Recombinant zymomonas for pentose fermentation

• US 5,514,583 A
• Filed: 04/15/1994
• Issued: 05/07/1996
• Est. Priority Date: 04/15/1994
• Status: Expired due to Term

• Current Assignee: Alliance For Sustainable Energy LLC
• Original Assignee: Midwest Research Institute
• Inventors: Eddy, Christina K., Deanda, Kristine A., Zhang, Min, Picataggio, Stephen K., Finkelstein, Mark
• Primary Examiner(s): Wax, Robert A.
• Assistant Examiner(s): PROUTY, REBECCA E
• Application Number:
  US08/228,303
• Time in Patent Office: 753 Days
• Field of Search: 435/252.3, 435/243, 435/320.1, 435/822, 435/161, 435/163, 435/165, 536/23.7, 536/23.2, 935/72, 935/60, 935/29
• US Class Current: 435/252.3
• CPC Class Codes: C12N 15/52 Genes encoding for enzymes or proenzymes
• CPC Class Codes: C12N 9/10 Transferases (2.) ribonucleases C12N9/22
• CPC Class Codes: C12N 9/90 Isomerases (5.)
• CPC Class Codes: C12P 7/065 with microorganisms other than yeasts
• CPC Class Codes: Y02E 50/10 Biofuels, e.g. bio-diesel
• CPC Class Codes: Y10S 435/822 using bacteria or actinomycetales

Abstract

The invention relates to microorganisms which normally do not ferment a pentose sugar and which are genetically altered to ferment this pentose to produce ethanol. A representative example is Zymomonas mobilis which has been transformed with E. coli xylose isomerase, xylulokinase, transaldolase and transketolase genes. Expression of the added genes are under the control of Zymomonas mobilis promoters. This newly created microorganism is useful for fermenting pentoses and glucose, produced by hydrolysis of hemicellulose and cellulose, to produce ethanol.

Claims

1. A microorganism of the genus Zymomonas containing exogenous genes encoding xylose isomerase, xylulokinase, transaldolase and transketolase and further comprising at least one promotor recognized by Zymomonas which regulates the expression of at least one of said genes, and wherein said microorganism is capable of growing on xylose as a sole carbon source and fermenting said xylose to ethanol at about 88% theoretical yield and, wherein said microorganism without said genes is incapable of growing on or fermenting said xylose to ethanol.

2. A microorganism according to claim 1, wherein the genes encoding xylose isomerase, xylulokinase, transaldolase and transketolase were obtained from bacteria selected from the group consisting of Xanthomonas, Klebsiella, E. coli, Rhodobacter, Flavobacterium, Acetobacter, Gluconobacter, Rhizobium, Agrobacterium, Salmonella and Pseudomonads.

3. A microorganism according to claim 1, wherein the genes are integrated into the host genome.

4. A microorganism according to claim 1, wherein said genes are contained on a vector.

5. A microorganism according to claim 4, wherein said xylose isomerase and xylulokinase genes are expressed under the control of a glyceraldehyde-3-phosphate dehydrogenase promoter recognized by Zymomonas and said transaldolase and transketolase genes are expressed under the control of a enolase promoter recognized by Zymomonas.

9. A microorganism according to claim 2, wherein the genes encoding xylose isomerase, xylulokinase,transaldolase and transketolase are obtained from E. coli.

6. A vector comprising genes encoding xylose isomerase, xylulokinase, transaldolase and transketolase and at least one promoter recognized by Zymomonas which regulates expression of said genes.

7. A vector according to claim 6, wherein the enzymes transaldolase and transketolase are regulated by an enolase promoter recognized by Zymomonas.

8. A vector according to claim 6, wherein the genes encoding xylose isomerase and xylulokinase are regulated by a glyceraldehyde-3-phosphate, dehydrogenase promoter recognized by Zyomomonas.