Ethanol production in Gram-positive microbes

• US 5,482,846 A
• Filed: 03/30/1994
• Issued: 01/09/1996
• Est. Priority Date: 08/31/1988
• Status: Expired due to Term

• Current Assignee: University of Florida Research Foundation, Incorporated (State University System of Florida)
• Original Assignee: University of Florida (State University System of Florida)
• Inventors: Ingram, Lonnie O'Neal, Barbosa-Alleyne, Maria D. F.
• Primary Examiner(s): Wax, Robert A.
• Assistant Examiner(s): GRIMES, ERIC BURTON
• Application Number:
  US08/220,072
• Time in Patent Office: 650 Days
• Field of Search: 435/252.3, 435/252.31, 435/161, 435/163, 435/320.1
• US Class Current: 435/161
• CPC Class Codes: C12N 15/01 Preparation of mutants without inserting foreign genetic material therein; Screening processes therefor
• CPC Class Codes: C12N 15/52 Genes encoding for enzymes or proenzymes
• CPC Class Codes: C12N 15/70 Vectors or expression systems specially adapted for E. coli
• CPC Class Codes: C12N 15/74 Vectors or expression systems specially adapted for prokaryotic hosts other than E. coli, e.g. Lactobacillus, Micromonospora
• CPC Class Codes: C12N 15/902 using homologous recombination
• CPC Class Codes: C12N 9/0006 acting on CH-OH groups as donors (1.1)
• CPC Class Codes: C12N 9/0008 acting on the aldehyde or oxo group of donors (1.2)
• CPC Class Codes: C12N 9/88 Lyases (4.)
• CPC Class Codes: C12P 7/065 with microorganisms other than yeasts
• CPC Class Codes: C12P 7/10 substrate containing cellulosic material
• CPC Class Codes: C12Y 101/01001 Alcohol dehydrogenase (1.1.1.1)
• CPC Class Codes: C12Y 102/04001 Pyruvate dehydrogenase (acetyl-transferring) (1.2.4.1)
• CPC Class Codes: G06T 1/00 General purpose image data processing
• CPC Class Codes: Y02E 50/10 Biofuels, e.g. bio-diesel

Abstract

The subject invention concerns the transformation of Gram-positive bacteria with heterologous genes which confer upon these microbes the ability to produce ethanol as a fermentation product. Specifically exemplified is the transformation of bacteria with genes, obtainable from Zymomonas mobilis, which encode pyruvate decarboxylase and alcohol dehydrogenase.

Claims

1. A Gram-positive bacterium selected from the group consisting of Bacillus subtilis and Bacillus polymyxa which has been transformed with Zymomonas mobilis genes encoding alcohol dehydrogenase and pyruvate decarboxylase, wherein said genes are expressed at sufficient levels to confer upon said Gram-positive bacterium transformant the ability to produce ethanol as a fermentation product.

2. A method for the production of ethanol, said method comprising:

a) transforming a Gram-positive bacterial host selected from the group consisting of Bacillus subtilis and Bacillus polymyxa with Zymomonas mobilis genes encoding alcohol dehydrogenase and pyruvate decarboxylase, wherein said genes are expressed at sufficient levels to result in production of ethanol as a fermentation product, and

b) growing said bacterial host so that ethanol is produced.