Compositions and methods for 3-hydroxypropionate bio-production from biomass

Abstract

Methods of obtaining mutant nucleic acid sequences that demonstrate elevated oxaloacetate a-decarboxylase activity are provided. Compositions, such as genetically modified microorganisms that comprise such mutant nucleic acid sequences, are described, as are methods to obtain the same.

What is claimed is: 
     
         1 . A method for producing an acrylic acid-based consumer product, said method comprising
 i) combining a carbon source and a microorganism cell culture to produce 3-hydroxypropionic acid;   ii) converting said 3-hydroxypropionic acid to acrylic acid; and   iii) processing said acrylic acid into a consumer product.   
     
     
         2 . The method of  claim 1 , wherein said cell culture comprises a genetically modified microorganism. 
     
     
         3 . The method of  claim 2 , wherein said microorganism is modified for increased tolerance to 3-hydroxypropionic acid. 
     
     
         4 . The method of  claim 3 , wherein said modification modulates one or more components of the chorismate superpathway. 
     
     
         5 . The method of  claim 2 , wherein said microorganism is modified for increased production of 3-hydroxypropionic acid. 
     
     
         6 . The method of  claim 5 , wherein said modification comprises reduction in activity of one or more enzymes selected from pyruvate kinase, phosphofructokinase, lactate dehydrogenase, phosphate acetyltransferase, pyruvate oxidase, pyruvate-formate lyase, and homologs thereof. 
     
     
         7 . The method of  claim 6 , wherein said pyruvate kinase is selected from pykA and pykF, said phosphofructokinase is selected from pfkA and pfkB, said lactate dehydrogenase is selected from ldhA, said phosphate acetyltransferase is selected from pta, said pyruvate oxidase is selected from poxB, said pyruvate-formate lyase is selected from pflB, and homologs thereof. 
     
     
         8 . The method of  claim 5 , wherein said modification comprises increase in activity of one or more enzymes selected from phosphoenolpyruvate carboxykinase, malonyl-CoA reductase, 3-hydroxypropionate dehydrogenase, malonate semialdehyde dehydrogenase A, alpha-ketoglutarate decarboxylase, oxaloacetate alpha-oxo-decarboxylase, and homologs thereof. 
     
     
         9 . The method of  claim 8 , wherein said phosphoenolpyruvate carboxykinase is selected from pck, said malonyl-CoA reductase is selected from mer, said malonate semialdehyde dehydrogenase A is selected from mmsA, said 3-hydroxypropionic acid dehydrogenase is selected from mmsB, said alpha-ketoglutarate decarboxylase is selected from kgd, said oxaloacetate alpha-oxo-decarboxylase is selected from oad, and homologs thereof. 
     
     
         10 . The method of  claim 8 , wherein said modification comprises an increase in activity in a malonyl-CoA reductase enzyme. 
     
     
         11 . The method of  claim 8 , wherein said modification comprises an increase in activity in an oxaloacetate alpha-oxo-decarboxylase enzyme. 
     
     
         12 . The method of  claim 2 , wherein said microorganism is modified for increased tolerance to 3-hydroxypropionic acid, and wherein said microorganism is modified for increased production of 3-hydroxypropionic acid. 
     
     
         13 . A method for producing an acrylic acid-based consumer product, said method comprising
 i) combining a carbon source and a genetically modified microorganism in cell culture to produce 3-hydroxypropionic acid;   ii) converting said 3-hydroxypropionic acid to acrylic acid; and   iii) processing said acrylic acid into a consumer product;   
       wherein said microorganism is modified for increased production of 3-hydroxypropionic acid via an increase in activity in an oxaloacetate alpha decarboxylase enzyme or homolog thereof. 
     
     
         14 . A method for producing acrylic acid, said method comprising
 i) combining a carbon source and a microorganism cell culture to produce 3-hydroxypropionic acid in a concentration of at least 10 g/L; and   ii) converting said 3-hydroxypropionic acid to acrylic acid.   
     
     
         15 . The method of  claim 14 , wherein the combining comprises combining a carbon source and a microorganism cell culture comprising a microorganism genetically modified to increase activity of one or more enzymes selected from phosphoenolpyruvate carboxykinase, malonyl-CoA reductase, malonate semialdehyde dehydrogenase A, 3-HP dehydrogenase, alpha-ketoglutarate decarboxylase, oxaloacetate alpha-oxo-decarboxylase, and homologs thereof. 
     
     
         16 . The method of  claim 15 , wherein said phosphoenolpyruvate carboxykinase is selected from pck, said malonyl-CoA reductase is selected from mer, said malonate semialdehyde dehydrogenase A is selected from mmsA, said 3-hydroxypropionic acid dehydrogenase is selected from mmsB, said alpha-ketoglutarate decarboxylase is selected from kgd, said oxaloacetate alpha-oxo-decarboxylase is selected from oad, and homologs thereof. 
     
     
         17 . The method of  claim 14 , wherein the combining comprises combining a carbon source and a microorganism cell culture comprising a microorganism genetically modified to increase activity of malonyl-CoA reductase enzyme increasing production of 3-hydroxypropionic acid. 
     
     
         18 . The method of  claim 14 , wherein the combining comprises combining a carbon source and a microorganism cell culture comprising a microorganism genetically modified to increase activity of an oxaloacetate alpha-oxo-decarboxylase enzyme, increasing production of 3-hydroxypropionic acid. 
     
     
         19 . Biologically-produced acrylic acid, wherein said acrylic acid is produced according to  claim 14 . 
     
     
         20 . A consumer product produced with acrylic acid according to  claim 19 .