Multifunctional Fatty Acid Derivatives And Biosynthesis Thereof
The disclosure relates to the field of specialty chemicals and methods for their synthesis. In embodiments, the disclosure provides novel multifunctional fatty acid derivative molecules such as e.g., fatty triols, fatty tetrols, dihydroxy fatty acids, etc. The disclosure further provides derivatives of the disclosed multifunctional molecules which are useful e.g., in the production of personal care products, surfactants, detergents, polymers, paints, coatings, and as emulsifiers, emollients, and thickeners in cosmetics and foods, as industrial solvents and plasticizers, etc. The disclosure further provides biochemical pathways, recombinant microorganisms and methods for the biological production of various multifunctional fatty acid derivatives.
1 . A multifunctional molecule having a chemical formula according to
wherein
R1=-OH; —O 2 H; —CH 2 ; —CH 2 OH; —CHO; —CH 2 NH 2 ; —CO 2 H; —CO 2 CH 3 ; —CO 2 C 2 H 5 ; —CO 2 C 3 H 7 ; —CO 2 C 2 H 3
R2=-H; —OH; —NH 2
R3=-H; —OH;
R4=-H; —OH;
R5=-CH 3 ; —CH 2 ; —CH 2 OH; —CHO; —CH 2 NH 2 ; —CO 2 H; —CO 2 CH 3 ; —CO 2 C 2 H 5 ; —CO 2 C 3 H 7 ; —CO 2 C 2 H 3 ;
wherein
m=1-10 and n=0-9 and
wherein
if R1 is OH or —O 2 H then 3<m+n≤10; and if R1 is other than OH or —O 2 H, then 2<m+n≤10, and
wherein
the multifunctional molecule has at least three functional groups comprising a heteroatom, and
wherein
at least two of R2, R3 and R4 are OH; or when R5 is other than CH 3 , CH 2 or C 3 H 7 CH 2 H 5 then at least one of R2, R3 and R4 are OH; and
wherein
when R5=CH 3 and R4=H then n=0 or 1; and
wherein
when R2=H, then R4 does not=H; and
wherein
when R3=H, then n+m=p and p=3-10; unless R1=-OH or —O 2 H in which case when R3=-H then n+m=p and p=4-10; and
wherein
when the multifunctional molecule comprises a double bond that is not terminal, the double bond is in a position corresponding to an omega7 (ω-7) position; and
wherein
the multifunctional molecule is not 1,3,12-dodecane triol; is not 3,11-dihydroxy-tetradecanoic acid; is not 3,11-dihydroxy-tetradecanoic acid methyl ester and is not naturally occurring.
2 . The multifunctional fatty acid derivative molecule of claim 1 , wherein the multifunctional fatty acid derivative molecule is a multifunctional alcohol.
3 . The multifunctional fatty acid derivative molecule of claim 2 ,
wherein
R1=CH 2 OH and R2=OH.
4 . The multifunctional fatty acid derivative molecule of claim 3 , wherein the multifunctional fatty acid derivative molecule is a member selected from the group consisting of 1,3,11-dodecane triol, 1,3,-10-dodecane triol, 1,3,9-dodecane triol, 1,3,12-dodecene triol, 1,3,11-dodecene triol, 1,3,10-dodecene triol, 1,3,9-dodecene triol, 1,3,11,12-dodecane tetrol, 1,3,10,12-dodecane tetrol, 1,3,9,12 dodecane tetrol, 1,3,7-decane triol, 1,3,8-decane triol, and 1,3,9-decane triol.
5 . The multifunctional fatty acid derivative molecule of claim 1 , wherein the multifunctional fatty acid derivative molecule is a multifunctional fatty acid ester.
6 . The multifunctional fatty acid derivative molecule of claim 5 , wherein
R1=CO 2 CH 3 and R2=OH.
7 . The multifunctional fatty acid derivative molecule of claim 6 , wherein the multifunctional fatty acid derivative molecule is a member selected from the group consisting of 3,12-dihydroxy dodecanoic acid methyl ester, 3,14-dihydroxy tetradecanoic acid methyl ester, 3,16-dihydroxy hexadecanoic acid methyl ester, 3,12-dihydroxy dodecenoic acid methyl ester, 3,14-dihydroxy tetradecenoic acid methyl ester, 3,16-dihydroxy hexadecenoic acid methyl ester, 3,11-dihydroxy dodecanoic acid methyl ester, 3,10-dihydroxy dodecanoic acid methyl ester, 3,9-dihydroxy dodecanoic acid methyl ester, 3,11-dihydroxy dodecenoic acid methyl ester, 3,10-dihydroxy dodecenoic acid methyl ester, 3,9-dihydroxy dodecenoic acid methyl ester, 3,13-dihydroxy tetradecanoic acid methyl ester, 3,12-dihydroxy tetradecanoic acid methyl ester, 3,13-dihydroxy tetradecenoic acid methyl ester, 3,12-dihydroxy tetradecenoic acid methyl ester, 3,11-dihydroxy tetradecenoic acid methyl ester, 3,15-dihydroxy hexadecanoic acid methyl ester, 3,14-dihydroxy hexadecanoic acid methyl ester, 3,13-dihydroxy hexadecanoic acid methyl ester, 3,15-dihydroxy hexadecenoic acid methyl ester, 3,14-dihydroxy hexadecenoic acid methyl ester, and 3,13-dihydroxy hexadecenoic acid methyl ester.
8 . The multifunctional fatty acid derivative molecule of claim 5 , wherein
R1=CO 2 CH 2 CH 3 and R2=OH.
9 . The multifunctional fatty acid derivative molecule of claim 8 , wherein the multifunctional fatty acid derivative molecule is a member selected from the group consisting of 3,12-dihydroxy dodecanoic acid ethyl ester, 3,14-dihydroxy tetradecanoic acid ethyl ester, 3,16-dihydroxy hexadecanoic acid ethyl ester, 3,12-dihydroxy dodecenoic acid ethyl ester, 3,14-dihydroxy tetradecenoic acid ethyl ester, 3,16-dihydroxy hexadecenoic acid ethyl ester, 3,11-dihydroxy dodecanoic acid ethyl ester, 3,10-dihydroxy dodecanoic acid ethyl ester, 3,9-dihydroxy dodecanoic acid ethyl ester, 3,11-dihydroxy dodecenoic acid ethyl ester, 3,10-dihydroxy dodecenoic acid ethyl ester, 3,9-dihydroxy dodecenoic acid ethyl ester, 3,13-dihydroxy tetradecanoic acid ethyl ester, 3,12-dihydroxy tetradecanoic acid ethyl ester, 3,11-dihydroxy tetradecanoic acid ethyl ester, 3,13-dihydroxy tetradecenoic acid ethyl ester, 3,12-dihydroxy tetradecenoic acid ethyl ester, 3,11-dihydroxy tetradecenoic acid ethyl ester, 3,15-dihydroxy hexadecanoic acid ethyl ester, 3,14-dihydroxy hexadecanoic acid ethyl ester, 3,13-dihydroxy hexadecanoic acid ethyl ester, 3,15-dihydroxy hexadecenoic acid ethyl ester, 3,14-dihydroxy hexadecenoic acid ethyl ester, and 3,13-dihydroxy hexadecenoic acid ethyl ester.
10 . The multifunctional fatty acid derivative molecule of claim 1 , wherein the multifunctional fatty acid derivative molecule is a multifunctional fatty acid.
11 . The multifunctional fatty acid derivative molecule of claim 10 , wherein n≠4.
12 . The multifunctional fatty acid derivative molecule of claim 11 , wherein
R2=H.
13 .- 36 . (canceled)
37 . A method for making a multifunctional fatty acid derivative molecule having a chemical formula according to:
wherein
R1=-OH; —O 2 H; —CH 2 ; —CH 2 OH; —CHO; —CH 2 NH 2 ; —CO 2 H; —CO 2 CH 3 ; —CO 2 C 2 H 5 ; —CO 2 C 3 H 7 ; —CO 2 C 2 H 3
R2=-H; —OH; —NH 2
R3=-H; —OH;
R4=-H; —OH;
R5=-CH 3 ; —CH 2 ; —CH 2 OH; —CHO; —CH 2 NH 2 ; —CO 2 H; —CO 2 CH 3 ; —CO 2 C 2 H 5 ; —CO 2 C 3 H 7 ; —CO 2 C 2 H 3 ;
wherein
m=1-10 and n=0-9 and
wherein
if R1 is OH or —O 2 H then 3<m+n≤10; and if R1 is other than OH or —O 2 H, then 2<m+n≤10, and
wherein
the multifunctional molecule has at least three functional groups comprising a heteroatom, and
wherein
at least two of R2, R3 and R4 are OH; or when R5 is other than CH 3 , CH 2 or C 3 H 7 CH 2 H 5 then at least one of R2, R3 and R4 are OH; and
wherein
when R5=CH 3 and R4=H then n=0 or 1; and
wherein
when R2=H, then R4 does not=H; and
wherein
when R3=H, then n+m=p and p=3-10; unless R1=-OH or —O 2 H in which case when R3=-H then n+m=p and p=4-10; and
wherein
when the multifunctional molecule comprises a double bond that is not terminal, the double bond is in a position corresponding to an omega7 (ω-7) position; and
wherein
the multifunctional molecule is not 1,3,12-dodecane triol; is not 3,11-dihydroxy-tetradecanoic acid; is not 3,11-dihydroxy-tetradecanoic acid methyl ester and is not naturally occurring,
the method comprising:
culturing
a recombinant microbe that comprises a heterologous enzyme pathway capable of producing a bifunctional fatty acid derivative molecule, and at least one heterologous hydroxylating enzyme,
in a culture medium comprising a simple carbon source,
wherein
the heterologous enzyme pathway capable of producing bifunctional fatty acid derivative molecule is selected from:
(i) a heterologous enzyme pathway capable of producing a 3-hydroxy fatty acid;
(ii) a heterologous enzyme pathway capable of producing a 3-hydroxy fatty ester;
(iii) a heterologous enzyme pathway capable of producing a 1,3-fatty diol
(iv) a heterologous enzyme pathway capable of producing a hydroxy fatty acid;
(v) a heterologous enzyme pathway capable of producing a hydroxy fatty ester; and
(vi) a heterologous enzyme pathway capable of producing a fatty diol; and
wherein
the at least one heterologous hydroxylating enzyme is selected from a heterologous hydroxylase enzyme and a heterologous hydratase enzyme or a combination thereof.
38 . The method of claim 37 ,
wherein
the heterologous hydroxylase enzyme is a member selected from the group consisting of omega-hydroxylases (o-hydroxylases), mid-chain hydroxylases, and subterminal hydroxylases.
39 . The method of claim 37 ,
wherein
the heterologous enzyme pathway capable of producing bifunctional fatty acid derivative molecule is the heterologous enzyme pathway capable of producing a 1,3-fatty diol.
40 . The method of claim 39 ,
wherein
the heterologous enzyme pathway capable of producing a 1,3-fatty diol comprises;
(i) a heterologous thioesterase and
(ii) a heterologous carboxylic acid reductase.
41 . The method of claim 40 ,
wherein
the heterologous enzyme pathway capable of producing a 1,3-fatty diol further comprises;
(iii) a heterologous alcohol dehydrogenase.
42 . The method of claim 41 ,
wherein
the heterologous enzyme pathway capable of producing a 1,3-fatty diol further comprises:
(i) a heterologous PhaG thioesterase from Pseudomonas putida,
(ii) a heterologous CarB carboxylic acid reductase from Mycobacterium smegmatis , and
(iii) a heterologous AlrA alcohol dehydrogenase from Acinetobacter baylyi.
43 . The method of claim 40 ,
wherein
the heterologous hydroxylase enzyme is a cyp102A subterminal-hydroxylase from Bacillus licheniformis , and
wherein
the method produces multifunctional molecules selected from the group consisting of 1,3,9 decanetriol, 1,3,8 decanetriol, 1,3,7 decanetriol, 1,3,11 dodecanetriol, 1,3,10 dodecanetriol, 1,3,9 dodecanetriol, (z5)1,3,11 dodecenetriol, (z5)1,3,10 dodecenetriol and (z5)1,3,9 dodecenetriol.
44 .- 60 . (canceled)