Microbial production of fatty diols
View Patent ↗The disclosure relates to fatty diols and recombinant microorganisms for producing them. More particularly, the disclosure relates to recombinant microorganisms engineered to produce fatty diols via fermentation. Further encompassed is a process that uses the microorganisms to produce fatty diols from a simple carbon source.
1. A recombinant Escherichia coli ( E. coli ) for producing a 1,3 fatty diol when grown in a fermentation broth with a simple carbon source, the E. coli engineered to express heterologous nucleic acid sequences encoding a thioesterase having the classification EC 3.1.2.2, EC 3.1.1.5, or EC 3.1.2.14; and a carboxylic acid reductase having the classification EC 1.2.99.6, wherein the thioesterase uses 3-hydroxy acyl-ACP as a substrate and is selected from the group consisting of TE_EEI82564, TE_CAD63310, and phaG and wherein the carboxylic acid reductase is selected from the group consisting of carB from Mycobacterium smegmatis , carA from Mycobacterium smegmatis , FadD9 from Mycobacterium smegmatis , car from Mycobacterium genavense , car from Nocardia lowensis , and car from Nocardia brasiliensis.
2. The recombinant E. coli of claim 1 , wherein said 1,3 fatty diol is produced in vivo.
3. The recombinant E. coli of claim 1 , wherein said 1,3 fatty diol is selected from the group consisting of a C 5 1,3 fatty diol, a C 6 1,3 fatty diol, a C 7 1,3 fatty diol, a C 8 1,3 fatty diol, a C 9 1,3 fatty diol, a C 10 1,3 fatty diol, a C 11 1,3 fatty diol, a C 12 1,3 fatty diol, a C 13 1,3 fatty diol, a C 14 1,3 fatty diol, a C 15 1,3 fatty diol, a C 16 1,3 fatty diol, a C 17 1,3 fatty diol, a C 18 1,3 fatty diol, and a C 19 1,3 fatty diol.
4. The recombinant E. coli of claim 1 , further expressing a nucleic acid sequence encoding an alcohol dehydrogenase having the classification EC 1.1.1.-.
5. The recombinant E. coli of claim 1 , wherein said simple carbon source is derived from a renewable feedstock.
6. The recombinant E. coli of claim 1 , wherein said carboxylic acid reductase is carB.
7. The recombinant E. coli of claim 4 , wherein said alcohol dehydrogenase is alrA.
8. A cell culture comprising the E. coli according to claim 1 .
9. The cell culture of claim 8 , wherein said cell culture produces 1,3 fatty diols.
10. The cell culture of claim 9 , wherein said 1,3 fatty diol is selected from the group consisting of a C 5 1,3 fatty diol, a C 6 1,3 fatty diol, a C 7 1,3 fatty diol, a C 8 1,3 fatty diol, a C 9 1,3 fatty diol, a C 10 1,3 fatty diol, a C 11 1,3 fatty diol, a C 12 1,3 fatty diol, a C 13 1,3 fatty diol, a C 14 1,3 fatty diol, a C 15 1,3 fatty diol, a C 16 1,3 fatty diol, a C 17 1,3 fatty diol, a C 18 1,3 fatty diol, and a C 19 1,3 fatty diol.
11. A method of producing a 1,3 fatty diol comprising culturing the E. coli of claim 1 .
12. A method of producing a 1,3 fatty diol comprising: (i) culturing a recombinant E. coli in a fermentation broth, wherein said E. coli expresses heterologous nucleic acid sequences encoding a thioesterase having the classification EC 3.1.2.2, EC 3.1.1.5, or EC 3.1.2.14; and a carboxylic acid reductase having the classification EC 1.2.99.6, and optionally an alcohol dehydrogenase having the classification EC 1.1.1.1 or 1.1.1.2; wherein the thioesterase uses 3-hydroxy acyl-ACP as a substrate and is selected from the group consisting of tesA (with leader sequence) from E. coli , tesA (without leader sequence) from E. coli , tes from Photbacterium profundum , tesB from E. coli , TE CAD63310 from Lactobacillus plantarum , TE EEI82564 from Anaerococcus tetraclius , fatB1 from Umbellularia californica , fatB2 from Cuphea hookeriana , fatB3 from Cuphea hookeriana , fatB from Cinnamomurn camphora , fatB from Arabidopsis thaliana , fatA1 from Helianthus animus, fatA from Brassica juncea , fatA from Cuphea hookeriana , fadM from Escherichia coli , yciA from Escherichia coli , ybgC from Escherichia coli , and phaG from Pseudomonas putida , and wherein the carboxylic acid reductase is selected from the group consisting of carB from Mycobacterium smegmatis , carA from Mycobacterium smegmatis , FadD9 from Mycobacterium smegmatis , car from Mycobacterium genavense , car from Nocardia lowensis , and car from Nocardia brasiliensis ; and (ii) isolating a 1,3 fatty diol from said fermentation broth, wherein said fermentation broth comprises a simple carbon source.
13. The method of claim 12 , wherein said 1,3 fatty diol is selected from the group consisting of a C 5 1,3 fatty diol, a C 6 1,3 fatty diol, a C 7 1,3 fatty diol, a C 8 1,3 fatty diol, a C 9 1,3 fatty diol, a C 10 1,3 fatty diol, a C 11 1,3 fatty diol, a C 12 1,3 fatty diol, a C 13 1,3 fatty diol, a C 14 1,3 fatty diol, a C 15 1,3 fatty diol, a C 16 1,3 fatty diol, a C 17 1,3 fatty diol, a C 18 1,3 fatty diol, and a C 19 1,3 fatty diol.