%, or 0.15-3 wt.%, or 0.2-3 wt.%, or 0.05-2 wt.%, or 0.1-2 wt.%, or 0.15-2 wt.%, or 0.2-2 wt.%, or 0.05-1 wt.%, or 0.1-1 wt.%, or 0.15-1 wt.%, or 0.2-1 wt.%, based on the total retinoid components.

[0031]

[0035] In another embodiment, the composition comprises, in place of the WSFRV defined above, a mixture of both cis and trans isomers of a retinoid component, preferably comprising retinyl acetate or retinol, the cis isomer being present in an amount of less than 1% by weight based on the weight of the total mixture of cis and trans isomers; and / or Based on the total weight of the retinoid component, the FARE is present in an amount of less than 3.5% by weight or less than 1.5% by weight.

[0032]

[0036] In various embodiments, the retinoid component comprises a mixture of cis and trans isomers, the cis isomers being present in an amount of: 0.1 to 3% by weight, or 0.2 to 3% by weight, or 0.4 to 3% by weight, or 0.5 to 3% by weight, or 0.1 to 2.5% by weight, or 0.2 to 2.5% by weight, or 0.4 to 2.5% by weight, or 0.5 to 2.5% by weight, or 0.1 to 2% by weight, or 0.2 to 2% by weight, or 0.4 to 2% by weight, or 0.5 to 2% by weight, or 0.1 to 1.5% by weight, or 0.2 to 1.5% by weight, or 0.4 to 1.5% by weight, or 0.5 to 1.5% by weight, or 0.1 to 1% by weight, or 0.2 to 1% by weight, or 0.4 to 1% by weight, or 0.5 to 1% by weight, or 0.1 to 0.9% by weight, or 0.2 to 0.9% by weight, or 0.4 to 0.9% by weight, or 0.5 to 0.9% by weight It exists in.

[0033]

[0037] With respect to the retinoid component (I), as mentioned above, it is understood that both cis and trans isomers of a particular retinoid exist. As used herein, terms such as "trans retinyl acetate," "trans retinol," and "trans retinal" are known to those skilled in the art and refer to the trans configuration of all double bonds in such retinyl acetate, retinol, and retinal compounds, including compounds according to formula (I), in accordance with the IUPAC-IUB nomenclature. The terms "trans retinyl ester," specifically "trans retinyl acetate," and "all-trans retinyl ester," specifically "all-trans retinyl acetate," are used interchangeably herein.

[0034]

[0038] Thus, if any one of several double bonds is in the cis configuration rather than the trans configuration, it will be understood herein that all such isomers are cis isomers (e.g., either cis-retinyl acetate or cis-retinol or cis-retinal, depending on the desired retinoid component (i)), and thus are included in determining "% cis isomer" or "cis isomer content." Conversely, only isomers having all double bonds in the trans configuration will be considered trans isomers herein.

[0035]

[0039] By way of example, cis-configured retinoid isomers of interest in the present invention include 9-cis retinol, 11-cis retinol, 13-cis retinol, 9,13-di-cis retinol, 11,13-di-cis retinol, 13-cis-3,4-didehydroretinol, 9-cis-3,4-didehydroretinol, 9,13-di-cis-3,4-didehydroretinol, 13-cis retinal, 11-cis retinal, 11,13-di-cis retinal, 9,13-di-cis retinal, 9-cis retinal, 13-cis-3,4-didehydroretinal, 11,13-di-cis-3,4-didehydroretinal, and the respective cis forms of retinyl acetate (for reviews, see, e.g., Gundersen and See Table 1 in Blomhoff, J. Chromatogr. A935:13-43, 2001).

[0036]

[0040] The inventors have discovered that retinoid-containing compositions having retinoid components configured to have specific cis- and trans-isomer contents can impart specific benefits to the compositions or (pre)products to which they are associated. Potential benefits include, but are not limited to, the use of such compositions as antimicrobial agents having a cis / trans ratio as defined herein. Specifically, it has been observed that when retinyl acetate is used as the retinoid component, the composition is particularly suitable as an antimicrobial agent against Cutibacterium acnes, Corynebacterium xerosis, and Malassezia, preferably M. furfur, while retinol is particularly suitable as an antimicrobial agent against Cutibacterium acnes.

[0037]

[0041] The cis / trans ratios given herein refer to the weight percent ratio of each all-trans isomer of a retinoid / retinoid mixture according to formula (I) to the sum of all cis isomers, as determined by HPLC, assuming equal response factors for all isomers. In preferred embodiments of the first aspect of the invention, the cis / trans ratio of the compound according to formula (I) is less than 0.03, or less than 0.02, or less than 0.01, or is less than 0.0099, 0.0095, 0.009, 0.0085, 0.008, 0.0075, 0.007, 0.0065, 0.006, 0.0055, 0.005, 0.0049, 0.0045, 0.004 3, 0.004, 0.0035, 0.003, 0.0028, 0.0025, 0.0022, 0.002, 0.0018, 0.0015, 0.001, 0.0005 or less, for example, 0.0001 or less, more preferably 0.0099 to 0.0001, 0.008 to 0.001, 0.007 to 0.002, 0.0099 to 0.001, 0.005 to 0.001, 0.00 5~0.0001, 0.005~0.0005, 0.006~0.001, 0.009~0.0001, 0.008~0.0001, 0.006~0.0001, 0.0055~0.0001, 0.005~0.002, 0.007~0.0001, 0.0045~0.0001, 0.004~0.0001, 0.0035~0.0001, 0.003~0.00 The cis / trans ratio is preferably in the range of 0.0049 to 0.0025, 0.004 to 0.0005, 0.006 to 0.0005, 0.006 to 0.0008, 0.006 to 0.0015, 0.006 to 0.002, 0.006 to 0.003, 0.006 to 0.0004, or 0.006 to 0.0025, and most preferably in the range of 0.0049 to 0.0025.

[0038]

[0042] In one preferred embodiment, the retinoid according to formula (I) has a concentration of, in particular, less than 0.03, or less than 0.02, or less than 0.01, preferably 0.0099, 0.0095, 0.009, 0.0085, 0.008, 0.0075, 0.007, 0.0065, 0.006, 0.0055, 0.005, 0.0049, 0.0045, 0.0043, 0.004, 0.005, 0.006 ... 0.0035, 0.003, 0.0028, 0.0025, 0.0022, 0.002, 0.0018, 0.0015, 0.001, 0.0005 or less, for example, 0.0001 or less, more preferably 0.0099 to 0.0001, 0.008 to 0.001, 0.007 to 0.002, 0.009 to 0.0001, 0.008 to 0.0001, 0.005 to 0. 001, 0.005~0.0001, 0.005~0.0005, 0.006~0.001, 0.006~0.0001, 0.0055~0.0001, 0.007~0.0001, 0.0045~0.0001, 0.005~0.002, 0.004~0.0001, 0.0035~0.0001, 0.003~0.0001, 0.005~0.002 5, 0.006 to 0.0005, 0.006 to 0.0008, 0.006 to 0.0015, 0.006 to 0.002, 0.006 to 0.003, 0.006 to 0.0004, 0.006 to 0.0025, 0.004 to 0.0005, and most preferably 0.0049 to 0.0025.

[0039]

[0043] In alternative embodiments, the cis / trans ratio may be described differently, such as the total percent cis isomer content. Thus, in various embodiments, the retinoid component comprises a mixture of cis and trans isomers, with the cis isomer accounting for: 0.1 to 3% by weight, or 0.2 to 3% by weight, or 0.4 to 3% by weight, or 0.5 to 3% by weight, or 0.1 to 2.5% by weight, or 0.2 to 2.5% by weight, or 0.4 to 2.5% by weight, or 0.5 to 2.5% by weight, or 0.1 to 2% by weight, or 0.2 to 2% by weight, or 0.4 to 2% by weight, or 0.5 to 2% by weight, or 0.1 to 1.5% by weight, or 0.2 to 1.5% by weight, or 0.4 to 1.5% by weight, or 0.5 to 1.5% by weight, or 0.1 to 1% by weight, or 0.2 to 1% by weight, or 0.4 to 1% by weight, or 0.5 to 1% by weight, or 0.1 to 0.9% by weight, or 0.2 to 0.9% by weight, or 0.4 to 0.9% by weight, or 0.5 to 0.9% by weight It exists in.

[0040]

[0044] Compositions containing retinyl acetate having such a cis / trans ratio may be useful in preventing, treating, or alleviating symptoms of acne, pruritus, tinea versicolor, dandruff, seborrheic dermatitis atopic dermatitis, psoriasis, and / or malodor caused by sweat, and may be useful as antimicrobial agents, particularly against Cutibacterium acnes, Corynebacterium xerosis, and Malassezia, preferably M. furfur.

[0041]

[0045] In one preferred embodiment, the retinoid according to formula (I) has a cis / trans ratio of less than 0.03, or less than 0.02, or less than 0.01, preferably 0.0099, 0.0095, 0.009, 0.0085, 0.008, 0.0075, 0.007, 0.0065, 0.006, 0.0055, 0.005, 0.0049, 0.0045, 0.0043 ... 0.004, 0.0035, 0.003, 0.0028, 0.0025, 0.0022, 0.002, 0.0018, 0.0015, 0.001, 0.0005 or less, for example, 0.0001 or less, more preferably 0.0099 to 0.0001, 0.008 to 0.001, 0.007 to 0.002, 0.0099 to 0.001, 0.005 to 0.001, 0.005 to 0.0001, 0.005~0.0005, 0.006~0.001, 0.009~0.0001, 0.008~0.0001, 0.006~0.0001, 0.0055~0.0001, 0.005~0.002, 0.007~0.0001, 0.0045~0.0001, 0.004~0.0001, 0.0035~0.0001, 0.003~0.000 The retinol may have a cis / trans ratio of 0.003, preferably 0.006-0.0025, 0.004-0.0005, 0.006-0.0005, 0.006-0.0008, 0.006-0.0015, 0.006-0.002, 0.006-0.003, 0.006-0.0004, or 0.006-0.0025, most preferably a cis / trans ratio of 0.003. Compositions containing retinol with such a cis / trans ratio may be useful for preventing, treating, and / or alleviating acne symptoms, and may be particularly useful as antimicrobial agents against Cutibacterium acnes.

[0042]

[0046] The mixture of cis and trans isomers of retinoids according to formula (I) of the present invention, particularly retinyl esters, preferably retinyl acetate and / or retinol, can be prepared by either mixing each all-trans isomer with one or more cis isomers obtained by chemical or biological processes.Methods for preparing such all-trans isomers and / or cis isomers are known to those skilled in the art.Alternatively, the mixture of the isomer ratios can be prepared by adjusting the process accordingly.

[0043]

[0047] Advantageously, in a preferred embodiment of the first aspect of the present invention, the retinoid, in particular the retinyl ester, such as retinyl acetate and / or retinol, is biologically produced through a fermentation process.

[0044]

[0048] In certain embodiments, mixtures and / or compositions comprising the aforementioned mixtures of cis- and trans-isomers of retinoids, particularly retinyl esters, preferably retinyl acetate and / or retinol, as defined herein, are produced in a fermentation process using suitable retinol-producing host cells, such as bacterial or fungal cells (see, for example, Sun et al., ACS Synth. Biol. 2019 Sep 20;8(9):2131-2140; Jang et al., Microbial Cell Facts 2011,10:59), which express the respective enzymes, such as trans-selective enzymes, i.e., β-carotene oxidase (BCO), involved in the biosynthesis of retinol by converting β-carotene to retinal, which can then be enzymatically converted to retinol and / or retinyl esters, particularly retinyl acetate. The fermentation is fed with ethanol, corn sugar, or corn oil, all of which are derived from agricultural production. Fermentation products containing retinyl esters, specifically retinyl acetate, can be extracted with the fatty phase and then purified into a crystalline form.

[0045]

[0049] Advantageously, fermentatively produced trans-retinyl esters, particularly trans-retinyl acetate and / or trans-retinol, can be treated with heat to form cis-retinyl esters, particularly cis-retinyl acetate and / or cis-retinol, to achieve the appropriate levels described in all embodiments of the present invention (see, e.g., McBee et al., JBC, Vol. 276, No. 51, pp. 48483-48493, 2001).

[0046]

[0050] As used herein, the terms "biologically produced" or "fermentatively produced" mean that a retinoid, particularly a retinyl ester, particularly retinyl acetate and / or retinol, is produced using a biotechnological process, such as a fermentation process, comprising culturing a suitable (carotenoid and / or retinoid-producing) host cell expressing the respective enzymes involved in the further conversion of a suitable carbon source to the retinyl esters, particularly retinyl acetate and / or retinol, as defined herein; the host cell may be selected from bacteria, fungi, particularly yeast, plants, or algae. "Bio-produced," "biologically derived," and "biologically produced" are used interchangeably herein. Thus, the aforementioned biologically produced retinoid according to formula (I) is composed of carbon derived from atmospheric carbon dioxide (also referred to as atmospheric carbon) converted by green plants into sugars and starch. This also includes the use of isolated and / or immobilized enzymes in the process to produce the retinoid mixtures defined herein, for example, the use of specific enzymes that can selectively catalyze the formation of retinoids, particularly retinyl esters, preferably retinyl acetate and / or retinol mixtures, having specific trans / cis ratios as defined herein.

[0047]

[0051] In one embodiment, as used herein, a "biobased" compound has a C-14 / C-12 isotope ratio ranging from 1:0 to greater than 0:1, as opposed to fossil-derived compounds which have a C-14 / C-12 isotope ratio of 0:1. The biobased content of a compound can be measured by known radiocarbon and isotope ratio mass spectrometry or accelerator mass spectrometry, such as, for example, ASTM test method D6866-05, which measures the C-14 / C-12 isotope ratio in a sample and indicates the percent biobased content of the sample compared to a standard 100% biobased material.

[0048]

[0052] As used herein, "atmospheric carbon" refers to carbon atoms in carbon dioxide molecules that have been recently released into the Earth's atmosphere in the past few decades. Such carbon mass is identifiable by the presence of certain radioactive isotopes, as described herein. "Green carbon," "atmospheric carbon," "green carbon," "life cycle carbon," "non-fossil carbon," "non-petroleum carbon," "atmospheric carbon," and "bio-based carbon" are used interchangeably herein.

[0049]

[0053] According to certain preferred embodiments of the present invention, advantageously, retinoids, particularly retinol and / or retinyl esters, preferably retinyl acetate, are produced solely from organic, renewable, biobased feedstocks, particularly fermentatively produced, and such retinol and / or retinyl esters, preferably retinyl acetate, have a zero anthropogenic CO2 emission profile upon biodegradation because all CO2 molecules released during degradation from such "fermentation-derived" or "fermentation-produced" retinoids have atmospheric origins. Thus, there is zero net CO2 emission to the atmosphere. Therefore, in preferred embodiments, the fermentation carbon source comprises, consists of, or essentially consists of a biogenic carbon source.

[0050]

[0054] In various embodiments of the first aspect, the biobased carbon content of the composition is greater than 60%, or greater than 70%, or greater than 80%, or greater than 90%, or greater than 99%, or even about 100%, and in still other embodiments, the biobased carbon content of the retinoid component (i) and its fermentation residue (ii) is greater than 60%, or greater than 70%, or greater than 80%, or greater than 90%, or greater than 99%, or even about 100%. Biobased content can be determined by any suitable method, such as via ASTM D6866-20 or the methods described elsewhere herein below. 14 It may be determined according to any suitable means, such as by C isotope characterization.

[0051]

[0055] As previously mentioned, the fermentation residue may contain one or more of a variety of substances, such as FARE, beta-carotene, retinol, retinal, and / or a fermentation carbon source. In other embodiments, particularly when the retinoid component is retinyl acetate, the fermentation residue may contain each of FARE, beta-carotene, retinol, retinal, and a fermentation carbon source.

[0052]

[0056] Various retinol isotopes, such as E-retinol, can be included as retinol.Similarly, various retinal isotopes, such as 9Z-retinal, can also be used.The present inventors have surprisingly observed that the composition containing 9Z-retinal can show improved oxidative stability when compared with that not containing such fermentation residue, and interestingly, oxidative stability seems to be inversely correlated with the amount of 9Z-retinal present in the retinoid-containing composition.

[0053]

[0057] Thus, in various embodiments of the first aspect, the fermentation residue contains 9Z-retinal, but the 9Z-retinal is present in an amount of less than 10% by weight, or less than 8% by weight, or less than 6% by weight, or less than 5% by weight, or less than 3% by weight, relative to the total weight of the retinoid components present. In other embodiments, the composition ultimately comprises 0.1 to 10 wt.%, or 0.5 to 10 wt.%, or 1 to 10 wt.%, or 1.5 to 10 wt.% of 9Z-retinal, or 0.1 to 8 wt.%, or 0.5 to 8 wt.%, or 1 to 8 wt.%, or 1.5 to 8 wt.% of 9Z-retinal, or 0.1 to 6 wt.%, or 0.5 to 6 wt.%, or 1 to 6 wt.%, or 1.5 to 6 wt.% of 9Z-retinal, or 0.1 to 5 wt.%, or 0.5 to 5 wt.%, or 1 to 5 wt.%, or 1.5 to 5 wt.% of 9Z-retinal, or 0.1 to 3 wt.%, or 0.5 to 3 wt.%, or 1 to 3 wt.%, or 1.5 to 3 wt.% of 9Z-retinal, based on the total weight of the retinoid components present.

[0054]

[0058] Yet another potential component of the fermentation residue is β-carotene. The presence of β-carotene is thought to contribute to the more orange or pink hue observed in fermentation-produced retinoids compared to their chemically synthesized analogs. β-Carotene confers its own health benefits, and therefore, the presence of β-carotene as a fermentation residue in a retinoid-containing composition is not necessarily considered undesirable and may be desirable depending on the intended end use. Thus, in one embodiment, the fermentation residue contains β-carotene, and the β-carotene is present in an amount of less than 4% by weight, or less than 3% by weight, or less than 2% by weight, or less than 1% by weight, or less than 0.5% by weight, based on the total weight of the retinoid components present.

[0055]

[0059] However, in other embodiments of the first aspect, the composition comprises 0-4 wt. %, or 0.2-4 wt. %, or 0.5-4 wt. %, or 1-4 wt. % β-carotene, or 0 to 3% by weight, or 0.2 to 3% by weight, or 0.5 to 3% by weight, or 1 to 3% by weight of β-carotene, or 0 to 2% by weight, or 0.2 to 2% by weight, or 0.5 to 2% by weight, or 1 to 2% by weight of β-carotene, or 0 to 1% by weight, or 0.2 to 1% by weight, or 0.5 to 1% by weight of β-carotene, or 0 to 0.5% by weight or 0.2 to 0.5% by weight of β-carotene Includes.

[0056]

[0060] Another potential fermentation residue component is farnesol. Farnesol has been identified as a potential skin irritant, and therefore its presence in retinoid-containing compositions is undesirable. Therefore, it is preferable to limit the amount of farnesol present in retinoid-containing compositions to as far as practical, for example, below the detection limit of many standard analytical techniques (e.g., less than 0.1% by weight of the total composition to which they relate). Thus, in one embodiment, the composition contains 0 to 10,000 ppm, or 2 to 10,000 ppm, or 5 to 10,000 ppm, or 10 to 10,000 ppm, or 100 to 10,000 ppm, of farnesol, or 0 to 5,000 ppm, or 2 to 5,000 ppm, or 5 to 5,000 ppm, or 10 to 5,000 ppm, or 100 to 5,000 ppm of farnesol; 0 to 1,000 ppm, or 2 to 1,000 ppm, or 5 to 1,000 ppm, or 10 to 1,000 ppm, or 100 to 1,000 ppm of farnesol; or Farnesol at 0 to 100 ppm, 2 to 100 ppm, 5 to 100 ppm, or 10 to 100 ppm In a particularly preferred embodiment, the composition is free or substantially free of farnesol.

[0057]

[0061] Another potential fermentation residue component is a dihydroretinoid, including dihydroretinol and / or dihydroretinyl acetate, particularly in the range of 0.2 to 0.01% by weight or less, e.g., 0.2, 0.18, 0.16, 0.15, 0.14, 0.12, 0.1, 0.05, or 0.01% or less, preferably 0.2 to 0.1, 0.17 to 0.06, 0.1 to 0.05, or 0.04 to 0.01%, and more preferably 0.1% or less, based on the total components in the fermentatively produced retinoid-containing composition as a whole. Also preferably, the proportion of dihydroretinyl acetate is about 0.05% or less, particularly about 0.01% or less, based on the total retinoids in the composition.

[0058]

[0062] Yet another potential fermentation residue component is unconsumed raw material or a fermentation carbon source. As noted above, it is desirable to maximize the recyclability of the retinoid composition from which such carbon sources are produced, and therefore it is desirable to utilize a biogenic carbon source. Various carbon sources suitable for retinoid fermentation are described in WO2022090548A1, which is incorporated herein by reference in its entirety as if fully set forth herein.

[0059]

[0063] Specifically, suitable carbon sources for use in the present invention may be selected from the group consisting of straight-chain alkanes, free fatty acids containing triglycerides, particularly vegetable oils, such as oils derived from corn, soybean, olive, sunflower, canola, cottonseed, rapeseed, sesame, safflower, grapeseed, or mixtures thereof, containing the respective free fatty acids, for example, oleic acid, palmitic acid, or linoleic acid. Suitable carbon sources may further be selected from ethanol, glycerol, or glucose, and mixtures of one or more of the above carbon sources. In one embodiment, the present invention relates to a process for the production of retinoids, particularly retinyl acetate, comprising culturing retinyl acetate-producing host cells, preferably oleaginous yeast cells, such as Yarrowia, in a two-phase culture system in the presence of a lipophilic solvent as defined herein, under suitable culture conditions. Preferably, the lipophilic solvent selected from solvents commercially available as Isopar™ liquids, in particular Isopar M, Isopar N, Isopar K, Isopar L, Isopar H or solvents with equivalent or identical properties obtained from other suppliers, is not consumed or evaporated during the fermentation process.

[0060]

[0064] In one embodiment, the amount of fermentable carbon source is present in an amount less than 2 wt%, or less than 1 wt%, or less than 0.75 wt%, based on the total weight of the retinoid component present. In various other embodiments, the fermentable carbon source is present at 0.05-2 wt%, or 0.1-2 wt%, or 0.2-2 wt%, or 0.05-1 wt%, or 0.1-1 wt%, or 0.2-1 wt%, or 0.05-0.5 wt%, or 0.1-0.5 wt%, or 0.2-0.5 wt%, based on the weight of the retinoid component.

[0061]

[0065] As an additional source of fermentation residues, so-called second phase solvents may be present. These, otherwise known as lipophilic solvents, may include isoparaffins, including mixtures of alkanes, cycloparaffins, isoalkanes, cycloalkanes, or dodecane. Solvents may be natural or synthetic. Examples of commercially available useful solvents may be selected from Total, e.g., sane® solvents, Shell, e.g., ShellSolTD or ShellSolT, ExxonMobile, e.g., Isopar™ fluids, in particular, e.g., Isopar M, Isopar N, Isopar H, Isopar K, Isopar L, or mixtures thereof or mixtures with isododecane isomers, such as those commercially available under the trade name AC365770010 (Acros Organics). Preferably, the second phase solvent is selected from isoparaffins including, for example, Isopar M, Isopar N, Isopar H, Isopar K, Isopar L and mixtures thereof, more preferably Isopar N, Isopar L and / or Isopar M.

[0062]

[0066] Further suitable lipophilic solvents, i.e., second phase solvents, as defined herein and used for the present invention, which are minimally lost and / or lost during fermentation, can be selected from lipophilic solvents including mixtures of n-alkanes, isoalkanes, and hydrocarbons. Solvents can be natural or synthetic. Examples of commercially available useful solvents can be selected from Exxon Mobil, such as those sold under the trade names Exxsol D60, D80, D95, or D110.

[0063]

[0067] It is understood that useful solvents include the commercially available solvents listed above as well as respective solvents having the same or equivalent properties but available from known / other suppliers.

[0064]

[0068] As used herein, solvents have properties comparable or identical to Isopar fluids, including Isopar M, Isopar H, Isopar K, and Isopar L, and are defined as branched isomers of linear alkanes containing 6 to 26 carbon atoms, preferably terminally methylated, possibly chemically bonded from smaller alkane precursors in strong acid, followed by hydrogenation with H2 and a catalyst such as nickel or platinum to remove unsaturation and trace aromatics. These are known to current industrial suppliers as Isopar (Exxon Mobil Chemical), Soltrol (Chevron Phillips Chemical Company), Shellsol OMS (Royal Dutch Shell), isooctane, and iso-dodecane. Specifically, the use of Isopar M is preferred. Their use in consumer products is outlined in a review by Johnson et al., Int J Toxicol. 2012 Nov-Dec;3l(6 Suppl):269S-95S. As used herein, the solvent has properties comparable to or identical to Exxsol D60, D80, D95, D110, which are narrow-boiling distillation cuts from cracked hydrocarbons that have been reduced by catalytic hydrogenation to remove aromatics and unsaturations.

[0065]

[0069] Thus, in one embodiment, the fermentation residue comprises an isoparaffinic fluid. In various related embodiments, the isoparaffinic fluid is present in an amount of less than 3 wt. %, or less than 2 wt. %, or less than 1.5 wt. %, or less than 1.25 wt. %, based on the total weight of the retinoid component (i) and its fermentation residue (ii), or the isoparaffinic fluid is present in an amount of 0.1-2 wt. %, or 0.2-2 wt. %, or 0.5-2 wt. %, or 0.1-1.5 wt. %, or 0.2-1.5 wt. %, or 0.5-1.5 wt. %, based on the total weight of the retinoid component (i) and its fermentation residue (ii). In a preferred embodiment, the isoparaffinic fluid, when present, comprises, consists of, or consists essentially of Isopar M.

[0066]

[0070] The fermentatively produced composition of the first aspect is further adjusted to have specific amounts of (i) and (ii) relative to each other. That is, in all embodiments of the first aspect, the retinoid-containing composition is configured so that the retinoid component is present in a weight ratio of greater than 4:1 relative to the fermentation residue. Thus, it is desirable to maintain a very high level of the "active" ingredient, i.e., the retinoid component, which is preferably retinol, retinyl acetate, or retinyl palmitate, relative to the fermentation residue. Thus, in other embodiments, the weight ratio of the retinoid component to the fermentation residue is adjusted to greater than 5:1, or greater than 6:1, or greater than 10:1, or greater than 20:1, or even greater than 25:1.

[0067]

[0071] However, the inventors have still surprisingly determined that the presence of the fermentation residues described elsewhere herein above, and in particular the specific types, amounts, and ratios of the fermentation residue components, are still associated with beneficial effects on the compositions to which they are associated, such as superior oxidative stability and / or antimicrobial efficacy. As described below, such advantages make the associated fermentatively produced retinoid-containing compositions particularly suitable for use in various formulation techniques, such as spray drying or extrusion. This, in turn, facilitates their use in various food, feed, pharmaceutical, or personal care products or pre-products (hereinafter "(pre)products"). Thus, in other embodiments, the retinoid component, preferably retinyl acetate, retinol, or retinyl palmitate, is present in a weight ratio relative to the fermentation residues of 4:1 to 100:1, or 5:1 to 100:1, or 6:1 to 100:1, or 10:1 to 100:1, or 20:1 to 100:1, or 25:1 to 100:1, or a weight ratio of 4:1 to 50:1, or 5:1 to 50:1, or 6:1 to 50:1, or 10:1 to 50:1, or 20:1 to 50:1, or 25:1 to 50:1, or a weight ratio of 4:1 to 35:1, or 5:1 to 35:1, or 6:1 to 35:1, or 10:1 to 35:1, or 20:1 to 35:1, or 25:1 to 35:1, or A weight ratio of 4:1 to 30:1, or 5:1 to 30:1, or 6:1 to 30:1, or 10:1 to 30:1, or 20:1 to 30:1, or 25:1 to 30:1 It exists in.

[0068]

[0072] While it has not previously been recognized that the contents, ratios, and types of the fermentation residue profile described herein are associated with one or more beneficial effects, those skilled in the art to which this invention pertains will recognize that such fermentation residue profiles can be tailored or configured as needed by numerous known methods and techniques. Specifically, the microorganisms utilized in the fermentation process can be selected and / or engineered to produce specific desired retinoid components and / or fermentation residues, as described elsewhere herein. Fermentation conditions can also be modified accordingly, including the selection and amount of various additives (i.e., antifoam agents) or the nature, type, and amount of carbon source and / or lipophilic solvent used. Furthermore, those skilled in the art can readily tailor the fermentation profile by the nature and number of downstream process steps, such as distillation, crystallization, filtration, and / or washing conditions. Fermentation and isolation conditions are further described elsewhere herein below.

[0069]

[0073] The retinoid-containing composition according to the first aspect can be provided in any suitable form, but is preferably produced in a manner that maximizes ease of handling and transport. In one embodiment, the retinoid-containing composition according to the first aspect is provided in a crystalline or predominantly crystalline form. As intended herein, crystalline form refers to a composition composed of solid materials or particles whose components are arranged in a highly ordered microscopic structure, forming a crystal lattice extending in all directions. The inventors have discovered that the fermentation residue profile can affect the crystal structure formed. Various retinoid-containing compositions in crystalline form are shown in Figures 1 to 5. Figures 1, 2, 3, and 4 show the crystal morphologies of the fermentatively produced retinoid-containing compositions of Examples 1 to 4, respectively, used in the following examples. The crystals were determined to be either elongated, rhomboid-shaped, with lengths on the order of 200 to 900 microns or more, as shown in Figure 1, 3, or 4, or more rectangular, with crystal lengths on the order of 100 to 200 microns, as shown in Figure 2. In contrast, Figure 5 shows the significantly different crystal structure that results when a chemically synthesized retinoid-containing composition without fermentation residue is used; the crystals in such cases are typically on the order of 100-200 microns or less. As can be seen, the average crystal size or length, as specifically determined by computer-automated microscopy analysis, varies between different fermentatively produced examples and chemical analogs. While not wishing to be bound by any theory, the inventors believe that crystal size may affect the oxidative stability of the relevant retinoid-containing composition.

[0070]

[0074] Thus, in certain embodiments, the fermentatively produced retinoid-containing composition is present in a crystalline form, the crystalline form having an average particle length D of greater than 100 microns, or greater than 200 microns, or greater than 300 microns, or greater than 400 microns, or between 100 and 1200 microns, or between 100 and 1000 microns, or between 100 and 800 microns, or between 200 and 1200 microns, or between 200 and 1000 microns, or between 200 and 800 micrometers, as determined by microscopic imaging methods. 50The crystals include a plurality of crystals having the formula:

[0071]

[0075] The form in which the fermentatively produced retinoid component appears can be so-called "dry crystalline" or "wet crystalline." In the dry crystalline form, the composition generally does not contain any liquid components and generally behaves and appears as a granular or powder composition. In the wet crystalline form, the composition contains a large amount of crystals but also contains a residual appreciable amount of liquid, such as associated solvent. As such, it may appear and behave as a slurry.

[0072]

[0076] In yet another embodiment, a composition containing a retinoid (i) and a fermentation residue (ii) is dissolved, mixed, and / or (most preferably) emulsified in oil. In a preferred embodiment, if not present in crystalline form, the retinoid component (i) and its fermentation residue (ii) are emulsified in a stabilized oil. This stabilized oil emulsion preferably contains any suitable oil, preferably a triglyceride, such as, but not limited to, vegetable oil and / or fat. In this sense, it will be understood that oil is not included as part of the fermentation residue component (ii).

[0073]

[0077] Suitable stabilizers for the stabilized emulsion preferably also include fat-soluble antioxidants, which may include, but are not limited to, ascorbic acid or its salts, tocopherol, butylated hydroxytoluene (BHT), butylated hydroxyanisole (BHA), propyl gallate, tert. butylhydroxyquinoline, ethoxyquin, and / or ascorbic acid esters of fatty acids. In this sense, it will be understood that stabilizers are not included as part of the fermentation residue component (ii).

[0074]

[0078] As noted above, compositions according to the first aspect are preferably configured to impart advantageous properties, such as excellent oxidative stability. Thus, when utilizing one or more compositions configured as described above, the amount of retinoid component (preferably retinyl acetate, retinol, or retinyl palmitate), measured relative to the weight of the total composition, is within 5 wt. %, or within 4 wt. %, or within 3 wt. % of the amount of said retinoid component present in the composition after subjecting the composition to a thermal stability test in which the composition is heated to 105° C. for 3 hours, according to the method described elsewhere herein.

[0075]

[0079] A second aspect of the present invention is a method for fermentatively producing a retinoid-containing composition, comprising: (a) culturing a microorganism under conditions that allow for the production of a fermentation product; (b) isolating the fermentation product to obtain an isolated retinoid-containing composition; the isolated retinoid-containing composition comprising: (i) a retinoid component comprising a mixture of cis and trans isomers; (ii) its fermentation residue; Including, The retinoid component is present in a weight ratio of more than 4:1 relative to the fermentation residue, The cis isomer is present in an amount of less than 3% by weight based on the weight of the total retinoid component.

[0076]

[0080] Embodiments according to the second aspect of the present invention relate to fermentation methods for producing retinoid-containing compositions. In all such embodiments, the method first comprises culturing a microorganism under conditions that allow for the production of a desired fermentation product, preferably comprising a retinoid component as described and used in all embodiments of the first aspect of the present invention.

[0077]

[0081] As will be appreciated by those skilled in the art, any suitable microorganism can be used. Particularly suitable microorganisms include fungi such as those of the genus Yarrowia, which are oleaginous in that they can accumulate lipids to at least 20% of their dry cell weight; furthermore, possibly as a result of genetic engineering, such microorganisms can produce the desired product, preferably accumulating it to at least 1% of their dry cell weight. A general method for culturing carotenoids (precursors of retinoids) is described in U.S. Patent No. 7,851,199, assigned to DSM IP Assets BV, which is incorporated herein by reference in its entirety as if fully set forth herein. The fermentative production of isoprenoids, a broader class of compounds that includes retinoids, is described in International Publication No. WO 2022090548, assigned to DSM IP Assets BV, which is also incorporated herein by reference in its entirety as if fully set forth herein.

[0078]

[0082] Those skilled in the art will readily appreciate that a variety of yeast and fungal strains exist that are naturally oleaginous or that naturally produce desired retinoids. Any of these strains can be utilized as host strains according to the second aspect of the present invention and can be engineered or otherwise manipulated to produce oleaginous, particularly retinoid-producing, strains of the present invention. Alternatively, strains that are not naturally oleaginous or that do not produce retinoids can be used. Furthermore, even if a particular strain has a natural ability to be oleaginous or to produce retinoids, that natural ability can be adjusted as described herein to alter the level of lipid and / or retinoid production. In certain embodiments, engineering or manipulating the strain results in altering the types of lipids and / or retinoids produced. For example, a strain may be naturally oleaginous and / or retinogenic, but engineering or manipulating the strain can be used to alter the types of lipids accumulated and the types of retinoids produced.

[0079]

[0083] When selecting a particular yeast or fungal strain for use in accordance with the second aspect of the present invention, it will generally be desirable to select a yeast or fungal strain whose cultural characteristics are suitable for commercial-scale production. For example, it will generally (though not always) be desirable to avoid filamentous organisms or organisms with particularly unusual or stringent requirements for growth conditions. However, filamentous organisms may be selected as host cells if conditions for commercial-scale production that allow their use are applicable. In some embodiments of the present invention, it will be desirable to utilize edible organisms as host cells, as they may optionally be incorporated directly into food or feed additives or dietary supplements, if desired. For ease of production, some embodiments of the invention utilize host cells that are genetically tractable, amenable to molecular genetics (e.g., capable of being efficiently transformed with particularly established or available vectors, optionally incorporating and / or integrating multiple genes, e.g., sequentially, and / or having known gene sequences), do not have complex growth requirements (e.g., a need for light), are mesophilic (e.g., preferring growth temperatures in the range of about 25-32°C), can assimilate a variety of carbon and nitrogen sources, and / or can grow to high cell densities. Alternatively or additionally, various embodiments of the invention utilize host cells that grow as unicellular rather than multicellular organisms (e.g., as mycelia).

[0080]

[0084] Generally, when it is desired to utilize a naturally occurring oleaginous organism in accordance with the second aspect of the present invention, any modifiable and culturable oleaginous organism may be used. In certain embodiments of the present invention, yeasts or fungi of the genera including, but not limited to, Blakeslea, Candida, Cryptococcus, Cunninghamella, Lipomyces, Mortierella, Mucor, Phycomyces, Rhodosporidium, Rhodotorula, Trichosporon, and Yarrowia are utilized.In certain embodiments, the strains selected from the group consisting of Blakeslea trispora, Candida pulcherrima, C. revkaufi, C. tropicalis, Cryptococcus curvatus, Cunninghamella echinulata, C. elegans, C. japonica, Lipomyces starkeyi, L. lipoferus, Mortierella alpina, and the like are .... Alpina, M. isabellina, M. ramanniana, M. vinacea, Mucor circinelloides, Phycomyces blakesleanus, Pythium irregulare, Rhodosporidium toruloides, Rhodotorula glutin, R. gracilis, R. graminis, R. mucilaginosa, R. pinicola, Trichosporon pullulans Species of organisms that have been used include T. pullans, T. cutaneum, and Yarrowia lipolytica.

[0081]

[0085] Those skilled in the art will understand that the selection of a particular host cell for use in accordance with embodiments of the second aspect of the invention will also influence various aspects such as, for example, the choice of expression sequences utilized with any heterologous polypeptides introduced into the cell, and will also influence culture conditions and the like. Much is known regarding the various gene regulatory, protein targeting sequence, and culture requirements of the various host cells that may be utilized in accordance with the second aspect of the present invention (e.g., for Yarrowia, see Barth et al. FEMS Microbiol Rev. 19:219, 1997; Madzak et al. J Biotechnol. 109:63, 2004; for Xanthophyllomyces, see Verdoes et al. Appl Environ Microbiol 69:3728-38, 2003; Visser et al. FEMS Yeast Res 4:221-31, 2003; Martinez et al. Antonie Van Leeuwenhoek. 73(2):147-53, 1998; Kim et al. Appl Environ Microbiol. 64(5):1947-9, 1998; Wery et al. Appl Environ Microbiol. 64(5):1947-9, 1998). (See, e.g., Guthrie and Fink, Methods in Enzymology 194:1-933, 1991, for Saccharomyces.) In certain embodiments, for example, a targeting sequence of the host cell (or a closely related analog) may be useful to include to direct the heterologous protein to subcellular localization. Thus, such useful targeting sequences may be added to the heterologous sequence for proper subcellular localization of activity. In other embodiments (e.g., addition of a mitochondrial targeting sequence), the heterologous targeting sequence may be removed or modified with the selected heterologous sequence (e.g., modification or removal of a plant source organism chloroplast targeting sequence).

[0082]

[0086] An embodiment of the second aspect also includes isolating the fermentation product to obtain an isolated retinoid-containing composition. As mentioned above, lipid body accumulation in oleaginous organisms is generally induced by growing the relevant organism in the presence of an excess carbon source, for example, by limiting nitrogen. Specific conditions that induce such accumulation have previously been established for several different oleaginous organisms (see, e.g., Wolf (ed.) Non-conventional yeasts in biotechnology Vol. 1, Springer-Verlag, Berlin, Germany, pp 313-338; Lipids 18(9):623, 1983; Indian J. Exp. Biol. 35(3):313, 1997; J. Ind. Microbiol. Biotechnol. 30(1):75, 2003; Bioresource Technol. 95(3):287, 2004, each of which is incorporated herein by reference in its entirety).

[0083]

[0087] Methods and systems for isolating lipid bodies have been established for a wide variety of oleaginous organisms (see, e.g., U.S. Patent Nos. 5,164,308, 5,374,657, 5,422,247, 5,550,156, 5,583,019, 6,166,231, 6,541,049, 6,727,373, 6,750,048, and 6,812,001, each of which is incorporated herein by reference in its entirety). Briefly, cells are typically harvested from culture, often by spray drying, filtration, or centrifugation. In some cases, the cells are homogenized and then subjected to supercritical fluid extraction or solvent extraction (e.g., using solvents such as chloroform, hexane, methylene chloride, methanol, isopropanol, ethyl acetate, etc.) to obtain a crude oil suspension. This oil suspension can optionally be refined as known in the art. The refined oil can be used directly as a feed or food additive. Alternatively, or in addition, retinoids can be isolated from the oil using conventional techniques.

[0084]

[0088] Given that retinoids are generally sensitive to oxidation, many embodiments of the present invention utilize oxidation stabilizers (e.g., tocopherol, vitamin C, ethoxyquin, vitamin E, BHT, BHA, TBHQ, etc., or combinations thereof) during and / or after retinoid isolation. Alternatively or additionally, microencapsulation, e.g., with proteins, may be used to add a physical barrier to oxidation and / or improve handling.

[0085]

[0089] Isolation of the fermentation product may further include one or more additional downstream processing steps, including, but not limited to, various processes and techniques for crystallization, distillation, filtration, and / or washing. Such processes are known and would be understood by those skilled in the art to which the present invention applies.

[0086]

[0090] Regardless of the particular type and nature of both the culturing (a) and isolation steps (b) according to embodiments of the second aspect, the process is configured to obtain a retinoid-containing composition having (i) a retinoid component comprising a mixture of cis- and trans-isomers, and (ii) a fermentation residue thereof, wherein the retinoid component is present in a weight ratio of greater than 4:1 relative to the fermentation residue, and the cis-isomer is present in an amount, by weight, of less than 3% by weight relative to the weight of the total retinoid component. It will be understood that the retinoid-containing composition produced in embodiments of the second aspect will preferably be according to any one of the embodiments described above with respect to the first aspect of the invention.

[0087]

[0091] Optionally, the method according to the second aspect also includes combining the isolated retinoid-containing composition, retinoid-containing composition, dry retinoid crystal composition, and / or isolated retinol product with one or more other additives, which may be done not only to create a stabilized formulation for subsequent shipping (further description of certain non-limiting processes is provided below), but also to create a (pre)product for food, feed, pharmaceutical, or personal care applications.

[0088]

[0092] A third aspect of the present invention is a method for preparing a food, feed, pharmaceutical or personal care (pre)product, comprising the steps of: (a) a composition comprising: (i) a retinoid component comprising a mixture of cis and trans isomers; (ii) its fermentation residue; wherein the retinoid component is present in a weight ratio of greater than 4:1 relative to the fermentation residue thereof, and the cis isomer is present in an amount of less than 3% by weight based on the weight of the total retinoid component, and optionally also wherein the biobased carbon content of (i) and (ii) is greater than 50%, or greater than 60%, or greater than 70%, or greater than 90%; (b) formulating the composition to produce a formulation, preferably for use as a food, feed, pharmaceutical or personal care (pre)product; The method includes:

[0089]

[0093] According to embodiments of the third aspect, a composition is provided that comprises, consists of, or consists essentially of (i) a retinoid component comprising a mixture of cis- and trans-isomers and (ii) a fermentation residue thereof, wherein the retinoid component is present in a weight ratio of greater than 4:1 with respect to the fermentation residue thereof, and the cis-isomer is present in an amount, by weight, of less than 3% by weight based on the weight of the total retinoid component, and optionally also wherein the biobased carbon content of (i) and (ii) is greater than 50%, or greater than 60%, or greater than 70%, or greater than 90%. Preferably, the composition provided is any of the fermentatively produced retinoid-containing compositions described in any of the embodiments of the first aspect of the invention, and is also preferably provided by any of the methods described in any of the embodiments of the second aspect.

[0090]

[0094] For provided compositions, it is often desirable to formulate such compositions to increase storage, handling and transportation, bioavailability, compatibility, or other properties for subsequent suitability, with or without one or more other additives (pre-products) for use in various end uses. Thus, the formulation techniques described according to embodiments of this third aspect preferably impart improved stability and / or suitability for use as food, feed, pharmaceutical, or personal care (pre-)products.

[0091]

[0095] Various formulation techniques are well known in the art to which the present invention is applied. The typical purpose of such techniques is to incorporate the main desired component (in this case, a retinoid component) into a stabilized emulsion or suspension in various ways. One known type of formulation technique involves microencapsulation. Microencapsulation techniques are known and commonly described, and may include, but are not limited to, spray drying, beadlet technology, coacervation, or spray chilling. A specific microencapsulation technique is described in U.S. Pat. No. 8,765,186 B2, which is incorporated herein by reference in its entirety as if fully set forth herein.

[0092]

[0096] In one embodiment, the method of the third aspect comprises a beadlet compounding process. Thus, in one embodiment, the method of preparing a food, feed, pharmaceutical or personal care (pre)product according to the third aspect comprises, in relation to compounding step (b), (1) a sub-step of dissolving a matrix component in water to prepare a matrix solution; (2) optionally, preferably also in oil, heating a composition according to any of the preceding claims to obtain an active phase; (3) a substep of emulsifying the active phase in the matrix solution to obtain a dispersion; (4) drying the dispersion in the presence of a capture medium to obtain at least one beadlet; Includes.

[0093]

[0097] In the dissolution substep, the matrix material is typically dissolved in water to produce a matrix solution. This forms the medium in which the microencapsulated active beadlets are formed. Any suitable matrix component, such as a lignosulfonate or hydrocolloid, can be used. The term "hydrocolloid," as used herein, includes gelatin, xanthan gum, acacia gum, pectin, guar, carob gum, alginate, cellulose, carboxymethylcellulose, and / or cellulose derivatives and / or modified polysaccharides. The term "modified polysaccharide," as used herein, refers to a polysaccharide containing a lipophilic moiety, e.g., a hydrocarbon moiety having a chain length of preferably 5 to 18 carbon atoms in a linear chain. Preferably, the modified polysaccharide should be acceptable for human consumption; i.e., preferred modified polysaccharides should be GRAS (Generally Recognized as Safe) or approved for food consumption, as determined by various regulatory authorities around the world. A preferred modified polysaccharide is modified food starch.

[0094]

[0098] As used herein, the term "modified food starch" refers to modified starch made from starch substituted with hydrophobic moieties by known chemical methods. For example, starch can be treated with cyclic dicarboxylic acid anhydrides, such as succinic anhydride and / or glutaric anhydride, substituted with alkyl or alkenyl hydrocarbon groups. According to certain embodiments, preferred starches include starch sodium octenyl succinate ("OSA").

[0095]

[0099] OSA-starch may further comprise hydrocolloids such as starch, maltodextrin, carbohydrates, gums, corn syrup, etc., and optionally typical emulsifiers (as co-emulsifiers), such as mono- and diglycerides of fatty acids, polyglycerol esters of fatty acids, lecithin, sorbitan monostearate, vegetable fibers, or sugars. OSA starches are available, for example, under the trade names HiCap 100, Capsul, Capsul HS, Purity Gum 2000, UNI-PURE, NYLON VII from National Starch, C from Rockete Freres, and C from CereStar. * It is commercially available under the trade name EmCap or from Tate & Lyle.

[0096]

[0100] The terms "modified polysaccharide", "modified starch" and "OSA starch" further encompass modified polysaccharides / modified starches / OSA starches that have been partially hydrolyzed enzymatically, e.g., by glycosylases (EC EC3.2; see http: / / www.chem.qmul.ac.uk / iubmb / enzyme / EC3.2 / ) and chemically by known methods.

[0097]

[0101] In other embodiments, the matrix component is selected from the group consisting of modified (food) starch, pectin, alginate, carrageenan, furcellaran, chitosan, maltodextrin, dextrin derivatives, cellulose and cellulose derivatives (e.g., cellulose acetate, methylcellulose, hydroxypropylmethylcellulose), lignosulfonates, polysaccharide gums (e.g., acacia gum, gum arabic, flaxseed gum, ghatti gum, tamarind gum, and arabinogalactan), gelatin (bovine, fish, porcine, poultry), vegetable proteins, and the like. The ingredients may include proteins (such as pea, soybean, castor bean, cotton, potato, sweet potato, manioc, rapeseed, sunflower, sesame, flaxseed, safflower, lentil, nuts, wheat, rice, corn, barley, rye, oat, lupin, and sorghum), animal proteins including milk or whey proteins, lecithin, polyglycerol esters of fatty acids, monoglycerides of fatty acids, diglycerides of fatty acids, sorbitan esters, PG esters, and sugar esters (and derivatives thereof).

[0098]

[0102] The heating sub-step involves heating the composition, preferably in oil, to obtain the active phase. The composition utilized in this step is preferably a fermentatively produced retinoid-containing composition according to any of the embodiments of the first aspect of the invention, as described elsewhere herein above.

[0099]

[0103] Any suitable oil can be used, although it is highly preferred that the oil used be compatible with the composition in which it is incorporated. In a preferred embodiment, the oil is a triglyceride, optionally selected from vegetable oils and / or fats, such as corn oil, sunflower oil, (hydrogenated) soybean oil, safflower oil, rapeseed oil, peanut oil, (hydrogenated) palm oil, palm kernel oil, cottonseed oil, and / or coconut oil, including fractions thereof. The triglyceride may preferably be a so-called MCT (medium-chain triglyceride), i.e., an ester of medium-chain fatty acids (preferably saturated fatty acids with a chain length of 6 to 12 carbon atoms) with glycerol. Preferred triglycerides are corn oil, sunflower oil, (hydrogenated) soybean oil, and / or (hydrogenated) palm oil.

[0100]

[0104] According to certain embodiments, it is useful to include a fat-soluble antioxidant to further limit degradation and / or oxidation of the active "retinoid ingredient" during and after the formulation process. The fat-soluble antioxidant can be incorporated at any suitable time, such as during substep (2) of heating the composition and oil. Any fat-soluble antioxidant can be utilized, such as ascorbic acid or its salts, tocopherol, butylated hydroxytoluene (BHT), butylated hydroxyanisole (BHA), propyl gallate, tert. butylhydroxyquinoline, ethoxyquin, and / or ascorbic acid esters of fatty acids.

[0101]

[0105] The amounts and ratios of the aforementioned matrix components, water, composition, oil and the fat-soluble antioxidant can be varied as needed and as understood by those skilled in the art to which the present invention applies.

[0102]

[0106] Once the matrix solution and active phase are provided, it is necessary to emulsify the active phase into the matrix solution to obtain a dispersion, which can be achieved by a variety of techniques and methods, including, but not limited to, those described in U.S. Pat. No. 8,765,186.

[0103]

[0107] The dispersion may optionally be further processed to ensure the desired droplet size or distribution, and to achieve this it may be fed, for example, through a homogenizer, after which it is preferably pumped to an apparatus or series of apparatuses to remove any solvent and dry the dispersion.

[0104]

[0108] The drying step can be carried out by any conventional drying process known to those skilled in the art, preferred are freeze drying, spray drying, spray drying in combination with fluidized bed granulation (commonly known as fluidized spray drying or FSD) and / or powder capture processes in which the sprayed emulsion or dispersion droplets are captured in a bed of an absorbent such as starch or calcium silicate or silicic acid or calcium carbonate or mixtures thereof and then dried.

[0105]

[0109] It is advantageous if the residual moisture content in the powder obtained by the drying sub-step is in the range of 0.5 to 7.0 wt. %, preferably 4 to 6 wt. %, each based on the total weight of the material. If employed, freeze-drying is preferably carried out for 10 to 48 hours at a temperature of -20°C to -50°C. In a preferred embodiment, the emulsion or dispersion obtained / obtainable by the process of such an embodiment of the third aspect of the present invention is spray-dried. In this case, it is preferable to select the spray-drying parameters as follows:

[0106]

[0110] Air inlet: about 100°C to about 250°C, preferably 150°C to about 200°C, more preferably about 160°C to about 190°C.

[0107]

[0111] Air outlet: about 45° C. to about 160° C., preferably about 55° C. to about 110° C., more preferably about 65° C. to about 95° C. The solid form may be further dried and / or granulated by a variety of other known techniques.

[0108]

[0112] Preferably, the drying process involves spraying the dispersion into a vessel containing a trapping medium, which traps and surrounds the dispersed particles, effectively facilitating further drying. An advantage of the trapping medium is that drying can be achieved at lower temperatures and with lower energy input than conventional spray drying techniques.

[0109]

[0113] The preferred trapping medium is cornstarch, but any other suitable trapping medium can be used according to the specific properties of active retinoid component and the desired end use.When this method comprises the sub-step of drying dispersion in the presence of trapping medium, the product obtained is known as beadlet.The technology of producing beadlet is known to produce the formulation with high level of stability, optimized handling and good dissolution properties.

[0110]

[0114] The inventors have discovered that compositions described in embodiments of the first aspect and / or those produced by embodiments of the second aspect are particularly suitable for use in such beadlet production processes because of the excellent oxidative stability properties they possess.

[0111]

[0115] Another compounding technique that can be used according to other embodiments of the third aspect of the present invention involves extrusion. Extrusion techniques are generally known and are described in International Publication No. WO 2014083065, which is incorporated herein by reference in its entirety as if fully set forth herein. Other exemplary publications include WO 2007 / 055815 A1, WO 2021 / 163836 A1, WO 2014 / 083065 A1, WO 2017 / 005622 A1, and WO 2019 / 175326 A1. The main advantage of using extrusion techniques is that highly viscous solutions can be formulated, reducing the amount of water available for dispersion and therefore reducing the need for drying. Furthermore, the extrusion process can be carried out as a continuous process. In the prior art, emulsions containing fat-soluble vitamins can be found to be extruded. US Patent Application Publication No. 2004 / 0201116 discloses pellets obtained by combining the generation of an emulsion using a device such as a high-pressure homogenizer with subsequent direct pelletization or extrusion as a second process step.

[0112]

[0116] According to an embodiment in which such a so-called extrusion method is used, the compounding step (b) of the third aspect comprises: (1) introducing a composition according to any of the preceding claims, a matrix component, water and oil into an extruder having an orifice; (2) a sub-step of extruding the composition, matrix component, water, and oil through an orifice to obtain an extrudate; Includes.

[0113]

[0117] The introduction substep (1) entails introducing a composition, preferably any of the fermentatively produced retinoid-containing compositions according to any of the embodiments of the first aspect and / or any of such compositions produced via any of the embodiments of the second aspect of the invention, together with a matrix component, oil, and water. The matrix component and oil may be selected from any suitable types, including those described above with respect to the beadlet technology embodiments of the third aspect. As described above with respect to other microencapsulation techniques, an oil-in-water / matrix emulsion or dispersion should be produced between the aforementioned components.

[0114]

[0118] The emulsion or dispersion can be formed prior to incorporation into the extruder as a premixed active phase, or alternatively, within the extruder apparatus itself, as described in WO2014083065. Thus, the introduction of the composition or "active," oil, water, and matrix can be carried out in up to four different steps, more robustly, or at up to four different locations, as deemed appropriate. The inventors have surprisingly discovered that retinoid-containing compositions produced by fermentation according to the first aspect and / or those produced by the method according to the second aspect are particularly suitable for extrusion methods in which the emulsion is formed within the extruder itself. Without wishing to be bound by any theory, the inventors theorize that this may be due to the superior oxidative stability of such compositions.

[0115]

[0119] In substep (2), the introduced material is extruded through the extruder and finally through an orifice to obtain an extrudate. In a non-limiting embodiment, the extruder device includes an extrusion die having a circular orifice for the material exit. Behind the die, the device typically includes one or more feeders for feeding the material into the barrel, and one, two, or more screws driven by a mechanical drive (e.g., an electric motor, a hydraulic motor, or a pneumatic motor) mix the material and convey it toward the die, building up pressure. Optionally, a means for heating and / or cooling the material can be disposed at a certain point in the barrel. The socket for holding the die can also be heated or cooled. Upon operation, the extruded material exits the orifice as a strand of a specific cross-section. The emulsion or suspension containing the composition, matrix component, water, and oil remains in a viscous state during extrusion and solidifies by cooling and / or drying after it exits the extrusion die.

[0116]

[0120] After extrusion, it is also preferred to incorporate substeps (3) and (4) of cutting the extrudate into individual particles, and optionally drying the individual particles. For the avoidance of doubt, these steps need not be performed sequentially but may be performed simultaneously, and indeed some amount of drying is preferably performed not only after cutting, but preferably before and during such processes as well.

[0117]

[0121] In a preferred embodiment, a cutter cuts pellets from the extrudate at the exit of the extrusion die and / or orifice. In one embodiment, a transverse flow of a transport fluid, typically transport air, carries the pellets away from the die. However, despite the cooling and drying effect of the transport fluid, the mass often remains viscous and sticky, especially at the core of the strand. This can lead to clogging of mechanical components, particularly the cutter and extrusion die, adversely affecting process stability and maintenance frequency. This can also lead to agglomeration of the product particles themselves, adversely affecting product quality. Depending on the type and shape and size of the extrudate mass, adjusting the temperature and flow rate of the transport air or cooling the cutter knives with a separate flow of cooling air may not always satisfactorily solve the problem.

[0118]

[0122] Thus, the drying step may further comprise means for providing an additional vertical flow of cooling air exiting the outlet to increase the rate of cooling and drying of the extrudate, particularly at and around the core of the extrudate, compared to cooling and drying by conveying air alone, which in turn has an additional cooling effect on the knives of the cutter compared to cooling by cooling air exiting the outlet alone.

[0119]

[0123] These effects of cooling air exiting such an outlet can be particularly advantageous in the extrusion of oil-in-water emulsions containing fat-soluble nutritional or pharmaceutical actives dissolved in the oil particles of the emulsion. Emulsions typically contain thickeners or gelling agents, which make them very viscous when they exit the die at high temperatures, and a perpendicular cooling air flow onto the face of the exiting strand of material reduces stickiness and the tendency for the device to clog. Thus, in one embodiment, the compounding step (b) further comprises directing a flow of cooling fluid over the orifice and the extrudate. In one embodiment, this directing step is performed such that the cooling fluid flows along a cooling flow axis and the extrudate is extruded through an exit axis, the cooling flow axis and the exit axis being substantially antiparallel and / or substantially collinear.

[0120]

[0124] A fourth aspect of the invention is a beadlet or extrudate produced by the method of any of the embodiments of the third aspect of the invention and / or incorporating a composition according to any of the embodiments of the first aspect of the invention and / or produced by any of the embodiments according to the second aspect of the invention.

[0121]

[0125] A fifth aspect of the invention is a food, feed, pharmaceutical or personal care product comprising any of the extrudates and / or beadlets according to the fourth aspect and / or produced according to any of the embodiments of the third aspect and / or comprising a composition according to any of the embodiments of the first aspect and / or produced by any of the methods of the second aspect.

[0122]

[0126] In particular, when used in personal care products, compositions according to the first aspect or products according to the fifth aspect containing them advantageously have antimicrobial activity. As used herein, the term "antimicrobial activity" or "antimicrobial effect" preferably refers to the ability to inhibit and / or eliminate the growth of cells of skin microorganisms, particularly Cutibacterium acnes, Corynebacterium xerosis, and / or Malassezia, preferably M. furfur, in a given sample or in a human or animal in need thereof, compared to an inoculum, respectively, as outlined in the Examples. As used herein, the terms "eliminate" and "kill" are used interchangeably. When used in relation to skin microorganisms, the term "growth" refers to the undesirable or unhealthy growth of microorganisms, as defined herein, resulting in an increase in microbial cells on the skin compared to a balanced, healthy skin microflora.

[0123]

[0127] Due to their antimicrobial activity against skin microorganisms such as Cutibacterium acnes, Corynebacterium xerosis and / or Malassezia, preferably M. furfur, compositions according to the first aspect and / or products of the fifth aspect incorporating them may be suitable for the treatment, prevention and / or alleviation of symptoms of adverse skin conditions associated with an overgrowth of the aforementioned microorganisms, such as in anti-acne applications (Cutibacterium acnes), deodorant applications (Corynebacterium xerosis) and hair care, especially dandruff applications (Malassezia, preferably M. furfur).

[0124]

[0128] For example, the antimicrobial effects described above can be used to balance the microbiome when applied, for example, to the external surfaces of the human or animal body.

[0125]

[0129] Thus, in another embodiment, compositions according to the first aspect and / or products of the fifth aspect incorporating them may be particularly useful as antimicrobial compounds in cosmetic or pharmaceutical products used to control / reduce acne, in particular to inhibit the overpopulation of Cutibacterium acnes on the skin, as an ingredient in deodorants to inhibit the overpopulation of Corynebacterium xerosis which causes malodour, in particular when sweating, or used to control / reduce dandruff, in particular to control / reduce the overproduction of Malassezia, preferably M. furfur.

[0126]

[0130] However, such antimicrobial action may also be used in the prevention and / or treatment of diseases associated with a dysbiosis or imbalance of the skin microbiota associated with an increase (or overgrowth) / proliferation of Cutibacterium acnes, Corynebacterium xerosis and / or Malassezia, preferably M. furfur.

[0127]

[0131] It will be appreciated that compositions according to the first aspect of the present invention and / or products of the fifth aspect incorporating them may be used in both a cosmetic and a pharmaceutical sense.

[0128]

[0132] Pharmaceutical uses are envisaged, for example, in the treatment, prevention and / or alleviation of the symptoms of tinea versicolor, dandruff, seborrheic dermatitis, atopic dermatitis, psoriasis and acne, e.g. in a human (e.g. patient) or animal in need thereof, in particular for the treatment, prevention and / or alleviation of any disorders and diseases associated with the aforementioned microorganisms, such as Cutibacterium acnes and / or Malassezia, preferably M. furfur.

[0129]

[0133] A cosmetic use is considered when the applicant intends to protect and / or enhance the beauty and / or youth of a human being, particularly in relation to the appearance of tissue or skin. Preferred cosmetic (non-therapeutic) uses according to the present invention include the treatment, prevention and / or relief of symptoms of itchy skin, maintaining skin homeostasis, providing a balance to the microflora and reducing malodour formation caused by sweat.

[0130]

[0134] To take advantage of their antimicrobial activity against Cutibacterium acnes, Corynebacterium xerosis and / or Malassezia, preferably M. furfur, compositions according to the first aspect and / or products incorporating them according to the fifth aspect may be used in a variety of applications, such as for example cosmetic or pharmaceutical compositions.

[0131]

[0135] Therefore, in another embodiment, the composition according to the present invention is a cosmetic or pharmaceutical composition further comprising a cosmetically or pharmaceutically acceptable carrier. The term "cosmetically or pharmaceutically acceptable carrier" refers to any carrier, excipient, and / or diluent conventionally used in cosmetic or pharmaceutical compositions. As used herein, it can be understood as a physiologically acceptable medium, i.e., a medium that is compatible with the intended application with keratinous materials, such as the skin, mucous membranes, and keratinous fibers.

[0132]

[0136] The term "cosmetic composition" or "cosmetic product" as used in this application refers to cosmetic compositions as defined in the "Kosmetika" section of Römpp Lexikon Chemie, 10th edition 1997, Georg Thieme Verlag Stuttgart, New York, and in A. Domsch, "Cosmetic Compositions", Verlag für chemische Industrie (ed. H. Ziolkowsky), 4 th edition, 1992, and refers to products / compositions used to treat, care for or improve the appearance of the skin and / or scalp.

[0133]

[0137] As used herein, the term "pharmaceutical composition" or "pharmaceutical product" refers to a composition used to treat, prevent, and / or alleviate symptoms of a disease and / or disorder.

[0134]

[0138] Thus, in another advantageous embodiment, the cosmetic or pharmaceutical composition / product according to the invention is an anti-acne composition / product comprising a composition according to the first aspect and / or a product of the fifth aspect incorporating same, which is potentially suitable for killing or significantly inhibiting Cutibacterium acnes. Particularly suitable anti-acne compositions are creams, lotions, etc., preferably in the form of an emulsion, such as an O / W emulsion, as disclosed herein.

[0135]

[0139] According to the present invention, the use of compositions according to the first aspect and / or products of the fifth aspect incorporating them as active compounds in anti-dandruff preparations is also advantageous according to the present invention, since they have antimicrobial activity against Malassezia, preferably M. furfur, even at very low concentrations of less than about 0.3% by weight, all definitions and priorities being as given herein.

[0136]

[0140] Therefore, in another advantageous embodiment, the cosmetic or pharmaceutical composition / product according to the invention is an anti-dandruff composition comprising a composition according to the fifth aspect in an amount of 0.1 to 0.3% by weight, based on the total weight of the composition, since these compositions are particularly suitable for significantly inhibiting the growth of and killing, respectively, Malassezia, preferably M. furfur. Particularly suitable anti-dandruff compositions are shampoos and hair tonics.

[0137]

[0141] The cosmetic or pharmaceutical compositions / products disclosed herein are preferably aqueous compositions, i.e., compositions that include water.

[0138]

[0142] It is further advantageous that the water content in the aforementioned cosmetic or pharmaceutical compositions / products according to the present invention is at least about 30% by weight, preferably at least about 40, 45, 50, 60, 70, 80, 90% by weight, such as in the range of 30-90% by weight, e.g., 40-90% by weight, 45-90% by weight or 50-90% by weight, based on the total weight of the composition. Further suitable ranges are 30-75% by weight, 30-70% by weight, 30-60% by weight and 40-60% by weight.

[0139]

[0143] In certain embodiments, the product of the fifth aspect is a cosmetic composition intended to be topically applied to mammalian keratinous tissue, for example, particularly human skin or human scalp. Such compositions are also called dermatological compositions. Therefore, preferably, in certain embodiments of the present invention, the cosmetic composition is a topical cosmetic (i.e., external skin) composition, having all the definitions and preferences described herein.

[0140]

[0144] Topical cosmetic compositions / products according to the present invention may be leave-on or rinse-off compositions and may include any cosmetic product applied to the human body primarily to improve appearance, for cleansing, deodorization, or overall beauty. Preferably, the cosmetic compositions of the present invention are leave-on compositions.

[0141]

[0145] It will be appreciated that cosmetic compositions / products according to the present invention intended for topical application comprise a physiologically acceptable medium, i.e. a medium compatible with keratinous materials such as the skin, mucous membranes and keratinous fibers. In particular, a physiologically acceptable medium is a cosmetically acceptable carrier. In an embodiment of the present invention, the carrier preferably comprises water.

[0142]

[0146] The term "cosmetically acceptable carrier" (also referred to herein as carrier) refers to all vehicles / carriers conventionally used in cosmetic compositions, i.e., those that are suitable for topical application to keratinous tissue, have good aesthetic properties, are compatible with the active ingredients present in the composition / product, and do not raise any unreasonable safety or toxicity concerns. Such carriers are well known to those skilled in the art and may include one or more compatible liquid or solid filler diluents, excipients, additives, or vehicles that are suitable for application to the skin.

[0143]

[0147] The exact amount of carrier will depend on the actual level of the active ingredient and any other optional ingredients that one of ordinary skill in the art would classify as different from a carrier (eg, other active ingredients).

[0144]

[0148] The fifth cosmetic composition of the present invention comprises a carrier in an amount of preferably 50% to 99.995%, such as 50 to 99.95%, more preferably 60% to 99.995%, such as 60 to 99.95%, even more preferably 75% to 99%, and most preferably 80% to 98%, such as 90% to 98%, by weight of the composition, based on the total weight of the composition.

[0145]

[0149] The cosmetic composition / product according to the fifth aspect of the invention may be in the form of a solution, lotion, thickened lotion, gel, cream, emulsion, ointment, paste, powder, make-up or solid tube stick, and may optionally be packaged as an aerosol and provided in the form of a mousse, spray, stick, e.g. an aerosol mousse, foam or spray foam.

[0146]

[0150] Preferably, the cosmetic compositions / products according to the fifth aspect of the present invention are in the form of lotions, creams, gels, and tonics. These product forms can be used for many applications, including, but not limited to, hand and body lotions, facial moisturizers, anti-aging formulations, makeup products including foundations, etc. Any additional ingredients required to formulate such products will vary depending on the type of product and can be routinely selected by those skilled in the art.

[0147]

[0151] When the cosmetic composition of the fifth aspect of the present invention is formulated as an aerosol and applied to the skin as a spray-on product, a propellant is added to the composition.

[0148]

[0152] The cosmetic composition / product according to the fifth aspect of the present invention can be prepared by conventional methods in the art, such as by mixing a mixture or composition comprising a mixture of cis and trans isomers of a retinoid as defined herein, having a cis / trans ratio of less than 0.01, with a cosmetically acceptable carrier, all definitions and priorities being as given herein.

[0149]

[0153] The cosmetic composition / product may contain further ingredients that may form part of the carrier. Such ingredients are, in particular, surfactants, emulsifiers, thickeners and oils. Such suitable surfactants, emulsifiers, thickeners and oils are well known to those skilled in the art.

[0150]

[0154] The cosmetic composition / product (including the carrier) of the fifth aspect of the present invention may comprise further conventional (cosmetic) adjuvants and additives such as preservatives / antioxidants, fatty substances / oils, water, organic solvents, silicones, thickeners, emollients, emulsifiers, antifoaming agents, aesthetic ingredients such as fragrances, surfactants, fillers, anionic, cationic, nonionic or amphoteric polymers or mixtures thereof, propellants, acidifying or basifying agents, dyes, colorants / colorants, abrasives, absorbents, chelating and / or sequestrants, essential oils, skin feel agents, astringents, pigments or any other ingredient normally incorporated into such compositions.

[0151]

[0155] Unless otherwise specified, the excipients, additives, diluents, etc. described below are suitable for the compositions according to the present invention. The required amounts of adjuvants and additives for cosmetic and dermatological applications can be easily determined by those skilled in the art based on the desired product.

[0152]

[0156] Additional ingredients can be added to either the oil phase, the water phase, or separately, as deemed appropriate. The mode of addition can be readily adapted by one skilled in the art.

[0153]

[0157] Examples of suitable cosmetic surfactants, emulsifiers, thickeners, oils, excipients, diluents, adjuvants, additives and active ingredients commonly used in the skin care industry that are suitable for use in the cosmetic compositions of the present invention are found, for example, but not limited to, in the International Cosmetic Ingredient Dictionary & Handbook by the Personal Care Product Council (http: / / www.personalcarecouncil.org / ), accessible via INFO BASE online at http: / / online.personalcarecouncil.org / jsp / Home.jsp.

[0154]

[0158] Cosmetic actives useful herein may in some cases provide more than one benefit or function by more than one mechanism of action.

[0155]

[0159] Of course, the skilled person will carefully select the above-mentioned optional additional components, adjuvants, diluents and additives and / or their amounts so that the advantageous properties inherently associated with the combination according to the invention are not adversely affected or are not substantially affected by the envisaged addition.

[0156]

[0160] The cosmetic compositions / products according to the invention may in particular be skin care, functional and / or hair care formulations, more particularly skin care or hair care formulations.

[0157]

[0161] Examples of skin care preparations are in particular photoprotective preparations (sun care preparations), anti-ageing preparations, preparations for treating photoageing, body oils, body lotions, body gels, treatment creams, skin protection ointments, moisturizing preparations such as moisturizing gels or moisturizing sprays, face and / or body moisturizers and skin whitening preparations.

[0158]

[0162] Examples of functional formulations are, but are not limited to, cosmetic compositions containing active ingredients such as hormone formulations, vitamin formulations, vegetable extract formulations, anti-aging formulations and / or antimicrobial (antibacterial or antifungal) formulations.

[0159]

[0163] Examples of suitable hair care compositions according to the invention include shampoos, hair conditioners (also called hair rinses), hair finishing compositions, hair tonics, hair regeneration compositions, hair lotions, water wave lotions, hair sprays, hair creams, hair gels, hair oils, hair pomades or hair brilliantines. These are therefore always compositions that are applied to the hair and scalp for a shorter or longer time depending on the actual purpose for which they are used.

[0160]

[0164] Preferably, in an embodiment of the present invention, the skin / functional and / or hair care formulation is a deodorant, antiperspirant, anti-dandruff or anti-acne composition comprising a composition according to the first aspect of the present invention.

[0161]

[0165] In a preferred embodiment, the cosmetic product / composition according to the present invention is in the form of an emulsion and / or a gel. Even more preferably, the cosmetic composition is an emulsion containing an oily phase and an aqueous phase, such as a multiple emulsion or Pickering emulsion, in particular O / W, W / O, Si / W, W / Si, O / W / O, or W / O / W, a PIT emulsion, a nanoemulsion, a microemulsion, or a multiple emulsion (e.g., O / W / O or W / O / W).

[0162]

[0166] The amount of oil phase (i.e., the phase containing all fats and oils, including polar oils) present in such emulsions, particularly O / W, W / O, Si / W, W / Si, O / W / O, W / O / W multiple or Pickering emulsions, is preferably at least about 10% by weight, for example in the range of 10 to 60% by weight, preferably in the range of 15 to 50% by weight, and most preferably in the range of 15 to 40% by weight, based on the total weight of the composition.

[0163]

[0167] The oil phase according to the invention is preferably composed of butylene glycol dicaprylat / -dicaprate, dimethicone, propylene glycol dicaprylat / -dicaprate, dicaprylyl ether, C 12~15 -Alkylbenzoat, C 18~38 - comprising an oil selected from fatty acid triglycerides, dibutyl adipate, cyclomethicone, 2-phenylethylbenzoat, isopropyl lauroyl sarkosinate, caprylic / capric triglyceride and mixtures thereof.

[0164]

[0168] The amount of aqueous phase present in such emulsions is preferably at least about 20% by weight, for example in the range of 20 to 90% by weight, preferably in the range of 30 to 80% by weight, most preferably in the range of 30 to 70% by weight, based on the total weight of the composition.

[0165]

[0169] Advantageously, in all emulsions according to the invention, the ratio of oily phase to aqueous phase is chosen in the range from 40:60 to 30:70.

[0166]

[0170] In one particularly advantageous embodiment, the cosmetic composition according to the fifth aspect of the invention is in the form of an oil-in-water (O / W) emulsion comprising an oily phase dispersed in an aqueous phase in the presence of an O / W emulsifier. The preparation of such O / W emulsions is known to those skilled in the art.

[0167]

[0171] When the cosmetic composition according to the fifth aspect of the invention is an O / W emulsion, it advantageously contains at least one O / W or Si / W emulsifier selected from the list: glyceryl stearate citrate, glyceryl stearate SE (self-emulsifying), stearic acid, salts of stearic acid, polyglyceryl-3-methylglycose distearate. Further suitable emulsifiers are phosphate esters and their salts, such as cetyl phosphate (e.g., as Amphisol® A from DSM Nutritional Products Ltd), diethanolamine cetyl phosphate (e.g., as Amphisol® DEA from DSM Nutritional Products Ltd), potassium cetyl phosphate (e.g., as Amphisol® K from DSM Nutritional Products Ltd), sodium cetearyl sulfate, sodium glyceryl oleate phosphate, hydrogenated vegetable glycerides phosphate, and mixtures thereof. Further suitable emulsifiers are sorbitan oleate, sorbitan sesquioleate, sorbitan isostearate, sorbitan trioleate, cetearyl glucoside, lauryl glucoside, decyl glucoside, sodium stearoyl glutamate, sucrose polystearate, and hydrated polyisobutene. Furthermore, one or more synthetic polymers may be used as emulsifiers, such as PVP eicosene copolymer, acrylates / C10-30 alkyl acrylate crosspolymer, and mixtures thereof.

[0168]

[0172] The at least one O / W or Si / W emulsifier is preferably used in an amount in the range of 0.5 to 10 wt. %, in particular in the range of 0.5 to 6 wt. %, such as more particularly in the range of 0.5 to 5 wt. %, for example most particularly in the range of 1 to 4 wt. %, based on the total weight of the cosmetic composition.

[0169]

[0173] Particular suitable O / W emulsifiers for use in the cosmetic composition according to the fifth aspect include phosphate ester emulsifiers, such as advantageously 8-10 alkyl ethyl phosphate, C9-15 alkyl phosphate, ceteareth-2 phosphate, ceteareth-5 phosphate, ceteth-8 phosphate, ceteth-10 phosphate, cetyl phosphate, C6-10 pareth-4 phosphate, C12-15 pareth-2 phosphate, C12-15 pareth-3 phosphate, DEA-ceteareth-2 phosphate, DEA-cetyl phosphate, DEA-oleth-3 phosphate, potassium cetyl phosphate, deceth-4 phosphate, deceth-6 phosphate and trilaureth-4 phosphate.

[0170]

[0174] A particularly suitable O / W emulsifier for use in the cosmetic composition according to the fifth aspect is, for example, potassium cetyl phosphate, commercially available as Amphisol® K from DSM Nutritional Products Ltd Kaiseraugst.

[0171]

[0175] Another particularly suitable class of O / W emulsifiers are non-ionic self-emulsifying systems derived from olive oil, for example known as (INCI name) cetearyl olivate and sorbitan olivate (chemical composition: sorbitan and cetearyl esters of olive oil fatty acids) and sold under the trade name OLIVEM 1000.

[0172]

[0176] In one particular embodiment of the fifth aspect, the present invention relates to a cosmetic composition / product, with all the definitions and preferences described herein, in the form of an O / W emulsion comprising an oily phase dispersed in an aqueous phase in the presence of an O / W emulsifier, the O / W emulsifier being potassium cetyl phosphate. The amount of oily phase in such an O / W emulsion is preferably at least about 10% by weight, more preferably in the range of 10-60% by weight, and most preferably in the range of 15-50% by weight, for example in the range of 15-40% by weight, based on the total weight of the composition.

[0173]

[0177] Preferably, the cosmetic composition according to the fifth aspect of the present invention further comprises at least one fatty alcohol (co-emulsifier), such as, in particular, cetyl alcohol, cetearyl alcohol and / or behenyl alcohol. The total amount of one or several fatty alcohols in the topical composition according to the present invention is preferably selected in the range of about 0.1 to about 10.0 wt. %, in particular in the range of about 0.5 to 6.0 wt. %, relative to the total weight of the topical composition.

[0174]

[0178] Preferably, the topical composition according to the fifth aspect comprises a thickener to help achieve the appropriate product consistency, particularly when the topical composition is in the form of an emulsion. Preferred thickeners are aluminum silicate, xanthan gum, hydroxypropyl methylcellulose, hydroxyethyl cellulose, polyacrylates such as carbopol (e.g., Carbopol® 980, 981, 1382, 2984, 5984) or mixtures thereof. Further preferred thickeners are acrylate / C 10~30 These include alkyl acrylate copolymers (such as Pemulen™ TR-1, Pemulen™ TR12, Carbopol™ 1328 by NOVEON) and Aristoflex® AVC (INCI: Ammonium acryloyldimethyltaurate / VP copolymer).

[0175]

[0179] The cosmetic composition according to the fifth aspect advantageously comprises a preservative, if present, which is preferably used in an amount of 0.1 to 2% by weight, more preferably 0.5 to 1.5% by weight, based on the total weight of the composition.

[0176]

[0180] The cosmetic composition according to the fifth aspect generally has a pH in the range of 3 to 10, preferably in the range of 4 to 8, and most preferably in the range of 4 to 7.5, for example in the range of 5 to 6.5. The pH is adjusted by methods known to those skilled in the art, for example by using a suitable acid, such as citric acid, or a base, such as sodium hydroxide (e.g., as an aqueous solution), triethanolamine (TEA Care), tromethamine (Trizma base), and aminomethylpropanol (AMP-Ultra PC2000), according to standard methods in the art.

[0177]

[0181] The amount of cosmetic composition applied to the skin is not critical and can be easily adjusted by one skilled in the art. Preferably, the amount is 0.1 to 3 mg / cm. 2 Skin area, e.g., preferably 0.1 to 2 mg / cm 2 Skin coverage, most preferably 0.5-2 mg / cm 2 Selected by skin area.

[0178]

[0182] The fermentatively produced retinoid-containing compositions of the present invention (and the food, feed, pharmaceutical and / or personal care products to which they relate) may be constructed through the selection of ingredients specified hereinabove, more readily by following the formulation guidelines herein and by extrapolating from the general approach taken in the embodiments illustrated in the following examples, which further illustrate the present invention but, of course, should not be construed as in any way limiting its scope.

[0179] [Example]

[0183] These examples illustrate embodiments of the present invention. Table 1 describes the characterization of various retinoid-containing compositions used in these examples. Table 2 shows the results of certain thermal stability performance tests performed on the examples so characterized in Table 1, while Table 3 demonstrates the biobased content of exemplary fermentatively produced retinoid-containing compositions. Finally, Table 4 describes antimicrobial performance tests performed on other retinoid-containing compositions formulated to have different isomer ratios.

[0180] [Overall Methods and Strains]

[0184] All basic molecular biology and DNA manipulation procedures described herein are generally performed according to Sambrook et al. (eds.), Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory Press: New York (1989) or Ausubel et al. (eds.), Current Protocols in Molecular Biology. Wiley: New York (1998). All exemplified genetic manipulations were performed in Yarrowia lipolytica.

[0181]

[0185] Shake plate assay. Typically, 200 μl of 0.075% yeast extract, 0.25% peptone (0.25X YP) was inoculated with 10 μl of freshly grown Yarrowia and overlaid with 200 μl of Drakeol 5 (Penreco, Karns City, PA, USA) mineral oil, silicone oil, or corn oil (containing 2% oleic acid or 2% glucose) as the carbon source. Clonal isolates of transformants were grown in 24-well plates (Multitron, 30°C, 800 RPM) in YPD medium containing one of the above overlays for 4 days. Overlay fractions were removed from the shake plate wells and analyzed by HPLC on a normal-phase column with a photodiode array detector.

[0182]

[0186] DNA transformation. Strains were transformed by overnight growth on YPD plates. 50 μl of cells were scraped from the plates and transformed by incubation in 500 μl of 1 μg of transforming DNA (typically linear DNA for integrative transformation), 40% PEG 3550MW, 100 mM lithium acetate, 50 mM dithiothreitol, 5 mM Tris-Cl pH 8.0, 0.5 mM EDTA for 60 minutes at 40°C and plated directly on selective medium, or for dominant antibiotic marker selection, cells were grown in YPD liquid medium for 4 hours at 30°C before plating on selective medium. Nourseothricin (Nat) selection was performed in YPD medium containing 100 μg / mL nourseothricin, and hygromycin (Hyg) selection was performed in YPD medium containing 100 μg / mL hygromycin. URA3 marker recycling was performed using 5-fluoroorotic acid (FOA). An episomal hygromycin resistance marker (Hyg) plasmid was passaged on nonselective medium, and Hyg-sensitive colonies were identified by replica plating colonies from the nonselective medium onto hygromycin-containing medium (100 μg / mL). The hygromycin resistance marker of the integrated hygromycin-tagged DNA was recycled using standard methods of cre-recombinase expression and identification of sensitive colonies by replica plating.

[0183]

[0187] List of Plasmids. The plasmids, strains and nucleotide sequences used are listed below and are set out in the sequence listing in Table 5 herein below. Generally, all unmodified sequences referred to herein are identical to the accession sequence in the database for the reference strain CLIB122 (Dujon B, et al, Nature. 2004 Jul 1;430(6995):35-44).

[0184] [Preparation of retinoid-producing Yarrowia strains and fermentation conditions]

[0188] Strain ML18743, described in WO2022090548, was sequentially transformed with HindIII+XbaI-digested plasmid MB8203 and HindIII+XbaI-digested plasmid MB9894 (listed in the table below) using the methods described in WO2022090548 to generate strain ML18743+MB8203+MB9894. This strain was transformed with SfiI-digested plasmid MB9523 to generate new strain ML18934. Strain ML18934 was then transformed with SfiI-digested plasmid MB7270 to generate new strain ML19284. This strain was made ku70- by CRISPR mutagenesis using plasmid MB9282. Into this strain, DNA41 and DNA53 were sequentially integrated by nuclease-directed integration using homologous recombination to generate the new strain ML19649. Strains ML18934, ML19284 and ML19649 were fermented using an ethanol feed with Isopar M as the second phase as described in WO2022090548 and further described below and in the sequence listing in Table 5.

[0185] [Table 1]

[0186]

[0189] The strains produced as described above were then used in fermentation processes to obtain Examples 1-4, as further described below.

[0187] [fermentation]

[0190] Fermentation products used in Example 1 Production reactor: New Brunswick Bioflo 610. Seed Train: Shakeflask (250ml) and New Brunswick Bioflo 310 Strain: ML18934 Culture medium: 6.13 g / l Tastone-154, 1.96 g / l MgSO4 7H2O, 0.2 g / l NaCl, 0.33 g / l CaCl2 2H2O, 8.18 g / l (NH4)2SO4, 8.47 g / l KH2PO4, 0.2 g / l thiamine, 8.13 g / l DF204 antifoam, trace metals, 50 g / l glucose, 9.86 g / l ethanol, and 197.5 g / l Isopar M. Seed medium: Flask: 30g / l Tastone-154, 22g / l glucose, 0.5g / l Basildon antifoam Bioreactor: 30g / l Tastone-154, 70g / l glucose, 10g / l Basildon antifoam Batch volume 45 kg (including inoculum) Fermentation time: 168 hours Supplied: 6N NH4OH, ethanol (200 proof), DF204 defoamer Process description: The batch phase ends when the initial glucose and ethanol are depleted. Ethanol feed fixed rate ramp down to 20% DO. DO is controlled at 20% by agitation, airflow and feed rate. pH is controlled at 5.5 by NH4OH.

[0188]

[0191] Fermentation products used in Example 2 Production reactor: New Brunswick Bioflo 610. Seed Train: Shakeflask (250ml) and New Brunswick Bioflo 310 Strain: ML19284 Culture medium: 1.96g / l MgSO4 7H2O, 0.2g / l NaCl, 8g / l (NH4)2SO4, 16g / l KH2PO4, 0.8g / l thiamine, 5g / l DF204 antifoam, 50g / l glucose, 9.86g / l ethanol and 296g / l Isopar M. Trace metals: 656mg / l citric acid, 58.8mg / l FeSO4·7H2O, 6.3mg / l CuSO4·5H2O, 9.45mg / l MnSO4·1H2O, 33.2mg / l ZnSO4·7H2O, 47.3mg / l CaCl2·2H2O. Seed medium: Flask: 30g / l Tastone-154, 22g / l glucose, 0.5g / l Basildon antifoam Bioreactor: 30g / l Tastone-154, 70g / l glucose, 10g / l Basildon antifoam Batch volume 40 kg (including inoculum) Fermentation time: 260 hours Supplied: 6N NH4OH, ethanol (200 proof), DF204 defoamer Process description: The batch phase ends when the initial glucose and ethanol are depleted. Ethanol feed fixed rate ramp down to 20% DO. DO is controlled at 20% by agitation, airflow and feed rate. pH is controlled at 5.5 by NH4OH.

[0189]

[0192] Fermentation products used in Example 3 Production reactor: New Brunswick Bioflo 610. Seed Train: Shakeflask (250ml) and New Brunswick Bioflo 310 Strain:ML19649 Culture medium: 1.96g / l MgSO4 7H2O, 0.2g / l NaCl, 8g / l (NH4)2SO4, 16g / l KH2PO4, 0.8g / l thiamine, 5g / l DF204 antifoam, 50g / l glucose, 9.86g / l ethanol and 296g / l Isopar M. Trace metals: 656mg / l citric acid, 58.8mg / l FeSO4·7H2O, 6.3mg / l CuSO4·5H2O, 9.45mg / l MnSO4·1H2O, 33.2mg / l ZnSO4·7H2O, 47.3mg / l CaCl2·2H2O. Seed medium: Flask: 30g / l Tastone-154, 22g / l glucose, 0.5g / l Basildon antifoam Bioreactor: 30g / l Tastone-154, 70g / l glucose, 10g / l Basildon antifoam Batch volume 40 kg (including inoculum) Fermentation time: 260 hours Supplied: 6N NH4OH, ethanol (200 proof), DF204 defoamer Process description: The batch phase ends when the initial glucose and ethanol are depleted. Ethanol feed fixed rate ramp down to 20% DO. DO is controlled at 20% by agitation, airflow and feed rate. pH is controlled at 5.5 by NH4OH.

[0190]

[0193] Fermentation products used in Example 4 Production reactor: New Brunswick Bioflo 610. Seed Train: Shakeflask (250ml) and New Brunswick Bioflo 310 Strain: ML18934 Culture medium: 6.13 g / l Tastone-154, 1.96 g / l MgSO4 7H2O, 0.2 g / l NaCl, 0.33 g / l CaCl2 2H2O, 8.18 g / l (NH4)2SO4, 8.47 g / l KH2PO4, 0.2 g / l thiamine, 8.13 g / l DF204 antifoam, trace metals, 50 g / l glucose, 9.86 g / l ethanol, and 197.5 g / l Isopar M. Seed medium: Flask: 30g / l Tastone-154, 22g / l glucose, 0.5g / l Basildon antifoam Bioreactor: 30g / l Tastone-154, 70g / l glucose, 10g / l Basildon antifoam Batch volume 45 kg (including inoculum) Fermentation time: 168 hours Supplied: 6N NH4OH, ethanol (200 proof), DF304 defoamer Process description: The batch phase ends when the initial glucose and ethanol are depleted. Ethanol feed fixed rate ramp down to 20% DO. DO is controlled at 20% by agitation, airflow and feed rate. pH is controlled at 5.5 by NH4OH. Comments: Fermentation tested at high dco2 concentrations. Contamination detected.

[0191] [DSP and preparation of dried crystals]

[0194] Each of Examples 1-4 was prepared into a dry crystalline form for subsequent characterization and performance testing via the following process. First, each of the fermentation products described above (each containing retinyl acetate as the retinoid component) was centrifuged in a laboratory-scale short-path evaporator (0.03 m²) at an absolute pressure of approximately 1 mbar and a jacket temperature of 80°C to provide a light-phase concentration such that the residue reached a retinyl acetate fraction of approximately 40-45% by weight. The concentrated residue was then mixed with absolute ethanol to obtain a solution containing 45-47% by weight ethanol at 20°C. This solution was then cooled to 14°C in a 250 ml double-jacket reactor with stirring, at which point seed crystals were added and the temperature was maintained at 14°C for an additional hour. The reactor was then cooled to -20°C with a cooling ramp of 5°C / hour and held at -20°C for approximately 16 hours. The resulting suspension was then filtered through a Buchner vacuum funnel, pre-cooled at −20° C., and washed twice with cold ethanol (−20° C.) to obtain wet crystals. Finally, these crystals were dried in a vacuum drying oven at room temperature and reduced pressure to remove residual ethanol and obtain dry crystals.

[0192] [Preparation of Example 5]

[0195] Sample 5 was provided as dried retinyl acetate crystals produced by chemical synthesis, which are readily available from DSM Nutritional Products AG, Kaiseraugst, Switzerland. It is used herein as a control representative of a non-fermentatively produced retinoid-containing composition.

[0193] [Preparation of Example 6]

[0196] Example 6 is a composition in crystalline form composed primarily (i.e., >80% by weight) of retinyl acetate and its fermentation residues, which was produced fermentatively generally according to the same techniques and methods used in connection with the preparation of each of Examples 1-4 above. In particular, the same ethanol used as the carbon source in the fermentation of each of Examples 1-4 was also used during the fermentation process leading to the production of Example 6.

[0194]

[0197] It can also be concluded that the biobased profile of Example 6 is representative of those of Examples 1-4 due to the fact that the same carbon source was used during fermentation.

[0195] [Preparation of Examples 7 to 11]

[0198] The compositions utilized in Examples 7-11 were prepared according to standard techniques in the art to which the present invention applies, as further described below. Retinoid mixtures containing retinyl acetate as defined herein can be prepared as described in WO2022090549 (Example 1). Samples were controlled as much as possible to ensure that only the cis / trans isomer ratio of the retinoid component for each sample series (i.e., 7-9 and 10-11) varied. The cis / trans ratio and specific properties of the retinoid component for each sample are shown in Table 4 below.

[0196] [Sample characterization]

[0199] Retinoid quantification: Retinoid analysis was performed using the C4 reverse phase retinoid method (see below). The addition of all added intermediates gives the total amount of retinoid.

[0197]

[0200] C4 Reversed-Phase Chromatography. For accurate measurement of discrete retinoids, a long-range reversed-phase system was used. Analytes were separated using an Agilent 1290 instrument equipped with a YMC Pro C4, 150 x 3.0 mm, 3 um column (YMC America, Allentown PA) stationary phase and a column and sample tray controlled at 23°C with a 5 μl injection loop volume and the gradient described in Table B. Analytes were detected at 210, 230 nm, and 325 nm, and peak identities were verified by LCMS. Analytes were separated as distinct peaks, which were assigned according to the table below.

[0198]

[0201] [Table 2]

[0199]

[0202] [Table 3]

[0200]

[0203] Calibration Method: The method is calibrated using high-purity retinyl acetate received from DSM Nutritional Products, Kaiseraugst, CH. Retinol and retinal are quantified relative to retinyl acetate. The dilutions listed in Table 3 are prepared as follows: 40 mg of retinyl acetate is weighed into a 100 mL volumetric flask and dissolved in ethanol to obtain a 400 μg / mL solution. The solution is sonicated as needed to ensure dissolution. Dilute 5 mL of this 400 μg / mL solution into 50 mL (1 / 10 dilution, final concentration 40 μg / mL), 5 mL into 100 mL (1 / 20 dilution, final concentration 20 μg / mL), 5 mL of 40 μg / mL into 50 mL (1 / 10 dilution, final concentration 4 μg / mL), and 5 mL of 20 μg / mL into 50 mL (1 / 10 dilution, 2 μg / mL) using 50 / 50 methanol / methyl tert-butyl ether (MTBE) as the diluent. All dilutions are performed in volumetric flasks. The purity of retinyl acetate is determined by further diluting the 400 μg / mL stock solution 100-fold in ethanol (using a 2 mL volumetric pipette and a 200 mL volumetric flask). The absorbance of this solution at 325 nm is blanked using ethanol and calculated using the equation (Abs * Dilution ratio (100) * Adjust the initial concentration using the molecular weight ((328.5) / 51180 = concentration in mg / mL). Lower concentrations are better due to the rapid maximization of UV absorbance of retinyl acetate.

[0201]

[0204] [Table 4]

[0202]

[0205] Sample preparation: 7 mg of accurately weighed crystals are dissolved in 10 mL tetrahydrofuran (THF) before injection.

[0203]

[0206] Isopar M quantification: Analysis of Isopar M was performed using the GC-FID method (see below).

[0204]

[0207] For IsoparM measurements, we used an Agilent 7890 instrument or similar equipped with an HP-5 Agilent column 30 m long, 0.25 mm internal diameter, and 0.25 um film thickness. H2 was used as the carrier gas at a flow rate of 1.7 mL / min. A 1 ul volume of sample was injected with a 50:1 split ratio into the FID detector at an inlet temperature of 250°C. The detector temperature was set at 325°C with an H2 flow rate of 40 mL / min and an air flow rate of 250 mL / min. The oven temperature program is shown in Table D.

[0205]

[0208] [Table 5]

[0206]

[0209] Calibration Method

[0207]

[0210] n-Decane was used as the internal standard (IS). The internal standard solution was prepared by diluting 500 μL of n-decane with isopropyl alcohol (IPA, HPLC grade) in a 50 mL volumetric flask. IsoparM standard solution was prepared by diluting 25 μL of an exact weight of IsoparM (Exxon Mobile) into 1 mL of internal standard solution.

[0208]

[0211] Isopar M was quantified in terms of the single point response factor for n-decane according to the formula set forth below.

number

[0209]

[0212] Table E. GC-FID retention times for Isoparm and n-decane The amount of Isopar M is determined by the following formula:

number

[0210]

[0213] Sample preparation: Dissolve 30 mg of accurately weighed crystals in 10 mL of internal standard solution injection.

[0211]

[0214] The values ​​measured by the above method are reported in Table 1 below. Below the row labeled "Total" in Table 1 below, there are three additional calculated values. "% cis isomer" represents the total amount of cis isomers (in this case, the row labeled "cis-retinyl acetate") relative to the total weight of the retinoid components (in this case, the row labeled "retinyl acetate (total)"). ("% cis isomer" is equal to the value in the row labeled "retinyl acetate (total)" divided by the value in the row labeled "retinyl acetate (total)," expressed as a percentage (i.e., multiplied by 100), rounded to the nearest whole number.) "FARE" is determined by adding the amounts of retinyl palmitate and retinyl oleate detected in the composition (no other fatty acid retinyl esters were detected). Finally, "WSFRV" represents the weighted selective fermentation residue value, which was determined by adding 10% of the determined "FARE" content to the value obtained from the "% cis isomer" content.

[0212] [Table 6]

[0213] [Oxidation stability and thermal stress test]

[0215] To determine the oxidative stability of Examples 1-5, each sample, the preparation of which is described above, was placed in its own small, amber glass vial under ambient conditions (preferably 20°C and 50% relative humidity), blanketed with argon, and placed in an oven preheated to 105°C. The samples were then left in the oven for three hours. After three hours, the samples were removed from the oven and analyzed by the same method used to determine the "initial" values ​​reported in Table 1 above. The value of each component was measured after the heat stress test and compared to the value determined before the test. The relative difference from the second measurement to the first measurement was then calculated, and the values ​​are reported in Table 2 below. For clarity, Example 1 exhibited a total retinoid content before the heat stress test of 90.53% (see the row in Table 1 above labeled "Total Retinyl Acetate (All Species)"), and the change shown in Table 2 of -1.31% means that the total retinoid content determined after the heat stress test was 89.22% (by weight, relative to the total sample weight).

[0214] [Table 7]

[0215] [Bio-based content]

[0216] To determine the biobased content, samples were sent to the Isolab GmbH Laboratorium für Stabil-Isotopenanalytik in Schweitenkirchen, Germany. Liquid scintillation counting was used to determine the biobased content. 14 In this laboratory, we analyzed the stable isotope ratios of plant foods and their components using the QMA-M-03 method. 13 C. 15 N, 34 S (using EA-IRMS), 18 O (equilibrated - using IRMS) and 2 H (using HTC-IRMS) is then determined. 14 The C content was calculated to determine the relative amount of carbon atoms present in the samples that were from recent natural materials (i.e., no carbon atoms from fossil-based sources such as petroleum). The results are reported as "C activity" expressed as a %. These results of this biobased testing for Example 6 are shown in Table 3 below.

[0216] [Table 8]

[0217] [Antimicrobial efficacy test] The antimicrobial efficacy of retinol and retinyl acetate with different cis / trans ratios was evaluated similarly to the regulatory challenge test method (NF EN ISO 11930). Therefore, solutions of each retinol and retinyl acetate mixture (reference sample with a purity of approximately 98% and the mixture of the present invention) were prepared in physiological serum containing 0.85 wt% NaCl under sterile conditions. The presence of cis and trans isomers in the retinoid component was calculated by reverse-phase C4 HPLC analysis, and the total cis isomer content in the retinoid component relative to the total weight of trans isomers detected in the retinoid component is reported in Table 4 below, in the row labeled "cis / trans ratio." For each sample, the specific retinoid used as the retinoid component (either retinyl acetate or retinol for all samples) was also reported.

[0218] Next, solutions of the active substances were transferred to 96-deep-well plates (1.6 ml / well) as specified in weight percent in Table 4 below. The wells were each contaminated with Cutibacterium acnes, Corynebacterium xerosis, and Malassezia furfur to obtain an initial contamination. The initial contamination count, listed as the "initial inoculum" for each test as shown in Table 4 below, was determined to be 100,000. After contamination, each well was thoroughly mixed to ensure uniform distribution of the microorganisms. Each plate was then incubated at 22°C for 24 hours. A count of the (remaining) population was performed 24 hours after contamination, as shown in Table 4 below by the row labeled "24 hours." The percentage change in count from the initial inoculum to the 24-hour count is reflected in the table below by the row labeled "ΔR." The results are shown in Table 4 below.

[0219] [Table 9]

[0220] [Discussion of results] Table 2 shows that various fermentatively produced retinoid-containing compositions (i.e., Examples 1-4) demonstrated comparable or superior oxidative stability of the retinoid component (retinyl acetate in each of such Examples) when compared to chemically synthesized analogs, as indicated by the relative loss of retinyl acetate throughout the thermal stability testing period. Specifically, the control (Example 5) exhibited a 5.50% loss of retinyl acetate, while Examples 2-4 performed similarly or slightly better, exhibiting losses of 4.10% to 5.00%. Example 1 performed best, losing only 1.31% of its retinyl acetate content after undergoing the thermal stability test. This result is particularly surprising and unexpected given that the compositions of Examples 1-4 each contain one or more residual fermentation components that would be expected to adversely contribute to the oxidative stability of such compositions.

[0221] As can be seen from Table 3, the carbon-14 activity of the composition represented by Example 6 suggests that the proportion of fossil carbon was not detectable within the error of the test. Therefore, according to this test, such a sample can be considered completely "biobased." This result can be similarly applied to each of Examples 1-4, which were prepared in a similar manner using the same carbon source during the fermentation process.

[0222] As can be seen from Table 4, each of the specific cis / trans mixtures of retinyl acetate and retinol according to the present invention has excellent efficacy in anti-acne applications, i.e., against the growth of Cutibacterium acnes. Retinyl acetate mixtures with 0.49% (Example 8) and 0.25% (Example 9) cis-isomer content were able to reduce Cutibacterium acnes by approximately 30%, while the reference with 3% cis-isomer content allowed 270% bacterial growth. Similar effects were observed with retinol-containing compositions. A similar composition with 1% cis-isomer content reduced bacteria by 99.0%, while a retinol composition with 0.3% cis-isomer content (Example 11) reduced bacteria by approximately 99.3%. The effectiveness of commercially available trans-retinol and trans-retinyl acetate (e.g., Dry Vitamin A-Acetate 500 or Retinol 50 C from BASF or Microvit® A from Adisseo), respectively, against the growth of Cutibacterium acnes is in the same range as the reference samples (not shown).

[0223] Similar trends can be observed with respect to activity against Corynebacterium xerosis and Malassezia furfur from Table 4. Notably, increasing the concentration of the retinoid-containing composition had the effect of further improving antimicrobial efficacy when tested against Corynebacterium xerosis, as observed in Examples 7, 8, and 9, respectively.

[0224] Finally, certain compositions containing retinol as a retinoid component have been demonstrated to have increased efficacy against the dandruff-causing bacteria Malassezia furfur. Specifically, compositions with a 0.1% cis-isomer content have been demonstrated to reduce activity by 60%, while compositions with a 1% cis-isomer content have been demonstrated to neither reduce nor allow growth of the bacteria.

[0225] Further Exemplary Embodiments

[0224] Please refer to the claims.

[0226]

[0225] Unless otherwise specified, the term wt% refers to the amount by mass of a particular component relative to the total liquid radiation curable composition in which it is incorporated.

[0227] The terms "a," "an," "the," and similar uses in connection with the description of the present invention (particularly in connection with the claims below) are to be construed as including both the singular and the plural, unless otherwise indicated herein or clearly contradicted by context. The terms "comprise," "have," "include," and "contain" are to be construed as open-ended terms (i.e., meaning "including, but not limited to"), unless otherwise specified. The recitation of ranges of values ​​herein is merely intended to serve as a shorthand method of individually referring to each separate value falling within the range, unless otherwise stated herein, and each separate value is incorporated herein as if it were individually set forth herein. All methods described herein may be performed in any suitable order unless otherwise indicated herein or clearly contradicted by context. Unless otherwise claimed, the use of any and all examples or exemplary language (e.g., "etc.") provided herein is intended merely to better clarify the invention and does not impose limitations on the scope of the invention. No language in the specification should be construed as indicating any non-claimed element essential to the practice of the invention.

[0228]

[0227] This specification describes preferred embodiments for carrying out the invention, including the best mode known to the inventors. Variations of these preferred embodiments will become apparent to those skilled in the art upon reading the foregoing description. The inventors anticipate that those skilled in the art will employ such variations as necessary, and the inventors intend the invention to be embodied in forms other than those specifically described herein. Accordingly, this invention includes all modifications and equivalents of the subject matter recited in the claims appended hereto as permitted by applicable law. Furthermore, this invention includes any combination of the above-described elements in all possible variations thereof unless otherwise indicated herein or clearly contradicted by context.

[0229]

[0228] Although the present invention has been described in detail and reference has been made to specific embodiments thereof, it will be apparent to those skilled in the art that various changes and modifications can be made without departing from the spirit and scope of the invention as claimed.

[0230] [Table 10]

[0231] [Table 11]

[0232] [Table 12]

[0233] [Table 13]

[0234] [Table 14]

[0235] Table 15

[0236] Table 16

[0237] Table 17

[0238] Table 18

[0239] Table 19

[0240] Table 20

[0241] Table 21

[0242] Table 22

[0243] Table 23

[0244] Table 24

[0245] Table 25

[0246] Table 26

[0247] Table 27

[0248] Table 28

[0249] Table 29

[0250] Table 30

[0251] Table 31

[0252] Table 32

[0253] Table 33

[0254] Table 34

[0255] Table 35

Claims

1. 1. A composition for use in a food, feed, pharmaceutical or cosmetic product, comprising: (i) a retinoid component comprising a mixture of cis and trans isomers; (ii) the fermentation residue thereof; comprising, consisting of, or consisting essentially of the retinoid component is present in a weight ratio of greater than 4:1 relative to the fermentation residue; the cis-isomer is present in an amount by weight of less than 3% by weight based on the total weight of the retinoid component; Optionally also, the biobased carbon content of (i) and (ii) is greater than 50%, or greater than 60%, or greater than 70%, or greater than 90%.

2. 2. The composition of claim 1, wherein the fermentation residue comprises fatty acid retinyl esters (FARE), retinal, retinol, farnesol, a fermentation carbon source, or β-carotene.

3. The composition has a Weighted Selected Fermentation Readiness Value (WSFRV) of from 0.1 to 2.5, or from 0.1 to 2.0, or from 0.1 to 1.0, or from 0.1 to 0.9, or from 0.4 to 2.5, or from 0.4 to 2.0, or from 0.4 to 1.0, or from 0.4 to 0.9, or from 0.6 to 2.5, or from 0.6 to 2.0, or from 0.6 to 1.0, or from 0.5 to 0.9, wherein the WSFRV is calculated according to the following formula: WSFRV=C+0.1×F (In the formula, C = weight percent of the cis isomer present relative to the total amount of retinoid component (i) and fermentation residue (ii), and F = weight percent of FARE present relative to the total amount of retinoid component (I) and fermentation residue (II) The composition of claim 1 or 2, determined according to

4. The retinoid component has the formula (I): 【Chemistry 1】 comprising, consisting of, or consisting essentially of a compound according to R is -CHO, -CH ２ OH, -COOH, -CH(R １ ) ２ , -CH ２ OR ２ , -COOR ３ , -CONHR ４ or -CO(NR ４ ) ２ and R1 is independently lower alkoxy or R １’ and R １’’ taken together is a lower alkylenedioxy; R ２ is alkanoyl or aroyl, R ３ is alkyl, aryl or aralkyl, and R ４ , R ４’ and R ４’’ are independently hydrogen, alkyl, aryl, or aralkyl; The term "lower alkoxy" refers to an alkoxy group having 1 to 6 carbon atoms, such as methoxy, ethoxy, or propoxy; The term "lower alkylenedioxy" similarly refers to groups containing 1 to 6 carbon atoms, such as methylenedioxy or ethylenedioxy, where the alkyl or alkylene moiety may be linear or branched depending on the number of carbon atoms; The term "alkanoyl" refers to any straight-chain or branched alkanoyl group having from 1 to 18 carbon atoms, such as formyl, acetyl, propionyl, butyryl, stearoyl, and palmitoyl; The term "aroyl" refers to aromatic carboxylic acids having 7 and 11 carbon atoms, including benzoyl or naphthoyl, respectively; The term "alkyl" refers to a straight or branched alkyl group having 1 to 18 carbon atoms, for example, methyl, ethyl, propyl, butyl, decyl, dodecyl, hexadecyl, or octadecyl; The term "aryl" by itself or as part of "aralkyl" is phenyl or naphthyl, and The composition of claim 1 or 2, wherein the term "aralkyl" includes groups having 1 to 4 carbon atoms in the aliphatic portion, such as benzyl and phenylpropyl.

5. 5. The composition of any one of claims 1 to 4, wherein the retinoid component comprises, consists of, or consists essentially of retinol or a retinyl ester, and the retinyl ester comprises, consists of, or consists essentially of retinyl acetate or retinyl palmitate.

6. The composition of any one of claims 1 to 5, wherein the retinoid component comprises, consists of, or consists essentially of retinyl acetate.

7. The retinoid component comprises a mixture of cis and trans isomers, the cis isomers being present in an amount, by weight, based on the total weight of the mixture: 0.1 to 3% by weight, or 0.2 to 3% by weight, or 0.4 to 3% by weight, or 0.5 to 3% by weight, or 0.1 to 2.5% by weight, or 0.2 to 2.5% by weight, or 0.4 to 2.5% by weight, or 0.5 to 2.5% by weight, or 0.1 to 2% by weight, or 0.2 to 2% by weight, or 0.4 to 2% by weight, or 0.5 to 2% by weight, or 0.1 to 1.5% by weight, or 0.2 to 1.5% by weight, or 0.4 to 1.5% by weight, or 0.5 to 1.5% by weight, or 0.1 to 1% by weight, or 0.2 to 1% by weight, or 0.4 to 1% by weight, or 0.5 to 1% by weight, or 0.1 to 0.9% by weight, or 0.2 to 0.9% by weight, or 0.4 to 0.9% by weight, or 0.5 to 0.9% by weight The composition of any one of claims 1 to 6, wherein

8. When the retinoid component is retinol, the cis isomers include 9-cis-retinol, 11-cis-retinol, 13-cis-retinol, 9,13-di-cis-retinol, 11,13-di-cis-retinol, 13-cis-3,4-didehydroretinol, 9-cis-3,4-didehydroretinol and / or 9,13-di-cis-3,4-didehydroretinol; or When the retinoid component is retinal, the cis isomers include 13-cis retinal, 11-cis-retinal, 11,13-di-cis retinal, 9,13-di-cis-retinal, 9-cis retinal, 13-cis-3,4-didehydroretinal and / or 11,13-di-cis-3,4-didehydroretinal; or 8. The composition of any one of claims 1 to 7, wherein when the retinoid component is retinyl acetate, the cis isomer comprises any one or more of the corresponding cis forms of retinyl acetate.

9. The composition according to any one of claims 1 to 8, wherein the fermentation residue comprises FARE, beta-carotene, retinol, retinal and a fermentation carbon source.

10. 10. The composition of any one of claims 1 to 9, wherein the fermentation residue comprises retinol, which retinol comprises, consists of, or consists essentially of E-retinol.

11. 11. The composition of any one of claims 1 to 10, wherein the fermentation residue comprises retinal, wherein the retinal comprises, consists of, or consists essentially of 9Z-retinal.

12. 12. The composition of any one of claims 1 to 11, wherein the fermentation residue comprises one or more FAREs, and wherein the one or more FAREs are present in an amount of less than 4% by weight, or less than 3% by weight, or less than 2% by weight, or less than 1% by weight, relative to the total weight of retinoid components present.

13. 0.05 to 4%, or 0.1 to 4%, or 0.15 to 4%, or 0.2 to 4% by weight of FARE based on the total weight of the retinoid components present; or 0.05 to 3 wt.%, or 0.1 to 3 wt.%, or 0.15 to 3 wt.%, or 0.2 to 3 wt.% FARE; or 0.05 to 2% by weight, or 0.1 to 2% by weight, or 0.15 to 2% by weight, or 0.2 to 2% by weight of FARE; or 0.05 to 1 wt. %, or 0.1 to 1 wt. %, or 0.15 to 1 wt. %, or 0.2 to 1 wt. % FARE The composition of any one of claims 1 to 12, comprising:

14. The composition of any one of claims 1 to 13, wherein the FARE comprises retinyl palmitate and / or retinyl oleate.

15. 15. The composition of any one of claims 1 to 14, wherein the fermentation residue comprises 9Z-retinal, and the 9Z-retinal is present in an amount of less than 10% by weight, or less than 8% by weight, or less than 6% by weight, or less than 5% by weight, or less than 3% by weight, relative to the total weight of the retinoid components present.

16. 0.1 to 10%, or 0.5 to 10%, or 1 to 10%, or 1.5 to 10% by weight of 9Z-retinal, based on the total weight of the retinoid components present; 0.1 to 8% by weight, or 0.5 to 8% by weight, or 1 to 8% by weight, or 1.5 to 8% by weight of 9Z-retinal; 0.1 to 6%, or 0.5 to 6%, or 1 to 6%, or 1.5 to 6% by weight of 9Z-retinal; 0.1 to 5% by weight, or 0.5 to 5% by weight, or 1 to 5% by weight, or 1.5 to 5% by weight of 9Z-retinal; 0.1 to 3% by weight, or 0.5 to 3% by weight, or 1 to 3% by weight, or 1.5 to 3% by weight of 9Z-retinal The composition of any one of claims 1 to 15, comprising:

17. 17. The composition of any one of claims 1 to 16, wherein the fermentation residue comprises β-carotene, and the β-carotene is present in an amount of less than 4% by weight, or less than 3% by weight, or less than 2% by weight, or less than 1% by weight, or less than 0.5% by weight, relative to the total weight of retinoid components present.

18. 0-4%, or 0.2-4%, or 0.5-4%, or 1-4% by weight of beta-carotene, based on the total weight of the retinoid components present; or 0-3 wt.%, or 0.2-3 wt.%, or 0.5-3 wt.%, or 1-3 wt.% β-carotene, or 0-2% by weight, or 0.2-2% by weight, or 0.5-2% by weight, or 1-2% by weight of β-carotene, or 0-1 wt. %, or 0.2-1 wt. %, or 0.5-1 wt. % β-carotene, or 0 to 0.5% by weight or 0.2 to 0.5% by weight of β-carotene The composition of any one of claims 1 to 17, comprising:

19. 19. The composition of any one of claims 1 to 18, wherein the fermentation residue comprises farnesol, and the farnesol is present in an amount of less than 10,000 parts per million (ppm), or less than 5,000 ppm, or less than 1,000 ppm, or less than 100 ppm, or less than 10 ppm, or less than 5 ppm, or less than 2 ppm, based on the total weight of the retinoid components present.

20. Farnesol, from 0 to 10,000 ppm, or from 2 to 10,000 ppm, or from 5 to 10,000 ppm, or from 10 to 10,000 ppm, or from 100 to 10,000 ppm, based on the total weight of the retinoid components present; or 0 to 5,000 ppm, or 2 to 5,000 ppm, or 5 to 5,000 ppm, or 10 to 5,000 ppm, or 100 to 5,000 ppm of farnesol; or 0 to 1,000 ppm, or 2 to 1,000 ppm, or 5 to 1,000 ppm, or 10 to 1,000 ppm, or 100 to 1,000 ppm of farnesol; or 0 to 100 ppm, or 2 to 100 ppm, or 5 to 100 ppm, or 10 to 100 ppm of farnesol The composition of any one of claims 1 to 19, comprising:

21. The composition of any one of claims 1 to 20, which is free of farnesol.

22. 22. The composition of any one of claims 1 to 21, wherein the fermentation carbon source comprises, consists of or consists essentially of a biogenic carbon source.

23. 23. The composition of any one of claims 1 to 22, wherein the fermentation carbon source comprises straight chain alkanes, free fatty acids, ethanol, glucose containing triglycerides, in particular vegetable oils containing the respective free fatty acids derived therefrom, such as oleic acid, palmitic acid or linoleic acid.

24. 24. The composition of any one of claims 1 to 23, wherein the fermentation residue comprises ethanol or glucose, and the ethanol or glucose is present in an amount of less than 2% by weight, or less than 1% by weight, or less than 0.75% by weight, relative to the total weight of the retinoid components present.

25. 0.05 to 2%, or 0.1 to 2%, or 0.2 to 2% by weight of ethanol or glucose based on the total weight of the retinoid components present; or 0.05 to 1% by weight, or 0.1 to 1% by weight, or 0.2 to 1% by weight of ethanol or glucose, or 0.05 to 0.5% by weight, or 0.1 to 0.5% by weight, or 0.2 to 0.5% by weight of ethanol or glucose The composition of any one of claims 1 to 24, comprising:

26. The composition of any one of claims 1 to 25, wherein the fermentation residue comprises an isoparaffinic fluid.

27. 27. The composition of any one of claims 1 to 26, wherein the isoparaffin fluid is present in an amount of less than 3 wt. %, or less than 2 wt. %, or less than 1.5 wt. %, or less than 1.25 wt. %, based on the total weight of the retinoid component (i) and its fermentation residue (ii).

28. the isoparaffin fluid is present in an amount of 0.1 to 2% by weight, or 0.2 to 2% by weight, or 0.5 to 2% by weight, or 0.1 to 1.5% by weight, or 0.2 to 1.5% by weight, or 0.5 to 1.5% by weight, based on the total weight of the retinoid component (i) and its fermentation residue (ii); and / or 28. The composition of any preceding claim, wherein the isoparaffinic fluid comprises, consists of, or consists essentially of Isopar M.

29. The composition of any one of claims 1 to 28, wherein the fermentation residue comprises the retinoid component in a dihydro form.

30. 30. The composition of any one of claims 1 to 29, wherein the dihydro form is present in an amount of less than 0.5 wt. %, or less than 0.25 wt. %, or less than 0.1 wt. %, or less than 0.05 wt. %, based on the total weight of the retinoid component (i) and the fermentation residue thereof (ii).

31. 31. The composition of any one of claims 1 to 30, wherein the dihydro form is present in an amount of 0.01 to 0.5 wt. %, or 0.01 to 0.25 wt. %, or 0.01 to 0.1 wt. %, or 0.01 to 0.05 wt. %, or 0.03 to 0.5 wt. %, or 0.03 to 0.25 wt. %, or 0.03 to 0.1 wt. %, or 0.03 to 0.05 wt. %, based on the total weight of the retinoid component (i) and the fermentation residue (ii) thereof.

32. 32. The composition of claim 31, wherein the dihydro form of the retinoid component comprises, consists of, or consists essentially of dihydro-retinyl acetate.

33. 33. The composition of any one of claims 1 to 32, wherein the fermentation residue comprises less than 0.5%, or less than 0.2%, or less than 0.1% by weight of rosafluene, phytoene, ergosterol, and dihydro-beta-ionone, or is substantially free thereof.

34. the cis isomer is present in an amount of less than 1% by weight based on the total weight of the mixture of cis and trans isomers, and / or 34. The composition of any one of claims 1 to 33, wherein FARE is present in an amount of less than 3.5% by weight or less than 1.5% by weight, based on the total weight of the retinoid component.

35. 35. The composition of any one of claims 1 to 34, wherein the retinoid component is present in a weight ratio to the fermentation residue of greater than 5:1, or greater than 6:1, or greater than 10:1, or greater than 20:1, or greater than 25:

1.

36. the retinoid component is present in a weight ratio relative to the fermentation residue of 4:1 to 100:1, or 5:1 to 100:1, or 6:1 to 100:1, or 10:1 to 100:1, or 20:1 to 100:1, or 25:1 to 100:1; or a weight ratio of from 4:1 to 50:1, or from 5:1 to 50:1, or from 6:1 to 50:1, or from 10:1 to 50:1, or from 20:1 to 50:1, or from 25:1 to 50:1; or a weight ratio of from 4:1 to 35:1, or from 5:1 to 35:1, or from 6:1 to 35:1, or from 10:1 to 35:1, or from 20:1 to 35:1, or from 25:1 to 35:1; or a weight ratio of 4:1 to 30:1, or 5:1 to 30:1, or 6:1 to 30:1, or 10:1 to 30:1, or 20:1 to 30:1, or 25:1 to 30:1 36. The composition of any one of claims 1 to 35, wherein

37. 37. The composition of any one of claims 1 to 36, wherein the retinoid component comprises, consists of, or consists essentially of retinyl acetate, and the retinyl acetate is present in a weight ratio to the fermentation residue of greater than 5:1, or greater than 6:1, or greater than 10:1, or greater than 20:1, or greater than 25:

1.

38. the retinoid component comprises, consists of, or consists essentially of retinyl acetate, the retinyl acetate being present in a weight ratio to the fermentation residue of from 4:1 to 100:1, or from 5:1 to 100:1, or from 6:1 to 100:1, or from 10:1 to 100:1, or from 20:1 to 100:1, or from 25:1 to 100:1; or a weight ratio of from 4:1 to 50:1, or from 5:1 to 50:1, or from 6:1 to 50:1, or from 10:1 to 50:1, or from 20:1 to 50:1, or from 25:1 to 50:1; or a weight ratio of from 4:1 to 35:1, or from 5:1 to 35:1, or from 6:1 to 35:1, or from 10:1 to 35:1, or from 20:1 to 35:1, or from 25:1 to 35:1; or a weight ratio of 4:1 to 30:1, or 5:1 to 30:1, or 6:1 to 30:1, or 10:1 to 30:1, or 20:1 to 30:1, or 25:1 to 30:1 38. The composition of any one of claims 1 to 37, wherein

39. 39. The composition of any one of the preceding claims, wherein the biobased carbon content of the composition is greater than 60%, or greater than 70%, or greater than 80%, or greater than 90%, or greater than 99%.

40. 40. The composition of any one of claims 1 to 39, wherein the bio-based carbon content of the retinoid component (i) and the fermentation residue thereof (ii) is greater than 60%, or greater than 70%, or greater than 80%, or greater than 90%, or greater than 99%.

41. The biobased carbon content is determined according to ASTM D6866-20 or １４ C and １３ 41. The composition of any one of claims 1 to 40, as determined by C isotope characterization.

42. 42. The composition of any one of claims 1-41, wherein the amount of retinyl acetate, measured relative to the weight of the total composition, is within 5 wt. %, or within 4 wt. %, or within 3 wt. % of the amount of retinyl acetate present in the composition after the composition has been subjected to a thermal stability test in which the composition is heated to 105°C for 3 hours.

43. 43. The composition of any one of claims 1 to 42, which is in crystalline form.

44. The crystalline morphology has an average particle length D of greater than 100 microns, or greater than 200 microns, or greater than 300 microns, or greater than 400 microns, or between 100 and 1200 microns, or between 100 and 1000 microns, or between 100 and 800 microns, or between 200 and 1200 microns, or between 200 and 1000 microns, or between 200 and 800 microns, as determined by microscopic imaging methods. ５０ 44. The composition of any one of claims 1 to 43, comprising a plurality of crystals having the formula:

45. The composition of any one of claims 1 to 44, further comprising a solvent.

46. The composition of any one of claims 1 to 45, wherein the solvent comprises ethanol or glucose.

47. 47. The composition of any one of claims 1 to 46, wherein the composition, the retinoid component (i) and the fermentation residue thereof (ii) are present as an emulsion in oil.

48. 48. The composition of any one of claims 1 to 47, wherein the emulsion in oil comprises a fat-soluble antioxidant and an oil.

49. 49. The composition of any one of claims 1 to 48, wherein the oil preferably comprises a triglyceride, such as a vegetable oil and / or fat, and the fat-soluble antioxidant comprises ascorbic acid or a salt thereof, tocopherol, butylated hydroxytoluene (BHT), butylated hydroxyanisole (BHA), propyl gallate, tert.butylhydroxyquinoline, ethoxyquin, and / or an ascorbic acid ester of a fatty acid.

50. A formulation comprising a composition according to any one of claims 1 to 49.

51. 51. The formulation of claim 50, which is an animal feed premix.

52. 52. The formulation of claim 51, wherein the animal feed premix comprises a matrix component and an additive, preferably comprising silica.

53. 53. A composition or formulation according to any one of the preceding claims, wherein the animal feed premix comprises at least one additional micronutrient.

54. A composition or formulation according to any one of claims 1 to 50 which is a cosmetic composition.

55. The cosmetic composition comprises: (a) a composition according to any one of claims 1 to 49; and (b) a pharmaceutically or cosmetically acceptable carrier, water, and at least one agent selected from the group consisting of surfactants, emulsifiers, thickeners, oils, and mixtures thereof; 55. The composition or formulation of claim 54, comprising:

56. 56. The cosmetic composition according to claim 55, which is a shampoo preparation, a hair conditioner, an O / W emulsion, a W / O emulsion or a gel.

57. 50. The composition of any one of claims 1 to 49, which is a food or drink for human consumption.

58. The composition of any one of claims 1 to 49, wherein the formulation is a pharmaceutical composition or a dietary supplement.

59. 1. A method for fermentatively producing a retinoid-containing composition, comprising: (a) culturing a microorganism under conditions that allow for the production of a fermentation product; (b) isolating the fermentation product to obtain an isolated retinoid-containing composition. wherein the isolated retinoid-containing composition comprises: (i) a retinoid component comprising a mixture of cis and trans isomers; (ii) the fermentation residue thereof; Including, the retinoid component is present in a weight ratio of greater than 4:1 relative to the fermentation residue; The method wherein the cis isomer is present in an amount of less than 3% by weight based on the total weight of the retinoid component.

60. 60. The method of claim 59, wherein the isolating step further comprises one or more steps of distilling, crystallizing, filtering and / or washing the produced fermentation product.

61. 61. The method of claim 59 or 60, further comprising drying the isolated retinoid-containing composition to obtain a dry retinoid crystalline composition.

62. 62. The method of any one of claims 59 to 61, further comprising saponifying the isolated retinoid-containing composition and / or the dried retinoid crystal composition to obtain a retinol product.

63. 63. The method of claim 62, further comprising isolating the retinol product to obtain an isolated retinol composition.

64. 64. The method of claim 63, wherein the step of isolating the retinol product further comprises one or more steps of distilling, crystallizing, filtering, and / or washing the isolated retinol product.

65. 65. The method of claim 64, wherein the isolated retinoid-containing composition, the dry retinoid crystal composition and / or the isolated retinol composition comprises, consists of, or consists essentially of the composition of any one of claims 1-49.

66. The isolated retinoid-containing composition, the dry retinoid crystal composition, or the isolated retinol composition, (i) a retinoid component comprising a mixture of cis and trans isomers; (ii) the fermentation residue thereof; Including, the retinoid component is present in a weight ratio of greater than 4:1 relative to the fermentation residue; 66. A method for fermentatively producing a retinoid-containing composition according to any one of claims 59 to 65, wherein the cis-isomer is present in an amount of less than 3% by weight based on the total weight of the retinoid component.

67. 67. The method for fermentatively producing a retinoid-containing composition according to claim 66, wherein the fermentation residue comprises fatty acid retinyl esters (FARE), retinal, retinol, farnesol, a fermentable carbon source, or β-carotene.

68. The composition has a Weighted Selected Fermentation Readiness Value (WSFRV) of from 0.1 to 2.5, or from 0.1 to 2.0, or from 0.1 to 1.0, or from 0.1 to 0.9, or from 0.4 to 2.5, or from 0.4 to 2.0, or from 0.4 to 1.0, or from 0.4 to 0.9, or from 0.6 to 2.5, or from 0.6 to 2.0, or from 0.6 to 1.0, or from 0.5 to 0.9, wherein the WSFRV is calculated according to the following formula: WSFRV=C+0.1×F (In the formula, C = weight percent of the cis isomer present relative to the total amount of retinoid component (i) and fermentation residue (ii), and F = weight percent of FARE present relative to the total amount of retinoid component (I) and fermentation residue (II) 68. A method for fermentatively producing the retinoid-containing composition of any one of claims 59 to 67, wherein the retinoid-containing composition is determined according to the following formula:

69. The retinoid component has the formula (I): 【Chemistry 2】 comprising, consisting of, or consisting essentially of a compound according to R is -CHO, -CH ２ OH, -COOH, -CH(R １ ) ２ , -CH ２ OR ２ , -COOR ３ , -CONHR ４ or -CO(NR ４ ) ２ and R1 is independently lower alkoxy or R １’ and R １’’ taken together is a lower alkylenedioxy; R ２ is alkanoyl or aroyl, R ３ is alkyl, aryl or aralkyl, and R ４ , R ４’ and R ４’’ are independently hydrogen, alkyl, aryl, or aralkyl; The term "lower alkoxy" refers to an alkoxy group having 1 to 6 carbon atoms, such as methoxy, ethoxy, or propoxy; The term "lower alkylenedioxy" similarly refers to groups containing 1 to 6 carbon atoms, such as methylenedioxy or ethylenedioxy, where the alkyl or alkylene moiety may be linear or branched depending on the number of carbon atoms; The term "alkanoyl" refers to any straight-chain or branched alkanoyl group having from 1 to 18 carbon atoms, such as formyl, acetyl, propionyl, butyryl, stearoyl, and palmitoyl; The term "aroyl" refers to aromatic carboxylic acids having 7 and 11 carbon atoms, including benzoyl or naphthoyl, respectively; The term "alkyl" refers to a straight or branched alkyl group having 1 to 18 carbon atoms, for example, methyl, ethyl, propyl, butyl, decyl, dodecyl, hexadecyl, or octadecyl; The term "aryl" by itself or as part of "aralkyl" is phenyl or naphthyl, and 69. The method of any one of claims 59 to 68, wherein the term "aralkyl" includes groups having 1 to 4 carbon atoms in the aliphatic portion, such as benzyl and phenylpropyl.

70. 70. The method of any one of claims 59 to 69, wherein the retinoid component comprises, consists of, or consists essentially of retinol or a retinyl ester, and the retinyl ester comprises, consists of, or consists essentially of retinyl acetate or retinyl palmitate.

71. 71. The method of any one of claims 59 to 70, wherein the retinoid component comprises, consists of, or consists essentially of retinyl acetate.

72. The retinoid component comprises a mixture of cis and trans isomers, the cis isomers being present in an amount, by weight, based on the total weight of the mixture: 0.1 to 3% by weight, or 0.2 to 3% by weight, or 0.4 to 3% by weight, or 0.5 to 3% by weight, or 0.1 to 2.5% by weight, or 0.2 to 2.5% by weight, or 0.4 to 2.5% by weight, or 0.5 to 2.5% by weight, or 0.1 to 2% by weight, or 0.2 to 2% by weight, or 0.4 to 2% by weight, or 0.5 to 2% by weight, or 0.1 to 1.5% by weight, or 0.2 to 1.5% by weight, or 0.4 to 1.5% by weight, or 0.5 to 1.5% by weight, or 0.1 to 1% by weight, or 0.2 to 1% by weight, or 0.4 to 1% by weight, or 0.5 to 1% by weight, or 0.1 to 0.9% by weight, or 0.2 to 0.9% by weight, or 0.4 to 0.9% by weight, or 0.5 to 0.9% by weight 72. The method of any one of claims 59 to 71, wherein

73. When the retinoid component is retinol, the cis isomers include 9-cis-retinol, 11-cis-retinol, 13-cis-retinol, 9,13-di-cis-retinol, 11,13-di-cis-retinol, 13-cis-3,4-didehydroretinol, 9-cis-3,4-didehydroretinol and / or 9,13-di-cis-3,4-didehydroretinol; or When the retinoid component is retinal, the cis isomers include 13-cis retinal, 11-cis-retinal, 11,13-di-cis retinal, 9,13-di-cis-retinal, 9-cis retinal, 13-cis-3,4-didehydroretinal and / or 11,13-di-cis-3,4-didehydroretinal; or 73. The method of any one of claims 59 to 72, wherein when the retinoid component is retinyl acetate, the cis isomer comprises any one or more of the corresponding cis forms of retinyl acetate.

74. 74. The method of any one of claims 59 to 73, wherein the fermentation residue comprises FARE, beta-carotene, retinol, retinal and a fermentation carbon source.

75. 75. The method of any one of claims 59 to 74, wherein the fermentation residue comprises retinol, which retinol comprises, consists of or consists essentially of E-retinol.

76. 76. The method of any one of claims 59 to 75, wherein the fermentation residue comprises retinal, wherein the retinal comprises, consists of, or consists essentially of 9Z-retinal.

77. 77. The method of any one of claims 59 to 76, wherein the fermentation residue comprises one or more FAREs, and wherein the one or more FAREs are present in an amount of less than 4% by weight, or less than 3% by weight, or less than 2% by weight, or less than 1% by weight, relative to the total weight of retinoid components present.

78. the composition comprises 0.05 to 4%, or 0.1 to 4%, or 0.15 to 4%, or 0.2 to 4% by weight of FARE, based on the total weight of the retinoid components present; 0.05 to 3 wt.%, or 0.1 to 3 wt.%, or 0.15 to 3 wt.%, or 0.2 to 3 wt.% FARE; or 0.05 to 2% by weight, or 0.1 to 2% by weight, or 0.15 to 2% by weight, or 0.2 to 2% by weight of FARE; or 0.05 to 1 wt. %, or 0.1 to 1 wt. %, or 0.15 to 1 wt. %, or 0.2 to 1 wt. % FARE 78. The method of any one of claims 59 to 77, comprising:

79. 79. The method of any one of claims 59 to 78, wherein the FARE comprises retinyl palmitate and / or retinyl oleate.

80. 80. The method of any one of claims 59 to 79, wherein the fermentation residue comprises 9Z-retinal, and the 9Z-retinal is present in an amount of less than 10% by weight, or less than 8% by weight, or less than 6% by weight, or less than 5% by weight, or less than 3% by weight, relative to the total weight of retinoid components present.

81. The composition comprises 0.1 to 10% by weight, or 0.5 to 10% by weight, or 1 to 10% by weight, or 1.5 to 10% by weight of 9Z-retinal, based on the total weight of the retinoid components present; 0.1 to 8% by weight, or 0.5 to 8% by weight, or 1 to 8% by weight, or 1.5 to 8% by weight of 9Z-retinal; 0.1 to 6%, or 0.5 to 6%, or 1 to 6%, or 1.5 to 6% by weight of 9Z-retinal; 0.1 to 5% by weight, or 0.5 to 5% by weight, or 1 to 5% by weight, or 1.5 to 5% by weight of 9Z-retinal; 0.1 to 3% by weight, or 0.5 to 3% by weight, or 1 to 3% by weight, or 1.5 to 3% by weight of 9Z-retinal 81. The method of any one of claims 59 to 80, comprising:

82. 82. The method of any one of claims 59 to 81, wherein the fermentation residue comprises β-carotene, and the β-carotene is present in an amount of less than 4 wt.%, or less than 3 wt.%, or less than 2 wt.%, or less than 1 wt.%, or less than 0.5 wt.%, relative to the total weight of retinoid components present.

83. the composition comprises 0-4%, or 0.2-4%, or 0.5-4%, or 1-4% by weight of beta-carotene, based on the total weight of the retinoid components present; 0-3 wt.%, or 0.2-3 wt.%, or 0.5-3 wt.%, or 1-3 wt.% β-carotene, or 0-2% by weight, or 0.2-2% by weight, or 0.5-2% by weight, or 1-2% by weight of β-carotene, or 0-1 wt. %, or 0.2-1 wt. %, or 0.5-1 wt. % β-carotene, or 0 to 0.5% by weight or 0.2 to 0.5% by weight of β-carotene 83. The method of any one of claims 59 to 82, comprising:

84. 84. The method of any one of claims 59 to 83, wherein the fermentation residue comprises farnesol, and wherein the farnesol is present in an amount of less than 10,000 parts per million (ppm), or less than 5,000 ppm, or less than 1,000 ppm, or less than 100 ppm, or less than 10 ppm, or less than 5 ppm, or less than 2 ppm, based on the total weight of retinoid components present.

85. the composition comprises from 0 to 10,000 ppm, or from 2 to 10,000 ppm, or from 5 to 10,000 ppm, or from 10 to 10,000 ppm, or from 100 to 10,000 ppm of farnesol, based on the total weight of the retinoid components present; 0 to 5,000 ppm, or 2 to 5,000 ppm, or 5 to 5,000 ppm, or 10 to 5,000 ppm, or 100 to 5,000 ppm of farnesol; or 0 to 1,000 ppm, or 2 to 1,000 ppm, or 5 to 1,000 ppm, or 10 to 1,000 ppm, or 100 to 1,000 ppm of farnesol; or 0 to 100 ppm, or 2 to 100 ppm, or 5 to 100 ppm, or 10 to 100 ppm of farnesol 85. The method of any one of claims 59 to 84, comprising:

86. 86. The method of any one of claims 59 to 85, wherein the composition does not contain farnesol.

87. 87. The method of any one of claims 59 to 86, wherein the fermentation carbon source comprises, consists of, or consists essentially of a biogenic carbon source.

88. 88. The method of any one of claims 59 to 87, wherein the fermentation carbon source comprises straight chain alkanes, free fatty acids, ethanol, glucose containing triglycerides, in particular vegetable oils containing the respective free fatty acids derived therefrom, such as oleic acid, palmitic acid or linoleic acid.

89. 89. The method of any one of claims 59 to 88, wherein the fermentation residue comprises ethanol or glucose, and the ethanol or glucose is present in an amount of less than 2% by weight, or less than 1% by weight, or less than 0.75% by weight, relative to the total weight of the retinoid components present.

90. the composition comprises 0.05 to 2%, or 0.1 to 2%, or 0.2 to 2% by weight of ethanol or glucose based on the total weight of the retinoid components present; 0.05 to 1% by weight, or 0.1 to 1% by weight, or 0.2 to 1% by weight of ethanol or glucose, or 0.05 to 0.5% by weight, or 0.1 to 0.5% by weight, or 0.2 to 0.5% by weight of ethanol or glucose 90. The method of any one of claims 59 to 89, comprising:

91. 91. The method of any one of claims 59 to 90, wherein the fermentation residue comprises an isoparaffinic fluid.

92. 92. The method of any one of claims 59 to 91, wherein the isoparaffin fluid is present in an amount of less than 3 wt. %, or less than 2 wt. %, or less than 1.5 wt. %, or less than 1.25 wt. %, based on the total weight of the retinoid component (i) and its fermentation residue (ii).

93. the isoparaffin fluid is present in an amount of 0.1 to 2% by weight, or 0.2 to 2% by weight, or 0.5 to 2% by weight, or 0.1 to 1.5% by weight, or 0.2 to 1.5% by weight, or 0.5 to 1.5% by weight, based on the total weight of the retinoid component (i) and its fermentation residue (ii); and / or 93. The method of any one of claims 59 to 92, wherein the isoparaffinic fluid comprises, consists of, or consists essentially of Isopar M.

94. 94. The method of any one of claims 59 to 93, wherein the fermentation residue comprises the retinoid component in a dihydro form.

95. 95. The method of any one of claims 59 to 94, wherein the dihydro form is present in an amount of less than 0.5 wt. %, or less than 0.25 wt. %, or less than 0.1 wt. %, or less than 0.05 wt. %, based on the total weight of the retinoid component (i) and the fermentation residue thereof (ii).

96. 96. The method of any one of claims 59 to 95, wherein the dihydro form is present in an amount of 0.01 to 0.5 wt. %, or 0.01 to 0.25 wt. %, or 0.01 to 0.1 wt. %, or 0.01 to 0.05 wt. %, or 0.03 to 0.5 wt. %, or 0.03 to 0.25 wt. %, or 0.03 to 0.1 wt. %, or 0.03 to 0.05 wt. %, based on the total weight of the retinoid component (i) and the fermentation residue (ii) thereof.

97. 97. The method of claim 96, wherein the dihydro form of the retinoid component comprises, consists of, or consists essentially of dihydro-retinyl acetate.

98. 98. The method of any one of claims 59 to 97, wherein the fermentation residue comprises less than 0.5%, or less than 0.2%, or less than 0.1% by weight of rosafluene, phytoene, ergosterol and dihydro-beta-ionone, or is substantially free thereof.

99. the cis isomer is present in an amount of less than 1% by weight based on the total weight of the mixture of cis and trans isomers, and / or 99. The method of any one of claims 59 to 98, wherein FARE is present in an amount of less than 3.5% by weight or less than 1.5% by weight, based on the total weight of the retinoid component.

100. 100. The method of any one of claims 59 to 99, wherein the retinoid component is present in a weight ratio relative to the fermentation residue of greater than 5:1, or greater than 6:1, or greater than 10:1, or greater than 20:1, or greater than 25:

1.

101. the retinoid component is present in a weight ratio relative to the fermentation residue of 4:1 to 100:1, or 5:1 to 100:1, or 6:1 to 100:1, or 10:1 to 100:1, or 20:1 to 100:1, or 25:1 to 100:1; or a weight ratio of from 4:1 to 50:1, or from 5:1 to 50:1, or from 6:1 to 50:1, or from 10:1 to 50:1, or from 20:1 to 50:1, or from 25:1 to 50:1; or a weight ratio of from 4:1 to 35:1, or from 5:1 to 35:1, or from 6:1 to 35:1, or from 10:1 to 35:1, or from 20:1 to 35:1, or from 25:1 to 35:1; or a weight ratio of 4:1 to 30:1, or 5:1 to 30:1, or 6:1 to 30:1, or 10:1 to 30:1, or 20:1 to 30:1, or 25:1 to 30:1 101. The method of any one of claims 59 to 100, wherein

102. 102. The method of any one of claims 59 to 101, wherein the retinoid component comprises, consists of, or consists essentially of retinyl acetate, and the retinyl acetate is present in a weight ratio to the fermentation residue of greater than 5:1, or greater than 6:1, or greater than 10:1, or greater than 20:1, or greater than 25:

1.

103. the retinoid component comprises, consists of, or consists essentially of retinyl acetate, the retinyl acetate being present in a weight ratio to the fermentation residue of from 4:1 to 100:1, or from 5:1 to 100:1, or from 6:1 to 100:1, or from 10:1 to 100:1, or from 20:1 to 100:1, or from 25:1 to 100:1; or a weight ratio of from 4:1 to 50:1, or from 5:1 to 50:1, or from 6:1 to 50:1, or from 10:1 to 50:1, or from 20:1 to 50:1, or from 25:1 to 50:1; or a weight ratio of from 4:1 to 35:1, or from 5:1 to 35:1, or from 6:1 to 35:1, or from 10:1 to 35:1, or from 20:1 to 35:1, or from 25:1 to 35:1; or a weight ratio of 4:1 to 30:1, or 5:1 to 30:1, or 6:1 to 30:1, or 10:1 to 30:1, or 20:1 to 30:1, or 25:1 to 30:1 103. The method of any one of claims 59 to 102, wherein

104. 104. The method of any one of claims 59 to 103, wherein the biobased carbon content of the composition is greater than 60%, or greater than 70%, or greater than 80%, or greater than 90%, or greater than 99%.

105. 105. The method of any one of claims 59 to 104, wherein the bio-based carbon content of the retinoid component (i) and the fermentation residue thereof (ii) is greater than 60%, or greater than 70%, or greater than 80%, or greater than 90%, or greater than 99%.

106. The biobased carbon content is determined according to ASTM D6866-20 or １４ C and １３ 106. The method of any one of claims 59 to 105, determined by C isotope characterization.

107. 107. The method of any one of claims 59-106, wherein the amount of retinyl acetate, measured relative to the weight of the total composition, is within 5 wt. %, or within 4 wt. %, or within 3 wt. % of the amount of retinyl acetate present in the composition after the composition has been subjected to a thermal stability test in which the composition is heated to 105°C for 3 hours.

108. 108. The method of any one of claims 59 to 107, wherein the composition is in crystalline form.

109. The crystalline morphology has an average particle length D of greater than 100 microns, or greater than 200 microns, or greater than 300 microns, or greater than 400 microns, or between 100 and 1200 microns, or between 100 and 1000 microns, or between 100 and 800 microns, or between 200 and 1200 microns, or between 200 and 1000 microns, or between 200 and 800 microns, as determined by microscopic imaging methods. ５０ 109. The method of any one of claims 59 to 108, comprising a plurality of crystals having:

110. The method of any one of claims 59 to 109, wherein the composition further comprises a solvent.

111. 111. The method of any one of claims 59 to 110, wherein the solvent comprises ethanol or glucose.

112. 112. The method of any one of claims 59 to 111, wherein the composition, the retinoid component (i) and the fermentation residue thereof (ii) are present as an emulsion in oil.

113. 113. The method of any one of claims 59 to 112, wherein the emulsion in oil comprises a fat-soluble antioxidant and an oil.

114. 114. The method of any one of claims 59 to 113, wherein the oil preferably comprises a triglyceride, such as a vegetable oil and / or fat, and the fat-soluble antioxidant comprises ascorbic acid or a salt thereof, tocopherol, butylated hydroxytoluene (BHT), butylated hydroxyanisole (BHA), propyl gallate, tert.butylhydroxyquinoline, ethoxyquin, and / or an ascorbic acid ester of a fatty acid.

115. 115. The method of any one of claims 59-114, further comprising combining the isolated retinoid-containing composition, the dried retinoid crystal composition, and / or the isolated retinol product with one or more other food, feed, pharmaceutical, or personal care additive ingredients.

116. 1. A method for preparing a food, feed, pharmaceutical or personal care (pre)product, comprising: (a) a composition comprising: (i) a retinoid component comprising a mixture of cis and trans isomers; (ii) the fermentation residue thereof; comprising, consisting of, or consisting essentially of the retinoid component is present in a weight ratio of greater than 4:1 relative to the fermentation residue; Optionally, the cis isomer is present in an amount by weight of less than 3% by weight based on the total weight of the retinoid component; Optionally also providing a composition wherein the biobased carbon content of (i) and (ii) is greater than 50%, or greater than 60%, or greater than 70%, or greater than 90%; (b) formulating said composition to produce a formulation, preferably for use as a food, feed, pharmaceutical or personal care (pre)product; A method comprising:

117. 117. The method for preparing a food, feed, pharmaceutical or personal care (pre)product of claim 116, wherein the fermentation residue comprises fatty acid retinyl esters (FARE), retinal, retinol, farnesol, a fermentation carbon source or β-carotene.

118. The composition has a Weighted Selected Fermentation Readiness Value (WSFRV) of from 0.1 to 2.5, or from 0.1 to 2.0, or from 0.1 to 1.0, or from 0.1 to 0.9, or from 0.4 to 2.5, or from 0.4 to 2.0, or from 0.4 to 1.0, or from 0.4 to 0.9, or from 0.6 to 2.5, or from 0.6 to 2.0, or from 0.6 to 1.0, or from 0.5 to 0.9, wherein the WSFRV is calculated according to the following formula: WSFRV=C+0.1×F (In the formula, C = weight percent of the cis isomer present relative to the total amount of retinoid component (i) and fermentation residue (ii), and F = weight percent of FARE present relative to the total amount of retinoid component (I) and fermentation residue (II) 118. A method for preparing a food, feed, pharmaceutical or personal care (pre)product according to claim 116 or 117, wherein the food, feed, pharmaceutical or personal care (pre)product is determined according to

119. The retinoid component has the formula (I): 【Transformation 3】 comprising, consisting of, or consisting essentially of a compound according to R is -CHO, -CH ２ OH, -COOH, -CH(R １ ) ２ , -CH ２ OR ２ , -COOR ３ , -CONHR ４ or -CO(NR ４ ) ２ and R1 is independently lower alkoxy or R １’ and R １’’ taken together is a lower alkylenedioxy; R ２ is alkanoyl or aroyl, R ３ is alkyl, aryl or aralkyl, and R ４ , R ４’ and R ４’’ are independently hydrogen, alkyl, aryl, or aralkyl; The term "lower alkoxy" refers to an alkoxy group having 1 to 6 carbon atoms, such as methoxy, ethoxy, or propoxy; The term "lower alkylenedioxy" similarly refers to groups containing 1 to 6 carbon atoms, such as methylenedioxy or ethylenedioxy, where the alkyl or alkylene moiety may be linear or branched depending on the number of carbon atoms; The term "alkanoyl" refers to any straight-chain or branched alkanoyl group having from 1 to 18 carbon atoms, such as formyl, acetyl, propionyl, butyryl, stearoyl, and palmitoyl; The term "aroyl" refers to aromatic carboxylic acids having 7 and 11 carbon atoms, including benzoyl or naphthoyl, respectively; The term "alkyl" refers to a straight or branched alkyl group having 1 to 18 carbon atoms, for example, methyl, ethyl, propyl, butyl, decyl, dodecyl, hexadecyl, or octadecyl; The term "aryl" by itself or as part of "aralkyl" is phenyl or naphthyl, and 119. A method for preparing a food, feed, pharmaceutical or personal care (pre)product according to any one of claims 116 to 118, wherein the term "aralkyl" includes groups having 1 to 4 carbon atoms in the aliphatic portion, such as benzyl and phenylpropyl.

120. 120. A method of preparing a food, feed, pharmaceutical or personal care (pre)product according to any one of claims 116 to 119, wherein the retinoid component comprises, consists of or consists essentially of retinol or a retinyl ester, and the retinyl ester comprises, consists of or consists essentially of retinyl acetate or retinyl palmitate.

121. 121. A method of preparing a food, feed, pharmaceutical or personal care (pre)product according to any one of claims 116 to 120, wherein the retinoid component comprises, consists of or consists essentially of retinyl acetate.

122. The retinoid component comprises a mixture of cis and trans isomers, the cis isomers being present in an amount, by weight, based on the total weight of the mixture: 0.1 to 3% by weight, or 0.2 to 3% by weight, or 0.4 to 3% by weight, or 0.5 to 3% by weight, or 0.1 to 2.5% by weight, or 0.2 to 2.5% by weight, or 0.4 to 2.5% by weight, or 0.5 to 2.5% by weight, or 0.1 to 2% by weight, or 0.2 to 2% by weight, or 0.4 to 2% by weight, or 0.5 to 2% by weight, or 0.1 to 1.5% by weight, or 0.2 to 1.5% by weight, or 0.4 to 1.5% by weight, or 0.5 to 1.5% by weight, or 0.1 to 1% by weight, or 0.2 to 1% by weight, or 0.4 to 1% by weight, or 0.5 to 1% by weight, or 0.1 to 0.9% by weight, or 0.2 to 0.9% by weight, or 0.4 to 0.9% by weight, or 0.5 to 0.9% by weight 122. A method for preparing a food, feed, pharmaceutical or personal care (pre)product according to any one of claims 116 to 121, wherein the product is present in a

123. When the retinoid component is retinol, the cis isomers include 9-cis-retinol, 11-cis-retinol, 13-cis-retinol, 9,13-di-cis-retinol, 11,13-di-cis-retinol, 13-cis-3,4-didehydroretinol, 9-cis-3,4-didehydroretinol and / or 9,13-di-cis-3,4-didehydroretinol; or When the retinoid component is retinal, the cis isomers include 13-cis retinal, 11-cis-retinal, 11,13-di-cis retinal, 9,13-di-cis-retinal, 9-cis retinal, 13-cis-3,4-didehydroretinal and / or 11,13-di-cis-3,4-didehydroretinal; or 123. A method of preparing a food, feed, pharmaceutical or personal care (pre)product according to any one of claims 116 to 122, wherein when the retinoid component is retinyl acetate, the cis isomer comprises any one or more of the corresponding cis forms of retinyl acetate.

124. 124. A method for preparing a food, feed, pharmaceutical or personal care (pre)product according to any one of claims 116 to 123, wherein the fermentation residue comprises FARE, beta-carotene, retinol, retinal and a fermentation carbon source.

125. 125. A method for preparing a food, feed, pharmaceutical or personal care (pre)product according to any one of claims 116 to 124, wherein the fermentation residue comprises retinol, which retinol comprises, consists of or consists essentially of E-retinol.

126. 126. A method for preparing a food, feed, pharmaceutical or personal care (pre)product according to any one of claims 116 to 125, wherein the fermentation residue comprises retinal, and wherein the retinal comprises, consists of or consists essentially of 9Z-retinal.

127. 127. A method for preparing a food, feed, pharmaceutical or personal care (pre)product according to any one of claims 116 to 126, wherein the fermentation residue comprises one or more FAREs, and wherein the one or more FAREs are present in an amount of less than 4% by weight, or less than 3% by weight, or less than 2% by weight, or less than 1% by weight, relative to the total weight of retinoid components present.

128. the composition comprises 0.05 to 4%, or 0.1 to 4%, or 0.15 to 4%, or 0.2 to 4% by weight of FARE, based on the total weight of the retinoid components present; 0.05 to 3 wt.%, or 0.1 to 3 wt.%, or 0.15 to 3 wt.%, or 0.2 to 3 wt.% FARE; or 0.05 to 2% by weight, or 0.1 to 2% by weight, or 0.15 to 2% by weight, or 0.2 to 2% by weight of FARE; or 0.05 to 1 wt. %, or 0.1 to 1 wt. %, or 0.15 to 1 wt. %, or 0.2 to 1 wt. % FARE 128. A method for preparing a food, feed, pharmaceutical or personal care (pre)product according to any one of claims 116 to 127, comprising:

129. 129. A method for preparing a food, feed, pharmaceutical or personal care (pre)product according to any one of claims 116 to 128, wherein the FARE comprises retinyl palmitate and / or retinyl oleate.

130. 130. A method for preparing a food, feed, pharmaceutical or personal care (pre)product according to any one of claims 116 to 129, wherein the fermentation residue comprises 9Z-retinal and wherein the 9Z-retinal is present in an amount of less than 10% by weight, or less than 8% by weight, or less than 6% by weight, or less than 5% by weight, or less than 3% by weight, relative to the total weight of retinoid components present.

131. The composition comprises 0.1 to 10% by weight, or 0.5 to 10% by weight, or 1 to 10% by weight, or 1.5 to 10% by weight of 9Z-retinal, based on the total weight of the retinoid components present; 0.1 to 8% by weight, or 0.5 to 8% by weight, or 1 to 8% by weight, or 1.5 to 8% by weight of 9Z-retinal; 0.1 to 6%, or 0.5 to 6%, or 1 to 6%, or 1.5 to 6% by weight of 9Z-retinal; 0.1 to 5% by weight, or 0.5 to 5% by weight, or 1 to 5% by weight, or 1.5 to 5% by weight of 9Z-retinal; 0.1 to 3% by weight, or 0.5 to 3% by weight, or 1 to 3% by weight, or 1.5 to 3% by weight of 9Z-retinal 131. A method for preparing a food, feed, pharmaceutical or personal care (pre)product according to any one of claims 116 to 130, comprising:

132. 132. A method for preparing a food, feed, pharmaceutical or personal care (pre)product according to any one of claims 116 to 131, wherein the fermentation residue comprises β-carotene, and wherein the β-carotene is present in an amount of less than 4 wt.%, or less than 3 wt.%, or less than 2 wt.%, or less than 1 wt.%, or less than 0.5 wt.%, relative to the total weight of retinoid components present.

133. the composition comprises 0-4%, or 0.2-4%, or 0.5-4%, or 1-4% by weight of beta-carotene, based on the total weight of the retinoid components present; 0-3 wt.%, or 0.2-3 wt.%, or 0.5-3 wt.%, or 1-3 wt.% β-carotene, or 0-2% by weight, or 0.2-2% by weight, or 0.5-2% by weight, or 1-2% by weight of β-carotene, or 0-1 wt. %, or 0.2-1 wt. %, or 0.5-1 wt. % β-carotene, or 0 to 0.5% by weight or 0.2 to 0.5% by weight of β-carotene 133. A method for preparing a food, feed, pharmaceutical or personal care (pre)product according to any one of claims 116 to 132, comprising:

134. 134. The method of preparing a food, feed, drug or personal care (pre)product according to any one of claims 116 to 133, wherein the fermentation residue comprises farnesol and wherein the farnesol is present in an amount of less than 10,000 parts per million (ppm), or less than 5,000 ppm, or less than 1,000 ppm, or less than 100 ppm, or less than 10 ppm, or less than 5 ppm, or less than 2 ppm, based on the total weight of retinoid components present.

135. the composition comprises from 0 to 10,000 ppm, or from 2 to 10,000 ppm, or from 5 to 10,000 ppm, or from 10 to 10,000 ppm, or from 100 to 10,000 ppm of farnesol, based on the total weight of the retinoid components present; 0 to 5,000 ppm, or 2 to 5,000 ppm, or 5 to 5,000 ppm, or 10 to 5,000 ppm, or 100 to 5,000 ppm of farnesol; or 0 to 1,000 ppm, or 2 to 1,000 ppm, or 5 to 1,000 ppm, or 10 to 1,000 ppm, or 100 to 1,000 ppm of farnesol; or 0 to 100 ppm, or 2 to 100 ppm, or 5 to 100 ppm, or 10 to 100 ppm of farnesol 135. A method for preparing a food, feed, pharmaceutical or personal care (pre)product according to any one of claims 116 to 134, comprising:

136. 136. A method for preparing a food, feed, pharmaceutical or personal care (pre)product according to any one of claims 116 to 135, wherein the composition does not contain farnesol.

137. 137. A method for preparing a food, feed, pharmaceutical or personal care (pre)product according to any one of claims 116 to 136, wherein the fermentation carbon source comprises, consists of or consists essentially of a biogenic carbon source.

138. 138. A method for preparing a food, feed, pharmaceutical or personal care (pre)product according to any one of claims 116 to 137, wherein the fermentation carbon source comprises straight chain alkanes, free fatty acids, ethanol, glucose with triglycerides, in particular vegetable oils with the respective free fatty acids derived therefrom, such as oleic acid, palmitic acid or linoleic acid.

139. 139. A method for preparing a food, feed, pharmaceutical or personal care (pre)product according to any one of claims 116 to 138, wherein the fermentation residue comprises ethanol or glucose, and the ethanol or glucose is present in an amount of less than 2% by weight, or less than 1% by weight, or less than 0.75% by weight, relative to the total weight of retinoid components present.

140. the composition comprises 0.05 to 2%, or 0.1 to 2%, or 0.2 to 2% by weight of ethanol or glucose based on the total weight of the retinoid components present; 0.05 to 1% by weight, or 0.1 to 1% by weight, or 0.2 to 1% by weight of ethanol or glucose, or 0.05 to 0.5% by weight, or 0.1 to 0.5% by weight, or 0.2 to 0.5% by weight of ethanol or glucose 140. A method for preparing a food, feed, pharmaceutical or personal care (pre)product according to any one of claims 116 to 139, comprising:

141. 141. A method for preparing a food, feed, pharmaceutical or personal care (pre)product according to any one of claims 116 to 140, wherein the fermentation residue comprises an isoparaffinic fluid.

142. 142. A method for preparing a food, feed, pharmaceutical or personal care (pre)product according to any one of claims 116 to 141, wherein the isoparaffin fluid is present in an amount of less than 3 wt%, or less than 2 wt%, or less than 1.5 wt%, or less than 1.25 wt%, based on the total weight of the retinoid component (i) and its fermentation residue (ii).

143. the isoparaffin fluid is present in an amount of 0.1 to 2% by weight, or 0.2 to 2% by weight, or 0.5 to 2% by weight, or 0.1 to 1.5% by weight, or 0.2 to 1.5% by weight, or 0.5 to 1.5% by weight, based on the total weight of the retinoid component (i) and its fermentation residue (ii); and / or 143. A method for preparing a food, feed, pharmaceutical or personal care (pre)product according to any one of claims 116 to 142, wherein the isoparaffinic fluid comprises, consists of or consists essentially of Isopar M.

144. 144. A method for preparing a food, feed, pharmaceutical or personal care (pre)product according to any one of claims 116 to 143, wherein the fermentation residue comprises the retinoid component in dihydro form.

145. 145. A method for preparing a food, feed, pharmaceutical or personal care (pre)product according to any one of claims 116 to 144, wherein the dihydro form is present in an amount of less than 0.5 wt.%, or less than 0.25 wt.%, or less than 0.1 wt.%, or less than 0.05 wt.%, based on the total weight of the retinoid component (i) and the fermentation residue thereof (ii).

146. 146. A method for preparing a food, feed, pharmaceutical or personal care (pre)product according to any one of claims 116 to 145, wherein the dihydro form is present in an amount of 0.01 to 0.5 wt. %, or 0.01 to 0.25 wt. %, or 0.01 to 0.1 wt. %, or 0.01 to 0.05 wt. %, or 0.03 to 0.5 wt. %, or 0.03 to 0.25 wt. %, or 0.03 to 0.1 wt. %, or 0.03 to 0.05 wt. % based on the total weight of the retinoid component (i) and the fermentation residue thereof (ii).

147. 147. The method of preparing a food, feed, drug or personal care (pre)product of claim 146, wherein the dihydro form of the retinoid component comprises, consists of or consists essentially of dihydro-retinyl acetate.

148. 148. A method for preparing a food, feed, pharmaceutical or personal care (pre)product according to any one of claims 116 to 147, wherein the fermentation residue contains less than 0.5% by weight, or less than 0.2% by weight, or less than 0.1% by weight of rosafluene, phytoene, ergosterol and dihydro-beta-ionone, or is substantially free thereof.

149. the cis isomer is present in an amount of less than 1% by weight based on the total weight of the mixture of cis and trans isomers, and / or 149. A method of preparing a food, feed, pharmaceutical or personal care (pre)product according to any one of claims 116 to 148, wherein FARE is present in an amount of less than 3.5% by weight or less than 1.5% by weight, based on the total weight of the retinoid component.

150. 150. A method of preparing a food, feed, pharmaceutical or personal care (pre)product according to any one of claims 116 to 149, wherein the retinoid component is present in a weight ratio to the fermentation residue of greater than 5:1, or greater than 6:1, or greater than 10:1, or greater than 20:1, or greater than 25:

1.

151. the retinoid component is present in a weight ratio relative to the fermentation residue of 4:1 to 100:1, or 5:1 to 100:1, or 6:1 to 100:1, or 10:1 to 100:1, or 20:1 to 100:1, or 25:1 to 100:1; or a weight ratio of from 4:1 to 50:1, or from 5:1 to 50:1, or from 6:1 to 50:1, or from 10:1 to 50:1, or from 20:1 to 50:1, or from 25:1 to 50:1; or a weight ratio of from 4:1 to 35:1, or from 5:1 to 35:1, or from 6:1 to 35:1, or from 10:1 to 35:1, or from 20:1 to 35:1, or from 25:1 to 35:1; or a weight ratio of 4:1 to 30:1, or 5:1 to 30:1, or 6:1 to 30:1, or 10:1 to 30:1, or 20:1 to 30:1, or 25:1 to 30:1 151. A method for preparing a food, feed, pharmaceutical or personal care (pre)product according to any one of claims 116 to 150, wherein the product is present in a

152. 152. A method of preparing a food, feed, pharmaceutical or personal care (pre)product according to any one of claims 116 to 151, wherein the retinoid component comprises, consists of or consists essentially of retinyl acetate, and the retinyl acetate is present in a weight ratio to the fermentation residue of greater than 5:1, or greater than 6:1, or greater than 10:1, or greater than 20:1, or greater than 25:

1.

153. the retinoid component comprises, consists of, or consists essentially of retinyl acetate, the retinyl acetate being present in a weight ratio to the fermentation residue of from 4:1 to 100:1, or from 5:1 to 100:1, or from 6:1 to 100:1, or from 10:1 to 100:1, or from 20:1 to 100:1, or from 25:1 to 100:1; or a weight ratio of from 4:1 to 50:1, or from 5:1 to 50:1, or from 6:1 to 50:1, or from 10:1 to 50:1, or from 20:1 to 50:1, or from 25:1 to 50:1; or a weight ratio of from 4:1 to 35:1, or from 5:1 to 35:1, or from 6:1 to 35:1, or from 10:1 to 35:1, or from 20:1 to 35:1, or from 25:1 to 35:1; or a weight ratio of 4:1 to 30:1, or 5:1 to 30:1, or 6:1 to 30:1, or 10:1 to 30:1, or 20:1 to 30:1, or 25:1 to 30:1 153. A method for preparing a food, feed, pharmaceutical or personal care (pre)product according to any one of claims 116 to 152, wherein the product is

154. 154. A method of preparing a food, feed, pharmaceutical or personal care (pre)product according to any one of claims 116 to 153, wherein the biobased carbon content of the composition is greater than 60%, or greater than 70%, or greater than 80%, or greater than 90%, or greater than 99%.

155. 155. The method for preparing a food, feed, pharmaceutical or personal care (pre)product according to any one of claims 116 to 154, wherein the bio-based carbon content of the retinoid component (i) and the fermentation residue thereof (ii) is greater than 60%, or greater than 70%, or greater than 80%, or greater than 90%, or greater than 99%.

156. The biobased carbon content is determined according to ASTM D6866-20 or １４ C and １３ 156. A method for preparing a food, feed, pharmaceutical or personal care (pre)product according to any one of claims 116 to 155, as determined by C isotope characterisation.

157. 157. A method of preparing a food, feed, pharmaceutical or personal care (pre)product according to any one of claims 116 to 156, wherein the amount of retinyl acetate, measured relative to the weight of the total composition, is within 5% by weight, or within 4% by weight, or within 3% by weight of the amount of retinyl acetate present in the composition after the composition has been subjected to a thermal stability test in which the composition is heated to 105°C for 3 hours.

158. 158. A method for preparing a food, feed, pharmaceutical or personal care (pre)product according to any one of claims 116 to 157, wherein the composition is present in crystalline form.

159. The crystalline morphology has an average particle length D of greater than 100 microns, or greater than 200 microns, or greater than 300 microns, or greater than 400 microns, or between 100 and 1200 microns, or between 100 and 1000 microns, or between 100 and 800 microns, or between 200 and 1200 microns, or between 200 and 1000 microns, or between 200 and 800 microns, as determined by microscopic imaging methods. ５０ 159. A method for preparing a food, feed, pharmaceutical or personal care (pre)product according to any one of claims 116 to 158, comprising a plurality of crystals having

160. 160. A method for preparing a food, feed, pharmaceutical or personal care (pre)product according to any one of claims 116 to 159, wherein the composition further comprises a solvent.

161. 161. A method for preparing a food, feed, pharmaceutical or personal care (pre)product according to any one of claims 116 to 160, wherein the solvent comprises ethanol.

162. 162. A method for preparing a food, feed, pharmaceutical or personal care (pre)product according to any one of claims 116 to 161, wherein the composition, the retinoid component (i) and the fermentation residue thereof (ii) are present as an emulsion in oil.

163. 163. A method for preparing a food, feed, pharmaceutical or personal care (pre)product according to any one of claims 116 to 162, wherein the emulsion in oil comprises a fat soluble antioxidant and oil.

164. 164. A method for preparing a food, feed, pharmaceutical or personal care (pre)product according to any one of claims 116 to 163, wherein the oil preferably comprises a triglyceride, such as a vegetable oil and / or fat, and the fat-soluble antioxidant comprises ascorbic acid or a salt thereof, tocopherol, butylated hydroxytoluene (BHT), butylated hydroxyanisole (BHA), propyl gallate, tert.butylhydroxyquinoline, ethoxyquin and / or ascorbic acid esters of fatty acids.

165. The blending step (b) (1) a sub-step of dissolving a matrix component in water to prepare a matrix solution; (2) optionally, preferably also in oil, heating the composition according to any one of claims 116 to 164 to obtain an active phase; (3) a substep of emulsifying the active phase in the matrix solution to obtain a dispersion; (4) drying the dispersion in the presence of a capture medium to obtain at least one beadlet; 165. A method for preparing a food, feed, pharmaceutical or personal care (pre)product according to any one of claims 116 to 164, comprising:

166. 166. A method for preparing a food, feed, pharmaceutical or personal care (pre)product according to any one of claims 116 to 165, wherein the blending step (b) further comprises the sub-step of (5) heating the dispersion and / or spray-dried dispersion.

167. 167. A method of preparing a food, feed, pharmaceutical or personal care (pre)product according to any one of claims 116 to 166, wherein the matrix component comprises, consists of or consists essentially of one or more hydrocolloids.

168. 168. A method of preparing a food, feed, pharmaceutical or personal care (pre)product according to any one of claims 116 to 167, wherein the one or more hydrocolloids comprise gelatin, xanthan gum, acacia gum, pectin, guar, carob gum, alginate, cellulose or cellulose derivatives or combinations thereof.

169. 169. A method for preparing a food, feed, pharmaceutical or personal care (pre)product according to any one of claims 116 to 168, wherein the cellulose derivative comprises carboxymethylcellulose and / or modified polysaccharides.

170. 170. A method for preparing a food, feed, pharmaceutical or personal care (pre)product according to any one of claims 116 to 169, wherein the modified polysaccharide comprises modified food starch.

171. The matrix components may be modified (food) starches, pectins, alginates, carrageenans, furcellarans, chitosan, maltodextrins, dextrin derivatives, cellulose and cellulose derivatives (e.g., cellulose acetate, methylcellulose, hydroxypropylmethylcellulose), lignosulfonates, polysaccharide gums (e.g., acacia gum, gum arabic, linseed gum, ghatti gum, tamarind gum, and arabinogalactans), gelatins (bovine, fish, porcine, poultry), vegetable proteins (e.g., pea, soybean, castor bean, cotton, potato, etc.), and the like.

171. A method for preparing a food, feed, pharmaceutical or personal care (pre)product according to any one of claims 116 to 170 comprising animal proteins including wheat, sweet potato, manioc, rapeseed, sunflower, sesame, linseed, safflower, lentil, nuts, wheat, rice, corn, barley, rye, oat, lupin and sorghum), animal proteins including milk or whey proteins, lecithin, polyglycerol esters of fatty acids, monoglycerides of fatty acids, diglycerides of fatty acids, sorbitan esters, PG esters and sugar esters (and derivatives thereof).

172. 172. A method for preparing a food, feed, pharmaceutical or personal care (pre)product according to any one of claims 116 to 171, wherein a fat-soluble antioxidant is added in the blending step (b).

173. 173. The method of preparing a food, feed, pharmaceutical or personal care (pre)product according to any one of claims 116 to 172, wherein the fat-soluble antioxidant comprises ascorbic acid or a salt thereof, tocopherol, butylated hydroxytoluene (BHT), butylated hydroxyanisole (BHA), propyl gallate, tert.butylhydroxyquinoline, ethoxyquin and / or ascorbic acid esters of fatty acids.

174. 174. A method for preparing a food, feed, pharmaceutical or personal care (pre)product according to any one of claims 116 to 173, wherein the fat-soluble antioxidant is added to the composition and the oil in the heating sub-step (2).

175. 175. A method for preparing a food, feed, pharmaceutical or personal care (pre)product according to any one of claims 116 to 174, wherein the oil comprises, consists or consists essentially of a triglyceride.

176. 176. A method for preparing a food, feed, pharmaceutical or personal care (pre)product according to any one of claims 116 to 175, wherein the triglycerides comprise vegetable oils and / or fats.

177. 177. A method of preparing a food, feed, pharmaceutical or personal care (pre)product according to any one of claims 116 to 176, wherein the vegetable oil and / or fat comprises corn oil, sunflower oil, (hydrogenated) soybean oil, safflower oil, rapeseed oil, peanut oil, (hydrogenated) palm oil, palm kernel oil, cottonseed oil and / or coconut oil, including fractional amounts thereof.

178. 178. A method for preparing a food, feed, pharmaceutical or personal care (pre)product according to any one of claims 116 to 177, wherein the entrapment medium comprises corn starch or an organic or inorganic anti-caking agent.

179. 179. A beadlet produced by the method of any one of claims 165 to 178.

180. 179. A method of preparing a food, feed, pharmaceutical or personal care (pre)product according to any one of claims 116 to 178, wherein the stabilised formulation comprises, consists or consists essentially of a beadlet according to claim 179 and / or is produced by a method according to any one of claims 165 to 178.

181. The blending step (b) (1) introducing the composition according to any one of claims 116 to 178 or 180, a matrix component, water and oil into an extruder having an orifice; (2) extruding the composition, matrix component, water, and oil through the orifice to obtain an extrudate; 181. A method for preparing a food, feed, pharmaceutical or personal care (pre)product according to any one of claims 116 to 178 or 180, comprising:

182. 182. The method of claim 181, wherein the extrudate comprises an emulsion of the composition and oil in the water and matrix component, the emulsion being formed inside the extruder.

183. 183. A method for preparing a food, feed, pharmaceutical or personal care (pre)product according to any one of claims 116 to 178 or 180 to 182, wherein the composition and oil are introduced together into the extruder as a premixed active phase.

184. The blending step (b) (3) cutting the extrudate into individual particles; (4) optionally, drying the individual particles; 184. A method for preparing a food, feed, pharmaceutical or personal care (pre)product according to any one of claims 116 to 178 or 180 to 183, further comprising:

185. 185. A method of preparing a food, feed, pharmaceutical or personal care (pre)product according to any one of claims 116 to 178 or 180 to 184, wherein the combining step (b) further comprises directing a flow of a cooling fluid over the orifice and the extrudate.

186. 186. A method for preparing a food, feed, pharmaceutical or personal care (pre)product according to any one of claims 116 to 178 or 180 to 185, wherein the cooling fluid is configured to flow along a cooling flow axis and the extrudate is configured to be extruded through an exit axis, and the cooling flow axis and the exit axis are substantially anti-parallel and / or substantially collinear.

187. 187. A method of preparing a food, feed, pharmaceutical or personal care (pre)product according to any one of claims 116 to 178 or 180 to 186, wherein the matrix component comprises, consists of or consists essentially of one or more hydrocolloids.

188. 188. A method of preparing a food, feed, pharmaceutical or personal care (pre)product according to any one of claims 116 to 178 or 180 to 187, wherein the one or more hydrocolloids comprise gelatin, xanthan gum, acacia gum, pectin, guar, carob gum, alginate, cellulose or cellulose derivatives or combinations thereof.

189. 189. A method for preparing a food, feed, pharmaceutical or personal care (pre)product according to any one of claims 116 to 178 or 180 to 188, wherein the cellulose derivative comprises carboxymethylcellulose and / or modified polysaccharides.

190. 190. A method for preparing a food, feed, pharmaceutical or personal care (pre)product according to any one of claims 116 to 178 or 180 to 189, wherein the modified polysaccharide comprises modified food starch.

191. The matrix components may be modified (food) starches, pectins, alginates, carrageenans, furcellarans, chitosan, maltodextrins, dextrin derivatives, cellulose and cellulose derivatives (e.g., cellulose acetate, methylcellulose, hydroxypropylmethylcellulose), lignosulfonates, polysaccharide gums (e.g., acacia gum, gum arabic, linseed gum, ghatti gum, tamarind gum, and arabinogalactans), gelatins (bovine, fish, porcine, poultry), vegetable proteins (e.g., pea, soybean, castor bean, cotton, potato, sweet potato, etc.), and the like.

191. A method for preparing a food, feed, pharmaceutical or personal care (pre)product according to any one of claims 116 to 178 or 180 to 190, comprising animal proteins including yams, manioc, rapeseed, sunflower, sesame, linseed, safflower, lentils, nuts, wheat, rice, corn, barley, rye, oats, lupin and sorghum), animal proteins including milk or whey proteins, lecithin, polyglycerol esters of fatty acids, monoglycerides of fatty acids, diglycerides of fatty acids, sorbitan esters, PG esters and sugar esters (and derivatives thereof).

192. 192. A method for preparing a food, feed, pharmaceutical or personal care (pre)product according to any one of claims 116 to 178 or 180 to 191, wherein a fat soluble antioxidant is added in the extruder.

193. 193. The method for preparing a food, feed, pharmaceutical or personal care (pre)product according to any one of claims 116 to 178 or 180 to 192, wherein the fat-soluble antioxidant comprises ascorbic acid or a salt thereof, tocopherol, butylated hydroxytoluene (BHT), butylated hydroxyanisole (BHA), propyl gallate, tert.butylhydroxyquinoline, ethoxyquin and / or ascorbic acid esters of fatty acids.

194. 194. A method for preparing a food, feed, pharmaceutical or personal care (pre)product according to any one of claims 116 to 178 or 180 to 193, wherein the oil comprises, consists of or consists essentially of a triglyceride.

195. 195. A method for preparing a food, feed, pharmaceutical or personal care (pre)product according to any one of claims 116 to 178 or 180 to 194, wherein the triglycerides comprise vegetable oils and / or fats.

196. 196. A method for preparing a food, feed, pharmaceutical or personal care (pre)product according to any one of claims 116 to 178 or 180 to 195, wherein the vegetable oil and / or fat comprises corn oil, sunflower oil, (hydrogenated) soybean oil, safflower oil, rapeseed oil, peanut oil, (hydrogenated) palm oil, palm kernel oil, cottonseed oil and / or coconut oil, including fractional amounts thereof.

197. 197. An extrudate produced by the method of any one of claims 181 to 196.

198. 197. A method of preparing a food, feed, pharmaceutical or personal care (pre)product as claimed in any one of claims 116 to 178 or 180 to 196, wherein the stabilised formulation comprises, consists or consists essentially of the extrudate of claim 197 and / or is produced by a method as claimed in any one of claims 181 to 196.

199. 199. A method of preparing the food, feed, drug or personal care (pre)product of any one of claims 116-178, 180-196 or 198, further comprising the step of: (c) combining the stabilized formulation with one or more additives to produce the food, feed, drug or personal care product.

200. 199. A method for preparing a food, feed, pharmaceutical or personal care (pre)product according to any one of claims 116 to 178, 180 to 196, 198 or 199, wherein the personal care product comprises a cosmetic liquid, lotion, thickened lotion, gel, cream, emulsion, ointment, paste, powder, make-up, solid tube stick, aerosol, mousse, foam and / or spray.

201. 197. Use of the beadlets of claim 179 and / or the extrudates of claim 197 in food, feed, pharmaceutical and personal care products.

202. 197. A food, feed, pharmaceutical or personal care product comprising the beadlet of claim 179 and / or the extrudate of claim 197.

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