US2015093794A1PendingUtilityA1

Cellular production of glucaric acid

55
Assignee: MASSACHUSETTS INST TECHNOLOGYPriority: Apr 4, 2008Filed: Aug 5, 2014Published: Apr 2, 2015
Est. expiryApr 4, 2028(~1.7 yrs left)· nominal 20-yr term from priority
Inventors:Tae Seok MoonSang-Hwal YoonKristala Lanett Jones Prather
C12P 7/58C12N 15/8245C12N 15/8243C12Y 113/99001C12N 9/0069C12P 19/00C12Y 101/01203C12Y 505/01004C12P 19/02C12N 9/0006C12N 9/90
55
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Claims

Abstract

The invention relates to the production of glucuronic and glucaric acid in cells through recombinant expression of myo-inositol 1-phosphate synthase, myo-inositol oxygenase and uronate dehydrogenase. Cloning and characterization of the gene encoding uronate dehydrogenase is also disclosed.

Claims

exact text as granted — not AI-modified
We claim: 
     
         1 . A cell that recombinantly expresses a gene encoding uronate dehydrogenase and recombinantly expresses a gene encoding myo-inositol oxygenase. 
     
     
         2 .- 6 . (canceled) 
     
     
         7 . The cell of  claim 1  wherein the cell recombinantly expresses a gene encoding myo-inositol 1-phosphate synthase. 
     
     
         8 . (canceled) 
     
     
         9 . (canceled) 
     
     
         10 . The cell of  claim 1 , wherein the cell is a prokaryotic cell. 
     
     
         11 . The cell of  claim 10  wherein the cell is a bacterial cell. 
     
     
         12 . (canceled) 
     
     
         13 . The cell of  claim 11  wherein the genes encoding myo-inositol oxygenase and/or myo-inositol 1-phosphate synthase have been modified by codon optimization for expression in bacteria. 
     
     
         14 . The cell of  claim 1 , wherein the cell is a eukaryotic cell, optionally a fungal cell, a yeast cell, an insect cell, a plant cell or a mammalian cell. 
     
     
         15 . (canceled) 
     
     
         16 . The cell of  claim 7  wherein the genes encoding uronate dehydrogenase, myo-inositol oxygenase and/or myo-inositol 1-phosphate synthase are expressed on plasmids or are integrated into the genome of the cell. 
     
     
         17 . (canceled) 
     
     
         18 . The cell of  claim 7  wherein the production of glucaric acid is increased by protein engineering of the uronate dehydrogenase, myo-inositol oxygenase and/or myo-inositol 1-phosphate synthase enzymes in the cell. 
     
     
         19 . The cell of  claim 1  wherein the production of glucaric acid is increased by mutating a component of the glucaric acid metabolism pathway in the cell. 
     
     
         20 . A method for producing glucaric acid comprising culturing the cell of  claim 1  to produce glucaric acid. 
     
     
         21 . The method of  claim 20 , further comprising recovering the glucaric acid from the cells. 
     
     
         22 . A genetically modified microorganism that comprises one or more recombinant nucleic acid molecules encoding uronate dehydrogenase, myo-inositol oxygenase and myo-inositol 1-phosphate synthase. 
     
     
         23 . A method for producing glucaric acid, the method comprising genetically modifying a cell to recombinantly express at least one of: uronate dehydrogenase, myo-inositol oxygenase and myo-inositol 1-phosphate synthase, culturing a population of said cells, and collecting glucaric acid from the population of cells that have been genetically modified to produce glucaric acid. 
     
     
         24 .- 33 . (canceled) 
     
     
         34 . The method of  claim 23 , wherein the cell is a prokaryotic cell. 
     
     
         35 . The method of  claim 34  wherein the cell is a bacterial cell, optionally an  E. coli  cell. 
     
     
         36 . The method of  claim 23  wherein the genes encoding myo-inositol oxygenase and/or myo-inositol 1-phosphate synthase have been modified by codon optimization for expression in bacteria. 
     
     
         37 . The method of  claim 23 , wherein the cell is a eukaryotic cell, optionally a fungal cell, a yeast cell, an insect cell, a plant cell or a mammalian cell. 
     
     
         38 . (canceled) 
     
     
         39 . The method of  claim 23  wherein the genes encoding uronate dehydrogenase, myo-inositol oxygenase and/or myo-inositol 1-phosphate synthase are expressed on plasmids or are integrated into the genome of the cell. 
     
     
         40 . (canceled) 
     
     
         41 . The method of  claim 23  wherein the production of glucaric acid is increased by protein engineering of the uronate dehydrogenase, myo-inositol oxygenase and/or myo-inositol 1-phosphate synthase enzymes in the cell. 
     
     
         42 . The method of  claim 23  wherein the production of glucaric acid is increased by mutating a component of the glucaric acid metabolism pathway in the cell. 
     
     
         43 - 74 . (canceled)

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