Microbial Production Of Fatty Diols

Abstract

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 .- 27 . (canceled) 
     
     
         28 . A recombinant microorganism for producing a 1,3 fatty diol when grown in a fermentation broth with a simple carbon source derived from a renewable feedstock, wherein the microorganism is engineered to express a nucleic acid sequence encoding:
 (a) a polypeptide comprising acyl-CoA reductase (EC 1.2.1.50) activity;   (b) a polypeptide comprising acyl-CoA synthetase (EC 6.2.1.3) activity; and   (c) a polypeptide comprising thioesterase (EC 3.1.2.-, or EC 3.1.1.5, or EC 3.1.2.14) activity.   
     
     
         29 . The recombinant microorganism of  claim 28 , wherein the 1,3 fatty diol is produced in vivo. 
     
     
         30 . The recombinant microorganism of  claim 28 , wherein the 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. 
     
     
         31 . The recombinant microorganism of  claim 28 , wherein the 1,3 fatty diol is a C 12  1,3 fatty diol. 
     
     
         32 . The recombinant microorganism of  claim 28 , further expressing a nucleic acid sequence encoding a polypeptide comprising alcohol dehydrogenase (EC 1.1.1.-) activity. 
     
     
         33 . The recombinant microorganism of  claim 28 , wherein the polypeptide comprising acyl-CoA reductase activity is encoded by acr1. 
     
     
         34 . The recombinant microorganism of  claim 28 , wherein the polypeptide comprising acyl-CoA synthetase activity is FadD. 
     
     
         35 . The recombinant microorganism of  claim 28 , wherein the polypeptide comprising thioesterase activity is selected from the group consisting of FatA, FatA1, FatB, FatB1, FatB2, FatB3, TesA with the leader sequence, TesA without the leader sequence, TesB, FadM, YciA, YbgC, TE_EEI82564, TE_CAD63310, and PhaG. 
     
     
         36 . The recombinant microorganism of  claim 32 , wherein the polypeptide comprising alcohol dehydrogenase activity is encoded by alrA, yqhD, yjgB, yahK, adhP, ydjL, yhdH, yafB (dkgB), yqhE (dkgA), ybbO, or gldA. 
     
     
         37 . The recombinant microorganism of  claim 28 , wherein the 1,3 fatty diol is unsaturated. 
     
     
         38 . The recombinant microorganism of  claim 28 , wherein the microorganism is a bacterium, cyanobacterium, yeast, fungus, or algae. 
     
     
         39 . A cell culture, comprising the recombinant microorganism of  claim 28 . 
     
     
         40 . The cell culture of  claim 39 , wherein the cell culture produces 1,3 fatty diols. 
     
     
         41 . The cell culture of  claim 40 , wherein the 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. 
     
     
         42 . The cell culture of  claim 40 , wherein the 1,3 fatty diol is unsaturated. 
     
     
         43 . A method of producing a 1,3 fatty diol, comprising:
 (a) providing a recombinant microorganism in a fermentation broth, wherein the recombinant microorganism expresses a nucleic acid sequence encoding a polypeptide comprising acyl-CoA reductase (EC 1.2.1.50) activity, a polypeptide comprising acyl-CoA synthetase (EC 6.2.1.3) activity, and a polypeptide comprising thioesterase (EC 3.1.2.-, or EC 3.1.1.5, or EC 3.1.2.14) activity; and   (b) isolating the 1,3 fatty diol from the fermentation broth,   wherein the fermentation broth comprises a simple carbon source derived from a renewable feedstock.   
     
     
         44 . The method of  claim 43 , wherein the recombinant microorganism further expresses a nucleic acid sequence encoding a polypeptide comprising alcohol dehydrogenase (EC 1.1.1.-) activity. 
     
     
         45 . The method of  claim 43 , wherein the 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. 
     
     
         46 . The method of  claim 43 , wherein the 1,3 fatty diol is a C 12  1,3 fatty diol. 
     
     
         47 . The method of  claim 43 , wherein the 1,3 fatty diol is unsaturated.