Aerobic succinate production in bacteria

• US 7,935,511 B2
• Filed: 06/14/2007
• Issued: 05/03/2011
• Est. Priority Date: 11/14/2003
• Status: Expired due to Fees

• Current Assignee: Rice University
• Original Assignee: Rice University
• Inventors: Bennett, George N., Lin, Henry, San, Ka-Yiu
• Primary Examiner(s): Kelly; Robert M
• Assistant Examiner(s): Hiriyanna; Kelaginamane T.
• Application Number:
  US11/763,424
• Publication Number:
  US 20070249028A1
• Time in Patent Office: 1,419 Days
• Field of Search: None
• US Class Current: 435/252.33
• CPC Class Codes: C12N 1/20 Bacteria; Culture media therefor
• CPC Class Codes: C12N 15/52 Genes encoding for enzymes or proenzymes
• CPC Class Codes: C12P 7/46 Dicarboxylic acids having four or less carbon atoms, e.g. fumaric acid, maleic acid

Abstract

Methods of increasing yields of succinate using aerobic culture methods and a multi-mutant E. coli strain are provided. Also provided is a mutant strain of E. coli that produces high amounts of succinic acid.

Claims

1. A mutant strain of E. coli bacteria wherein said mutant strain comprises inactivation of sdhAB, ackA-pta, and poxB.

2. The mutant strain of claim 1, wherein said mutant strain of E. coli bacteria further comprises inactivation of i) ptsG, or ii) iclR, or iii) icd, or iv) both ptsG and iclR, or v) both icd and iclR, or v) ptsG, iclR, and icd.

3. The mutant strain of claim 1, wherein said mutant strain of E. coli bacteria further comprises overexpression of pepc.

4. The mutant strain of claim 1, wherein said mutant strain of E. coli bacteria further comprises inactivation of iclR and overexpresses pepc.

5. The mutant strain of claim 1, wherein said mutant strain of E. coli bacteria further comprises inactivation of ptsG and iclR, and comprises overexpression of pepc.

6. The mutant strain of claim 1, wherein the bacteria produces succinate at a ratio of at least 0.6:

1 moles succinate per mole glucose.

7. A mutant strain of E. coli bacteria, wherein said mutant strain comprises:

i) inactivation of sdhAB, ackA-pta, poxB, and icd, and

ii) overexpression of pepc.

8. The mutant strain of claim 7, wherein the bacteria produces succinate at a ratio of at least 0.6:

1 moles succinate per mole glucose.

9. A mutant strain of E. coli bacteria, wherein said mutant strain comprises:

i) inactivation of sdhAB, ackA-pta, poxB, ptsG, icd, and iclR, and

ii) overexpression of pepc.

10. The mutant strain of claim 9, wherein the bacteria produces succinate at a ratio of at least 0.9:

1 moles succinate per mole glucose.