LIGNOCELLULOSIC HYDROLYSATES AS FEEDSTOCKS FOR ISOBUTANOL FERMENTATION

• US 20130035515A1
• Filed: 06/15/2012
• Published: 02/07/2013
• Est. Priority Date: 06/17/2011
• Status: Abandoned Application

• Current Assignee: Butamax(TM) Advanced Biofuels LLC (BP PLC)
• Original Assignee: Butamax(TM) Advanced Biofuels LLC (BP PLC)
• Inventors: Kruckeberg, Arthur Leo, Dobson, Ian David
• Application Number:
  US13/525,032
• Publication Number:
  US 20130035515A1
• Time in Patent Office: Days
• US Class Current: 568/840
• CPC Class Codes: C12P 7/16 Butanols
• CPC Class Codes: Y02E 50/10 Biofuels, e.g. bio-diesel

Abstract

The invention relates generally to the field of industrial microbiology and butanol production from sources of 5-carbon sugars such as lignocellulosic hydrolysates. More specifically, the invention relates to the use of an xylulose or xylulose-5-phosphate-producing enzyme and micro-aerobic or anaerobic conditions to increase butanol production from such sugars and recovery of said butanol through ins situ product recovery methods.

Claims

1. A method of producing butanol comprising:

(a) providing a composition comprising (i) a microorganism capable of producing butanol and (ii) an enzyme or combination of enzymes capable of converting a 5-carbon sugar to xylulose or xylulose-5-phosphate;

(b) contacting the composition with a carbon substrate comprising mixed sugars; and

(c) culturing the microorganism under conditions of limited oxygen utilization, whereby butanol is produced.

4. The method of claim 1, wherein the enzyme or combination of enzymes capable of converting a 5-carbon sugar to xylulose or xylulose-5-phosphate are recombinantly expressed by a microorganism in the composition.

5. The method of claim 4, wherein the enzyme or combination of enzymes capable of converting a 5-carbon sugar to xylulose or xylulose-5-phosphate are recombinantly expressed by the microorganism capable of producing butanol.

6. The method of claim 4, wherein the enzyme or combination of enzymes capable of converting a 5-carbon sugar to xylulose or xylulose-5-phosphate are not produced by the microorganism capable of producing butanol.

7. The method of claim 1, wherein the enzyme or combination of enzymes capable of converting a 5-carbon sugar to xylulose or xylulose-5-phosphate are not produced by a microorganism in the composition.

8. The method of claim 1, wherein the enzyme or combination of enzymes capable of converting a 5-carbon sugar to xylulose or xylulose-5-phosphate is selected from the group consisting of:

(i) xylose isomerase;

(ii) xylose reductase;

(iii) xylitol dehydrogenase;

(iv) arabinose isomerase;

(v) ribulokinase;

(vi) ribulose-phosphate-5-epimerase;

(vii) arabinose reductase;

(viii) arabitol dehydrogenase;

(ix) xylulose reductase;

(x) xylulokinase;

(xi) aldose reductase; and

(xii) combinations thereof.

17. The method of claim 1, wherein the source of 5-carbon sugars is a source of xylose.

18. The method of claim 1, wherein the source of 5-carbon sugars is a source of arabinose.

21. The method of claim 1, additionally comprising contacting the composition with a source of 6-carbon sugars.

22. The method of claim 21, wherein 5-carbon and 6-carbon sugars are cumulatively consumed at a rate of at least about 1.5 g/gdcw/h.

25. The method of claim 21, wherein 5-carbon sugars and 6-carbon sugars are consumed, and the rate of 5-carbon sugar consumption is at least about 1% the rate of 6-carbon sugar consumption.

26. The method of claim 1, wherein the source of 5-carbon sugars is lignocellulosic biomass.

28. The method of claim 26, wherein the lignocellulose biomass is pretreated with ammonia.

39. The method of claim 1, wherein an inhibitor of respiration is added to the composition.

41. The method of claim 1, wherein the composition and/or the source of 5-carbon sugars further comprises a microorganism that is not capable of producing butanol, and the microorganism capable of producing butanol is present at a concentration that is at least equal to 1 g/l.

43. The method of claim 1, wherein the specific butanol production is at least about 0.4 g/g/h.

45. The method of claim 1, further comprising purifying butanol from the culture.

47. A butanol composition obtained by the method of claim 1

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49. A composition for producing butanol comprising:

(a) a microorganism capable of producing butanol;

(b) an enzyme or combination of enzymes capable of converting a 5-carbon sugar to xylulose or xylulose-5-phosphate;

(c) a source of 5-carbon sugars; and

(d) a fermentation media.

50. The composition of claim 49, wherein the butanol is isobutanol.

51. The composition of claim 49, wherein the microorganism is a yeast.

54. The composition of claim 49, wherein the source of 5-carbon sugars is present at a concentration of at least about 20 g/L.

57. The composition of claim 49, wherein the microorganism capable of producing butanol comprises polynucleotides encoding polypeptides that catalyze the conversion of:

(a) pyruvate to acetolactate;

(b) acetolactate to 2,3-dihydroxyisovalerate;

(c) 2,3-dihydroxyisovalerate to 2-ketoisovalerate;

(d) 2-ketoisovalerate to isobutyraldehyde; and

(e) isobutyraldehyde to isobutanol.

61. The composition of claim 49, wherein the microorganism capable of producing butanol is Saccharomyces cerevisiae.

63. The composition of claim 49, further comprising a microorganism that does not produce butanol.

65. The composition of claim 49, wherein the composition is capable of producing butanol and ethanol.

66. The composition of claim 49, wherein the composition is capable of producing butanol at least about 0.4 g/g % theoretical yield.

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