Method for construction of bacterial strains with increased succinic acid production

• US 6,159,738 A
• Filed: 04/28/1998
• Issued: 12/12/2000
• Est. Priority Date: 04/28/1998
• Status: Expired due to Fees

• Current Assignee: U Chicago Argonne, LLC
• Original Assignee: University of Chicago
• Inventors: Chatterjee, Ranjini, Sanville-Millard, Cynthia, Donnelly, Mark I.
• Primary Examiner(s): Prouty, Rebecca E.
• Assistant Examiner(s): Hutson, Richard
• Application Number:
  US09/067,931
• Time in Patent Office: 959 Days
• Field of Search: 435/145, 435/252.3, 435/471, 435/140, 435/252.31, 435/252.33, 435/252.9, 435/440, 435/472, 536/23.2, 536/23.1, 536/23.7
• US Class Current: 435/471
• CPC Class Codes: C12N 15/70 Vectors or expression systems specially adapted for E. coli
• CPC Class Codes: C12N 15/902 using homologous recombination
• CPC Class Codes: C12P 7/46 Dicarboxylic acids having four or less carbon atoms, e.g. fumaric acid, maleic acid

Abstract

A fermentation process for producing succinic acid is provided comprising selecting a bacterial strain that does not produce succinic acid in high yield, disrupting the normal regulation of sugar metabolism of said bacterial strain, and combining the mutant bacterial strain and selected sugar in anaerobic conditions to facilitate production of succinic acid. Also provided is a method for changing low yield succinic acid producing bacteria to high yield succinic acid producing bacteria comprising selecting a bacterial strain having a phosphotransferase system and altering the phosphotransferase system so as to allow the bacterial strain to simultaneously metabolize different sugars.

Claims

1. A method for changing fermentative bacteria to succinic acid producing bacteria comprising:

a) selecting a bacterial strain having a phosphotransferase system; and

b) altering the phosphotransferase system so as to allow the bacterial strain to simultaneously metabolize different sugars, wherein the alteration of said phosphotransferase system involves homologous recombination of a defective ptsG gene from strain LA-12G.

2. The method as recited in claim 1 wherein the bacteria are selected from the group consisting of E. coli Klebsiella, Erwinia, and Lactobacillus.

3. The method as recited in claim 1 wherein the step of altering the phosphotransferase system further includes the step of homologously recombining the defective phosphotransferase gene with a portion of the bacteria'"'"'s chromosome.

4. The method as recited in claim 3 wherein the recombination occurs via phage transduction.

5. The method as recited in claim 1 wherein the sugars are selected from the group consisting of glucose, sorbitol, xylose, arabinose, mannose, lactose, glucuronic acid, and combinations thereof.

6. A method for increasing succinic acid production in a bacterium, comprising:

a) selecting a bacteria having a phosphotransferase system containing a ptsG gene;

b) creating a mutation in the ptsG gene which allows the bacteria to metabolize sugars other than glucose in the presence of glucose; and

c) selecting those bacteria with increased succinic acid production.

7. The method as recited in claim 6 wherein the bacteria are selected from the group consisting of E. coli, Klebsiella, Erwinia, and Lactobacillus.

8. The method as recited in claim 6 wherein the sugar is selected from the group consisting of glucose, sorbitol, xylose, arabinose, mannose, lactose, glucuronic acid, and combinations thereof.

9. The method as recited in claim 7 wherein the ptsG mutation resides in the 10-30 minute region of the E. coli chromosome.