Methods of Using Natural and Engineered Organisms to Produce Small Molecules for Industrial Application
Aspects of the invention relate to methods of producing small molecules for industrial application using natural organisms and engineered organisms.
1 .- 63 . (canceled)
64 . A method for producing and harvesting amino acids, comprising:
culturing in growth medium comprising a carbon-containing gas a bacterial cell that can grow in the presence of the carbon-containing gas,
wherein the bacterial cell is of the genus Cupriavidus or of the genus Rhodococcus,
wherein said carbon-containing gas is CO 2 ,
wherein said bacterial cell chemoautotrophically converts the CO 2 into amino acid products,
wherein said bacterial cell does not form poly-β-hydroxybutyrate (PHB) under chemoautotrophic growth conditions, and
wherein said bacterial cell secretes said amino acids products into the growth medium; and
separating the secreted amino acid products from the growth medium.
65 . The method of claim 64 , wherein the secreted amino acid products comprise lysine, tyrosine or phenylalanine.
66 . The method of claim 64 , wherein the bacterial cell is recombinant.
67 . The method of claim 66 , 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.
68 . The method of claim 67 , wherein the enzyme involved in lysine biosynthesis is an aspartate kinase or a dihydrodipicolinate synthase.
69 . The method of claim 67 , wherein the enzyme involved in lysine secretion is a lysine exporter.
70 . The method of claim 66 , 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.
71 . The method of claim 70 , wherein the enzyme in the citric acid cycle is succinyl-CoA synthase.
72 . The method of claim 66 , wherein the bacterial cell exhibits increased expression or activity relative to a wild type cell of an enzyme involved in the Shikimate pathway.
73 . The method of claim 72 , wherein the enzyme involved in the Shikimate pathway is chorismate synthase.
74 . The method of claim 64 , wherein the bacterial cell is a Cupriavidus necator cell or a Rhodococcus opacus cell.
75 . The method of claim 64 , wherein the bacterial cell is a Cupriavidus necator DSM 541 cell.
76 . The method of claim 64 , wherein the bacterial cell is an engineered cell that secretes at least 125 fold higher level of an amino acid than the corresponding wild type cell.
77 . The method of claim 76 , wherein the said amino acid is lysine.
78 . The method of claim 64 , wherein the bacterial cell is a mutant and/or engineered cell that secretes at least 2.5 fold higher level of an amino acid than the corresponding wild type cell.
79 . The method of claim 78 , wherein the said amino acid is an aromatic amino acid.
80 . The method of claim 79 , wherein the said aromatic amino acid is phenylalanine, tryptophan, tyrosine, and/or histidine.