Method for producing 3-hydroxypropionic acid and other products

Abstract

This invention relates to metabolically engineered microorganism strains, such as bacterial strains, in which there is an increased utilization of malonyl-CoA for production of a chemical product, which includes 3-hydroxypropionic acid.

1 . (canceled) 
     
     
         2 . A method for producing a product having malonyl-CoA as a substrate in a microbial production pathway of the product, the method comprising combining a carbon source and a microorganism with a cell culture to produce said product, wherein:
 a) said microorganism is genetically modified for reduced enzymatic activity of an enzyme wherein the enzyme is selected from the group consisting of beta-ketoacyl-ACP synthase, beta-ketoacyl-ACP reductase, beta-hydroxyacyl-ACP dehydratase, and enoyl-ACP reductase;   b) the enzymatic activity of the enzyme is reduced after fermentation is initiated; and either:   c1) said microorganism is further genetically modified for overexpression of udhA or pntAB; or   c2) said microorganism is genetically modified for overexpression of accA, accB, accC, accD, or any combination thereof.   
     
     
         3 . The method of  claim 2 , wherein said carbon source is glucose, sucrose, fructose, dextrose, lactose, or a combination thereof. 
     
     
         4 . The method of  claim 2 , wherein said microorganism is genetically modified for increased enzymatic activity in the organism's malonyl-CoA reductase (mcr) pathway by introduction of a heterologous nucleic acid sequence coding for a polypeptide having bi-functional malonyl-CoA reductase activity or mono-functional malonyl-CoA reductase activity. 
     
     
         5 . The method of  claim 4 , wherein said polypeptide has at least 70% homology with a sequence selected from the group consisting of SEQ ID NOs: 783-789. 
     
     
         6 . The method of  claim 2 , wherein said microorganism is further genetically modified for overexpression of udhA or pntAB, and
 wherein said microorganism is genetically modified for overexpression of accA, accB, accC, accD, or any combination thereof.   
     
     
         7 . The method of  claim 2 , wherein said overexpression of udhA or pntAB occurs by introduction of a heterologous nucleic acid sequence having at least 90% homology with a sequence selected from the group consisting of SEQ ID NOs. 886, 779, and 781. 
     
     
         8 . The method of  claim 2 , wherein said microorganism is further genetically modified for increased intracellular bicarbonate levels by introduction of a heterologous nucleic acid sequence coding for a polypeptide having cyanase and/or carbonic anhydrase activity. 
     
     
         9 . The method of  claim 8 , wherein said heterologous nucleic acid sequence is a sequence having at least 90% homology with SEQ ID NO. 337. 
     
     
         10 . The method of  claim 2 , wherein said overexpression of accA, accB, accC, accD, or any combination thereof occurs by introduction of a heterologous nucleic acid sequence coding for a polypeptide having at least 90% homology with a sequence selected from the group consisting of SEQ ID NO. 772, 774, 776, and 778. 
     
     
         11 . The method of  claim 2 , wherein said microorganism is further genetically modified to decrease activity of lactate dehydrogenase, phosphate acetyltransferase, pyruvate oxidase, pyruvate-formate lyase, or a combination thereof. 
     
     
         12 . The method of  claim 2 , wherein said microorganism cell culture comprises increased intracellular bicarbonate levels by supplementation with bicarbonate or carbonate.