Production Of Fatty Acids And Derivatives Thereof

Abstract

Compositions and methods for production of fatty alcohols using recombinant microorganisms are provided as well as fatty alcohol compositions produced by such methods.

1 .- 20 . (canceled) 
     
     
         21 . A microorganism comprising one or more exogenous nucleic acid sequences encoding at least one wax synthase (EC 2.3.1.75), one thioesterase (3.1.2-, 3.1.1.-), and a fatty acyl-CoA synthase (EC 2.3.1.86). 
     
     
         22 . The microorganism of  claim 21 , further comprising an acyl-CoA reductase (EC 1.2.1.50). 
     
     
         23 . The microorganism of  claim 21 , wherein the microorganism further expresses one or more exogenous nucleic acid sequences encoding accABCD, aceE (EC 1.2.4.1, 2.3.1.61, 2.3.1.12), aceF (EC 1.2.4.1, 2.3.4.16, 2.3.1.12), acpP (AAC74178), cer1 (EC 4.1.99.5), fabA (EC4.2.1.60), fabB (EC 2.3.1.41), fabD (EC 2.3.1.39), fabG (EC 1.1.1.100), fabH (EC 2.3.1.180), fab1 (EC 1.3.1.9), fabZ (EC 4.2.1.-), lipase (EC 3.1.1.3), malonyl-CoA decarboxylase (EC 4.1.1.9, 4.1.1.41), panD (EC 4.1.1.11), panK (EC 2.7.1.33), pdh (EC 1.2.4.1), or udhA (EC 1.6.1.1). 
     
     
         24 . The microorganism of  claim 21 , comprising one or more attenuated endogenous nucleic acid sequences selected from ackA (EC 2.7.2.1), ackB (EC 2.7.2.1), adhE (EC 1.1.1.1, 1.2.1.10), fabF (EC 2.3.1.179), fabR (accession NP_418398), fadE (EC 1.3.99.3, 1.3.99.-), GST (EC 6.3.2.3), gpsA (EC 1.1.1.94), IdhA (EC 1.1.1.28), pflB (EC 2.3.1.54), plsB (EC 2.3.1.15), poxb (EC 1.2.2.2), pta (EC 2.3.1.8), glutathione synthase (EC 6.3.2.3), and combinations thereof. 
     
     
         25 . The microorganism of  claim 21 , wherein the microorganism is  E. coli.    
     
     
         26 . The microorganism of  claim 21 , wherein the microorganism additionally comprises a fatty acid derivative. 
     
     
         27 . The microorganism of  claim 26 , wherein the microorganism additionally comprises an exogenous nucleic sequence encoding ACP, Sfa, or a combination thereof; or expresses one or more exogenous nucleic acid sequences encoding an enzyme selected from one or more components of the branch chain keto acid dehydrogenase complex (EC1.2.4.4), Ilve (EC 2.6.1.42), Ipd (EC 1.8.1.4), Ccr (EC 1.1.19), IcmA (EC 5.4.99.2), IcmB (5.4.99.13), fabH (EC 2.3.1.180), fabF (EC 2.3.1.179), fabH3 (EC 2.3.1.180), fabC3 (NP_823468), beta-ketoacyl-ACP synthase II (EC 2.3.1.180), enoyl-CoA reductase (EC 1.3.1.34), enoyl-CoA isomerase (EC 4.2.1.-), and a combination thereof, wherein the fatty acid derivative is branched; or expresses one or more exogenous nucleic acid sequences encoding an enzyme selected from fabB (EC 2.3.1.41), fabK (EC 1.2.1.9), fabL (EC 1.2.1.9), fabM (5.3.3.14), fadE (EC 1.3.99.3, 1.3.99.-) and a combination thereof and wherein the fatty acid derivative is unsaturated. 
     
     
         28 . The microorganism of  claim 21 , wherein the microorganism is in a vessel comprising a fermentation broth comprising at least 10 mg/L fatty acid ester, 10 mg/L fatty alcohol, 10 mg/L hydrocarbon or at least 10 mg/L wax. 
     
     
         29 . The microorganism of  claim 26 , wherein the fatty acid derivative comprises from about 1 to about 5 double bonds; or wherein the fatty acid derivative comprises a carbon chain length of from about 8 to about 30; or wherein the fatty acid derivative comprises from about 1 to about 5 branch points. 
     
     
         30 . The microorganism of  claim 21 , wherein the microorganism additionally comprises a fatty acid ester or wax having an A side and a B side. 
     
     
         31 . The microorganism of  claim 30 , wherein the A side and the B side are produced by the microorganism; or wherein the A side, B side, or both the A side and the B side, comprise from about 1 to about 5 double bonds; or wherein the A side, B side, or both the A side and the B side, comprise a carbon chain length of from about 1 to about 26; or wherein the A side, B side, or both the A side and the B side, comprise from about 1 to about 5 carbon branch points; or wherein the A side, B side, or both the A side and the B side, comprise between 1 and 5 cyclopropyl moieties. 
     
     
         32 . A method of producing a fatty acid ester comprising culturing the microorganism of  claim 21 , under conditions sufficient to produce a fatty acid ester; and separating the fatty acid ester. 
     
     
         33 . A method of producing a wax comprising culturing the microorganism of  claim 21 , under conditions sufficient to produce a wax; and separating the wax. 
     
     
         34 . A method of obtaining a purified fatty acid derivative comprising,
 a) culturing the microorganism of  claim 21 , under conditions sufficient to produce a fatty acid derivative;   b) allowing the fatty acid derivative to separate into an organic phase; and   c) purifying the fatty acid derivative from the organic phase.