Methods of Using Natural and Engineered Organisms to Produce Small Molecules for Industrial Application

Abstract

Aspects of the invention relate to methods of producing small molecules for industrial application using natural organisms and engineered organisms.

What is claimed is: 
     
         1 .- 8 . (canceled) 
     
     
         9 . A method for harvesting amino acids, comprising:
 culturing in growth medium comprising a carbon-containing gas a bacterial cell that can grow in the presence of a carbon-containing gas and that secretes amino acids into the growth medium; and   separating the secreted amino acids from the growth medium.   
     
     
         10 . The method of  claim 9 , wherein the amino acid is lysine, tyrosine or phenylalanine. 
     
     
         11 . The method of  claim 9 , wherein the bacterial cell is recombinant. 
     
     
         12 . The method of  claim 11 , wherein the bacterial cell exhibits increased expression or activity relative to a wild type cell of an enzyme involved in lysine biosynthesis or in lysine secretion. 
     
     
         13 . The method of  claim 12 , wherein the enzyme involved in lysine biosynthesis is an aspartate kinase or a dihydrodipicolinate synthase. 
     
     
         14 . The method of  claim 12 , wherein the enzyme involved in lysine secretion is a lysine exporter. 
     
     
         15 . The method of  claim 11 , wherein the bacterial cell exhibits decreased expression or activity relative to a wild type cell of one or more enzymes in the citric acid cycle. 
     
     
         16 . The method of  claim 15 , wherein the enzyme in the citric acid cycle is succinyl-CoA synthase. 
     
     
         17 . The method of  claim 11 , wherein the bacterial cell exhibits increased expression or activity relative to a wild type cell of an enzyme involved in the Shikimate pathway. 
     
     
         18 . The method of  claim 17 , wherein the enzyme involved in the Shikimate pathway is chorismate synthase. 
     
     
         19 . The method of  claim 9 , wherein the bacterial cell is of the genus  Ralstonia  or of the genus  Rhodococcus.    
     
     
         20 . The method of  claim 19 , wherein the bacterial cell is a  Ralstonia eutropha  cell or a  Rhodococcus opacus  cell. 
     
     
         21 . A method for producing putrescine, comprising:
 culturing in growth medium comprising a carbon-containing gas a bacterial cell that can grow in the presence of a carbon-containing gas and that secretes putrescine into the growth medium; and   separating the putrescine from the growth medium.   
     
     
         22 . The method of  claim 21 , wherein the bacterial cell is recombinant. 
     
     
         23 . The method of  claim 22 , wherein the bacterial cell exhibits increased expression or activity relative to a wild type cell of an arginine decarboxylase and/or an ornithine decarboxylase. 
     
     
         24 . A method for producing caprolactam, comprising:
 culturing in growth medium comprising a carbon-containing gas a recombinant bacterial cell that can grow in the presence of a carbon-containing gas and that secretes caprolactam into the growth medium; and   separating the caprolactam from the growth medium.   
     
     
         25 . The method of  claim 24 , wherein the bacterial cell exhibits increased expression or activity relative to a wild type cell of an enzyme selected from a carbon nitrogen lyase, an α-β-enoate reductase, an amidohydrolase and a tyrosine phenol lyase. 
     
     
         26 . A method for producing styrene, comprising:
 culturing in growth medium comprising a carbon-containing gas a recombinant bacterial cell that can grow in the presence of a carbon-containing gas and that secretes styrene into the growth medium; and   separating the styrene from the growth medium.   
     
     
         27 . The method of  claim 26 , wherein the bacterial cell exhibits increased expression or activity relative to a wild type cell of a phenylalanine ammonium lyase enzyme and/or an oxylate decarboxylase enzyme. 
     
     
         28 . A method for producing 1,3-butanediol, comprising:
 culturing in growth medium comprising a carbon-containing gas a bacterial cell that can grow in the presence of a carbon-containing gas and that secretes 1,3-butanediol into the growth medium; and   separating the 1,3-butanediol from the growth medium.   
     
     
         29 . The method of  claim 28 , wherein the bacterial cell exhibits increased expression or activity relative to a wild type cell of an oleate hydratase enzyme. 
     
     
         30 . A recombinant cell that exhibits increased expression or activity of:
 (i) one or more of an aspartate kinase, a dihydrodipicolinate synthase or a lysine exporter, wherein the recombinant cell can produce a cell culture that contains lysine;   (ii) a chorismate synthase enzyme, wherein the recombinant cell can produce a cell culture that contains lysine or phenylalanine;   (iii) an arginine decarboxylase and/or an ornithine decarboxylase, wherein the recombinant cell can produce a cell culture that contains putrescine;   (iv) an enzyme selected from a carbon nitrogen lyase, an α-β-enoate reductase, an amidohydrolase and a tyrosine phenol lyase, wherein the recombinant cell can produce a cell culture that contains caprolactam;   (v) a phenylalanine ammonium lyase enzyme and/or an oxylate decarboxylase enzyme, wherein the recombinant cell can produce a cell culture that contains styrene; or   (vi) an oleate hydratase enzyme, wherein the recombinant cell can produce a cell culture that contains 1,3-butadiene.   
     
     
         31 . The recombinant cell of  claim 30 , wherein the recombinant cell is of the genus  Ralstonia  or of the genus  Rhodococcus.    
     
     
         32 . The recombinant cell of  claim 31 , wherein the recombinant cell is a  Ralstonia eutropha  cell or a  Rhodococcus opacus  cell.