METHODS AND COMPOSITIONS FOR PRODUCING CHEMICAL PRODUCTS FROM C. PHYTOFERMENTANS

• US 20110183382A1
• Filed: 12/15/2010
• Published: 07/28/2011
• Est. Priority Date: 12/15/2009
• Status: Abandoned Application

• Current Assignee: Qteros, Inc.
• Original Assignee: Qteros, Inc.
• Inventors: Ju, Chelsea, Verhoff, Francis H., Coil, Gregory S., Schmalisch, Matthias
• Application Number:
  US12/969,582
• Publication Number:
  US 20110183382A1
• Time in Patent Office: Days
• US Class Current: 435/109
• CPC Class Codes: C12N 1/205 Bacterial isolates
• CPC Class Codes: C12P 13/14 Glutamic acid; Glutamine
• CPC Class Codes: C12P 13/20 Aspartic acid; Asparagine
• CPC Class Codes: C12P 3/00 Preparation of elements or inorganic compounds except carbon dioxide recovery of carbon dioxides as by-products C12F3/02
• CPC Class Codes: C12P 7/10 substrate containing cellulosic material
• CPC Class Codes: C12P 7/16 Butanols
• CPC Class Codes: C12P 7/40 containing a carboxyl group including Peroxycarboxylic acids
• CPC Class Codes: C12P 7/42 Hydroxy-carboxylic acids
• CPC Class Codes: C12P 7/46 Dicarboxylic acids having four or less carbon atoms, e.g. fumaric acid, maleic acid
• CPC Class Codes: C12P 7/54 Acetic acid vinegar C12J
• CPC Class Codes: C12P 7/56 Lactic acid
• CPC Class Codes: C12R 2001/145 Clostridium
• CPC Class Codes: Y02E 50/10 Biofuels, e.g. bio-diesel

Abstract

This invention provides systems and methods for the production of compounds by C. phytofermentans. C. phytofermentans is genetically-engineered for hydrolysis and fermentation of carbonaceous biomass to synthesize compounds of commercial value.

Claims

1. A composition for producing first and second fermentation end-products comprising:

a. a carbonaceous biomass;

b. a microorganism that hydrolyses and ferments pentose and hexose saccharides from said biomass;

c. a first fermentation end-product, wherein said first fermentation end-product is not ethanol, propanol, isopropanol, n-butanol, hydrogen, formic acid, lactic acid, acetic acid, formate, lactate or acetate; and

d. a second fermentation end-product.

3. The composition of claim 1, wherein said microorganism is genetically modified to produce a higher concentration of said first fermentation end-product compared to a non-genetically modified form of said microorganism.

5. The composition of claim 1, wherein said microorganism is a bacterium.

6. The composition of claim 1, wherein said microorganism is a species of Clostridia.

7. The composition of claim 1, wherein said microorganism is Clostridium phytofermentans.

8. The composition of claim 1, wherein said microorganism is Clostridium sp. Q.D.

9. The composition of claim 1, wherein the microorganism is non-recombinant.

10. The composition of claim 1, wherein the microorganism is recombinant.

11. The composition of claim 1, wherein said microorganism is genetically modified to enhance production of a fermentation end-product.

12. The composition of claim 1, wherein said microorganism is genetically modified to express a protein encoded by a heterologous polynucleotide.

13. The composition of claim 1, wherein said microorganism is genetically modified to enhance expression of a protein by deregulation of an endogenous promoter or by expression of an additional copy of an endogenous polynucleotide encoding said protein.

15. The composition of claim 1, wherein said first fermentation end-product is aspartic acid, aspartate, glutamic acid, glutamate, malic acid, or malate.

17. The composition of claim 1, wherein said second fermentation end-product is not ethanol, propanol, isopropanol, n-butanol, hydrogen, formic acid, lactic acid, acetic acid, formate, lactate or acetate.

18. The composition of claim 1, wherein said second fermentation end-product is ethanol, propanol, isopropanol, n-butanol, hydrogen, formic acid, lactic acid, acetic acid, formate, lactate or acetate and is present at a lower amount than said first fermentation product.

19. The composition of claim 1, wherein said second fermentation end-product is ethanol, propanol, isopropanol, n-butanol, hydrogen, formic acid, lactic acid, acetic acid, formate, lactate or acetate and is present at an amount less than 99%, 75%, 50%, 30%, 20%, or 10% of the amount of said first fermentation product.

27. The composition of claim 1, wherein said first fermentation end-product is a 1,4 diacid (succinic, fumaric or malic), 2,5 furan dicarboxylic acid, 3 hydroxy propionic acid, aspartic acid, aspartate, glucaric acid, glutamic acid, glutamate, malate, itaconic acid, levulinic acid, 3-hydroxybutyrolactone, glycerol, sorbitol, xylitol/arabinitol, butanediol, an isoprenoid, or a terpene.

29. The composition of claim 1, wherein said carbonaceous biomass comprises one or more of xylan, cellulose, hemicellulose, fructose, glucose, mannose, rhamnose, or xylose.

30. The composition of claim 1, wherein said carbonaceous biomass is plant matter.

31. The composition of claim 1, wherein said carbonaceous biomass comprises woody plant matter, non-woody plant matter, cellulosic material, lignocellulosic material, hemicellulosic material, carbohydrates, pectin, starch, inulin, fructans, glucans, corn, corn stover, sugar cane, grasses, switch grass, sorghum, bamboo, distillers grains, Distillers Dried Solubles (DDS), Distillers Dried Grains (DDG), Condensed Distillers Solubles (CDS), Distillers Wet Grains (DWG), Distillers Dried Grains with Solubles (DDGS), peels, citrus peels, bagasse, poplar, or algae.

32. The composition of claim 1, wherein said carbonaceous biomass comprises a higher concentration of oligomeric carbohydrates relative to monomeric carbohydrates.

33. The composition of claim 1, wherein said hydrolysis provides results in a greater concentration of cellobiose relative to monomeric carbohydrates.

34. The composition of claim 32, wherein said monomeric carbohydrates comprise xylose and arabinose.

35. The composition of claim 1, wherein said carbonaceous biomass is pre-treated with an acid, alkali or heat prior to contact with said microorganism.

36. The composition of claim 1, further comprising a second species of microorganism.

37. The composition of claim 36, wherein said second species of microorganism is selected from the group consisting of a yeast, an other fungus, and a bacterium.

38. The composition of claim 35, wherein said second species of microorganism is S. cerevisiae or Aspergillus niger.

2. (canceled)

4. (canceled)

14. (canceled)

16. (canceled)

20-26. -26. (canceled)

28. (canceled)

39. A fermentation composition comprising:

a. a carbonaceous biomass;

b. a strain of Clostridium phytofermentans; and

c. a fermentation end-product comprising aspartic acid, aspartate, glutamic acid, glutamate, malic acid, or malate,

wherein said strain of Clostridium phytofermentans produces said fermentation end-product from said carbonaceous biomass.

40. (canceled)

41. A process for producing a first and second fermentation end-product comprising:

a. contacting a carbonaceous biomass with;

a microorganism that hydrolyses and ferments pentose and hexose saccharides from said carbonaceous biomass to produce said first and second fermentation end-products, wherein said first fermentation end-product is not ethanol, propanol, isopropanol, n-butanol, hydrogen, formic acid, lactic acid, acetic acid, formate, lactate or acetate; and

b. allowing sufficient time for said hydrolysis and fermentation to produce said first and second fermentation end-products.

42. The process of claim 41, wherein said microorganism is genetically modified to produce a higher concentration of said first fermentation end-product compared to a non-genetically modified or non-mutagenized form of said microorganism.

44. The process of claim 41, wherein said microorganism is a bacterium.

45. The process of claim 41, wherein said microorganism is a species of Clostridia.

46. The process of claim 41, wherein said microorganism is Clostridium phytofermentans.

47. The process of claim 41, wherein said microorganism is Clostridium sp. Q.D.

50. The process of claim 41, wherein said microorganism is genetically modified to enhance production of a fermentation end-product.

54. The process of claim 41, wherein said first fermentation end-product is aspartic acid, aspartate, glutamic acid, glutamate, malic acid, or malate.

56. The process of claim 41, wherein said first fermentation end-product is a 1,4 diacid (succinic, fumaric or malic), 2,5 furan dicarboxylic acid, 3 hydroxy propionic acid, aspartic acid, aspartate, glucaric acid, glutamic acid, glutamate, malate, itaconic acid, levulinic acid, 3-hydroxybutyrolactone, glycerol, sorbitol, xylitol/arabinitol, butanediol, an isoprenoid, or a terpene.

59. The process of claim 41, wherein said second fermentation end-product is not ethanol, propanol, isopropanol, n-butanol, hydrogen, formic acid, lactic acid, acetic acid, formate, lactate or acetate.

60. The process of claim 41, wherein said second fermentation end-product is ethanol, propanol, isopropanol, n-butanol, hydrogen, formic acid, lactic acid, acetic acid, formate, lactate or acetate and is present at a lower amount than said first fermentation product.

146. A fermentation end-product produced by the process of claim 41, wherein said microorganism is Clostridium phytofermentans or mutant thereof.

147. The fermentation end-product of claim 146, wherein said fermentation end-product is aspartic acid, aspartate, glutamic acid, glutamate, malic acid, or malate.

43. (canceled)

48. (canceled)

49. (canceled)

51-53. -53. (canceled)

55. (canceled)

57. (canceled)

58. (canceled)

61-105. -105. (canceled)

106. A system for producing a fermentation end-product comprising a fermentation vessel comprising:

a. a carbonaceous biomass; and

b. a microorganism that hydrolyses and ferments pentose and hexose saccharides from said carbonaceous biomass, wherein said microorganism produces a first and second fermentation end-products, wherein said first fermentation end-product is not ethanol, propanol, isopropanol, n-butanol, hydrogen, formic acid, lactic acid, acetic acid, formate, lactate or acetate, wherein said fermentation vessel is adapted to provide suitable conditions for fermentation of pentose and hexose saccharides into said first and second fermentation end-products.

108. The system of claim 106, wherein said microorganism is genetically modified to produce a higher concentration of said first fermentation end-product compared to a non genetically modified or mutagenized form of said microorganism, wherein said fermentation vessel is adapted to provide suitable conditions for fermentation of pentose and hexose saccharides into said fermentation end-product.

110. The system of claim 106, wherein said microorganism is a bacterium.

111. The system of claim 106, wherein said microorganism is a species of Clostridia.

112. The system of claim 106, wherein said microorganism is Clostridium phytofermentans.

113. The system of claim 106, wherein said microorganism is Clostridium sp. Q.D.

121. The system of claim 106, wherein said first fermentation end-product is aspartic acid, aspartate, glutamic acid, glutamate, malic acid, or malate.

124. The system of claim 106, wherein said first fermentation end-product is a 1,4 diacid (succinic, fumaric or malic), 2,5 furan dicarboxylic acid, 3 hydroxy propionic acid, aspartic acid, aspartate, glucaric acid, glutamic acid, glutamate, malate, itaconic acid, levulinic acid, 3-hydroxybutyrolactone, glycerol, sorbitol, xylitol/arabinitol, butanediol, an isoprenoid, or a terpene.

125. The system of claim 106, wherein said second fermentation end-product is not ethanol, propanol, isopropanol, n-butanol, hydrogen, formic acid, lactic acid, acetic acid, formate, lactate or acetate.

126. The system of claim 106, wherein said second fermentation end-product is ethanol, propanol, isopropanol, n-butanol, hydrogen, formic acid, lactic acid, acetic acid, formate, lactate or acetate and is present at a lower amount than said first fermentation product.

107. (canceled)

109. (canceled)

114-120. -120. (canceled)

122. (canceled)

123. (canceled)

127-143. -143. (canceled)

144. A composition for producing first and second fermentation end-products comprising:

a. a carbonaceous biomass;

b. a Clostridium phytofermentans strain that hydrolyses and ferments pentose and hexose saccharides from said biomass;

c. a first fermentation end-product, wherein said first fermentation end-product is a 1,4 diacid (succinic, fumaric or malic), 2,5 furan dicarboxylic acid, 3 hydroxy propionic acid, aspartic acid, aspartate, glucaric acid, glutamic acid, glutamate, malate, itaconic acid, levulinic acid, 3-hydroxybutyrolactone, glycerol, sorbitol, xylitol/arabinitol, butanediol, an isoprenoid, or a terpene; and

d. a second fermentation end-product, wherein said second fermentation end-product is not ethanol, propanol, isopropanol, n-butanol, hydrogen, formic acid, lactic acid, acetic acid, formate, lactate or acetate.

145. A composition for producing first and second fermentation end-products comprising:

a. a carbonaceous biomass;

b. a Clostridium phytofermentans strain that hydrolyses and ferments pentose and hexose saccharides from said biomass;

c. a first fermentation end-product, wherein said first fermentation end-product is a 1,4 diacid (succinic, fumaric or malic), 2,5 furan dicarboxylic acid, 3 hydroxy propionic acid, aspartic acid, aspartate, glucaric acid, glutamic acid, glutamate, malate, itaconic acid, levulinic acid, 3-hydroxybutyrolactone, glycerol, sorbitol, xylitol/arabinitol, butanediol, an isoprenoid, or a terpene; and

d. a second fermentation end-product, wherein said second fermentation end-product is ethanol, propanol, isopropanol, n-butanol, hydrogen, formic acid, lactic acid, acetic acid, formate, lactate or acetate and is present at a lower amount than said first fermentation product.