1,3-Fatty diol compounds and their derivatives

Abstract

Provided is a 1,3-fatty diol compound that is useful as an emulsifier, etc. [Solution] A compound of formula (II) is a 12-carbon unbranched unsaturated fatty diol having a single Δ5 double bond and hydroxy groups at carbon number 1 (C-1) and carbon number 3 (C-3), with the chiral center located at C-3, and a compound of formula (IV) is a 16-carbon unbranched unsaturated fatty diol having a single Δ9 double bond and hydroxy groups at carbon number 1 (C-1) and carbon number 3 (C-3), with the chiral center located at C-3. TIFF2022092019000049.tif60170

Description

[Technical field]

[0001] Field The present disclosure relates generally to the field of specialty chemicals suitable for use, for example, as industrial substances and process components in the production of detergents and surfactants, as emulsifiers, emollients, and thickeners in cosmetics and foods, as industrial solvents and plasticizers, and the like.

[0002] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of U.S. Provisional Application No. 62 / 394,537, filed Sep. 14, 2016, the entirety of which is incorporated herein by reference. [Background technology]

[0003] background Fatty alcohols, especially fatty diols (or aliphatic diols), are amphiphilic molecules with commercial and industrial applications. For example, fatty alcohols find use as softeners and thickeners in cosmetics and foods, as industrial solvents, plasticizers, lubricants, emulsifiers, building blocks of polymers, etc. (see, for example, H. Maag (1984) Journal of the American Oil Chemists' Society 61(2):259-267). Particularly useful are 1,3-fatty diols.

[0004] 1,3-fatty diols are useful as lubricants and as linking molecules between other molecules (e.g., examples in the production of polymers). 1,3-fatty diols are also useful as surfactants and surfactant precursors, for example, 1,3-fatty diols can be used to prepare "gemini" surfactants in which both alcohol moieties are chemically modified (e.g., ethoxylated, glycosylated, sulfated, etc.). Gemini surfactants or gemini-like surfactants exhibit superior properties compared to those of similar conventional surfactants (e.g., see Gemini Surfactants: Synthesis, Interfacial and Solution-Phase Behavior, and Applications, Vol. 117, Zana, R.; Xia, J., Eds.; Marcel Dekker: New York, 2004).

[0005] The 3-hydroxy moiety of 1,3-fatty diols forms a chiral center at the third carbon (C-3), making them useful as synthons for the generation of chirally important compounds such as pharmaceuticals, nutraceuticals, pesticides, herbicides, flavors, fragrances, solvents, and biologically active compounds.

[0006] In addition to the functionality of the hydroxyl groups, variations in the structure of the carbon chains of 1,3-fatty diols provide the molecules with additional chemical properties and / or potentially new properties that can be used to address existing problems in an improved way and / or find entirely new applications. For example, unsaturated fatty diols have additional functional groups in the form of double bonds that are available for chemical reactions.

[0007] Unsaturated fatty alcohols and diols are especially valuable because the presence of double bonds contributes many advantages to the molecule. For example, compared to their saturated counterparts, unsaturated fatty diols have lower melting points, higher water solubility, and offer the possibility of introducing functional groups to the C=C double bonds (Egan, R., et al., (1984) Journal of the American Oil Chemists' Society, Vol. 61(2): 324-329 (Non-Patent Document 3)). Therefore, unsaturated fatty alcohols are important intermediates for many products in the chemical industry (see, for example, U. Ploog et al. in Seifen-Ole-Fette-Wachse 109, 225 (1983) (Non-Patent Document 4)).

[0008] Unlike saturated fatty diols, which are readily produced from petroleum precursors, for example using the Prins reaction (see, e.g., E. Arundale, L.A. Mikeska Chem. Rev.; 1952; 51(3); 505-555), unsaturated fatty diols are much more difficult to produce from petrochemical materials and processes.

[0009] Typically, unsaturated fatty alcohols are produced by subjecting fatty acid methyl ester mixtures derived from oils such as sunflower oil, palm oil, palm kernel, and coconut to high pressure hydrogenation in the presence of chromium and / or zinc-containing mixed oxide catalysts (see, for example, Ullmann's Encyclopedia of Industrial Chemistry, Vol. 7, No. 1, 2003, pp. 111-115, 2003). th Edition, Vol. 14:117. John Wiley and Sons, Inc. 2011 (Non-Patent Document 6). Zinc chromite-type catalysts promote the selective hydrogenation of carbonyl groups instead of C=C double bonds (see, for example, Adkins and Sauer, (1937) Journal of the American Chemical Society, Vol. 59(1):1-3 (Non-Patent Document 7)).

[0010] Unfortunately, unsaturated fatty alcohols and unsaturated fatty diols can only be produced from a limited number of natural oils, so the supply of raw materials can be unstable and variable. In addition, the reliance on natural oils and their inherent structures for the production of fatty diols limits the variety of chemical structures that can be produced.

[0011] Thus, because 1,3-fatty diols, and especially unsaturated 1,3-fatty diols, are such useful molecules, it is apparent that what is needed in the art are new 1,3-fatty diols.

[0012] Fortunately, the present invention addresses this and other needs, as will become apparent from the detailed description that follows. [Prior art documents] [Non-patent literature]

[0013] [Non-Patent Document 1] H. Maag (1984) Journal of the American Oil Chemists' Society61(2):259-267 [Non-Patent Document 2] Gemini Surfactants:Synthesis,Interfacial and Solution-Phase Behavior,and Applications,Vol.117,Zana,R.;Xia,J.,Eds.;Marcel Dekker:New York,2004 [Non-Patent Document 3] Egan, R., et al., (1984) Journal of the American Oil Chemists' Society, Vol. 61(2): 324-329 [Non-Patent Document 4] U. Ploog et al. in Seifen-Ole-Fette-Wachse109,225(1983) [Non-Patent Document 5] E.Arundale,LAMikeskaChem.Rev.;1952;51(3);505-555 [Non-Patent Document 6] Ullmann's Encyclopedia of Industrial Chemistry7th Edition,Vol.14:117.John Wiley and Sons,Inc.2011 [Non-Patent Document 7] Adkins and Sauer,(1937)Journal of the American Chemical Society,Vol.59(1):1-3 Summary of the Invention

[0014] overview One aspect of the present disclosure provides a 12-carbon unbranched unsaturated fatty diol having a single Δ5 double bond, a hydroxy group at carbon number 1 (C-1) and a hydroxy group at carbon number 3 (C-3), wherein a chiral center is present at C-3, and the fatty diol has the general chemical formula of Formula II. TIFF0007601403000001.tif18128

[0015] In one exemplary embodiment, the double bond is in the (Z) configuration. In another exemplary embodiment, the double bond is in the (E) configuration. In one exemplary embodiment, the chiral center at C-3 has an R configuration. In one exemplary embodiment, the chiral center at C-3 has an S configuration. In another exemplary embodiment, the double bond is in the (Z) configuration and the chiral center at C-3 has an R configuration.

[0016] One aspect of the present disclosure provides a 14 carbon unbranched unsaturated fatty diol having a single Δ7 double bond, a hydroxy group at carbon number 1 (C-1) and a hydroxy group at carbon number 3 (C-3), wherein a chiral center is present at C-3, and the fatty diol has the general chemical formula of Formula III. TIFF0007601403000002.tif19128

[0017] In one exemplary embodiment, the double bond is in the (Z) configuration. In another exemplary embodiment, the double bond is in the (E) configuration. In one exemplary embodiment, the chiral center at C-3 has an R configuration. In one exemplary embodiment, the chiral center has an S configuration. In another exemplary embodiment, the double bond is in the (Z) configuration and the chiral center at C-3 has an R configuration.

[0018] One aspect of the present disclosure provides a 16 carbon unbranched unsaturated fatty diol having a single Δ9 double bond, a hydroxy group at carbon number 1 (C-1) and a hydroxy group at carbon number 3 (C-3), wherein a chiral center is present at C-3, and the fatty diol has the general chemical formula of Formula IV. TIFF0007601403000003.tif18128

[0019] In one exemplary embodiment, the double bond is in the (Z) configuration. In another exemplary embodiment, the double bond is in the (E) configuration. In one exemplary embodiment, the chiral center at C-3 has an R configuration. In one exemplary embodiment, the chiral center at C-3 has an S configuration. In another exemplary embodiment, the double bond is in the (Z) configuration and the chiral center at C-3 has an R configuration.

[0020] One aspect of the present disclosure provides a 1,3-fatty diol derivative having the chemical formula of Formula V: TIFF0007601403000004.tif19128, where n is an integer selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, and 14; R 1 and R 2 are each independently selected from the group consisting of H, a monosaccharide linked at the anomeric carbon of a monosaccharide, a disaccharide linked at the anomeric carbon of a disaccharide, a trisaccharide linked at the anomeric carbon, and a polysaccharide linked at the anomeric carbon. 1 and R 2In another exemplary embodiment, R 1 Also 2 is not H. In another exemplary embodiment, the monosaccharide is selected from pentose and hexose sugars. In another exemplary embodiment, the hexose sugar is selected from allose, altrose, glucose, mannose, gulose, iodose, galactose, or talose. In another exemplary embodiment, the disaccharide, trisaccharide, and polysaccharide comprise a sugar selected from a pentose sugar, a hexose sugar, or a mixture of any two or more thereof. In another exemplary embodiment, the hexose sugar is selected from allose, altrose, glucose, mannose, gulose, iodose, galactose, talose, or a mixture of any two or more thereof. In another exemplary embodiment, R 1 and R 2 are different monosaccharides. In another exemplary embodiment, R 1 and R 2 are the same monosaccharide. In another exemplary embodiment, the monosaccharides, disaccharides, trisaccharides, and polysaccharides comprise hexose sugars selected from allose, altrose, glucose, mannose, gulose, iodos, galactose, talose, or a mixture of any two or more thereof. In another exemplary embodiment, the monosaccharides, disaccharides, trisaccharides, and polysaccharides comprise sugars selected from furanose sugars, pyranose sugars, or a mixture of any two or more thereof.

[0021] One aspect of the present disclosure provides a 1,3-fatty diol derivative having the chemical formula of Formula VI: TIFF0007601403000005.tif20128, where m is an integer selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, and 18; R 3 and R 4are each independently selected from the group consisting of H, a monosaccharide linked at the anomeric carbon of a monosaccharide, a disaccharide linked at the anomeric carbon of a disaccharide, a trisaccharide linked at the anomeric carbon, and a polysaccharide linked at the anomeric carbon. 3 and R 4 In another exemplary embodiment, R 3 Also 4 is not H. In another exemplary embodiment, the monosaccharide is selected from pentose and hexose sugars. In another exemplary embodiment, the hexose sugar is selected from allose, altrose, glucose, mannose, gulose, iodose, galactose, or talose. In another exemplary embodiment, the disaccharide, trisaccharide, and polysaccharide comprise a sugar selected from a pentose sugar, a hexose sugar, or a mixture of any two or more thereof. In another exemplary embodiment, the hexose sugar is selected from allose, altrose, glucose, mannose, gulose, iodose, galactose, talose, or a mixture of any two or more thereof. In another exemplary embodiment, R 3 and R 4 are different monosaccharides. In another exemplary embodiment, R 3 and R 4 are the same monosaccharide. In another exemplary embodiment, the monosaccharides, disaccharides, trisaccharides, and polysaccharides comprise hexose sugars selected from allose, altrose, glucose, mannose, gulose, iodos, galactose, talose, or a mixture of any two or more thereof. In another exemplary embodiment, the monosaccharides, disaccharides, trisaccharides, and polysaccharides comprise sugars selected from furanose sugars, pyranose sugars, or a mixture of any two or more thereof. [The present invention 1001] A 12-carbon unbranched unsaturated fatty diol having a single Δ5 double bond, a hydroxy group at carbon number 1 (C-1) and a hydroxy group at carbon number 3 (C-3), wherein the chiral center is at C-3, said fatty diol being represented by formula (II): TIFF0007601403000006.tif18128 The fatty diol having the general chemical formula: [The present invention 1002] The fatty diol of the present invention, wherein the double bond is in the (Z) configuration. [The present invention 1003] 1001. The fatty diol of claim 1001, wherein the double bond is in the (E) configuration. [The present invention 1004] 1001. The fatty diol of claim 1001, wherein said chiral center at C-3 has an R configuration. [The present invention 1005] 1001. The fatty diol of claim 1001, wherein said chiral center at C-3 has an S configuration. [The present invention 1006] 1001. The fatty diol of claim 1001, wherein said double bond is of the (Z) configuration and said chiral center at C-3 has an R configuration. [The present invention 1007] A 16-carbon unbranched unsaturated fatty diol having a single Δ double bond, a hydroxy group at carbon number 1 (C-1) and a hydroxy group at carbon number 3 (C-3), wherein the chiral center is at C-3, said fatty diol being represented by formula (IV): TIFF0007601403000007.tif18128 The fatty diol having the chemical formula: [The present invention 1008] The fatty diol of the present invention 1007, wherein the double bond is in the (Z) configuration. [The present invention 1009] The fatty diol of the present invention 1007, wherein the double bond is in the (E) configuration. [The present invention 1010] 1007. The fatty diol of claim 10, wherein said chiral center at C-3 has an R configuration. [The present invention 1011] 1007. The fatty diol of claim 10, wherein said chiral center at C-3 has an S configuration. [The present invention 1012] 1007. The fatty diol of claim 10, wherein said double bond is of the (Z) configuration and said chiral center at C-3 has an R configuration. [Brief description of the drawings]

[0022] [Figure 1A] Schematically illustrates fragment ions formed from fragmentation of 1,3-dodecanediol trimethylsilyl ether. [Figure 1B] Figure 1 shows the experimental mass spectral fragmentation pattern of 1,3-dodecanediol trimethylsilyl ether. Note the characteristic ions at 331 (MW-15), 229, and 219 for the saturated diol adduct. [Figure 2A]Figure 1 illustrates a schematic of fragment ions formed from the fragmentation of 5-dodecene-1,3-diol trimethylsilyl ether. In the schematic of the fragment ions, the double bond is shown as (E). However, a mixture is present of both (Z) and (E). In fact, fermentation produces primarily 5-(Z)-dodecene-1,3-diol as a result of specific fatty acid metabolism. [Figure 2B] Figure 1 shows the experimental mass spectral fragmentation pattern of 5-dodecene-1,3-diol trimethylsilyl ether. Note the characteristic ions at 329 (MW-15), 227, and 219 for the unsaturated diol adduct. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0023] Detailed Description definition As used in this specification and the appended claims, singular articles such as "a" and "an" and "the" and similar referents in the context of describing elements are to be construed to encompass both the singular and the plural, unless otherwise indicated herein or the context clearly contradicts.

[0024] As used herein, "about" is understood by those of ordinary skill in the art and can vary to some extent depending on the context in which it is used. If there are uses of the term that are not clear to those of ordinary skill in the art, "about" will mean up to plus or minus 10% of the specific term, taking into account the context in which the term is used.

[0025] As will be understood by one of ordinary skill in the art, for all purposes, all ranges disclosed herein also encompass all possible subranges and combinations of those subranges. Moreover, as will be understood by one of ordinary skill in the art, a range includes each individual member. Thus, for example, a group having 1-3 atoms refers to groups having 1, 2, or 3 atoms. Similarly, a group having 1-5 atoms refers to groups having 1, 2, 3, 4, or 5 atoms, and so on.

[0026] Unless otherwise defined, technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. In particular, this disclosure makes use of conventional techniques in the fields of recombinant genetics, organic chemistry, fermentation, and biochemistry. Basic texts disclosing common terms in molecular biology and genetics include, for example, Lackie, Dictionary of Cell and Molecular Biology, Elsevier (5 thed. 2013). A basic text disclosing general methods and terminology in biochemistry is, for example, Lehninger Principles of Biochemistry sixth edition, David L. Nelson and Michael M. Cox eds. WH Freeman (2012). A basic text disclosing general methods and terminology in fermentation science is, for example, Principles of Fermentation Technology, 3rd Edition by Peter F Stanbury, Allan Whitaker and Stephen J Hall. Butterworth-Heinemann (2016). Basic texts disclosing general methods and terminology in organic chemistry include, for example, Favre, Henri A. and Powell, Warren H. Nomenclature of Organic Chemistry. IUPAC Recommendations and Preferred Names 2013. Cambridge, UK: The Royal Society of Chemistry, 2013; Practical Synthetic Organic Chemistry: Reactions, Principles, and Techniques, Stephane Caron ed., John Wiley and Sons Inc. (2011); Organic Chemistry, 9th Edition-Francis Carey and Robert Giuliano, McGraw Hill (2013).

[0027] Those skilled in the art will understand that the compounds of the present technology may exhibit the phenomenon of tautomerism, conformational isomerism, geometric isomerism, and / or stereoisomerism.The depiction of formulas in this specification and claims may represent only one of possible tautomerism, conformational isomerism, stereoisomerism, or geometric isomerism, so it should be understood that the present technology encompasses any tautomerism, conformational isomerism, stereoisomerism, or geometric isomerism of the compounds having one or more of the utilities described herein, and mixtures of their various different forms.

[0028] "Tautomer" refers to the isomeric forms of a compound that are in equilibrium with each other.The presence and concentration of isomeric forms depends on the environment in which the compound is found, and may vary depending on, for example, whether the compound is solid or in organic or aqueous solution.For example, in aqueous solution, quinazolinone may exhibit the following isomeric forms, which are called tautomers of each other: TIFF0007601403000008.tif28128

[0029] As another example, in protic organic solution, guanidine can exhibit the following isomeric forms, also referred to as tautomers of one another: TIFF0007601403000009.tif29128

[0030] Because of the limitation of representing compounds by structural formulas, all chemical formulas of compounds described herein represent all tautomeric forms of the compounds and are understood to be within the scope of the present technology.

[0031] The stereoisomers (also known as optical isomers) of a compound include all chiral, diastereomeric, and racemic forms of a structure unless a specific stereochemistry is explicitly indicated.Thus, the compounds used in the present technology include enriched optical isomers or resolved optical isomers at any or all asymmetric atoms that are evident from the depiction.Both racemic and diastereomeric mixtures and individual optical isomers can be isolated or synthesized to be substantially free of their enantiomeric or diastereomeric partners, and all of these stereoisomers are within the scope of the present technology.

[0032] Geometric isomers can be represented by the symbols representing bonds that can be single, double, or triple bonds as described herein. Provided herein are various geometric isomers and mixtures thereof resulting from the arrangement of substituents around a carbon-carbon double bond. Substituents around a carbon-carbon double bond are designated as being in the "Z" or "E" configuration, and the terms "Z" or "E" are used according to IUPAC standards. Unless otherwise specified, structures depicting double bonds encompass both the "Z" and "E" isomers.

[0033] Substituents around a carbon-carbon double bond may alternatively be referred to as "cis" or "trans", with "cis" representing the substituents on the same side of the double bond and "trans" representing the substituents on opposite sides of the double bond. The term "cis" represents the substituents on the same side of the plane of the ring and the term "trans" represents the substituents on opposite sides of the plane of the ring. Mixtures of compounds in which the substituents are located on both the same and opposite sides of the plane of the ring are designated "cis / trans".

[0034] In certain embodiments, the pharma- ceutically acceptable form is an isomer. "Isomers" are different compounds that have the same molecular formula. "Stereoisomers" are isomers that differ only in the way the atoms are arranged in space. As used herein, the term "isomer" includes all geometric and stereoisomers. For example, "isomer" includes cis and trans isomers, E and Z isomers, R and S enantiomers, diastereoisomers, (d) isomers, (l) isomers, racemic mixtures thereof, and other mixtures thereof within the scope of this disclosure.

[0035] "Enantiomers" are pairs of stereoisomers that are non-superimposable mirror images of each other. A mixture of a pair of enantiomers in any ratio may be known as a "racemic" mixture. The term "(+-)" is used to designate a racemic mixture when necessary. "Diastereoisomers" are stereoisomers that have at least two asymmetric atoms but are not mirror images of each other. Absolute stereochemistry is specified according to the Cahn-Ingold-Prelog RS system. When a compound is a pure enantiomer, the stereochemistry of each chiral carbon may be specified by either R or S. Resolved compounds whose absolute configuration is unknown may be specified as (+) or (-) depending on the direction (dextrorotatory or levorotatory) they rotate plane polarized light at the wavelength of the sodium D line. Some of the compounds described herein contain one or more asymmetric centers and may thus result in enantiomers, diastereoisomers, and other stereoisomeric forms that can be defined in terms of absolute stereochemistry as (R) or (S). The chemical compounds, pharmaceutical compositions, and methods are intended to include all such possible isomers, including racemic mixtures, optically pure forms, and intermediate mixtures. Optically active (R) and (S) isomers can be prepared, for example, using chiral synthons or chiral reagents, or resolved using conventional techniques. The optical activity of a compound can be analyzed via any suitable method, including, but not limited to, chiral chromatography and polarimetry, to determine the degree of dominance of one stereoisomer over the other.

[0036] As used herein, the term "fatty diol" or "fatty diol" refers to an aliphatic di-alcohol having a carbon chain length of at least 5 carbons, which contains two hydroxy (-OH) groups covalently bonded to the carbon chain. In an exemplary embodiment, the "fatty diol" is a "1,3-fatty diol". As used herein, the expression "1,3-fatty diol" refers to an aliphatic di-alcohol having a carbon chain length of at least 8 carbons, with the alcohol moieties located at the first carbon (C-1) and the third carbon (C-3). A "1,3-fatty diol" may be saturated or unsaturated. Thus, as generally used herein, the term "1,3-fatty diol" refers to a molecule having the structural formula shown in Formula IA or Formula IB.

[0037] TIFF0007601403000010.tif18128In formula IA, m is an integer selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, and 18, and the C-3 carbon is either the (R) enantiomer or and the (S) enantiomer.

[0038] TIFF0007601403000011.tif18128In formula IB, n is an integer selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, and 14, the double bond may be in either the (Z) or (E) configuration, and the C-3 carbon may be either the (R) enantiomer or and (S) enantiomer.In an exemplary embodiment, the 1,3 fatty diol is 5-dodecene-1,3-diol.In another exemplary embodiment, the 1,3 fatty diol is 9-hexadecene-1,3-diol.

[0039] As used herein, the term "5-dodecene-1,3-diol" or equivalently "1,3-dodec-5-enediol" or equivalently "5-dodecene-1,3-diol" or equivalently "dodec-5-ene-1,3-diol" refers to a novel 12-carbon unbranched unsaturated 1,3-diol having a double bond between the 5th and 6th carbons (i.e., a Δ5 double bond that may be in either the (Z) or (E) configuration), a hydroxy group at carbon number 1 (C-1), and a hydroxy group at carbon number 3 (C-3), with a chiral center at C-3, which may be either the (R) enantiomer or and (S) enantiomer. Such molecules have the general structure shown in formula II. TIFF0007601403000012.tif18128

[0040] As used herein, the term "7-tetradecene-1,3-diol" or equivalently "1,3-tetradec-7-enediol" or equivalently "7-tetradecene-1,3-diol" or equivalently "tetradec-7-ene-1,3-diol" refers to a novel 14-carbon unbranched unsaturated 1,3-diol having a double bond between the 7th and 8th carbons (i.e., a Δ7 double bond that may be in either the (Z) or (E) configuration), a hydroxy group at carbon number 1 (C-1), and a hydroxy group at carbon number 3 (C-3), with a chiral center at C-3, which may be either the (R) enantiomer or and (S) enantiomer. Such molecules have the general structure shown in formula III. TIFF0007601403000013.tif18128

[0041] As used herein, the term "9-hexadecene-1,3-diol" or equivalently "1,3-hexadec-9-enediol" or equivalently "9-hexadecene-1,3-diol" or equivalently "hex-9-ene-1,3-diol" refers to a novel 16-carbon unbranched unsaturated 1,3-diol having a double bond between the 9th and 10th carbons (i.e., a Δ9 double bond that may be in either the (Z) or (E) configuration), a hydroxy group at carbon number 1 (C-1), and a hydroxy group at carbon number 3 (C-3), with a chiral center at C-3, which may be either the (R) enantiomer or and (S) enantiomer. Such molecules have the general structure shown in formula IV. TIFF0007601403000014.tif18128

[0042] As used herein, the term "polyol" refers to a compound having two or more hydroxy groups, typically a fatty alcohol. Thus, as referred to herein, a polyol may have two hydroxy groups, three hydroxy groups, four hydroxy groups, etc. In general, a "polyol" having two hydroxy groups is referred to herein as a "diol," a "polyol" having three hydroxy groups is referred to herein as a "thiol," a "polyol" having four hydroxy groups is referred to herein as a "tetrol," etc.

[0043] The terms "hydroxy group," "hydroxyl group," and "alcohol group" are used interchangeably herein and refer to a chemical functional group that contains an oxygen atom covalently bonded to a hydrogen atom (-OH).

[0044] The term "Enzyme Classification (EC) number" refers to a number that represents a specific polypeptide sequence or enzyme. EC numbers classify enzymes according to the reaction they catalyze. EC numbers have been established by the Nomenclature Commission of the International Union of Biochemistry and Molecular Biology (IUBMB), and their descriptions are available on the World Wide Web at the IUBMB Enzyme Nomenclature website.

[0045] As used herein, the term "isolated" with respect to a product (such as, for example, 5-dodecene-1,3-diol) refers to a product that is separated from cellular components, cell culture medium, or chemical or synthetic precursors.

[0046] As used herein, the terms "purify", "purified" or "purification" refer to the removal or isolation of a molecule from its environment, for example, by isolation or separation. "Substantially purified" molecules are at least about 60% free of other components with which they are associated (e.g., at least about 65% free, at least about 70% free, at least about 75% free, at least about 80% free, at least about 85% free, at least about 90% free, at least about 95% free, at least about 97% free, at least about 98% free, at least about 99% free). As used herein, these terms also refer to the removal of contaminants from a sample. For example, the removal of contaminants can result in an increase in the percentage of fatty diols in a sample. For example, when 1,3-diols are produced in recombinant host cells, the 1,3-diols can be purified by the removal of host cell proteins. After purification, the percentage of 1,3-diols in a sample is increased. The terms "purify," "purified," and "purification" are relative terms that do not require absolute purity. Thus, for example, when a 1,3-diol is produced in a recombinant host cell, a purified 1,3-diol is a 1,3-diol that is substantially separated from other cellular components (e.g., nucleic acids, polypeptides, lipids, carbohydrates, or other hydrocarbons).

[0047] I. 1,3-Fatty diol compounds and their derivatives A. General Method This disclosure utilizes conventional techniques in recombinant genetics. Basic texts that disclose common methods and terms in molecular biology and genetics include, for example, Sambrook et al., Molecular Cloning, a Laboratory Manual, Cold Spring Harbor Press 4th edition (Cold Spring Harbor, NY 2012), Current Protocols in Molecular Biology Volumes 1-3, John Wiley & Sons, Inc. (1994-1998). This disclosure also utilizes conventional techniques in the field of biochemistry. Basic texts that disclose common methods and terms in biochemistry include, for example, Lehninger Principles of Biochemistry sixth edition, David L. Nelson and Michael M. Cox eds. WH Freeman (2012). This disclosure also utilizes conventional techniques in industrial fermentation. Basic texts disclosing common methods and terminology in fermentation include, for example, Principles of Fermentation Technology, 3rd Edition by Peter F. Stanbury, Allan Whitaker and Stephen J. Hall. Butterworth-Heinemann (2016), Fermentation Microbiology and Biotechnology, 2nd Edition, EMT El-Mansi, CF A Bryce, Arnold L. Demain and AR Allman eds. CRC Press (2007). This disclosure also utilizes conventional techniques in the field of organic chemistry.Basic texts disclosing common methods and terminology in organic chemistry include, for example, Practical Synthetic Organic Chemistry: Reactions, Principles, and Techniques, Stephane Caron ed., John Wiley and Sons Inc. (2011), The Synthetic Organic Chemist's Companion, Michael C. Pirrung, John Wiley and Sons Inc. (2007), and Organic Chemistry, 9th Edition-Francis Carey and Robert Giuliano, McGraw Hill (2013).

[0048] B. 1,3-Fatty Diols and Their Derivatives As discussed above, 1,3-fatty diols are particularly useful molecules. In addition to providing two hydroxyl functional groups for reaction to form derivatives, the 3-hydroxy moiety of 1,3-fatty diols forms a chiral center at the third carbon (C-3), making them useful as synthons for the production of chirally important compounds, such as pharmaceuticals, nutraceuticals, pesticides, herbicides, flavors, fragrances, solvents, and biologically active compounds.

[0049] In addition to the functionality of the hydroxyl groups, variations in the structure of the carbon chains of 1,3-fatty diols provide the molecules with additional chemical properties and / or potentially new properties that can be used to address existing problems in an improved way and / or find entirely new applications. For example, unsaturated fatty alcohols have an additional functional group in the form of a double bond that is available for chemical reactions.

[0050] Unsaturated fatty alcohols are particularly valuable because the presence of double bonds contributes to a number of advantageous properties on the molecule. For example, compared to their saturated counterparts, unsaturated fatty alcohols have lower melting points, higher water solubility, and offer the possibility of introducing functional groups to C=C double bonds (Egan, R., et al., (1984) Journal of the American Oil Chemists' Society, Vol. 61 (2): 324-329). Therefore, unsaturated fatty alcohols are important intermediates for many products in the chemical industry (see, for example, U. Ploog et al. in Seifen-Ole-Fette-Wachse 109, 225 (1983)).

[0051] In an exemplary embodiment, the present disclosure provides a novel unsaturated unbranched 1,3-fatty diol. In an exemplary embodiment, the novel unsaturated unbranched 1,3-fatty diol has the systematic name: 5-dodecene-1,3-diol or dodec-5-ene-1,3-diol. In some exemplary embodiments, the double bond is in the (Z) configuration. In other exemplary embodiments, the double bond of 5-dodecene-1,3-diol is in the (E) configuration. 5-dodecene-1,3-diol has the general structure of formula II: TIFF0007601403000015.tif18128

[0052] In formula II, the chiral C-3 carbon may be either the (R) enantiomer or the (S) enantiomer.

[0053] In another exemplary embodiment, the novel unsaturated unbranched 1,3-fatty diol has the systematic name: 7-hexadecene-1,3-diol or hexadec-7-ene-1,3-diol. In some exemplary embodiments, the double bond is in the (Z) configuration. In other exemplary embodiments, the double bond is in the (E) configuration. 7-hexadecene-1,3-diol has the general structure of formula (III): TIFF0007601403000016.tif18128

[0054] In formula III, the chiral C-3 carbon may be either the (R) enantiomer or the (S) enantiomer.

[0055] In another exemplary embodiment, the novel unsaturated unbranched 1,3-fatty diol has the systematic name: 9-hexadecene-1,3-diol or hexadec-9-ene-1,3-diol. In some exemplary embodiments, the double bond is in the (Z) configuration. In other exemplary embodiments, the double bond is in the (E) configuration. 9-hexadecene-1,3-diol has the general structure of formula (IV). TIFF0007601403000017.tif20128

[0056] In formula IV, the chiral C-3 carbon may be either the (R) enantiomer or the (S) enantiomer.

[0057] 1. Physical properties of 1,3-fatty diols All of the 1,3-fatty diols disclosed herein contain two alcohol groups and a chiral center at the C-3 carbon. In addition, the unsaturated 1,3-fatty diols disclosed herein, such as 5-dodecene-1,3-diol, also contain a double bond. Thus, the 1,3-fatty diols disclosed herein can undergo a variety of chemical reactions to form a wide variety of molecules. Thus, in addition to the value (e.g., as a surfactant) of any of the 1,3-fatty diols disclosed herein, in exemplary embodiments, the 1,3-fatty diols disclosed herein, such as 5-dodecene-1,3-diol, serve as building blocks for an infinite number of unique and useful molecules.

[0058] a.Double bond The unsaturated 1,3-fatty diols disclosed herein, such as 5-dodecene-1,3-diol, contain a double bond. Depending on the method by which the 1,3-fatty diol is produced and / or processed (see below in this specification), the double bond can be either (Z) or (E). Thus, the unsaturated 1,3-fatty diols disclosed herein, such as 5-dodecene-1,3-diol, can participate in chemical reactions involving the double bond, including, for example, polymerization, alkylation, metathesis, etc. Chemical reactions utilizing carbon-carbon double bonds are known in the art (see, for example, Practical Synthetic Organic Chemistry: Reactions, Principles, and Techniques, Stephane Caron ed. (above)).

[0059] Thus, in exemplary embodiments, the carbon-carbon double bonds of the unsaturated 1,3-fatty diols disclosed herein, such as 5-dodecene-1,3-diol, 9-hexadecene-1,3-diol, etc., participate in an addition reaction. Exemplary addition reactions include, for example, halogenation, hydrogenation, hydration, epoxidation, radical polymerization, hydroxylation, etc. Thus, in some exemplary embodiments, the carbon-carbon double bonds of the unsaturated 1,3-fatty diols disclosed herein, such as 5-dodecene-1,3-diol, 9-hexadecene-1,3-diol, etc., participate in a hydrogenation reaction, thus forming the corresponding alkane. In other exemplary embodiments, the carbon-carbon double bonds of the unsaturated 1,3-fatty diols disclosed herein, such as 5-dodecene-1,3-diol, 9-hexadecene-1,3-diol, etc., participate in a hydration reaction to add water across the double bond, thereby resulting in a polyol.

[0060] In other exemplary embodiments, the carbon-carbon double bonds of the unsaturated 1,3-fatty diols disclosed herein, such as 5-dodecene-1,3-diol, 9-hexadecene-1,3-diol, and the like, participate in polymerization reactions to yield industrially valuable polymers (e.g., specialty plastics).

[0061] B hydroxyl group All of the 1,3-fatty diols disclosed herein, such as 5-dodecene-1,3-diol, 9-hexadecene-1,3-diol, etc., contain two hydroxyl functional groups that are available for chemical reactions.

[0062] As is generally known in the art, the chemistry of diols is almost the same as that of alcohols (see, for example, Organic Chemistry ninth edition Francis Carey and Robert Giuliano (2013) (above)). Thus, due to the polar nature of the -OH bond, the 1,3-fatty diols disclosed herein, such as 5-dodecene-1,3-diol, 9-hexadecene-1,3-diol, etc., readily form hydrogen bonds with other 1,3-fatty diol molecules or hydrogen bonding systems (e.g., water). Thus, 1,3-fatty diols generally have relatively high melting and boiling points, as well as high solubility in aqueous media, compared to similar alkanes.

[0063] Hydroxyl functional group can participate in a number of chemical reactions characteristic of hydroxyl group.Thus, in one exemplary embodiment, hydroxyl functional group of 1,3-fatty diol disclosed herein, such as 5-dodecene-1,3-diol, 9-hexadecene-1,3-diol, etc., participates in nucleophilic substitution reaction, in which hydroxyl functions as a leaving group, or -OH or -O- functions as a nucleophile (e.g., substitution with halide).

[0064] In other exemplary embodiments, the hydroxyl functional groups of the 1,3-fatty diols disclosed herein, such as 5-dodecene-1,3-diol, 9-hexadecene-1,3-diol, and the like, participate in nucleophilic addition reactions in which the hydroxyl groups function as nucleophiles, thereby forming acetals or ketones. Exemplary nucleophilic addition reactions include, for example, glycosylation reactions, which are discussed in more detail herein below.

[0065] In yet other exemplary embodiments, the hydroxyl functional groups of the 1,3-fatty diols disclosed herein, such as 5-dodecene-1,3-diol, 9-hexadecene-1,3-diol, and the like, participate in nucleophilic acyl substitution reactions in which the hydroxyl groups function as nucleophiles to form esters with carboxylic acids and carboxylic acid derivatives (e.g., nucleophilic acyl substitution of 5-dodecene-1,3-diol with fatty acids, e.g., to form fatty esters).

[0066] In yet other exemplary embodiments, the hydroxyl functional groups of the 1,3-fatty diols disclosed herein, such as 5-dodecene-1,3-diol, 9-hexadecene-1,3-diol, and the like, participate in an elimination reaction in which the hydroxyl group is eliminated as water and a carbon double bond (alkene) is formed.

[0067] In yet another exemplary embodiment, the hydroxyl functional groups of the 1,3-fatty diols disclosed herein, such as 5-dodecene-1,3-diol, 9-hexadecene-1,3-diol, etc., participate in an oxidation reaction in which the hydroxyl groups are converted to carbonyl groups (C=O), thus producing carbonyl compounds. In the oxidation reaction, the resulting carbonyl compounds can be aldehydes, ketones, or carboxylic acids depending on the oxidizing agent used (see, for example, Organic Chemistry 9th Edition, Francis Carey and Robert Giuliano (2013) (supra)).

[0068] Thus, the multiple hydroxyl functional groups of the 1,3-fatty diols disclosed herein, such as 5-dodecene-1,3-diol, 9-hexadecene-1,3-diol, etc., provide a wide range of possible reactions for the disclosed 1,3-fatty diols. In turn, this provides the possibility of a large number of derivatives with unique and useful properties. Some exemplary hydroxyl derivatives of the 1,3-fatty diols disclosed herein, such as 5-dodecene-1,3-diol, 9-hexadecene-1,3-diol, etc., are discussed below.

[0069] C. Chirality Chiral molecules, such as 1,3-fatty diols (e.g., 5-dodecene-1,3-diol), which have a chiral center at the C-3 carbon, are building blocks for the synthesis of stereochemically influenced compounds (e.g., pharmaceuticals, nutraceuticals, etc.). Chirality is an important property to consider, for example, in drug design, since most isomers of chiral drugs exhibit significant differences in biological activities, such as, for example, pharmacology, toxicity, pharmacokinetics, biorecognition, metabolism, etc. Indeed, the selection of the appropriate enantiomer can have a significant impact on the biological properties of the molecule.

[0070] 2.Nonionic surfactants In some exemplary embodiments, nonionic surfactants are prepared using the hydroxyl functional groups of the 1,3-fatty diols disclosed herein, such as 5-dodecene-1,3-diol, 9-hexadecene-1,3-diol, and the like.

[0071] i. Glycosylated Derivatives In exemplary embodiments, the hydroxyl moiety of the 1,3-fatty diol disclosed herein, such as 5-dodecene-1,3-diol, is available for reaction with sugar to provide alkyl polyglycosides. Alkyl polyglycosides are a class of non-ionic surfactants derived from sugars and fatty alcohols, and are well known in the art (see, for example, Alkyl Polyglycosides: Technology, Properties, Applications, Karlheinz Hill; Wolfgang von Rybinski; Gerhard Stoll, eds. Wiley (2008); Novel Surfactants: Preparation Applications And Biodegradability, Surfactant Science Series. Volume 114 Second Edition, Krister Holmberg ed. Marcel Dekker: New York and Basel, Switzerland. 2003). Thus, in an exemplary embodiment, a 1,3-fatty diol disclosed herein, such as 5-dodecene-1,3-diol, is reacted with, for example, xylose, galactose, mannose, glucose, etc., to form alkyl polyglycosides with new and useful properties.

[0072] When derived from glucose, alkyl polyglycoside is referred to as alkyl polyglucoside. Thus, in an exemplary embodiment, 1,3-fatty diol disclosed herein, such as 5-dodecene-1,3-diol, 9-hexadecene-1,3-diol, etc., is reacted with glucose to form alkyl polyglucoside. Alkyl polyglucosides are known in the art (see, for example, Nonionic Surfactants: Alkyl Polyglucosides. Surfactant Science Series. Volume 91 Dieter Balzer and Harald Luders, eds. Marcel Dekker: New York and Basel, Switzerland. 2000; Rather and Mishra: β-Glycosidases: An alternative enzyme based method for synthesis of alkyl-glycosides. Sustainable Chemical Processes 2013 1:7).

[0073] The alkyl polyglycosides can be prepared by any method known in the art. Exemplary methods suitable for preparing the glycosylated molecules disclosed herein include those described, for example, in U.S. Pat. No. 5,449,763, U.S. Pat. No. 3,547,828, U.S. Pat. No. 3,839,318, Boge, K., Tietze, L. "Synthesis of alkyl polyglycosides: Effect of catalyst-type on reaction rate and product composition" Eur. J. Lipid Sci. Tech. (1998) 100, 38-41, Rybinski, W., Hill, K. "Alkyl Polyglycosides-Properties and Applications of a new Class of Surfactants" Angew. Chem. Int. Ed. (1998) 37(10), 1328-1345, "Handbook of Chemical Glycosylation: Advances in Stereoselectivity and Therapeutic Relevance" Ed. AV Demchenko, John Wiley&Sons, Apr9, 2008, Luley-Goedl, C. et al. Carbohydrate Research (2010) 345, 1736-1740, Rather, MY, Mishra, S. (2013) (above), Ojha, S. et al. “Synthesis of hexyl α-glucoside and α-polyglucosdies by a novel Microbacterium isolate” Appl. Microbiol. Biotechnol. (2013) 97, 5293-5301, Wooten, J. “Glycosylation of Amino Acids and Efficient Synthesis of Glycosphingolipids” Thesis, Georgia State University, 2015).

[0074] Thus, in an exemplary embodiment, the disclosure provides alkyl polyglycosides having the general formula of Formula V and / or Formula VI.

[0075] TIFF0007601403000018.tif20128In formula (V), n is an integer selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, and 14, the double bond may be in either the (Z) or the (E) configuration, and the C-3 carbon may be either the (R) enantiomer or and (S) enantiomer.

[0076] Further, in formula (V), R 1 and R 2 are each independently H, a monosaccharide linked at the anomeric carbon of a monosaccharide, a disaccharide linked at the anomeric carbon of a disaccharide, a trisaccharide linked at the anomeric carbon, or a polysaccharide linked at the anomeric carbon. 1 and R 2 cannot both be H. In another exemplary embodiment, R 1 Also 2 It's not H either.

[0077] In exemplary embodiments, the monosaccharides are selected from pentose and hexose sugars. In some exemplary embodiments, the hexose sugars are selected from allose, altrose, glucose, mannose, gulose, iodose, galactose, or talose. In other exemplary embodiments, the disaccharides, trisaccharides, and / or polysaccharides comprise sugars selected from pentose sugars, hexose sugars, or mixtures of any two or more thereof. In some exemplary embodiments, the hexose sugars are selected from allose, altrose, glucose, mannose, gulose, iodose, galactose, talose, or mixtures of any two or more thereof. In some exemplary embodiments, R 1 and R 2 are different monosaccharides. In another exemplary embodiment, R 1 and R 2are the same monosaccharide.

[0078] In exemplary embodiments, the monosaccharides, disaccharides, trisaccharides, and / or polysaccharides comprise hexose sugars selected from allose, altrose, glucose, mannose, gulose, iodos, galactose, talose, or mixtures of any two or more thereof. In other exemplary embodiments, the monosaccharides, disaccharides, trisaccharides, and polysaccharides comprise sugars selected from furanose sugars, pyranose sugars, or mixtures of any two or more thereof.

[0079] TIFF0007601403000019.tif20128In formula (VI), m is an integer selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, and 18, and the C-3 carbon is either the (R) enantiomer or and the (S) enantiomer.

[0080] Further, in formula (VI), R 3 and R 4 are each independently H, a monosaccharide linked at the anomeric carbon of a monosaccharide, a disaccharide linked at the anomeric carbon of a disaccharide, a trisaccharide linked at the anomeric carbon, or a polysaccharide linked at the anomeric carbon. 3 and R 4 cannot both be H. In another exemplary embodiment, R 3 Also 4 It's not H either.

[0081] In exemplary embodiments, the monosaccharides are selected from pentose and hexose sugars. In other exemplary embodiments, the disaccharides, trisaccharides, and / or polysaccharides include sugars selected from pentose sugars, hexose sugars, or mixtures of any two or more thereof. In some exemplary embodiments, R 3 and R 4 are different monosaccharides. In another exemplary embodiment, R 3 and R 4 are the same monosaccharide.

[0082] In exemplary embodiments, the monosaccharides, disaccharides, trisaccharides, and / or polysaccharides comprise sugars selected from allose, altrose, glucose, mannose, gulose, iodos, galactose, talose, or mixtures of any two or more thereof. In yet other exemplary embodiments, the monosaccharides, disaccharides, trisaccharides, and polysaccharides comprise sugars selected from furanose sugars, pyranose sugars, or mixtures of any two or more thereof.

[0083] The term "anomeric carbon" as applied herein to any compound of the present technology is well known to those of skill in the art, and one example of attachment at the anomeric carbon of a sugar is exemplified by the following glycosylation variants: TIFF0007601403000020.tif46128

[0084] The anomeric carbon of a sugar (here a monosaccharide) is indicated by an open arrow. Similarly, glucose disaccharides include those illustrated in the two schemes below.

[0085] Scheme 1: α(1→4) disaccharide (α-maltose) with α-glycosidic linkage to the non-glycosylated chain TIFF0007601403000021.tif72159

[0086] Scheme 2: α(1→6) disaccharide (α-isomaltose) with α-glycosidic linkage to the non-glycosylated chain TIFF0007601403000022.tif78146

[0087] As illustrated in Schemes 1 and 2, for example, R 1is "attached at the anomeric carbon" of the disaccharide maltose or isomaltose, this should be understood to refer not to the anomeric carbon(s) involved in the bond between the sugars of disaccharides, trisaccharides, and polysaccharides (hereinafter "glycosidic bond") as shown by the dashed arrow, but to the anomeric carbon of the free disaccharide having a free hydroxyl group susceptible to such bond formation, for example with compounds of formulae IA, IB, II, III, IV, as shown by the open arrow above. This applies to the meaning of "attached at the anomeric carbon" in relation to trisaccharides and polysaccharides. Thus, in any embodiment herein, R 1 , R 2 , R 3 , and / or R 4 is a monosaccharide, disaccharide, trisaccharide, and / or polysaccharide, R 1 , R 2 , R 3 , and / or R 4 can be linked through α-glycosidic or β-glycosidic bonds. Additionally, the disaccharides, trisaccharides, and / or polysaccharides can be saccharides containing α-glycosidic bonds, β-glycosidic bonds, or (in the case of trisaccharides and / or polysaccharides) mixtures thereof.

[0088] In exemplary embodiments, the monosaccharides are selected from pentose and hexose sugars. In other exemplary embodiments, the disaccharides, trisaccharides, and / or polysaccharides include sugars selected from pentose sugars, hexose sugars, or mixtures thereof. In some exemplary embodiments, R 3 and R 4 are different monosaccharides. In another exemplary embodiment, R 3 and R 4 are the same monosaccharide.

[0089] In exemplary embodiments, the monosaccharides, disaccharides, trisaccharides, and / or polysaccharides comprise sugars selected from allose, altrose, glucose, mannose, gulose, iodos, galactose, talose, or mixtures of any two or more thereof. In yet other exemplary embodiments, the monosaccharides, disaccharides, trisaccharides, and polysaccharides comprise sugars selected from furanose sugars, pyranose sugars, or mixtures thereof.

[0090] Thus, in any embodiment herein, monosaccharides, disaccharides, trisaccharides, and polysaccharides may be pentose sugars, hexose sugars, or (R 1 and R 2 Or R 3 and R 4 are different monosaccharides, and / or in the case of any one or more disaccharides, trisaccharides, or polysaccharides, may include mixtures of any two or more thereof. The monosaccharides, disaccharides, trisaccharides, and polysaccharides of any embodiment herein may include allose, altrose, glucose, mannose, gulose, iodose, galactose, talose, or (R 1 and R 2 Or R 3 and R 4 are different monosaccharides, and / or in the case of any one or more disaccharides, trisaccharides, or polysaccharides, may include mixtures of any two or more thereof. Further, in any embodiment herein, monosaccharides, disaccharides, trisaccharides, and polysaccharides may include furanose sugars, pyranose sugars, or (R 1 and R 2 Or R 3 and R 4 are different monosaccharides, and / or in the case of any one or more disaccharides, trisaccharides, or polysaccharides, may include mixtures of any two or more thereof. 1 , R 2 , R 3 , and R 4 are each independently H or TIFF0007601403000023.tif46128, R 5Each occurrence of is independently H or TIFF0007601403000024.tif45128, r is an integer of 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, and s is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. 1 , R 2 , R 3 , R 4 , and R 5 are each independently allose, altrose, glucose, mannose, gulose, iodose, galactose, talose, or (R 1 , R 2 , R 3 , R 4 , and R 5 are different monosaccharides, and / or in the case of any one or more disaccharides, trisaccharides, or polysaccharides, mixtures of any two or more thereof. 1 , R 2 , R 3 , R 4 , and / or R 5 is a monosaccharide, disaccharide, trisaccharide, and / or polysaccharide, R 1 , R 2 , R 3 , R 4 , and / or R 5 can be linked via an α-glycosidic bond or a β-glycosidic bond. 1 , R 2 , R 3 , R 4 , and / or R 5 The disaccharides, trisaccharides, and / or polysaccharides may be saccharides containing α-glycosidic linkages, β-glycosidic linkages, or (in the case of trisaccharides and / or polysaccharides) mixtures of any two or more thereof.

[0091] a. Compositions containing alkyl polyglucosides Alkyl polyglycosides are particularly useful molecules that can be advantageously incorporated into numerous industrial materials and purposes.Indeed, alkyl polyglycosides find use, for example, as emulsifiers, emollients, and thickeners in cosmetics and foods, in agricultural formulations, for example, in pesticide formulations to deliver active ingredients to targets (e.g., the waxy surface of leaves), as industrial solvents, in the oil and gas industry to enhance oil recovery, as plasticizers, as surfactants and detergents, and in the production of surfactants and detergents, etc.

[0092] Alkyl polyglycoside surfactants are particularly valuable in that they enjoy advantages in terms of dermatological properties, compatibility with standard products, and favorable environmental profile compared to other surfactants.Therefore, they are widely used in a variety of household and industrial applications, for example, in pharmaceuticals for solubilizing hydrophobic drugs in aqueous media, as a component of emulsions or surfactant self-assembly vehicles for oral and transdermal drug delivery, as plasticizers in semi-solid delivery systems, as agents for improving drug absorption and penetration, etc.Therefore, in exemplary embodiments, alkyl polyglycosides are incorporated into personal care products, for example, as biological system components with reduced toxicity profile and less irritating to the skin than other surfactants and solubilizers.

[0093] Thus, in an exemplary embodiment, a personal care composition is provided that includes a compound disclosed herein and a cosmetically acceptable carrier.

[0094] Personal care products are known in the art, see, for example, Surfactants in Personal Care Products and Decorative Cosmetics, Third Edition, Linda D. Rhein, Mitchell Schlossman, Anthony O'Lenick, P. Somasundaran, eds. CRC Press (2006). In one exemplary embodiment, the personal care product is a skin care composition. Skin care compositions are known in the art, see, for example, Cosmetic Formulation of Skin Care Products, Zoe Diana Draelos, Lauren A. Thaman, eds. CRC Press (2005).

[0095] In an exemplary embodiment, the cosmetically acceptable carrier in the personal care composition is selected from the group consisting of water, emollients, fatty acids, fatty alcohols, thickeners, and combinations thereof.

[0096] In one exemplary embodiment, the personal care composition comprises water at a concentration of about 40 weight percent (wt %) to about 96 wt %.

[0097] In another exemplary embodiment, the personal care composition comprises an emollient selected from the group consisting of silicone oils, natural or synthetic esters, hydrocarbons, alcohols, fatty acids, and combinations thereof. In some exemplary embodiments, the emollient is present at a concentration of about 0.1% to about 60% by weight of the personal care composition. In other exemplary embodiments, the emollient is present at a concentration of about 30% to about 50% by weight.

[0098] In another exemplary embodiment, the personal care composition comprises a silicone oil. Silicone oils can be divided into volatile and non-volatile varieties. As used herein, the term "volatile" refers to materials that have a measurable vapor pressure at ambient temperature. Volatile silicone oils are selected from cyclic polydimethylsiloxanes (cyclomethicones) or linear polydimethylsiloxanes, preferably containing 3, 4, 5, 6, 7, 8, or 9 silicon atoms, preferably 5 to 6 silicon atoms. Non-volatile silicone oils include polyalkylsiloxanes, polyalkylarylsiloxanes, and polyether siloxane copolymers. Non-volatile polyalkylsiloxanes useful herein include, for example, polyalkylsiloxanes having a viscosity of 5×10-6 m at 25° C. 2 / sec ~ approx. 0.1m 2 / sec, preferably about 1×10-5 m at 25°C 2 / sec ~ approx. 4×10-4m 2 Polydimethylsiloxanes having a viscosity of 10 ...

[0099] In another exemplary embodiment, the personal care composition includes an ester emollient. In another exemplary embodiment, the ester emollient is an alkyl ester of a saturated fatty acid having from 10 to 24 carbon atoms. In an exemplary embodiment, the alkyl ester is a member selected from the group consisting of behenyl neopentanoate, isononyl isononanoate, isopropyl myristate, and octyl stearate.

[0100] In another exemplary embodiment, the personal care composition comprises an ether-ester (fatty acid ester) of an ethoxylated saturated fatty alcohol.

[0101] In another exemplary embodiment, the personal care composition comprises a polyhydric alcohol ester such as an acceptable polyhydric alcohol ester, such as ethylene glycol mono- and di-fatty acid esters, diethylene glycol mono- and di-fatty acid esters, polyethylene glycol (200-6000) mono- and di-fatty acid esters, propylene glycol mono- and di-fatty acid esters, polypropylene glycol 2000 monostearate, ethoxylated propylene glycol monostearate, glyceryl mono- and di-fatty acid esters, polyglycerin polyfatty esters, ethoxylated glyceryl monostearate, 1,3-butylene glycol monostearate, 1,3-butylene glycol distearate, polyoxyethylene polyol fatty acid esters, sorbitan fatty acid esters, and polyoxyethylene sorbitan fatty acid esters. In another exemplary embodiment, the polyhydric alcohol ester is selected from the group consisting of pentaerythritol, trimethylolpropane, and neopentyl glycol esters of C1-C30 alcohols.

[0102] In another exemplary embodiment, the personal care composition includes wax esters such as beeswax, spermaceti, and tribehenin wax.

[0103] In another exemplary embodiment, the personal care composition includes sugar esters of fatty acids, such as, for example, sucrose polybehenate and sucrose polycottonseedate.

[0104] In some exemplary embodiments, the personal care composition includes a natural ester emollient. The natural ester emollient is primarily based on monoglycerides, diglycerides, and triglycerides. Representative examples include sunflower seed oil, coconut oil, cottonseed oil, borage oil, borage seed oil, primrose oil, castor oil and hydrogenated castor oil, rice bran oil, soybean oil, olive oil, safflower oil, shea butter, jojoba oil, and combinations of any two or more thereof. Animal-derived emollients include, for example, lanolin oil and lanolin derivatives. In exemplary embodiments, the amount of natural ester is present in a concentration ranging from about 0.1% to about 20% by weight of the personal care composition.

[0105] In some exemplary embodiments, the personal care composition comprises a hydrocarbon. Suitable cosmetically acceptable carriers include, for example, petrolatum, mineral oil, C8-C30 n-alkanes, C8-C30 n-alkanes, C11-C13 isoparaffins, polybutenes, and in particular isohexadecane (commercially available as Permethyl 101A from Presperse Inc.).

[0106] In some exemplary embodiments, the personal care composition comprises a fatty acid. Typically, a fatty acid having 6 to 30 carbon atoms is provided as the cosmetically acceptable carrier. In some exemplary embodiments, a fatty acid having 10 to 30 carbon atoms is provided as the cosmetically acceptable carrier. In other exemplary embodiments, a fatty acid having 8 to 24 carbon atoms is provided as the cosmetically acceptable carrier. In yet other exemplary embodiments, a fatty acid having 6 to 24 carbon atoms is provided as the cosmetically acceptable carrier.

[0107] Some exemplary fatty acids having 1 to 30 carbons include pelargonic acid, lauric acid, myristic acid, palmitic acid, isostearic acid, oleic acid, linoleic acid, linolenic acid, hydroxystearic acid, and behenic acid, and mixtures of any two or more thereof. Fatty alcohols having 10 to 30 carbon atoms are another useful category of cosmetically acceptable carriers. Illustrative of this category are stearyl alcohol, lauryl alcohol, myristyl alcohol, oleyl alcohol, cetyl alcohol, and mixtures of any two or more thereof.

[0108] In some exemplary embodiments, the personal care composition comprises a thickening agent. Exemplary thickening agents include crosslinked acrylates (e.g., Carbopol 982®), hydrophobically modified acrylates (e.g., Carbopol 1382®), polyacrylamides (e.g., Sepigel 305®), acryloylmethylpropanesulfonic acid / salt polymers and copolymers (e.g., Aristoflex HMB® and AVC®), cellulosic derivatives, natural gums, and combinations of any two or more thereof. Useful cellulosic derivatives include sodium carboxymethylcellulose, hydroxypropylmethylcellulose, hydroxypropylcellulose, hydroxyethylcellulose, ethylcellulose, hydroxymethylcellulose, and combinations of any two or more thereof. Natural gums include, but are not limited to, guar, xanthan, sclerotium, carrageenan, pectin, and combinations of any two or more thereof. Combinations of any two or more of inorganic materials, particularly clays (such as bentonite and hectorite), fumed silica, talc, calcium carbonate, and silicates (such as magnesium aluminum silicate (Veegum®)), as well as organic materials, may be utilized as thickening agents. Combinations of any two or more thickening agents are also useful in the personal care compositions of the present technology. The amount of thickening agent may range from about 0.0001% to about 10%, preferably from about 0.001% to about 1%, optimally from about 0.01% to about 0.5% by weight of the personal care composition.

[0109] In some exemplary embodiments, the personal care composition includes a humectant. In exemplary embodiments, a humectant of the polyhydric alcohol type is used as the cosmetically acceptable carrier. Exemplary polyhydric alcohols include, for example, glycerol, polyalkylene glycols (more preferably alkylene polyols and their derivatives, including propylene glycol, dipropylene glycol, polypropylene glycol, polyethylene glycol, and their derivatives), sorbitol, hydroxypropyl sorbitol, hexylene glycol, 1,3-butylene glycol, isoprene glycol, 1,2,6-hexanetriol, ethoxylated glycerol, propoxylated glycerol, and mixtures thereof. In exemplary embodiments, the amount of humectant is present in a concentration of about 0.5% to about 50% by weight of the personal care composition. In exemplary embodiments, the amount of humectant present in the personal care composition is about 0.5% to about 50% by weight. In some exemplary embodiments, when a humectant is included, it is included in an amount that is about 1% to 15% by weight of the personal care composition.

[0110] In some exemplary embodiments, a skin moisturizer is included as a cosmetically acceptable carrier. Thus, in an exemplary embodiment, hyaluronic acid and / or its precursor N-acetylglucosamine is included as a cosmetically acceptable carrier. In some exemplary embodiments, N-acetylglucosamine is derived from shark cartilage or shiitake mushrooms and is commercially available from Maypro Industries, Inc. (New York). In other exemplary embodiments, the skin moisturizer included as a cosmetically acceptable carrier is hydroxypropyltri(C1-C3 alkyl)ammonium salt. In some exemplary embodiments, hydroxypropyltri(C1-C3 alkyl)ammonium salt is obtained from a synthetic procedure, for example, from hydrolysis of chlorohydroxypropyltri(C1-C3 alkyl)ammonium salt. In yet other exemplary embodiments, particularly when used in combination with the above-mentioned ammonium salts, humectants include, for example, substituted ureas such as hydroxymethylurea, hydroxyethylurea, hydroxypropylurea, and the like; bis(hydroxymethyl)urea; bis(hydroxyethyl)urea; bis(hydroxypropyl)urea; N,N'-dihydroxymethylurea; N,N'-di-hydroxyethylurea; N,N'-di-hydroxypropylurea; N,N,N'-tri-hydroxyethylurea; tetra(hydroxymethyl)urea; tetra(hydroxyethyl)urea; tetra(hydroxypropylurea; N-methyl-N'-hydroxyethylurea; N-ethyl-N'-hydroxyethylurea; N-hydroxypropyl-N'-hydroxyethylurea, N,N'-dimethyl-N-hydroxyethylurea.

[0111] In another exemplary embodiment, the personal care composition has a pH of about 4 to about 8. In some exemplary embodiments, the pH of the personal care composition is about 4, about 4.5, about 5, about 5.5, about 6, about 6.5, about 7, about 7.5, about 8, or any range including and between any two of these values. In one exemplary embodiment, the pH of the personal care composition is about 5 to about 7. In other exemplary embodiments, the pH of the personal care composition is about 5 to about 6.

[0112] In another exemplary embodiment, the personal care composition comprises an inorganic sunscreen. In one exemplary embodiment, the amount of inorganic sunscreen is present at a concentration of about 0.1% to about 10% by weight of the personal care composition. Inorganic sunscreens are well known in the art and include, but are not limited to, zinc oxide, iron oxide, silica (e.g., fumed silica), and titanium dioxide.

[0113] In some exemplary embodiments, the personal care composition comprises a cosmetically beneficial ingredient. Exemplary cosmetically beneficial ingredients include, but are not limited to, skin lightening ingredients, retinoids, herbal extracts, antifungals, resveratrol, alpha-lipoic acid, ellagic acid, kinetin, retinoxytrimethylsilane, ceramides, pseudoceramides, colorants, opacifiers, abrasives, and combinations thereof.

[0114] ii. Ethoxylated derivatives In an exemplary embodiment, the hydroxyl moieties of 5-dodecene-1,3-diol are available for ethoxylation to prepare novel alcohol ethoxylates, ethoxy sulfates, propoxylates and butoxylates, fatty alcohol polyglycol ethers, etc. Alcohol ethoxylates are known in the art (see, for example, U.S. Pat. No. 4,223,163, Surfactants. In Elvers, Barbara, et al. Ullmann's Encyclopedia of Industrial Chemistry. Weinheim, GER: Wiley-VCH).

[0115] Fatty alcohol ethoxylates were the first nonionic surfactants produced on a technical scale. They are widely used in cosmetics and other commercial products (e.g. detergents, cleaning agents, etc.).

[0116] 3. Polyurethane In some exemplary embodiments, the hydroxyl functional groups of 5-dodecene-1,3-diol are used to prepare polyurethanes.

[0117] The double bond of 5-dodecene-1,3-diol serves as a site for epoxidation, for example, as shown in Formula VII. TIFF0007601403000025.tif22128

[0118] As is known in the art, standard epoxidation chemistry involves hydrogen peroxide and heat at 30° C. to 80° C. using a Jacobsen class catalyst or a TS-1 zeolite catalyst.

[0119] In exemplary embodiments, the present invention uses a method for the preparation of synthetic organic compounds, which are known in the art (e.g., Practical Synthetic Organic Chemistry, Stephane Caron ed. (2011) (supra), Organic Chemistry, 9 thEdition, Carey and Giuliano (2013) (supra)), and for example, the alcohol group of 5-dodecene-1,3-diol is prevented from reacting itself at the epoxy ring by protecting group chemistry, such as an acetyl group, as shown in Formula VIII. TIFF0007601403000026.tif20128

[0120] In an exemplary embodiment, the epoxy ring on the derivatized 5-dodecene-1,3-diol is opened using methods known in the art, for example, using water, as shown in formula IX, to provide the tetraol. TIFF0007601403000027.tif26128

[0121] In another exemplary embodiment, the epoxy ring is opened using methods known in the art, for example, using a hydrogen reactant, as shown in formula X, to provide the triol. TIFF0007601403000028.tif28128

[0122] In an exemplary embodiment, the use of selective ring opening with hydrogen on dodecane-1,3,6-triol is preferred. Epoxide chemistry is by far the most widely used, and other diols, alcohols, and functionalized moieties are reacted at this site (Y. Li et al., Bio-based Polyols and Polyurethanes, Springer Briefs in Green Chemistry for Sustainability, DOI10.1007 / 978-3-319-21539-6_2).

[0123] In another exemplary embodiment, the alcohol group on 5-dodecene-1,3-diol is used to open the ring with an epoxy using a one-step "single-pot" reaction known in the art (e.g., Monteavaro LL, et al. J Am. Oil Chem. Soc. 82:365-371, 2005), resulting in a structure such as that shown in formula XI. TIFF0007601403000029.tif45131

[0124] The alcohol groups on 5-dodecene-1,3-diol are used to ring open with an epoxy to provide a branched polyol with a higher viscosity than the original 5-dodecene-1,3-diol. High viscosity polyols are useful in applications such as, for example, oil exploration and recovery, paints and coatings, and personal care.

[0125] In another exemplary embodiment, dodecane-1,3,6-triol of formula X is reacted with an epoxide of formula VII to provide a branched polyol of formula XII. TIFF0007601403000030.tif46128

[0126] In an exemplary embodiment, the polyol produced from 5-dodecene-1,3-diol is further derivatized, for example, by copolymerization with ethylene oxide, to provide a polyether polyol. The resulting polyether polyol can be used as is in a variety of applications, for example, as a building block for polyurethanes.

[0127] In an exemplary embodiment, 5-dodecene-1,3-diol, related triols or tetraols disclosed herein, or related branched polyols or other polyols produced using 5-dodecene-1,3-diol disclosed herein can proceed with standard chemistry with isocyanate compounds to form polyurethanes. These reactions can be accelerated by UV light or by catalysts such as dibutyltin dilaurate or bismuth octanoate, as known in the art (see, for example, Y. Li et al., Bio-based Polyols and Polyurethanes, Springer Briefs in Green Chemistry for Sustainability, DOI10.1007 / 978-3-319-21539-6_2). Many different isocyanates can be used, ranging from linear to aromatic, and techniques for preparing the polyols may or may not go through a prepolymer phase (e.g., preparation of diols, triols, polyols with isocyanate groups) (see, e.g., U.S. Pat. No. 4,532,316).

[0128] In an exemplary embodiment, carbamates are used as intermediates for synthesizing isocyanates and for the direct conversion of diols to prepare non-isocyanate polyurethanes (NIPUs, see, for example, Maisonneuve, L. et al. (2015) Chem. Rev. 115:12407-12439). Non-isocyanate polyurethanes are particularly useful worldwide because they allow the production of polyurethane performance and properties used in a variety of applications, from building materials to medical devices, without the use of carcinogenic isocyanates. This allows for safer working conditions for producers, commercial users, and even everyday consumers who may be exposed during the use of polyurethane products such as coatings and adhesives. Thus, the use of 5-dodecene-1,3-diol in the preparation of NIPUs allows access to new carbonate structures with improved reactivity compared to 1,2-diols (1,3-diol on a 6-membered ring versus 1,2-diol on a 5-membered ring), combined with the flexibility of crosslinking or rearrangement afforded by the double bond, to provide new non-isocyanate polyurethane products.

[0129] To prepare NIPUs, 5-dodecene-1,3-diol can be used as a diol or converted to a polyol as discussed above and used to react with a wide range of carbamates (see, for example, Rokocki, G, et al. Polym. Adv. Technol. 26, 707-761, 2015). The 1,3-diol configuration in 5-dodecene-1,3-diol has the advantage over 1,2-diols in that there is less steric hindrance from the alcohol reactive center.

[0130] The hydroxyl moieties of 5-dodecene-1,3-diol are available for reaction with dimethyl carbonate or carbon dioxide to prepare a 6-membered cyclic carbonate ring, which is then reacted with a primary amine to provide a novel "non-isocyanate" polyurethane (NIPU). If the carbonate is bi-terminated, that is, if it is first reacted with itself in a crosslinking or metathesis reaction to provide two ends for the polymer chain to continue (this can be done before or after the carbonate structure is formed), then the carbonate is available for further reaction to the NIPU. Thus, the branched polyol of formula XII may also be converted, for example, to a 2-carbonate polymer building block of formula XIII. TIFF0007601403000031.tif42128

[0131] Exemplary catalysts that can be used to make carbonate derivatives from 5-dodecene-1,3-diol or to make tetraols generated from 5-dodecene-1,3-diol include, for example, 1,5,7-triazabicyclo[4.4.0]dec-5-ene with dimethyl carbonate (Mutlu et al, Green Chem., 2012); various imidazolium or thiazolium carbene catalysts in the presence of cesium carbonate, dibromomethane, and CO2 at atmospheric pressure (Bobbink et al, Chem. Commun., 2016); and CeO2 with 2-cyanopyridine in the presence of CO2 (Honda et al, ACS Catal., 2014).

[0132] Six-membered cyclic carbonates derived from 1,3-diol moieties, such as those produced from 5-dodecene-1,3-diol, are 30 times more reactive than five-membered cyclic carbonates derived from 1,2-diol moieties and are therefore preferred for use (Maisonneuve et al, Chem. Rev., 2015 (supra)).

[0133] In an exemplary embodiment, self-metathesis to form the "two-end" structure employs a metathesis catalyst known in the art, such as a first or second generation Grubbs catalyst, one specific example being bis(tricyclohexylphosphine)dichlororuthenium(II) benzylidene, [(PCy)Cl]Ru=CHPh (see, e.g., International Application No. PCT / US95 / 09655).

[0134] The metathesis reaction is driven under vacuum to strip off the internal olefin by-products. In some exemplary embodiments, the metathesis reaction is carried out before the carbonate is formed (i.e., diol vs. tetraol), see formula XIV. Formula XIV has utility as a polyol, as discussed above. TIFF0007601403000032.tif42128

[0135] Alternatively, the carbonate may be first generated from a diol and then self-metathesised to both terminal carbonates. That is, a 6-membered cyclic carbonate molecule generated from 5-dodecene-1,3-diol is then self-metathesised to a molecule having two 6-membered cyclic carbonates, thus providing a molecule see formula XV. The metathesis reaction may result in a mixture of cis and trans double bonds. TIFF0007601403000033.tif33128

[0136] In some exemplary embodiments, 5-dodecene-1,3-diol or related 6-membered cyclic carbonates are first reacted by ethylene metathesis to form the terminal alkene counterpart, as shown below. TIFF0007601403000034.tif63128

[0137] Subsequent self-metathesis of compounds of formula XVI and formula XVII, respectively, provides both terminal compounds of formula XIV and XV, respectively.

[0138] Further reactions of formula XV can be carried out across the double bond to add rigidity, explore increasing the glass transition temperature (Tg) of the subsequent polymer, or to add pendant groups. In one exemplary embodiment, formula XV is reacted with butadiene and a Lewis acid in a standard Diels-Alder reaction to provide formula XVIII. TIFF0007601403000035.tif33128

[0139] 4. Polyester As noted above, 1,3-diol structures, for example, are less sterically hindered than 1,2-diols of similar chain length, aiding in polymer formation.

[0140] The chemistry for forming polyesters has been studied for over 100 years and is well known in the art. Exemplary chemistries include, but are not limited to, heat and acid catalyzed reactions, lipase enzyme catalyzed polycondensation, the use of scandium triflate as a catalyst, etc. (see, for example, Diaz, A. et al., Macromolecules 2005, 38, 1048-1050).

[0141] In an exemplary embodiment, 5-dodecene-1,3-diol is reacted with a diacid, such as, for example, adipic acid, to form a "bushy" polyester, such as, for example, the molecule shown below as Formula XIX, where n is an integer from 1 to 1000. TIFF0007601403000036.tif72128

[0142] The "bush-like" character of the polyester structure may result in lower crystallinity and improved performance in micelle formation in surfactants as compared to, for example, polyesters made with alpha-omega diols (1,2-diols), as well as hydrophobicity in other polymer applications.

[0143] The structure of 5-dodecene-1,3-diol functions to affect crystallinity (e.g., promoting lower crystallinity) and glass transition temperature (Tg), polymer phase interactions, extrusion properties, and solubility and surface interactions with water, solvents, and composite formulations. Thus, in an exemplary embodiment, 5-dodecene-1,3-diol is used as an additive to increase hydrophobicity and provide a plasticizer effect. Thus, in an exemplary embodiment, 5-dodecene-1,3-diol is used at a concentration of about 0.001% to about 45% of the diol or polyol species to affect the structure of known polyesters. Thus, in an exemplary embodiment, 5-dodecene-1,3-diol is incorporated into polybutyrate, a copolyester of adipic acid, 1,4-butanediol, and dimethyl terephthalate, at a lower percentage than the major monomer, to provide a molecule such as that shown below as formula XX, where m and n are each independently an integer between 1 and 1000. TIFF0007601403000037.tif48153

[0144] II. Preparation of (Z)-5-dodecene-1,3-diol A. General pathways of biological synthesis 5-dodecene-1,3-diol can be made by any method known in the art.As discussed above, unsaturated fatty alcohols are difficult to produce from petroleum.Rather, unsaturated fatty alcohols are typically produced from the processing of non-petroleum sources, such as fats and oils of vegetable and animal origin (see, for example, EF Hill, et al. (1954) Ind. Eng. Chem., 46(9):1917-1921).Such processes are messy, can cause pollution, and typically produce only a limited variety of unsaturated fatty alcohol products.

[0145] Fortunately, however, biological methods are available for the direct production of 5-dodecene-1,3-diol. Thus, in an exemplary embodiment, 5-dodecene-1,3-diol is made using the biological method disclosed, for example, in WO2016 / 011430A1 and disclosed in detail in Example 1 herein below.

[0146] Briefly, the preparation of 5-dodecene-1,3-diol can be carried out in recombinant host cells (e.g., bacterial cells) engineered to produce 1,3-diol by utilizing nucleic acids and their corresponding polypeptides with enzymatic functions to modify an enzymatic pathway for the in vivo production of a desired compound, such as 5-dodecene-1,3-diol. Enzyme polypeptides are identified herein below by Enzyme Accession Numbers (EC Numbers).

[0147] WO2016 / 011430A1 discloses an enzymatic pathway engineered to produce 1,3-diols. One exemplary pathway for the production of 5-dodecene-1,3-diol utilizes 3' hydroxyacyl carrier protein (ACP) (e.g., acyl-ACP or 3-hydroxyacyl-ACP) carrying an acyl intermediate, which is converted to 1,3-diols with 3' hydroxy fatty acid (3'-OH FA) and 3' hydroxy fatty aldehyde (3'-OH fatty aldehyde) as intermediates. Without being bound by theory, it is believed that a simple carbon source such as glucose is first converted to 3' hydroxyacyl-ACP by a microorganism (e.g., Escherichia, Bacillus, Lactobacillus, etc.). Thus, in one embodiment, the acyl-ACP or 3' hydroxyacyl-ACP that starts the engineered enzymatic pathway is produced by a natural pathway of the microorganism. In an exemplary embodiment, the 3' hydroxyacyl-ACP is converted to an intermediate such as a 3'-OH FA by an enzyme with thioesterase (TE) activity (EC 3.1.2.- or EC 3.1.2.14 or EC 3.1.1.5). The intermediate 3'-OH FA is then converted to another intermediate such as a 3'OH aldehyde by an enzyme with carboxylic acid reductase (CAR) activity (EC 1.2.99.6). An enzyme with alcohol dehydrogenase (ADH) or aldehyde reductase (AR) activity (EC 1.1.1.1 or EC 1.1.1.2) then converts the 3'OH aldehyde to a 1,3-diol.

[0148] In other embodiments, 3' hydroxyacyl-ACP is converted to intermediates such as 3'-OH fatty aldehydes by an enzyme with acyl-ACP reductase activity (AAR, EC 1.2.1.42). The production of fatty alcohols and / or fatty aldehydes by AAR can be enhanced through heterologous expression of a gene called accABCD, which encodes acetyl-CoA carboxylase. An enzyme with alcohol dehydrogenase (ADH) or aldehyde reductase (AR) activity (EC 1.1.1.1 or EC 1.1.1.2) can then convert the 3'-OH aldehyde to a fatty diol, such as 1,3-diol. Thus, the present disclosure provides recombinant microorganisms that can efficiently and selectively produce 1,3-fatty diols, such as 5-dodecene-1,3-diol, in vivo. It should be noted that most cells can be cytotoxic, so they produce enzymes that can reduce aldehydes originally. Thus, heterologous expression of AR and ADH may not be required for the production of fatty alcohols and diols, although they can improve the efficiency of production of fatty diols.

[0149] Thus, using the methods disclosed herein, saturated and unsaturated 1,3-fatty diols are produced. Exemplary 1,3-fatty diols include, for example, C5 1,3 fatty diols (e.g., 1,3-pentanediol), C6 1,3 fatty diols (e.g., 1,3-hexanediol), C7 1,3 fatty diols (e.g., 1,3-heptanediol), C8 1,3 fatty diols (e.g., 1,3-octanediol), C9 1,3 fatty diols (e.g., 1,3-nonanediol), C1,3 fatty diols (e.g., 1,3-nonanediol), C2,3 fatty diols (e.g., 1,3-nonanediol), C3,3 fatty diols (e.g., 1,3-nonanediol), C4,3 fatty diols (e.g., 1,3-nonanediol), C5,3 fatty diols (e.g., 1,3-pentanediol), C6,3 fatty diols (e.g., 1,3-hexanediol), C7,3 fatty diols (e.g., 1,3-heptanediol), C8,3 fatty diols (e.g., 1,3-octanediol), C9,3 fatty diols (e.g., 1,3-nonanediol), C1,3 fatty diols (e.g., 1,3-nonanediol), C1,3 fatty diols (e.g., 1,3-nonanediol), C2,3 fatty diols (e.g., 1,3-nonanediol), C3,3 fatty diols (e.g., 1,3-nonanediol), C4,3 fatty diols (e.g., 1,3-nonane 10 1,3 fatty diols (e.g., 1,3-decanediol), C 11 1,3 fatty diols (e.g., 1,3-undecanediol), C 12 1,3 fatty diols (e.g., 1,3-dodecanediol, 5-dodecene-1,3-diol), C 13 1,3 fatty diols (e.g., 1,3-tridecanediol), C 141,3 fatty diols (e.g., 1,3-tetradecanediol, 7-tetradecene-1,3-diol), C 15 1,3 fatty diols (e.g., 1,3-pentadecanediol), C 16 1,3 fatty diols (e.g., 1,3-hexadecanediol, 9-hexadecene-1,3-diol), C 17 1,3 fatty diols (e.g., 1,3-heptadecanediol), C 18 1,3 fatty diols (e.g., 1,3-octadecanediol, 11-octadecene-1,3-diol), C 19 and 1,3 fatty diols (e.g., 1,3-nonadecanediol). Although primarily even-chain 1,3-fatty diols are disclosed herein, odd-chain 1,3-fatty diols are also found, such as those having 7 to 21 carbons, and more preferably 5 to 19 carbons.

[0150] Generally, in exemplary embodiments, the unsaturated 1,3-fatty diols produced using the microorganisms disclosed herein above carry a double bond in the (Z) configuration. However, as discussed herein below, methods are available for rearranging the (Z) double bond of the unsaturated 1,3-fatty diol, such as the double bond of (Z)-5-dodecene-1,3-diol, so that the double bond is produced in the (E) configuration.

[0151] For further guidance regarding the biological synthesis of 1,3-fatty diols, one of skill in the art can refer, for example, to Examples 1-3 herein below and / or International Patent Application Publication No. WO2016 / 011430A1.

[0152] b. Chirality of 5-dodecene-1,3-diol The 3-hydroxy functionality of the 1,3-aliphatic diols disclosed herein, such as 5-dodecene-1,3-diol, forms a stereocenter at C-3 and provides a point of chirality for the molecule. As noted above, chirality can be a useful molecular attribute in defining molecular applications, including, for example, polymer performance, biological activity, pharmaceutical efficacy, and the like.

[0153] The stereoisomers of 1,3-fatty diols produced by microorganisms depend on the selectivity of the fatty acid biosynthetic pathway (FAS) in which they are produced. By manipulating which FAS enzymes are involved in the synthesis of 1,3-fatty diols, such as 5-dodecene-1,3-diol, the chirality of the resulting 1,3-fatty diol can be controlled.

[0154] For example, in one exemplary embodiment, native E. coli FAS is utilized to produce the (R) enantiomer of an unsaturated 1,3-fatty diol, such as 5-dodecene-1,3-diol. In this embodiment, the chiral center of the unsaturated 1,3-fatty diol is created by the activity of 3-ketoacyl-ACP reductase, an enzyme encoded by the FabG gene in E. coli. The activity of 3-ketoacyl-ACP reductase produces (R)-3-hydroxyl acyl ACP, which can then enter the engineered enzyme pathway(s) discussed above in Section IIa.

[0155] In another exemplary embodiment, the beta-oxidation pathway is utilized to generate the (S) enantiomer of an unsaturated 1,3-fatty diol, such as 5-dodecene-1,3-diol. In this embodiment, the (S) enantiomer of the unsaturated 1,3-fatty diol is prepared by causing the accumulation of (S)-3-hydroxyacyl CoA, which is an intermediate in the breakdown of fatty acids through the beta-oxidation pathway. The excess (S)-3-hydroxy-acyl CoA is then converted to the (S) enantiomer of the unsaturated 1,3-fatty diol through the action of a fatty alcohol-forming polypeptide.

[0156] Thus, in one exemplary embodiment, to prepare the (S) enantiomer of an unsaturated 1,3-fatty diol, e.g., 5-dodecene-1,3-diol, an available free fatty acid is first converted to acyl-CoA by acyl-CoA synthase, a reaction catalyzed by FadD in E. coli (and homologs in other microorganisms). The resulting acyl-CoA is then oxidized to trans-2-enoyl-CoA by fatty acyl-CoA dehydrogenase, a reaction catalyzed by FadE in E. coli (and homologs in other microorganisms). The resulting trans-2-enoyl-CoA is then hydrated to (S)-3-hydroxy-acyl-CoA by 2-trans-enoyl-CoA hydratase / (S)-3-hydroxy-acyl-CoA dehydratase, a reaction catalyzed by FadB in E. coli (and homologs in other microorganisms).

[0157] In the wild-type beta-oxidation pathway, (S)-3-hydroxy-acyl-CoA is further oxidized to 3-keto-acyl-CoA by 3-keto-acyl-CoA dehydrogenase, a reaction also catalyzed by FadB in E. coli (and homologues in other microorganisms). The resulting 3-keto-acyl-CoA is thiolyzed to acyl-CoA and acetyl-CoA by 3-ketoacyl-CoA thiolase, a reaction catalyzed by FadA in E. coli (and homologues in other microorganisms).

[0158] In one exemplary embodiment, accumulation of (S)-3-hydroxy-acyl-CoA is caused by selectively blocking the dehydrogenase activity of 3-keto-acyl-CoA dehydrogenase (FadB) to prevent oxidation of (S)-3-hydroxy-acyl-CoA to 3-keto-acyl-CoA. In an exemplary embodiment, selective blocking of the (S)-3-hydroxy-acyl-CoA dehydrogenase activity of FadB is achieved by mutation of histidine 450 in the FadB gene of E. coli (see, e.g., He XY and Yang SY (1996) Biochemistry 35(29):9625-9630). (S)-3-hydroxy-acyl-CoA that accumulates in the cell is converted to the (S) enantiomer of an unsaturated 1,3-fatty diol, such as (S)-5-dodecene-1,3-diol, through the action of a fatty alcohol-forming polypeptide, such as those disclosed in WO2016 / 011430A1.

[0159] The determination / confirmation of the resulting enantiomeric configuration is achieved by any method known in the art, for example, by non-chromatographic techniques such as polarimetry, nuclear magnetic resonance, isotope dilution, calorimetry, and enzymatic techniques. These techniques require pure samples and do not involve separation of enantiomers. Quantification and separation of enantiomers (without the need for pure samples) can be performed simultaneously by chiral chromatography, such as gas chromatography (GC) or high performance liquid chromatography (HPLC), using a chiral column (see, for example, Stereochemistry of Organic Compounds, Ernest L. Elil and Sanuel H. Wilen, 1994, John Wiley & Sons, Inc.). The chiral purity of the product can be identified using chiral chromatographic methods, such as chiral HPLC or LC / MS (see, for example, U.S. Patent Application Publication Nos. US2008 / 0248539A1 and US2013 / 0052699A1).

[0160] c. Fermentation and production of 1,3-fatty diols As used herein, fermentation broadly refers to the conversion of organic material to a target substance by recombinant host cells. For example, this includes the conversion of a carbon source to a fatty acid derivative (such as a 1,3-fatty diol) by recombinant host cells by growing a culture of the recombinant host cells in a medium containing a carbon source. Conditions that allow the production of a target substance, such as a 1,3-fatty diol, e.g., 5-dodecene-1,3-diol, are any conditions that allow the host cells to produce a desired product (such as a 1,3-fatty diol composition). Suitable conditions include, for example, typical fermentation conditions, see, for example, Principles of Fermentation Technology, 3rd Edition (2016) (supra), Fermentation Microbiology and Biotechnology, 2nd Edition (2007) (supra).

[0161] Fermentation conditions may include many parameters, including but not limited to temperature range, pH level, aeration level, feed rate, and medium composition. Each of these conditions, individually and in combination, allows for the growth of the host cells. Fermentation may be aerobic, anaerobic, or variations thereof (such as microaerobic). Exemplary culture media include broth (liquid) or gel (solid). Generally, the medium includes a carbon source (e.g., a simple carbon source from a renewable feedstock) that can be directly metabolized by the host cells. In addition, enzymes may be used in the medium to facilitate mobilization (e.g., depolymerization of starch or cellulose into fermentable sugars) and subsequent metabolism into the carbon source.

[0162] For small scale production, host cells engineered to produce 1,3-fatty diols may be grown and fermented in batches of, for example, about 100 μL, 200 μL, 300 μL, 400 μL, 500 μL, 1 mL, 5 mL, 10 mL, 15 mL, 25 mL, 50 mL, 75 mL, 100 mL, 500 mL, 1 L, 2 L, 5 L, or 10 L, etc., and induced to express a desired polynucleotide sequence, such as a polynucleotide encoding a polypeptide having a particular enzymatic activity (e.g., thioesterase (TE), carboxylic acid reductase (CAR), alcohol dehydrogenase (ADH), fatty acyl-CoA / ACP reductase (FAR), acyl-CoA reductase (ACR), acyl-CoA carboxylase (ACC), and / or acyl-ACP / CoA reductase (AAR) enzymatic activity). For large-scale production, engineered host cells may be grown, fermented, and induced to express any desired polynucleotide in cultures having volumetric batches of about 10 L, 100 L, 1000 L, 10,000 L, 100,000 L, 1,000,000 L, or more. The 1,3-fatty diol compositions described herein can be found in the extracellular environment of recombinant host cell cultures and can be easily isolated from the culture medium. The 1,3-fatty diols, e.g., fatty acid derivatives such as 5-dodecene-1,3-diol, and / or fatty alcohols, may be secreted by recombinant host cells and transported to the extracellular environment, or passively introduced into the extracellular environment of recombinant host cell cultures. The 1,3-fatty diol compositions may be isolated from recombinant host cell cultures using routine methods known in the art (see, e.g., Example 2 herein below).

[0163] Exemplary microorganisms suitable for use as production host cells include, for example, bacteria, cyanobacteria, yeast, algae, or filamentous fungi. To produce 1,3-fatty diols, the production host cell (or equivalently the host cell) is engineered to contain a fatty acid biosynthetic pathway that is modified compared to an unengineered or native host cell, for example, engineered as discussed above in section II.a and disclosed in WO2016 / 011430A1. The production host engineered to contain a modified fatty acid biosynthetic pathway can efficiently convert glucose or other renewable feedstocks to fatty acid derivatives, including fatty alcohols and 1,3-fatty diols, such as 5-dodecene-1,3-diol. High density fermentation protocols and procedures for producing a variety of compounds have been established (see, e.g., U.S. Pat. Nos. 8,372,610, 8,323,924, 8,313,934, 8,283,143, 8,268,599, 8,183,028, 8,110,670, 8,110,093, and 8,097,439).

[0164] In some exemplary embodiments, the production host cells are cultured in a culture medium (e.g., fermentation medium) that includes an initial concentration of a carbon source (e.g., a simple carbon source) of about 20 g / L to about 900 g / L. In other embodiments, the culture medium includes an initial concentration of a carbon source of about 2 g / L to about 10 g / L, about 10 g / L to about 20 g / L, about 20 g / L to about 30 g / L, about 30 g / L to about 40 g / L, or about 40 g / L to about 50 g / L. In some embodiments, the level of available carbon source in the culture medium may be monitored during the fermentation process. In some embodiments, the method further includes adding additional carbon source to the culture medium when the level of the initial carbon source in the medium is less than about 0.5 g / L.

[0165] In some exemplary embodiments, the additional carbon source is added to the culture medium when the initial carbon source level in the medium is less than about 0.4 g / L, less than about 0.3 g / L, less than about 0.2 g / L, or less than about 0.1 g / L. In some embodiments, the additional carbon source is added to maintain a carbon source level of about 1 g / L to about 25 g / L. In some embodiments, the additional carbon source is added to maintain a carbon source level of about 2 g / L or more (e.g., about 2 g / L or more, about 3 g / L or more, about 4 g / L or more). In certain embodiments, the additional carbon source is added to maintain a carbon source level of about 5 g / L or less (e.g., about 5 g / L or less, about 4 g / L or less, about 3 g / L or less). In some embodiments, additional carbon source is added to maintain a carbon source level of about 2 g / L to about 5 g / L, about 5 g / L to about 10 g / L, or about 10 g / L to about 25 g / L.

[0166] In an exemplary embodiment, the carbon source for fermentation is derived from a renewable feedstock. In some embodiments, the carbon source is glucose. In other embodiments, the carbon source is glycerol. Other possible carbon sources include, but are not limited to, fructose, mannose, galactose, xylose, arabinose, starch, cellulose, pectin, xylan, sucrose, maltose, cellobiose, and turanose; cellulosic materials and variants (such as hemicellulose, methylcellulose, and sodium carboxymethylcellulose); saturated or unsaturated fatty acids, succinates, lactates, and acetates; alcohols (such as ethanol, methanol, and glycerol), or mixtures thereof. In an embodiment, the carbon source is derived from corn, sugarcane, sorghum, sugar beet, switchgrass, ensilage, straw, timber, pulp, sewage, trash, cellulosic municipal waste, flue gas, synthesis gas, or carbon dioxide. Simple carbon sources may also be products of photosynthesis, such as glucose or sucrose. In one embodiment, the carbon source is derived from waste products such as glycerol, flue gas, or syngas; reformulation of organic materials such as biomass; natural gas or methane, or reformulation of those materials into syngas; photosynthetically fixed carbon dioxide (e.g., 1,3 diols can be produced by recombinant cyanobacteria growing photosynthetically using CO2 as a carbon source). In a particular embodiment, the carbon source is derived from biomass. One exemplary source of biomass is plant matter or vegetation, such as corn, sugarcane, or switchgrass. Another exemplary source of biomass is metabolic waste products, such as animal matter (e.g., cow manure). Further exemplary sources of biomass include algae and other marine plants. Biomass also includes waste products from industry, agriculture, forestry, and households, including, but not limited to, fermentation waste, ensilage, straw, timber, sewage, garbage, cellulosic municipal waste, municipal solid waste, and food waste.

[0167] In some exemplary embodiments, the 1,3-fatty diol, e.g., 5-dodecene-1,3-diol, is produced at a concentration of about 0.5 g / L to about 40 g / L. In some embodiments, the 1,3-fatty diol is produced at a concentration of about 1 g / L or more (e.g., about 1 g / L or more, about 10 g / L or more, about 20 g / L or more, about 50 g / L or more, about 100 g / L or more). In some embodiments, the 1,3-fatty diol is produced at a concentration of about 1 g / L to about 170 g / L, about 1 g / L to about 10 g / L, about 40 g / L to about 170 g / L, about 100 g / L to about 170 g / L, about 10 g / L to about 100 g / L, about 1 g / L to about 40 g / L, about 40 g / L to about 100 g / L, or about 1 g / L to about 100 g / L.

[0168] In other exemplary embodiments, the 1,3-fatty diol is about 25 mg / L, about 50 mg / L, about 75 mg / L, about 100 mg / L, about 125 mg / L, about 150 mg / L, about 175 mg / L, about 200 mg / L, about 225 mg / L, about 250 mg / L, about 275 mg / L, about 300 mg / L, about 325 mg / L, about 350 mg / L, about 375 mg / L, about 400 mg / L, about 425 mg / L, about 450 mg / L, about 475 mg / L, about 500 mg / L, about 525 mg / L, about 550 mg / L, Approximately 575mg / L, approximately 600mg / L, approximately 625mg / L, approximately 650mg / L, approximately 675mg / L, approximately 700mg / L, approximately 725mg / L, approximately 750mg / L, approximately 775mg / L, approximately 800mg / L, approximately 825mg / L, approximately 850mg / L, approximately 87 5mg / L, approximately 900mg / L, approximately 925mg / L, approximately 950mg / L, approximately 975mg / L, approximately 1000mg / L, approximately 1050mg / L, approximately 1075mg / L, approximately 1100mg / L, approximately 1125mg / L, approximately 1150mg / L, approximately 1175mg / L , about 1200mg / L, about 1225mg / L, about 1250mg / L, about 1275mg / L, about 1300mg / L, about 1325mg / L, about 1350mg / L, about 1375mg / L, about 1400mg / L, about 1425mg / L, about 1450mg / L, Approx. 1475mg / L, approx. 1500mg / L, approx. 1525mg / L, approx. 1550mg / L, approx. 1575mg / L, approx. 1600mg / L, approx. 1625mg / L, approx. The present invention is produced in a titer of about 1750 mg / L, about 1775 mg / L, about 1800 mg / L, about 1825 mg / L, about 1850 mg / L, about 1875 mg / L, about 1900 mg / L, about 1925 mg / L, about 1950 mg / L, about 1975 mg / L, about 2000 mg / L (2 g / L), 3 g / L, 5 g / L, 10 g / L, 20 g / L, 30 g / L, 40 g / L, 50 g / L, 60 g / L, 70 g / L, 80 g / L, 90 g / L, 100 g / L, or a range bounded by any two of the aforementioned values.In other embodiments, 1,3-fatty diols (e.g., 1,3-diols) are produced in titers of greater than 100 g / L, greater than 200 g / L, greater than 300 g / L, or more (e.g., 500 g / L, 700 g / L, 1000 g / L, 1200 g / L, 1500 g / L, or 2000 g / L). Preferred titers of 1,3-fatty diols, such as 1,3-diols, produced by recombinant host cells according to the methods of the present disclosure are 5 g / L to 200 g / L, 10 g / L to 150 g / L, 20 g / L to 120 g / L, and 30 g / L to 100 g / L, 100 g / L to 150 g / L, and 120 g / L to 180 g / L. In one embodiment, the preferred titer of 1,3-fatty diols, such as 1,3-diols produced by recombinant host cells according to the methods of the present disclosure, is from about 1 g / L to about 250 g / L, and more specifically, from 90 g / L to about 120 g / L. The titer can refer to a particular 1,3-diol or a combination of 1,3-diols of different chain lengths or different functionality, such as a mixture of saturated and unsaturated 1,3-fatty diols produced by a given recombinant host cell culture.

[0169] In other exemplary embodiments, host cells engineered to produce 1,3-fatty diols, such as, for example, 5-dodecene-1,3-diol according to the methods of the present disclosure have a yield of at least 1%, at least 2%, at least 3%, at least 4%, at least 5%, at least 6%, at least 7%, at least 8%, at least 9%, at least 10%, at least 11%, at least 12%, at least 13%, at least 14%, at least 15%, at least 16%, at least 17%, at least 18%, at least 19%, at least 20%, at least 21%, at least 22%, at least 23%, at least 24%, at least 25%, at least 26%, at least 27%, at least 28%, at least 29%, or at least 30%, or at least 40%, or a range bounded by any two of the above values. In other embodiments, 1,3-fatty diols, such as 5-dodecene-1,3-diol, are produced in yields of greater than 30%, greater than 40%, greater than 50%, greater than 60%, greater than 70%, greater than 80%, greater than 90%, or greater. Alternatively, or in addition, the yield is about 30% or less, about 27% or less, about 25% or less, or about 22% or less. Thus, the yield can be bounded by any two of the above endpoints. For example, the yield of 1,3-fatty diols, such as 1,3-diols, produced by recombinant host cells according to the methods of the present disclosure can be 5% to 15%, 10% to 25%, 10% to 22%, 15% to 27%, 18% to 22%, 20% to 28%, or 20% to 30%. In one particular embodiment, the yield of 1,3-fatty diol, such as 1,3-diol, produced by the recombinant host cell is about 10% to about 40%. In another particular embodiment, the yield of 1,3-fatty diol, such as 1,3-diol, produced by the recombinant host cell is about 25% to about 30%. The yield refers to a particular 1,3-fatty diol, such as 5-dodecene-1,3-diol, or combination of 1,3-diols produced by a given recombinant host cell culture. In addition, the yield will also depend on the feedstock used.

[0170] In some exemplary embodiments, the productivity of a host cell engineered to produce a 1,3-fatty diol, such as, for example, 5-dodecene-1,3-diol according to the methods of the present disclosure, is at least 100 mg / L / hr, at least 200 mg / L / hr, at least 300 mg / L / hr, at least 400 mg / L / hr, at least 500 mg / L / hr, at least 600 mg / L / hr, at least 700 mg / L / hr, at least 800 mg / L / hr, at least 900 mg / L / hr, at least 1000 mg / L / hr, at least 1000 mg / L / hr, at least 15 ...1000 mg / L / hr, at least 1500 mg / L / hr, at least 1000 mg / L / hr, at least 1500 mg / L / hr, at least 1000 mg / L / hr, at least 1500 mg / L / hr, at least 1000 mg / L / hr, at least 1500 mg / L / hr, at least 1000 mg / L / hr, at least 1500 mg / L / hr, at least 1000 mg / L / hr, at least 1500 mg / L / hr, at least 1000 mg / L / hr, at least 15 100 mg / L / hr, at least 1200 mg / L / hr, at least 1300 mg / L / hr, at least 1400 mg / L / hr, at least 1500 mg / L / hr, at least 1600 mg / L / hr, at least 1700 mg / L / hr, at least 1800 mg / L / hr, at least 1900 mg / L / hr, at least 2000 mg / L / hr, at least 2100 mg / L / hr, at least 2200 mg / L / hr, at least 2300 mg / L / hr, at least 2400 mg / L / hr, or at least 2500 mg / L / hr. For example, the productivity of a host cell engineered to produce a 1,3-fatty diol, such as a 1,3-diol produced by a recombinant host cell according to the methods of the present disclosure, can be between 500 mg / L / hr and 2500 mg / L / hr, or between 700 mg / L / hr and 2000 mg / L / hr. In one exemplary embodiment, the productivity is from about 0.7 mg / L / hr to about 3 g / L / hr. As used herein, productivity refers to the specific 1,3-fatty diol, such as, for example, 5-dodecene-1,3-diol, produced by a given recombinant host cell culture.

[0171] In some exemplary embodiments, the host cells used in the fermentation procedures discussed herein (above) are mammalian cells, plant cells, insect cells, yeast cells, fungal cells, filamentous fungal cells, algae cells, cyanobacterial cells, and bacterial cells. In certain embodiments, the host cells are selected from the group consisting of Escherichia, Bacillus, Pseudomonas, Lactobacillus, Rhodococcus, Synechococcus, Synechoystis, Pseudomonas, Aspergillus, Trichoderma, Neurospora, Fusarium, Humicola, Rhizomucor, Kluyveromyces, Pichia, and the like. hia, Mucor, Myceliophtora, Penicillium, Phanerochaete, Pleurotus, Trametes, Chrysosporium, Saccharomyces, Stenotrophamonas, Schizosaccharomyces, Yarrowia, or Streptomyces.In other exemplary embodiments, the host cell is a Bacillus lentus cell, a Bacillus brevis cell, a Bacillus stearothermophilus cell, a Bacillus licheniformis cell, a Bacillus alkalophilus cell, a Bacillus coagulans cell, a Bacillus circulans cell, a Bacillus pumilis cell, a Bacillus thuringiensis cell, a Bacillus clausii cell, a Bacillus megaterium cell, a Bacillus subtilis cell, or a Bacillus spp. In another exemplary embodiment, the host cell is a Bacillus subtilis cell, or a Bacillus amyloliquefaciens cell. In another exemplary embodiment, the host cell is a Pseudomonas putida cell.In certain embodiments, the host cell is selected from the group consisting of Synechococcus (spp. PCC7002, Synechococcus elongatus PCC7942, Synechocystis spp. PCC6803, Synechococcus elongatus PCC6301, Prochlorococcus marinus CCMP1986 (MED4), Anabaena variabilis ATCC29413, Nostoc punctiforme ATCC29133 (PCC73102), Gloeobacter violaceus, and the like. violaceus ATCC 29082 (PCC 7421), Nostoc spp. ATCC 27893 (PCC 7120), Cyanothece spp. PCC 7425 (29141), Cyanothece spp. ATCC 51442, or Synechococcus spp. ATCC 27264 (PCC 7002).In other exemplary embodiments, the host cell is a Trichoderma koningii cell, a Trichoderma viride cell, a Trichoderma reesei cell, a Trichoderma longibrachiatum cell, an Aspergillus awamori cell, an Aspergillus fumigates cell, an Aspergillus foetidus cell, an Aspergillus nidulans cell, an Aspergillus niger cell, an Aspergillus oryzae cell, a Humicola insolens cell, or a combination thereof. In another exemplary embodiment, the host cell is an Actinomycetes cell. In yet another exemplary embodiment, the host cell is a Streptomyces lividans cell or a Streptomyces murinus cell. In another exemplary embodiment, the host cell is a Saccharomyces cerevisiae cell.

[0172] In yet other exemplary embodiments, the host cell is a cell derived from a eukaryotic plant, algae, cyanobacteria, green sulfur bacteria, green non-sulfur bacteria, purple sulfur bacteria, purple non-sulfur bacteria, extremophiles, yeast, fungi, engineered versions thereof, or synthetic organisms. In some exemplary embodiments, the host cell is selected from the group consisting of Arabidopsis thaliana, Panicum virgatum, Miscanthus giganteus, Zea mays, botryococcus braunii, Chalamydomonas reinhardtii, Dunaliela salina, Thermosynechococcus elongatus, Synechococcus elongatus, Synechococcus spp., Synechocystis spp., Chlorobium tepidum, Chloroflexus auranticus, Chromatium vinosum, and the like. vinosum, Rhodospirillum rubrum, Rhodobacter capsulatus, Rhodopseudomonas palusris, Clostridium ljungdahlii, Clostridium thermocellum, or Penicillium chrysogenum.In some other exemplary embodiments, the host cell is a cell derived from Pichia pastoris, Saccharomyces cerevisiae, Yarrowia lipolytica, Schizosaccharomyces pombe, Pseudomonas fluorescens, Pseudomonas putida, or Zymomonas mobilis. In still further exemplary embodiments, the host cell is a cell derived from Synechococcus sp. PCC7002, Synechococcus sp. PCC7942, or Synechocystis sp. PCC6803. In some exemplary embodiments, the host cell is a CHO cell, a COS cell, a VERO cell, a BHK cell, a HeLa cell, a Cv1 cell, an MDCK cell, a 293 cell, a 3T3 cell, or a PC12 cell. In some exemplary embodiments, the host cell is an E. coli cell. In some exemplary embodiments, the E. coli cell is a B, C, K, or W strain E. coli cell.

[0173] D. Metathesis As discussed above, the double bonds of unsaturated 1,3-fatty diols, such as 5-dodecene-1,3-diol, produced by recombinant host cells engineered to produce 1,3-fatty diols, are predominantly in the (Z) configuration.

[0174] US Patent No. 9,163,267 teaches a method for producing olefins by contacting a composition comprising at least one omega-7-olefin fatty acid or its derivative with a cross-metathesis catalyst under conditions that allow cross-metathesis transformation, wherein the at least one omega-7-olefin fatty acid or its derivative is produced in a genetically engineered microorganism. Thus, in an exemplary embodiment, a method such as that disclosed in US Patent No. 9,163,267 is used to prepare the (E) isomer of an unsaturated (Z)-1,3-fatty diol, such as the (E) isomer of 5-dodecene-1,3-diol, which is produced using the engineered microorganism disclosed hereinabove. As is well known in the art, in cross-metathesis reactions, (Z)-(E) selectivity is typically biased towards the formation of the (E) isomer (see, for example, Naeimeh Bahri-Laleh et al., (2011) Beilstein J. Org. Chem. 7:40-45).

[0175] III. Compositions and Formulations of 1,3-Fatty Diols Bioproducts, e.g., compositions including 5-dodecene-1,3-diol produced utilizing the engineered microorganisms discussed above in Sections IIa-IId., are produced from renewable sources (e.g., from simple carbon sources derived from renewable feedstocks) and are therefore novel compositions of matter. These novel bioproducts can be identified by dual carbon isotope fingerprinting or 14 Based on C dating, petrochemical carbon can be distinguished from organic compounds. In addition, the specific source of biogenic carbon (e.g., glucose vs. glycerol) can be determined by dual carbon isotope fingerprinting by methods known in the art (see, e.g., U.S. Pat. No. 7,169,588, WO2016 / 011430A1, etc.).

[0176] The following examples are offered to illustrate, but not to limit, the invention. EXAMPLES

[0177] Example 1: The following example illustrates the production of 5-dodecene-1,3-diol by fermentation. 5-dodecene-1,3-diol was produced by fermentation using an E. coli strain engineered for the production of fatty alcohols and 1,3-diols. These 1,3-fatty diols were then purified from the resulting fermentation broth.

[0178] For example, as disclosed in WO2016 / 011430A1, the stNH1282 strain, a derivative of the MG1655 strain modified to attenuate the activity of FadE (involved in fatty acid degradation) and engineered to overexpress thioesterases specific for C12 chain length (such as FatB1), EntD (phosphopantethienyltransferase), CarB (carboxylate reductase), and AlrA-ADP1 (alcohol dehydrogenase), was inoculated from 1 mL of glycerol freezer stock into LB medium (100 mL) containing spectinomycin (115 mg / L) and shaken at 32° C. for 6-8 hours until the culture reached an OD of 3-6.

[0179] Two seed bioreactors were prepared with 4 liters of seed medium 2 (Table I) and then inoculated with this LB culture at 1.0% (vol / vol) and cultivated using the following bioreactor parameters: pH = 6.9 with NH4OH addition, airflow = 0.5 vol / vol / min, (dissolved oxygen) DO = 30% of saturation, and temperature = 33 °C. These bioreactors were run overnight (approximately 16 hours) until glucose was exhausted and the optical density (OD) of the culture was between 20 and 30 absorbance units (AU).

[0180] Table I: Bioreactor seed medium composition TIFF0007601403000038.tif92128

[0181] A 700 liter (L) production bioreactor was first batched with 250 L of production medium (Table II) and inoculated with 7.5 L of seed bioreactor (3% by volume (v / v)). This bioreactor was cultivated using the following bioreactor parameters: pH = 6.9 with NH4OH addition, airflow = 0.5 volume / volume / min (v / v / m), DO = 15% of saturation (at 1 bar back pressure), and temperature = 33 °C.

[0182] When the OD was above 10 AU, the culture was induced by bringing the medium to 0.5 mM IPTG. After exhaustion of the initial glucose in the tank, the bioreactor was fed with glucose solution (62% by weight (w / w)) at a rate of 10 g / L (initial volume) / h for 1 h. Upon glucose exhaustion caused by the rise of dissolved oxygen in the tank, glucose feeding was started for the next 1 h. This was continued throughout the fermentation. Any foaming was controlled by the automated addition of a Xiameter 1410® (Dow Corning). The run was terminated after 72 h and the culture broth was harvested by centrifugation.

[0183] Table II: Production bioreactor medium composition TIFF0007601403000039.tif104128

[0184] Table III: Trace Vitamin Solutions TIFF0007601403000040.tif53128

[0185] Table IV: Trace Vitamin Solutions TIFF0007601403000041.tif53128

[0186] Example 2: The following example illustrates the purification of 1,3 diols from centrifuged fermentation broth.

[0187] The product of the fermentation described in Example 1 was a mixture of three phases: an organic phase rich in fatty alcohols and 1,3 diols, an aqueous phase containing the spent fermentation medium, and a solid phase containing the E. coli biomass. Upon centrifugation, the light organic phase was collected. This material was then deacidified using alkaline refining and water drying. The resulting deacidified oil was then fractionated by distillation, and the fractions rich in 5-dodecane-1,3-diol and 5-dodecene-1,3 diol were collected and pooled. The composition of fatty alcohols and 1,3 diols in the enriched pool was determined by gas chromatography and mass spectrometry as described in Example 3 herein below and shown in Table V. The mass spectra of the major components 5-dodecane-1,3 diol and 5-dodecene-1,3 diol are shown in Figures 1 and 2, respectively.

[0188] Table V: Analysis of 1,3-diol-enriched distillation fractions TIFF0007601403000042.tif129164

[0189] Example 3: The following example illustrates one exemplary method for the analysis of fatty alcohols and 1,3 diols using gas chromatography (GC) and mass spectrometry (MS).

[0190] A sample, such as the 1,3-diol-enriched distillation fraction described in Example 2, is reacted with a 1:1 mixture of (N,O-bis(trimethylsilyl)trifluoroacetamide (BSTFA)+(1% trimethylchlorosilane (TMCS)):toluene to form silyl ethers of the alcohols. Hydrocarbons, methyl esters, and aldehydes are not derivatized. The sample is then analyzed by fast temperature gradient method using a small bore column. The GC program calculates the weight percentage of the sample components by comparing the response factor of the sample to that of the standard using tridecanoic acid methyl ester as an internal standard. The results of the oil analysis of the 1,3-diol-enriched fraction of Example 2 are shown in Table V. The identity of each compound eluting from the GC is identified by its retention time and mass spectrum. Graphical schemes showing the mass spectrum of 1,3-diols and the possible silyl ether derivatized ions are shown in Figures 1 and 2.

[0191] Example 4: The following example illustrates the synthesis of (S)-2-[2-(benzyloxy)ethyl]oxirane, useful for the preparation of 5-dodecene-1,3-diol via the chemical synthesis route disclosed herein below in Example 5. This is a prophetic example. TIFF0007601403000043.tif15128

[0192] (S)-2-[2-(benzyloxy)ethyl]oxirane is prepared via literature procedures (see, for example, Cink, RD; Forsyth, CJJ Org. Chem 1995, 60, 8122). Briefly, to a magnetically stirred solution of S-4-(benzyloxy)butane-1,2-diol (207 mg, 1.05 mmol, CAS number 69985-32-6) in THF (10.6 mL) at 0° C., NaH (63 mg, 2.6 mmol) is added. The resulting mixture is warmed to room temperature and stirred for 1 h, then cooled to 0° C. before adding N-tosylimidazole (237 mg, 1.07 mmol) in three equal portions over 20 min. The mixture is warmed to room temperature and stirred for 45 min, then recooled to 0° C. A saturated aqueous solution of NH4Cl and Et2O (65 mL) is then added, the separated organic phase is washed with H2O (25 mL) and brine (25 mL), and the combined organic layers are extracted with Et2O (2 x 25 mL). The combined organic layers are then dried (Na2SO4), filtered, and concentrated by rotary evaporation. Chromatography of the residue (silica gel; hexane / EtOAc 5:1) is expected to give the desired product.

[0193] Example 5: The following example illustrates a method for producing (R,Z)-dodec-5-ene-1,3-diol (i.e., (R,Z)-5-dodecene-1,3-diol) using a synthetic chemistry approach. This is a prophetic example. TIFF0007601403000044.tif57165

[0194] A solution of oct-1-yne (1.8 equiv.) in THF (0.5 M) at −78 °C is added to 2.5 M n-BuLi (1.81 equiv.) in hexanes, and the resulting solution is stirred for 15 min before being added to a solution of (S)-2-[2-(benzyloxy)ethyl]oxirane (1 equiv.) in THF (0.2 M) at −78 °C. BF3·OEt2 (2 equiv.) is then added dropwise, and the resulting mixture is stirred at −78 °C until completion. If necessary, the temperature is slowly raised to −30 °C. A saturated aqueous solution of NH4Cl and Et2O is then added, the separated organic phase is washed successively with H2O and then with brine, and the combined aqueous layer is extracted with Et2O. The combined organic layers are then dried (Na2SO4), filtered, and concentrated by rotary evaporation. The residue is then chromatographed (silica gel; hexane / EtOAc) to give (R)-1-(benzyloxy)dodec-5-yn-3-ol.

[0195] (R)-1-(benzyloxy)dodec-5-yn-3-ol is then taken up in hexane to produce a 0.2M mixture in a flask to which is added freshly prepared Lindlar's catalyst (see Lindlar, H.; Dubuis, R. Org. Synth. 1966, 46, 89, using freshly precipitated calcium carbonate) at 20% by weight relative to (R)-1-(benzyloxy)dodec-5-yn-3-ol or 5% palladium on calcium carbonate (10% by weight relative to (R)-1-(benzyloxy)dodec-5-yn-3-ol) and freshly distilled quinolone (10% by weight relative to (R)-1-(benzyloxy)dodec-5-yn-3-ol). The flask is flushed with hydrogen three times, after which the mixture is stirred under H2 and monitored by H2 consumption and / or by gas chromatography. Upon indication of reaction completion (e.g., when 1 equivalent of H2 has been consumed), the suspension is filtered through a pad of silica gel and the pad is then washed with diethyl ether. The organic phase is concentrated and purified by chromatography (silica gel; hexanes / EtOAc) to give (R,Z)-1-(benzyloxy)dodec-5-en-3-ol. [Note: it is well known that the benzyl protecting group usually survives Lindlar hydrogenation. However, to the extent that these conditions result in some degree of debenzylation to give (R,Z)-dodec-5-en-1,3-diol, this can be obtained via a chromatographic step].

[0196] (R,Z)-1-(benzyloxy)dodec-5-en-3-ol is taken up in CHCl to produce a 2M solution and stirred at 0° C. To this solution, 1.3 equivalents of neat trimethylsilyl iodide are added via a dry syringe. The reaction progress is monitored by TLC. Upon completion, the reaction is quenched by slow addition of 4 equivalents of MeOH. A saturated aqueous solution of NHCl and EtO is then added, the separated organic phase is washed with H0 and brine, and the combined aqueous layer is extracted with EtO (2×25 mL). The combined organic layers are then dried (NaSO), filtered, and concentrated by rotary evaporation. The residue is then chromatographed (silica gel; hexanes / EtOAc) to give (R,Z)-dodec-5-en-1,3-diol.

[0197] Alternatively, the secondary alcohol of (R,Z)-1-(benzyloxy)dodec-5-en-3-ol may be protected by a tert-butyldimethylsilyl ("TBS") group prior to cleavage of the benzyl group, and the TBS group may then be removed after debenzylation. Thus, to a 0.1 M solution of (R,Z)-1-(benzyloxy)dodec-5-en-3-ol (1 eq.) in CHCl, 2,6-lutidine (1.3 eq.) is added, the mixture is then cooled to 0° C., and tert-butyldimethylsilyl triflate (1.1 eq.) is then added dropwise with stirring. The solution is stirred at 0° C. until completion, as monitored by thin layer chromatography. A saturated aqueous solution of NHCl and EtO is then added, the separated organic phase is washed successively with HO and then with brine, and the combined aqueous layers are extracted with EtO. The combined organic layers are then dried (Na2SO4), filtered, and concentrated by rotary evaporation. The residue is then chromatographed (silica gel; hexanes / EtOAc) to give (R,Z)-(1-(benzyloxy)dodec-5-en-3-yl)oxy)(tert-butyl)dimethylsilane.

[0198] (R,Z)-((1-(benzyloxy)dodec-5-en-3-yl)oxy)(tert-butyl)dimethylsilane is taken up in CHCl to produce a 2.0 M solution and stirred at 0° C. To this solution is added 1.3 equivalents of neat trimethylsilyl iodide via a dry syringe. The reaction progress is monitored by thin layer chromatography. Upon completion, the reaction is quenched by slow addition of 4 equivalents of MeOH. A saturated aqueous solution of NHCl and EtO is then added, the separated organic phase is washed with H0 and brine, and the combined aqueous layers are extracted with EtO (2×25 mL). The combined organic layers are then dried (NaSO), filtered, and concentrated by rotary evaporation. The residue is then chromatographed (silica gel; hexanes / EtOAc) to give (R,Z)-3-(tert-butyldimethylsilyl)oxy)dodec-5-en-1-ol.

[0199] To a stirred 2.0 M solution of (R,Z)-3-((tert-butyldimethylsilyl)oxy)dodec-5-en-1-ol in THF at 0° C., tetrabutylammonium fluoride (1.0 M in THF, 1.2 eq.) is added, and the mixture is then allowed to warm slowly to room temperature, allowing the reaction to proceed to completion (monitored by thin layer chromatography). Upon completion, saturated aqueous solutions of NH4Cl and Et2O are added, the separated organic phase is washed successively with H2O and then with brine, and the combined aqueous layers are extracted with Et2O. The combined organic layers are then dried (Na2SO4), filtered, and concentrated by rotary evaporation. The residue is then chromatographed (silica gel; hexanes / EtOAc) to give (R,Z)-dodec-5-ene-1,3-diol.

[0200] Example 6: The following example illustrates an exemplary chemical synthesis method for the preparation of (R,Z)-tetradec-7-ene-1,3-diol. This is a prophetic example. TIFF0007601403000045.tif58164

[0201] 1-Bromo-3-yne may be obtained from commercial sources or via reaction of oct-1-yne with oxirane in the presence of BF3·OEt2 (see Example 5 above) to produce dec-3-yn-1-ol, followed by conversion of dec-3-yn-1-ol to 1-bromo-3-yne (e.g., via reaction with thionyl bromide as described in U.S. Pat. No. 9,353,090). Specifically, to a stirred solution of dec-3-yn-1-ol (1 eq.) in CHCl2 (0.2 M) at 0° C., freshly distilled dimethylformamide (0.5 eq.) is added, followed by thionyl bromide (1.3 eq.). The mixture is allowed to continue stirring and slowly warm to 20° C. Upon reaction completion (as indicated, for example, by thin layer chromatography and / or gas chromatography), a saturated aqueous solution of Et2O and then NH4Cl is added, the separated organic phase is washed with H2O and brine, and the combined aqueous layers are extracted with Et2O. The combined organic layers are then dried (Na2SO4), filtered, and concentrated by rotary evaporation. The residue is then chromatographed (silica gel; petroleum ether / Et2O) to give 1-bromodeca-3-yne.

[0202] To a 0.1 M solution of (S)-2-[2-(benzyloxy)ethyl]oxirane at room temperature in THF, CuCN (0.5 eq.) is added, and the mixture is stirred for 5 minutes and then cooled to -40°C. To this stirred solution, 3.3 eq. of freshly prepared dec-3-yn-1-yl magnesium bromide (1.0 M, prepared from 1-bromodec-3-yne) in THF at -20°C is added. The resulting mixture is maintained at -40°C for approximately 1 hour and then warmed to 0°C over 75 minutes. Upon completion of the reaction, a saturated aqueous solution of NH4Cl and Et2O is then added, the separated organic phase is washed with H2O and brine, and the combined aqueous layer is extracted with Et2O. The combined organic layers are then dried (Na2SO4), filtered, and concentrated by rotary evaporation. The residue is then chromatographed (silica gel; hexanes / EtOAc) to give (R)-1-(benzyloxy)tetradec-7-yn-3-ol.

[0203] (R)-1-(benzyloxy)tetradec-7-yn-3-ol is then taken up in hexane to produce a 0.2M mixture in a flask to which is added freshly prepared Lindlar's catalyst (see Lindlar, H.; Dubuis, R. Org. Synth. 1966, 46, 89, using freshly precipitated calcium carbonate) at 20% by weight relative to (R)-1-(benzyloxy)tetradec-7-yn-3-ol or 5% palladium on calcium carbonate (10% by weight relative to (R)-1-(benzyloxy)tetradec-7-yn-3-ol) and freshly distilled quinolone (10% by weight relative to (R)-1-(benzyloxy)tetradec-7-yn-3-ol). [Note: solubility may be enhanced by utilizing EtOAc as a co-solvent or in place of hexane]. The flask is flushed with hydrogen three times, after which the mixture is stirred under H2 and monitored by H2 consumption and / or by gas chromatography. Upon indication of reaction completion (e.g., when 1 equivalent of H2 has been consumed), the suspension is filtered through a pad of silica gel, and the pad is then washed with diethyl ether. The organic phase is concentrated and purified by chromatography (silica gel; hexanes / EtOAc), which is expected to give (R,Z)-1-(benzyloxy)tetradec-7-en-3-ol. [Note: it is well known that benzyl protecting groups usually survive Lindlar hydrogenation. However, to the extent that these conditions result in some degree of debenzylation leading to (R,Z)-tetradec-7-en-1,3-diol, which can be isolated via a chromatographic step].

[0204] (R,Z)-1-(benzyloxy)tetradec-7-en-3-ol is taken up in CHCl to produce a 2M solution, which is stirred at 0°C. To this solution, 1.5 equivalents of EtN are added via dry syringe, followed by 2.3 equivalents of neat trimethylsilyl iodide. The reaction progress is monitored by TLC. Upon completion, the reaction is quenched by slow addition of 4 equivalents of MeOH. An aqueous solution of NaHSO (1.0 M) is then added, and the mixture is stirred for 15 minutes, followed by addition of EtO. The separated organic phase is then washed with H0 and brine, and the combined aqueous layers are extracted with EtO (2 x 25 mL). The combined organic layers are then dried (NaSO), filtered, and concentrated by rotary evaporation. The residue is then chromatographed (silica gel; hexanes / EtOAc) to give (R,Z)-tetradec-7-en-1,3-diol.

[0205] Alternatively, the secondary alcohol of (R,Z)-1-(benzyloxy)tetradec-7-en-3-ol may be TBS protected, followed by deprotection of the benzyl group and finally TBS deprotection via the alternative procedure disclosed in Example 5 herein above.

[0206] Example 7: The following example illustrates an exemplary chemical synthesis method for the preparation of (R,Z)-tridec-6-ene-1,3-diol. This is a prophetic example. TIFF0007601403000046.tif68160

[0207] The 1-bromonona-2-yne (CAS number 5921-74-4) used in the production of nona-2-yn-1-yl magnesium bromide can be obtained from commercial sources.

[0208] To a 0.1 M solution of (S)-2-[2-(benzyloxy)ethyl]oxirane at room temperature in THF, CuCN (0.5 eq.) is added, and the mixture is stirred for 5 minutes and then cooled to -40°C. To this stirred solution, 3.3 eq. of freshly prepared non-2-yn-1-yl magnesium bromide (1.0 M, prepared from 1-bromonona-2-yne) in THF at -20°C is added. The resulting mixture is maintained at -40°C for approximately 1 hour and then warmed to 0°C over 75 minutes. Upon completion of the reaction, a saturated aqueous solution of NH4Cl and Et2O is then added, the separated organic phase is washed with H2O and brine, and the combined aqueous layer is extracted with Et2O. The combined organic layers are then dried (Na2SO4), filtered, and concentrated by rotary evaporation. The residue is then chromatographed (silica gel; hexanes / EtOAc) to give (R)-1-(benzyloxy)tridec-6-yn-3-ol.

[0209] (R)-1-(benzyloxy)tridec-6-yn-3-ol is taken up in hexane to produce a 0.2M mixture in a flask to which is added freshly prepared Lindlar catalyst (see Lindlar, H.; Dubuis, R. Org. Synth. 1966, 46, 89, using freshly precipitated calcium carbonate) at 20% by weight relative to (R)-1-(benzyloxy)tridec-6-yn-3-ol or 5% palladium on calcium carbonate (10% by weight relative to (R)-1-(benzyloxy)tridec-6-yn-3-ol) and freshly distilled quinolone (10% by weight relative to (R)-1-(benzyloxy)tridec-6-yn-3-ol). [Note: solubility may be enhanced by utilizing EtOAc as a co-solvent or in place of hexane]. The flask is flushed with hydrogen three times, after which the mixture is stirred under H2 and monitored by H2 consumption and / or by gas chromatography. Upon indication of reaction completion (e.g., when 1 equivalent of H2 has been consumed), the suspension is filtered through a pad of silica gel and the pad is then washed with diethyl ether. The organic phase is concentrated and purified by chromatography (silica gel; hexanes / EtOAc) to give (R,Z)-1-(benzyloxy)tridec-6-en-3-ol. [Note: it is well known that the benzyl protecting group usually survives Lindlar hydrogenation. However, to the extent that these conditions result in some degree of debenzylation to give (R,Z)-tridec-6-en-1,3-diol, which can be isolated via a chromatographic step].

[0210] (R,Z)-1-(benzyloxy)tridec-6-en-3-ol is taken up in CHCl to produce a 2M solution, which is stirred at 0° C. To this solution, 1.5 equivalents of EtN are added via dry syringe, followed by 2.3 equivalents of neat trimethylsilyl iodide. The reaction progress is monitored by TLC. Upon completion, the reaction is quenched by slow addition of 4 equivalents of MeOH. An aqueous solution of NaHSO (1.0 M) is then added, and the mixture is stirred for 15 minutes, followed by addition of EtO. The separated organic phase is then washed with H0 and brine, and the combined aqueous layers are extracted with EtO (2×25 mL). The combined organic layers are then dried (NaSO), filtered, and concentrated by rotary evaporation. The residue is then chromatographed (silica gel; hexanes / EtOAc) to give (R,Z)-tridec-6-en-1,3-diol.

[0211] Alternatively, the secondary alcohol of (R,Z)-1-(benzyloxy)tridec-6-en-3-ol may be TBS protected, followed by deprotection of the benzyl group and finally TBS deprotection via the alternative procedure disclosed in Example 5 herein above.

[0212] Example 8: The following example illustrates the chemical synthesis of (R,E)-dodec-5-ene-1,3-diol. This is a prophetic example. TIFF0007601403000047.tif60144

[0213] To a 0.1 M solution of (S)-2-[2-(benzyloxy)ethyl]oxirane in THF at room temperature, CuCN (0.5 equiv.) is added, and the mixture is stirred for 5 minutes and then cooled to -40°C. To this stirred solution, 3.3 equiv. of a freshly prepared (E)-oct-1-en-1-yl magnesium bromide THF solution (1.0 M) at -20°C is added. The resulting mixture is maintained at -40°C for approximately 1 hour and then warmed to 0°C over 75 minutes. Upon completion of the reaction, a saturated aqueous solution of NH4Cl and Et2O is then added, the separated organic phase is washed with H2O and brine, and the combined aqueous layer is extracted with Et2O. The combined organic layers are then dried (Na2SO4), filtered, and concentrated by rotary evaporation. The residue is then chromatographed (silica gel; hexanes / EtOAc) to give (R,E)-1-(benzyloxy)dodec-5-en-3-ol.

[0214] (R,E)-1-(benzyloxy)dodec-5-en-3-ol is taken up in CHCl to produce a 2M solution and stirred at 0° C. To this solution, 1.5 equivalents of EtN are added via dry syringe, followed by 2.3 equivalents of neat trimethylsilyl iodide. The reaction progress is monitored by TLC. Upon completion, the reaction is quenched by slow addition of 4 equivalents of MeOH. An aqueous solution of NaHSO (1.0 M) is then added and the mixture is stirred for 15 minutes, followed by addition of EtO. The separated organic phase is then washed with H0 and brine, and the combined aqueous layers are extracted with EtO (2×25 mL). The combined organic layers are then dried (NaSO), filtered, and concentrated by rotary evaporation. The residue is then chromatographed (silica gel; hexane / EtOAc) to give (R,E)-dodec-5-en-1,3-diol.

[0215] Alternatively, the secondary alcohol of (R,E)-1-(benzyloxy)dodec-5-en-3-ol may be TBS protected, followed by deprotection of the benzyl group and finally TBS deprotection via the alternative procedure described in Example 2.

[0216] Example 9. The following example illustrates the chemical synthesis of (R)-dodecane-1,3-diol. This is a prophetic example.

[0217] A flask is charged with (R)-1-(benzyloxy)dodec-5-yn-3-ol (1 eq), Pd / C (0.01 eq based on Pd content), and degassed EtOAc to form a 0.1 M solution based on (R)-1-(benzyloxy)dodec-5-yn-3-ol. The flask is flushed with hydrogen three times, after which the mixture is stirred under H2 and monitored by gas chromatography and / or NMR. Upon completion, the suspension is filtered through a pad of silica gel, which is then washed with diethyl ether. The organic phase is concentrated and purified by chromatography (silica gel; hexanes / EtOAc), which is expected to give (R)-dodecane-1,3-diol.

[0218] Example 10. Exemplary two-step synthesis of glycosylated (R,Z)-dodec-5-ene-1,3-diol. The following example illustrates one exemplary two-step synthesis of glycosylated (R,Z)-dodec-5-ene-1,3-diol. This is a prophetic example.

[0219] The synthesis of glycosylated (R,Z)-dodec-5-ene-1,3-diols is also accomplished via a two-step method similar to that described by El-Sukkary et al. “Synthesis and Characterization of some Alkyl Polyglycosides Surfactants” J. Surfact Deterg, 2008, 11, 129-137. Briefly, to prepare glycosylated (R,Z)-dodec-5-ene-1,3-diols using the process disclosed in the El-Sukkary reference, glucose is first reacted with butanol in the presence of p-toluenesulfonic acid to form butyl polyglucosides. The now more organophilic butyl polyglucosides are then reacted with long chain alcohols under elevated temperature and vacuum to remove butanol and thus form the desired alkyl polyglucosides. The reported yields of this procedure were 35-45%. In this adaptation, a sugar (e.g., glucose) is reacted with butanol in the presence of p-toluenesulfonic acid to form a butyl sugar, which is then reacted with (R,Z)-dodec-5-ene-1,3-diol to obtain glycosolated (R,Z)-dodec-5-ene-1,3-diol. When glucose is used as the sugar, mono- and / or bis-glucosylated (R,Z)-dodec-5-ene-1,3-diol can be achieved based on the selection of conditions. 1 The product yield is determined and the product is characterized by 1 H-NMR, FTIR, and LC-MS.

[0220] Example 11: The following example illustrates an exemplary one-step process for the preparation of (R,Z)-dodec-5-ene-1,3-diol. This is a prophetic example.

[0221] As exemplified in the El-Sukkary reference, the direct synthesis of glycosylated long chain aliphatic hydrocarbonols is limited due to the immiscibility of the starting reagents (e.g., water-soluble glucose and organic-soluble 1-octanol). This solubility problem can be alleviated by enhancing the solubility of compounds of formulas IA, IB, II, III, and / or IV, thus enabling a one-step synthesis of compounds of formulas V and / or VI.

[0222] In one exemplary embodiment, comparison of the water solubility of compounds is accomplished by comparing their respective log P values. One of ordinary skill in the art will appreciate that log P is the logarithm of the ratio of the concentration of a compound between n-octanol and water, as exemplified by Equation 1. logP=log([compound] n-オクタノール / ([compound] 水 ) Equation 1

[0223] Thus, compounds with higher log P are more lipophilic and less water soluble, and compounds with lower log P values ​​are less lipophilic and more water soluble. As exemplified by the calculated log P values ​​for (R,Z)-dodec-5-en-1,3-diol, (Z)-dodec-5-en-1-ol, and dodecan-1-ol, (R,Z)-dodec-5-en-1,3-diol is more than an order of magnitude more water soluble than (Z)-dodec-5-en-1-ol and nearly two orders of magnitude more water soluble than dodecan-1-ol. TIFF0007601403000048.tif89128

[0224] The synthesis of glycosylated compounds of formula V and / or VI via one-step reaction is carried out on a 25 g initial glucose scale (total reaction volume of about 240 mL). The molar ratio of aliphatic diol (e.g., one or more compounds of formula IA, IB, II, and / or III) to glucose is set at 6:1, and the reaction temperature is set at about 120° C. Varying the reaction conditions to generate different glycosylated products of formula V and / or VI includes varying the catalyst concentration of p-toluenesulfonic acid and reaction time. 1 The product yield is determined and the product is characterized by 1 H-NMR, FTIR, and LC-MS.

[0225] As would be apparent to one skilled in the art, various modifications and variations of the above aspects and embodiments can be made without departing from the spirit and scope of the present disclosure, and therefore such modifications and variations are within the scope of the present disclosure.

Claims

1. Formula V: (In the formula, R １ and R ２ is independently selected from H, a monosaccharide, a disaccharide, a trisaccharide, and a polysaccharide; each of the monosaccharides, disaccharides, trisaccharides, or polysaccharides is linked at an anomeric carbon; n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14; the double bond is in the (Z) or (E) configuration; A chiral center is present at C-3; The chiral center at C-3 has an R or S configuration.

2. 2. The compound of claim 1, wherein n is 1, 3 or 5.

3. n is 3; R １ and R ２ is H; The compound is a 14 carbon unbranched unsaturated fatty diol having a single Δ7 double bond, a hydroxy group at carbon number 1 (C-1) and a hydroxy group at carbon number 3 (C-3); The following formula III:

2. The compound of claim 1 having the structure shown in

4. A compound represented by the following formula VIII: A compound having a structure represented by the following formula:

5. R １ , or R ２ , or R １ and R ２ are both monosaccharides, disaccharides, trisaccharides, or polysaccharides; 3. The compound of claim 1 or 2, wherein the monosaccharide, disaccharide, trisaccharide, or polysaccharide comprises a pentose or hexose sugar, or a mixture thereof.

6. 6. The compound of any one of claims 1 to 5, wherein the double bond is in the (Z) configuration and the chiral center at C-3 is in the R configuration.

7. 7. The compound of any of claims 1, 2, 5, and 6, wherein the monosaccharide, disaccharide, trisaccharide, or polysaccharide is linked at the anomeric carbon via an α-glycosidic or β-glycosidic bond.

8. R １ and R ２ are different monosaccharides, or R １ and R ２ is the same monosaccharide, A compound according to any one of claims 1, 2, 5 to 7.

9. The compound according to any one of claims 1, 2, 5 to 8, wherein the monosaccharide, disaccharide, trisaccharide, or polysaccharide comprises allose, altrose, glucose, mannose, gulose, iodose, galactose, talose, furanose, pyranose, xylose, or a mixture of any two or more thereof.

10. R １ The compound according to any one of claims 1, 2, 5 to 9, wherein is glucose.

11. R １ The compound according to any one of claims 1, 2, and 5 to 9, wherein is maltose or isomaltose.

12. A composition for use as a personal care product, pharmaceutical, food, or agricultural formulation comprising a compound according to any one of claims 1, 2, 5 to 11.

13. The following formula VI: (In the formula, m is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, or 18; R ３ and R ４ are each independently selected from H, a monosaccharide, a disaccharide, a trisaccharide, and a polysaccharide, each of the monosaccharides, disaccharides, trisaccharides, or polysaccharides being linked at an anomeric carbon; R ３ and R ４ one or both of are not H; A chiral center is present at C-3; The chiral center at C-3 has an R or S configuration.

14. A method for preparing a polyol comprising the steps of: (a) converting a double bond of the compound of claim 1 into an epoxide, R １ and R ２ is H and the compound of claim 1 is an unsaturated 1,3-fatty diol; and (b) opening the epoxy ring with water or hydrogen; wherein the step of opening the epoxy rings with water produces a polyol which is a tetraol, and the step of opening the epoxy rings with hydrogen produces a polyol which is a triol; Here, prior to step (a), the hydroxy group of the 1,3-aliphatic diol may be protected with an acetyl group.

15. n is 1 and the epoxide has formula VII: The method of claim 14 , having a structure represented by:

16. The polyol is represented by the following formula IX: The method according to claim 14 or 15, wherein the tetraol has a structure represented by:

17. Selective ring opening of the epoxy ring with hydrogen gives the compound of formula X: The method of claim 14 or 15, wherein the triol produced is dodecane-1,3,6-triol having a structure represented by:

18. A method for preparing a branched polyol comprising the steps of: (a) converting a double bond of the compound of claim 1 into an epoxide, R １ and R ２ is H and the compound of claim 1 is an unsaturated 1,3-fatty diol; and (b) opening the epoxy ring using a hydroxy group of a 1,3-aliphatic diol or polyol; Here, prior to step (a), the hydroxy group of the 1,3-aliphatic diol may be protected with an acetyl group.

19. n is 1, and the epoxide in step (a) is represented by the following formula VII: having a structure represented by The 1,3-fatty diol in step (b) is 5-dodecene-1,3-diol; The branched polyol has the following formula XI:

20. The method of claim 18 having a structure represented by:

20. n is 1, and the epoxide in step (a) is represented by the following formula VII: having a structure represented by The polyol in step (b) is dodecene-1,3,6-triol; The branched polyol has the following formula XII:

20. The method of claim 18 having a structure represented by:

21. The following steps: (c) reacting the polyol or branched polyol with a catalyst which converts the polyol or branched polyol into its carbonate derivative; The method of any one of claims 14 to 20, further comprising:

22. The branched polyol of claim 1, wherein the branched polyol has the following formula XII: having a structure represented by The carbonate derivative of the branched polyol has the following formula XIII:

22. The method of claim 21 , having a structure represented by:

23. reacting the polyol or branched polyol with an isocyanate to prepare a polyurethane; or reacting the polyol or branched polyol with a carbamate to prepare a non-isocyanate polyurethane (NIPU); The method of any one of claims 14 to 22, further comprising:

24. A method for preparing a polyurethane or non-isocyanate polyurethane (NIPU) comprising the steps of: reacting the compound of claim 1 with an isocyanate to prepare a polyurethane; or reacting the compound of claim 1 with a carbamate to prepare NIPU; Here, R １ and R ２ is H; and the compound of claim 1 is an unsaturated 1,3-fatty diol.

25. 23. The method of claim 21 or 22, further comprising reacting the carbonate with a primary amine to prepare a non-isocyanate polyurethane (NIPU).

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