Human gut microbiome-derived biosynthetic enzymes for production of fatty acid amides

Abstract

Disclosed herein, in some embodiments, are vectors encoding biosynthetic enzymes from gut microbiome-derived bacterium (e.g., Clostridia enzymes), engineered cells comprising the vectors, and methods of using biosynthetic enzymes from gut microbiome-derived bacterium (e.g., Clostridia enzymes) to produce fatty acid amides.

1 - 38 . (canceled) 
     
     
         39 . A vector for the production of fatty acid amides, comprising 
 genes, wherein each of the genes is under the control of a regulatory element, wherein each of the genes is derived from a gut microbiome-derived bacterium, and wherein the genes encode at least a fatty acyl transferase, an acyl carrier protein, and a fatty acyl-ACP synthetase.   
     
     
         40 . The vector of  claim 39 , wherein the regulatory element is an inducible promoter. 
     
     
         41 . The vector of  claim 39 , wherein the regulatory element is a constitutive promoter. 
     
     
         42 . The vector of  claim 39 , wherein the regulatory element is a repressible promoter. 
     
     
         43 . The vector of  claim 39 , wherein the regulatory element comprises a T7 promoter. 
     
     
         44 . The vector of  claim 43 , wherein the vector further encodes a T7 RNA polymerase. 
     
     
         45 . The vector of  claim 39 , further comprising a ribosomal binding site. 
     
     
         46 . The vector of  claim 39 , wherein the genes are codon-optimized. 
     
     
         47 . The vector of  claim 39 , wherein the genes are open reading frames. 
     
     
         48 . The vector of  claim 39 , wherein the genes are codon-optimized for production in E.  coli . 
     
     
         49 . The vector of  claim 39 , wherein the gut microbiome-derived bacterium is from Clostridia. 
     
     
         50 . The vector of  claim 39 , wherein the genes also encode a hydrolase. 
     
     
         51 . The vector of  claim 39 , wherein the genes also encode a lipid transfer protein. 
     
     
         52 . The vector of  claim 39 , wherein the genes also encode a glycosyltransferase. 
     
     
         53 . The vector of  claim 39 , wherein the genes encode Sfp 4′-phosphopantetheinyl transferase. 
     
     
         54 . The vector of  claim 39 , wherein the genes are in a single operon. 
     
     
         55 - 74 . (canceled) 
     
     
         75 . An engineered cell comprising the vector of  claim 39 . 
     
     
         76 . A method of producing fatty acid amides, the method comprising:
 contacting a composition, comprising one or more exogenous fatty acids and one or more amines, with a set of biosynthetic enzymes which are human gut microbiome-derived clostridia biosynthetic enzymes in an effective amount to produce a fatty acid amide, wherein the set of biosynthetic enzymes comprise a fatty acyl transferase, an acyl carrier protein, and a fatty acyl-ACP synthetase.