Method for the production of glycerol by recombinant organisms

• US 6,358,716 B1
• Filed: 05/11/1999
• Issued: 03/19/2002
• Est. Priority Date: 11/13/1996
• Status: Expired due to Term

• Current Assignee: E. I. du Pont de Nemours and Company (Corteva, Inc.), Genencor International Incorporated (International Flavors & Fragrances Incorporated)
• Original Assignee: E. I. du Pont de Nemours and Company (Corteva, Inc.)
• Inventors: Hsu, Amy Kuang-Hua, Gatenby, Anthony Arthur, Haynie, Sharon Loretta, Bulthuis, Ben A., Lareau, Richard D.
• Primary Examiner(s): Achutamurthy, Ponnathapu
• Assistant Examiner(s): BRAGDON, KATHLEEN KERR
• Application Number:
  US09/297,928
• Time in Patent Office: 1,043 Days
• Field of Search: 435/159, 435/252.33
• US Class Current: 435/159
• CPC Class Codes: C12P 7/20 Glycerol

Abstract

Recombinant organisms are provided comprising genes encoding a glycerol-3-phosphate dehydrogenase and/or a glycerol-3-phosphatase activity useful for the production of glycerol from a variety of carbon substrates.

Claims

1. A method for the production of glycerol from a recombinant microorganism comprising:

(i) transforming a suitable host cell with an expression cassette comprising

(a) a gene encoding a NADH-dependent glycerol-3-phosphate dehydrogenase enzyme or a NADPH-dependent glycerol-3-phosphate dehydrogenase enzyme; and

(b) a gene encoding a polypeptide encoding a glycerol-3-phosphatase (EC 3.1.3.21) enzyme;

(ii) culturing the transformed host cell of (i) in the presence of at least one carbon source selected from the group consisting of monosaccharides, oligosaccharides, polysaccharides, and single-carbon substrates, whereby glycerol is produced; and

(iii) recovering the glycerol produced in (ii).

2. A method according to claim 1 wherein the suitable host cell is selected from the group consisting of bacteria and fungi.

3. A method according to claim 2 wherein the suitable host cell is selected from the group consisting of Citrobacter, Enterobacter, Clostridium, Klebsiella, Lactobacillus, Aspergillus, Saccharomyces, Schizosaccharomyces, Zygosaccharomyces, Pichia, Kluyveromyces, Candida, Mucor, Torulopsis, Methylobacter, Escherichia, Salmonella, Bacillus, Streptomyces, and Pseudomonas.

4. A method according to claim 3 wherein the suitable host cell is E. coli or Saccharomyces.

5. A method according to claim 1 wherein the carbon source is glucose.

6. A method according to claim 1 wherein the gene encoding a NADH-dependent glycerol-3-phosphate dehydrogenase enzyme or NADPH-dependent glycerol-3-phosphate dehydrogenase enzyme encodes the amino acid sequence given in SEQ ID NO:

7, SEQ ID NO;

8, or SEQ ID NO;

10 and wherein the amino acid sequence encompasses amino acid substitutions, deletions or insertions that do not alter the functional properties of the enzyme.