Methods and materials for the biosynthesis of diol alcohols and related compounds

2:

• The process of claim 1 wherein the organism is C. necator or an organism with properties similar thereto.

3:

• The process of claim 1 wherein the organism is altered to express one or more enzymes of glycerol dehydrogenase, 1,2-propanediol oxidoreductase, methylglyoxal reductase, methylglyoxal synthase and/or dihydroxyacetone kinase.

4-7. (canceled)

8:

• The process of claim 3 wherein the glycerol dehydrogenase and/or 1,2-propanediol oxidoreductase is from E. coli, the methylglyoxal reductase is from S. Cerevisiae, the methylglyoxal synthase is from Clostridium acetobutylicum or C. necator, and/or the dihydroxyacetone kinase is from Citrobacter freundii.

9:

• The process of claim 3 wherein the glycerol dehydrogenase comprises SEQ ID NO:2 or a polypeptide with similar enzymatic activities exhibiting at least about 50% sequence identity to an amino acid sequence set forth in SEQ ID NO:
• 2 or a functional fragment thereof or is encoded by a nucleic acid sequence comprising SEQ ID NO:1 or SEQ ID NO:3 or a nucleic acid sequence encoding a polypeptide with similar enzymatic activities exhibiting at least about 50% sequence identity to the nucleic acid sequence set forth in SEQ ID NOs:
• 1 or 3 or a functional fragment thereof.

10-11. (canceled)

12:

• The process of claim 3 wherein the 1,2-propanediol oxidoreductase comprises SEQ ID NO:5 or a polypeptide with similar enzymatic activities exhibiting at least about 50% sequence identity to an amino acid sequence set forth in SEQ ID NO:
• 5 or a functional fragment thereof or is encoded by a nucleic acid sequence comprising SEQ ID NO:4 or a nucleic acid sequence encoding a polypeptide with similar enzymatic activities exhibiting at least about 50% sequence identity to the nucleic acid sequence set forth in SEQ ID NO:
• 4 or a functional fragment thereof.

13-14. (canceled)

15:

• The process of claim 3 wherein the methylglyoxal reductase comprises SEQ ID NO:7 or a polypeptide with similar enzymatic activities exhibiting at least about 50% sequence identity to an amino acid sequence set forth in SEQ ID NO:
• 7 or a functional fragment thereof or is encoded by a nucleic acid sequence comprising SEQ ID NO:6 or a nucleic acid sequence encoding a polypeptide with similar enzymatic activities exhibiting at least about 50% sequence identity to the nucleic acid sequence set forth in SEQ ID NO:6 or a functional fragment thereof.

16-17. (canceled)

18:

• The process of claim 3 wherein the methylglyoxal synthase comprises SEQ ID NO:9, 10, 12 or 13 or a polypeptide with similar enzymatic activities exhibiting at least about 50% sequence identity to an amino acid sequence set forth in SEQ ID NOs:
• 9, 10, 12 or 13 or a functional fragment thereof or is encoded by a nucleic acid sequence comprising SEQ ID NO:8 or SEQ ID NO:11 or a nucleic acid sequence encoding a polypeptide with similar enzymatic activities exhibiting at least about 50% sequence identity to the nucleic acid sequence set forth in SEQ ID NO:8 or 11 or a functional fragment thereof.

19-20. (canceled)

21:

• The process of claim 3 wherein the dihydroxyacetone kinase comprises SEQ ID NO:15 or a polypeptide with similar enzymatic activities exhibiting at least about 50% sequence identity to an amino acid sequence set forth in SEQ ID NO:
• 15 or a functional fragment thereof or is encoded by a nucleic acid sequence comprising SEQ ID NO:14 or a nucleic acid sequence encoding a polypeptide with similar enzymatic activities exhibiting at least about 50% sequence identity to the nucleic acid sequence set forth in SEQ ID NO:14 or a functional fragment thereof.

(canceled)

23:

• The process of claim 1 wherein the organism is further altered to channel the carbon flux toward the intermediates of the 1,2-PD pathway.

24:

• The process of claim 1 wherein one or more genes encoding for D-lactate dehydratases and/or D-lactate dehydrogenases and/or lactoylglutathione lyases are eliminated.

25:

• The process of claim 1 wherein the organism is further altered to eliminate phaCAB, involved in PHBs production and/or H16-A0006-9 encoding endonucleases thereby improving transformation efficiency.

(canceled)

27:

• An altered organism capable of producing more 1,2-PD, derivatives thereof and/or compounds related thereto as compared to an unaltered organism.

28:

• The altered organism of claim 27 which is C. necator or an organism with properties similar thereto.

29:

• The altered organism of claim 27 which expresses one or more of a glycerol dehydrogenase, 1,2-propanediol oxidoreductase, methylglyoxal reductase, methylglyoxal synthase and/or dihydroxyacetone kinase.

30-33. (canceled)

34:

• The altered organism of claim 29 wherein the glycerol dehydrogenase and/or 1,2-propanediol oxidoreductase is from E. coli, the methylglyoxal reductase is from S. Cerevisiae, the methylglyoxal synthase is from Clostridium acetobutylicum or C. necator, and/or the dihydroxyacetone kinase is from Citrobacter freundii.

35:

• The altered organism of claim 29 wherein the glycerol dehydrogenase comprises SEQ ID NO:2 or a polypeptide with similar enzymatic activities exhibiting at least about 50% sequence identity to an amino acid sequence set forth in SEQ ID NO:
• 2 or a functional fragment thereof or is encoded by a nucleic acid sequence comprising SEQ ID NO:1 or SEQ ID NO:3 or a nucleic acid sequence encoding a polypeptide with similar enzymatic activities exhibiting at least about 50% sequence identity to the nucleic acid sequence set forth in SEQ ID NOs:
• 1 or 3 or a functional fragment thereof.

36-37. (canceled)

38:

• The altered organism of claim 29 wherein the 1,2-propanediol oxidoreductase comprises SEQ ID NO:5 or a polypeptide with similar enzymatic activities exhibiting at least about 50% sequence identity to an amino acid sequence set forth in SEQ ID NO:
• 5 or a functional fragment thereof or is encoded by a nucleic acid sequence comprising SEQ ID NO:4 or a nucleic acid sequence encoding a polypeptide with similar enzymatic activities exhibiting at least about 50% sequence identity to the nucleic acid sequence set forth in SEQ ID NO:
• 4 or a functional fragment thereof.

39-40. (canceled)

41:

• The altered organism of claim 29 wherein the methylglyoxal reductase comprises SEQ ID NO:7 or a polypeptide with similar enzymatic activities exhibiting at least about 50% sequence identity to an amino acid sequence set forth in SEQ ID NO:
• 7 or a functional fragment thereof or is encoded by a nucleic acid sequence comprising SEQ ID NO:6 or a nucleic acid sequence encoding a polypeptide with similar enzymatic activities exhibiting at least about 50% sequence identity to the nucleic acid sequence set forth in SEQ ID NO:6 or a functional fragment thereof.

42-43. (canceled)

44:

• The altered organism of claim 29 wherein the methylglyoxal synthase comprises SEQ ID NO:9, 10, 12 or 13 or a polypeptide with similar enzymatic activities exhibiting at least about 50% sequence identity to an amino acid sequence set forth in SEQ ID NOs:
• 9, 10, 11 or 12 or a functional fragment thereof or is encoded by a nucleic acid sequence comprising SEQ ID NO:8 or SEQ ID NO:11 or a nucleic acid sequence encoding a polypeptide with similar enzymatic activities exhibiting at least about 50% sequence identity to the nucleic acid sequence set forth in SEQ ID NO:8 or 11 or a functional fragment thereof.

45-46. (canceled)

47:

• The altered organism of claim 29 wherein the dihydroxyacetone kinase comprises SEQ ID NO:15 or a polypeptide with similar enzymatic activities exhibiting at least about 50% sequence identity to an amino acid sequence set forth in SEQ ID NO:
• 15 or a functional fragment thereof or is encoded by a nucleic acid sequence comprising SEQ ID NO:14 or a nucleic acid sequence encoding a polypeptide with similar enzymatic activities exhibiting at least about 50% sequence identity to the nucleic acid sequence set forth in SEQ ID NO:14 or a functional fragment thereof.

(canceled)

49:

• The altered organism of claim 27 wherein the organism is further altered to channel the carbon flux toward the intermediates of the 1,2-PD pathway.

50:

• The altered organism of claim 27 wherein one or more genes encoding for D-lactate dehydratases and/or D-lactate dehydrogenases and/or lactoylglutathione lyase are eliminated.

51:

• The altered organism of claim 27 wherein the organism is further altered to eliminate phaCAB, involved in PHBs production and/or H16-A0006-9 encoding endonucleases thereby improving transformation efficiency.

(canceled)

54:

• A bio-derived, bio-based or fermentation derived product produced in accordance with the central metabolism depicted in FIG. 1.

55:

• An exogenous genetic molecule of the altered organism of claim 27.

56:

• The exogenous genetic molecule of claim 55 comprising a codon optimized nucleic acid sequence or an expression construct or synthetic operon of a glycerol dehydrogenase, 1,2-propanediol oxidoreductase, methylglyoxal reductase, methylglyoxal synthase and/or dihydroxyacetone kinase.

57:

• The exogenous genetic molecule of claim 56 codon optimized for C. necator.

58:

• The exogenous genetic molecule of claim 55 comprising a nucleic acid sequence encoding a glycerol dehydrogenase, a 1,2-propanediol oxidoreductase, a methylglyoxal reductase, a methylglyoxal synthase or a dihydroxyacetone kinase.

59:

• The exogenous genetic molecule of claim 55 comprising SEQ ID NO:1 or SEQ ID NO:3, a nucleic acid sequence exhibiting at least about 50% sequence identity to the nucleic acid sequence set forth in SEQ ID NO:
• 1 or 3 or a functional fragment thereof, or a nucleic acid sequence encoding a polypeptide with similar enzymatic activities and exhibiting at least about 50% sequence identity to the polypeptide encoded by the nucleic acid sequence set forth in SEQ ID NO:
• 1 or 3 or a functional fragment thereof.

(canceled)

61:

• The exogenous genetic molecule of claim 55 comprising SEQ ID NO:4, a nucleic acid sequence exhibiting at least about 50% sequence identity to the nucleic acid sequence set forth in SEQ ID NO:4 or a functional fragment thereof, or a nucleic acid sequence encoding a polypeptide with similar enzymatic activities and exhibiting at least about 50% sequence identity to the polypeptide encoded by the nucleic acid sequence set forth in SEQ ID NO:
• 4 or a functional fragment thereof.

(canceled)

63:

• The exogenous genetic molecule of claim 55 comprising SEQ ID NO:6, a nucleic acid sequence exhibiting at least about 50% sequence identity to the nucleic acid sequence set forth in SEQ ID NO:6 or a functional fragment thereof, or a nucleic acid sequence encoding a polypeptide with similar enzymatic activities and exhibiting at least about 50% sequence identity to the polypeptide encoded by the nucleic acid sequence set forth in SEQ ID NO:6 or a functional fragment thereof.

(canceled)

65:

• The exogenous genetic molecule of claim 55 comprising SEQ ID NO:8 or 11, a nucleic acid sequence exhibiting at least about 50% sequence identity to the nucleic acid sequence set forth in SEQ ID NO:8 or 11 or a functional fragment thereof, or a nucleic acid sequence encoding a polypeptide with similar enzymatic activities and exhibiting at least about 50% sequence identity to the polypeptide encoded by the nucleic acid sequence set forth in SEQ ID NO:8 or 11 or a functional fragment thereof.

(canceled)

67:

• The exogenous genetic molecule of claim 55 comprising SEQ ID NO:14, a nucleic acid sequence exhibiting at least about 50% sequence identity to the nucleic acid sequence set forth in SEQ ID NO:14 or a functional fragment thereof, or a nucleic acid sequence encoding a polypeptide with similar enzymatic activities and exhibiting at least about 50% sequence identity to the polypeptide encoded by the nucleic acid sequence set forth in SEQ ID NO:14 or a functional fragment thereof.

68-69. (canceled)

70:

• A process for the biosynthesis of 1,2-propanediol (1,2-PD), derivatives thereof and/or compounds related thereto, said process comprising:
• providing a means for producing 1,2-PD, derivatives thereof and/or compounds related thereto, and;
• producing 1,2-PD, derivatives thereof and/or compounds related thereto with said means.

72-73. (canceled)