TH41437A - Method for producing 1,3-propediol by recombinant organisms - Google Patents

Abstract

DC60 (24/06/42) provided recombinant organisms containing encoding receptors for glycerol-3-phosphate dehydrogenase, glycerol-3-phosphatase, glycerol dehydratase, and 1,3-propanediol oxidoreductase activity, which are useful for the production of 1,3-propanediol from various carbon substrates.

Claims (14)

1. Methods for producing 1,3-propanediol from recombinant organisms containing (i) transformation of a suitable host cell with a transformation cassette containing one of (a) the gene encoding glycerol-3-phosphate dehydrogenase activity, (b) the gene encoding glycerol-3-phosphate phosphatase activity, (c) the gene encoding dehydratase activity, and (d) the gene encoding 1,3-propanediol oxydoreductase activity, provided that if the transformation cassette contains fewer genes than (a) - (d), then the suitable host will contain at least one gene in the transformed host cell containing each gene of (a) - (d). (ii) Culture of the transformed host cell with at least one carbon source selected from the group containing with monosaccharides, oligosaccharides, polysaccharides or single carbon substrates that produce 1,3-propanediol and (iii) storage of 1,3-propanediol 2. Method according to claim 1, where the transformation disk contains all of the digits of (a) - (d). 3. Method according to Claim 1, where a suitable host cell is selected from a group consisting of bacteria and hyphae. 4. Method according to claim 3, where a suitable host cell is selected from a group consisting of Citrobacter, Enterobacter, Clostridium, Klebsiella, Aerobacter, Lactobacillus, Aspergillus, Saccharomyces, Schizosaccharomyces, Zygosaccharomyces, Pichia, Kluyveromyces, Candida, Hansenula, Debaryomyces, Mucor, Torulopisis, Methylobacter, Escherichia, Salmonella, Bacillus, Streptomyces, and Pseudomonas. 5. According to claim 4, the appropriate host cell is selected from a group consisting of E. coli or Saccharomyces spp. 6. Method according to claim 1, where the transformed host organism is Saccharomyces spp. transformed by a transformation deslate containing the genes dha1, dha2, dha3, and dhaT, which are permanently inserted into the genome of Saccharomyces cerevisiae. 7. Method according to Claim 1, where the transformed host organism is Klebsiella spp. transformed by a transformation cassette containing the genes GPD1 and GPD2. 8. Method according to claim 1, where the carbon source is glucose. 9. The method under claim 1 in which the Chinese encoding of the enzyme glycerol-3-phosphate dehydrogenase is determined by Chinese sequences corresponding to the amino acid sequences provided in SEQ ID NO: 11, SEQ ID NO: 12, and SEQ ID NO: 13, encompasses amino acid substitutions, cleavage, or insertion that do not alter the functional properties of the enzyme glycerol-3-phosphate dehydrogenase. 10. According to Claim 1, China's sequencing of the enzyme glycerol-3-phosphatase corresponds to the amino acid sequences provided in SEQ ID NO:33 and SEQ ID NO:F17. These amino acid sequences include amino acid substitutions, cleavage, or insertion that do not alter the functional properties of glycerol-3-phosphatase. 11. According to claim 1, China's method of encoding the enzyme glycerol-3-phosphatase corresponds to the amino acid sequence provided in SEQ ID NO:18, and where the amino acid sequence encompasses amino acid substitution, cleavage, or insertion without altering the functional properties of the glycerol-3-phosphatase enzyme. 12. The method under Claim 1 of the Chinese law for computation of the dehydratase enzyme sequences containing dha1, dha2, dha3, and dhaT corresponding to the amino acid sequences provided in SEQ ID NO: 34, SEQ ID NO: 35, and SEQ ID NO: 36, includes amino acid substitutions, elimination, or insertion that do not alter the functional properties of the dehydratase enzyme. 13. The method under Chinese claim 1 that encodes the enzyme 1,3-propanediol oxidoreductase corresponding to the amino acid sequence given in SEQ ID NO: 37, shall encompass amino acid substitution, truncation, or insertion that does not alter the functional properties of the 1,3-propanediol oxidoreductase enzyme. 14. Transformed host cells containing (a) a group of genes comprising (1) a gene encoding the enzyme glycerol-3-phosphate hydrogenase corresponding to the amino acid sequence given in SEQ ID NO: 11, (2) a gene encoding the enzyme glycerol-3-phosphate phosphatase corresponding to the amino acid sequence given in SEQ ID NO: 17, (3) a gene encoding the enzyme glycerol-3-phosphate hydrogenase corresponding to the amino acid sequence given in SEQ ID NO: 34, (4) a gene encoding the B subunit of the enzyme glycerol dehydrase corresponding to the amino acid sequence given in SEQ ID NO: 35, (5) a gene encoding the T subunit of the enzyme glycerol dehydrase corresponding to the amino acid sequence given in SEQ ID NO: 36, (6) a gene encoding the enzyme 1,3-Occhidoreductase corresponding to the amino acid sequences given in SEQ ID NO: 37, the amino acid sequences of (a) (1) - (6) that cover amino acids with substitution, deletion, addition of amino acids that do not change the enzyme activity of genes (1) - (6), and (b) host cells transformed with the group of genes of (a) by host cells transformed to produce 1,3-propanediol in at least one substrate selected from the group of monosaccharides, oligosaccharides and polysaccharides or from single-carbon substrates (14 claims, 3 pages, 0 figures).