CA2173793A1 - Pentose fermentation by recombinant zymomonas - Google Patents
Pentose fermentation by recombinant zymomonasInfo
- Publication number
- CA2173793A1 CA2173793A1 CA002173793A CA2173793A CA2173793A1 CA 2173793 A1 CA2173793 A1 CA 2173793A1 CA 002173793 A CA002173793 A CA 002173793A CA 2173793 A CA2173793 A CA 2173793A CA 2173793 A1 CA2173793 A1 CA 2173793A1
- Authority
- CA
- Canada
- Prior art keywords
- microorganism
- transketolase
- transaldolase
- gene
- exogenous
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 150000002972 pentoses Chemical class 0.000 title claims abstract 14
- 238000000855 fermentation Methods 0.000 title claims abstract 4
- 230000004151 fermentation Effects 0.000 title claims abstract 4
- 241000588901 Zymomonas Species 0.000 title claims 4
- 108090000623 proteins and genes Proteins 0.000 claims abstract 37
- 244000005700 microbiome Species 0.000 claims abstract 32
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract 26
- 102100028601 Transaldolase Human genes 0.000 claims abstract 15
- 108020004530 Transaldolase Proteins 0.000 claims abstract 15
- 102000014701 Transketolase Human genes 0.000 claims abstract 15
- 108010043652 Transketolase Proteins 0.000 claims abstract 15
- 241000588902 Zymomonas mobilis Species 0.000 claims abstract 14
- 108010018080 L-arabinose isomerase Proteins 0.000 claims abstract 10
- 108090000416 L-ribulose-5-phosphate 4-epimerases Proteins 0.000 claims abstract 10
- 108020002667 ribulokinase Proteins 0.000 claims abstract 10
- 108700040099 Xylose isomerases Proteins 0.000 claims abstract 5
- 241000588724 Escherichia coli Species 0.000 claims abstract 4
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims abstract 4
- 102100029089 Xylulose kinase Human genes 0.000 claims abstract 4
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 claims abstract 4
- 229920002678 cellulose Polymers 0.000 claims abstract 4
- 239000001913 cellulose Substances 0.000 claims abstract 4
- 239000008103 glucose Substances 0.000 claims abstract 4
- 108091022915 xylulokinase Proteins 0.000 claims abstract 4
- 238000000034 method Methods 0.000 claims 17
- 102000004190 Enzymes Human genes 0.000 claims 9
- 108090000790 Enzymes Proteins 0.000 claims 9
- 229940088598 enzyme Drugs 0.000 claims 9
- PYMYPHUHKUWMLA-UHFFFAOYSA-N arabinose Natural products OCC(O)C(O)C(O)C=O PYMYPHUHKUWMLA-UHFFFAOYSA-N 0.000 claims 7
- SRBFZHDQGSBBOR-UHFFFAOYSA-N beta-D-Pyranose-Lyxose Natural products OC1COC(O)C(O)C1O SRBFZHDQGSBBOR-UHFFFAOYSA-N 0.000 claims 7
- SRBFZHDQGSBBOR-IOVATXLUSA-N Xylose Natural products O[C@@H]1COC(O)[C@H](O)[C@H]1O SRBFZHDQGSBBOR-IOVATXLUSA-N 0.000 claims 5
- 108010059892 Cellulase Proteins 0.000 claims 3
- 230000003301 hydrolyzing effect Effects 0.000 claims 3
- 102000012288 Phosphopyruvate Hydratase Human genes 0.000 claims 2
- 108010022181 Phosphopyruvate Hydratase Proteins 0.000 claims 2
- PYMYPHUHKUWMLA-WDCZJNDASA-N arabinose Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)C=O PYMYPHUHKUWMLA-WDCZJNDASA-N 0.000 claims 2
- 125000000089 arabinosyl group Chemical group C1([C@@H](O)[C@H](O)[C@H](O)CO1)* 0.000 claims 2
- 102000006602 glyceraldehyde-3-phosphate dehydrogenase Human genes 0.000 claims 2
- 108020004445 glyceraldehyde-3-phosphate dehydrogenase Proteins 0.000 claims 2
- 239000013612 plasmid Substances 0.000 claims 1
- 125000000969 xylosyl group Chemical group C1([C@H](O)[C@@H](O)[C@H](O)CO1)* 0.000 claims 1
- 229920002488 Hemicellulose Polymers 0.000 abstract 1
- 230000007062 hydrolysis Effects 0.000 abstract 1
- 238000006460 hydrolysis reaction Methods 0.000 abstract 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/02—Preparation of oxygen-containing organic compounds containing a hydroxy group
- C12P7/04—Preparation of oxygen-containing organic compounds containing a hydroxy group acyclic
- C12P7/06—Ethanol, i.e. non-beverage
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02E—REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
- Y02E50/00—Technologies for the production of fuel of non-fossil origin
- Y02E50/10—Biofuels, e.g. bio-diesel
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Life Sciences & Earth Sciences (AREA)
- Genetics & Genomics (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Biotechnology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Microbiology (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Tropical Medicine & Parasitology (AREA)
- Virology (AREA)
- Medicinal Chemistry (AREA)
- Physics & Mathematics (AREA)
- Biophysics (AREA)
- Molecular Biology (AREA)
- Plant Pathology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention relates to microorganisms which normally do not ferment pentose sugar and which are genetically altered to ferment pentose sugar to produce ethanol, and fermentation processes utilizing the same. Examples include Zymomonas mobilis which has been transformed with combinations of E. coli genes for xylose isomerase, xylulokinase, transaldolase, transketolase, L-arabinose isomerase, L-ribulokinase, and L-ribulose 5-phosphate 4-epimerase. Expression of the added genes are under the control of Zymomonas mobilis promoters. These newly created microorganisms are useful for fermenting pentoses and glucose, produced by hydrolysis of hemicellulose and cellulose, to produce ethanol.
Claims (34)
1. A microorganism of the genus Zymomonas containing one or more exogenous genes and is capable of fermenting a pentose sugar to ethanol, wherein said microorganism without said gene is incapable of fermenting said pentose sugar to ethanol.
2. The microorganism of claim 1, wherein said pentose sugar is arabinose.
3. The microorganism of claim 2, wherein said exogenous gene encodes an enzyme selected from the group consisting of: L-arabinose isomerase, L-ribulokinase, L-ribulose 5-phosphate 4-epimerase, transaldolase and transketolase.
4. The microorganism of claim 1, wherein said exogenous gene encodes an enzyme selected from the group consisting of: transaldolase and transketolase, and is expressed under the control of the Z.mobilis enolase promoter.
5. The microorganism of claim 2, wherein said exogenous gene encodes an enzyme selected from the group consisting of:
L-arabinose isomerase, L-ribulokinase, and L-ribulose 5-phosphate 4-epimerase, and is expressed under the control of the Z. mobilis glyceraldehyde-3-phosphate dehydrogenase promoter.
L-arabinose isomerase, L-ribulokinase, and L-ribulose 5-phosphate 4-epimerase, and is expressed under the control of the Z. mobilis glyceraldehyde-3-phosphate dehydrogenase promoter.
6. The microorganism of claim 3, wherein said exogenous genes are obtained from E. coli.
7. The microorganism of claim 3, wherein said exogenous gene is expressed under the control of one or more promoters located on said gene, whereby said gene is expressed in said microorganism.
8. The microorganism of claim 6, wherein the gene is integrated into the host genome.
9. The microorganism of claim 6, wherein said gene is contained on a vector.
10. The microorganism of claim 9, wherein said vector is a plasmid.
11. A vector comprising one or more genes encoding an enzyme selected from the group consisting of: L-arabinose isomerase, L-ribulokinase, L-ribulose 5-phosphate 4-epimerase, transaldolase and transketolase, and at least one promotor recognized by Zymomonas which regulates the expression of the gene.
12. The vector of claim 11, comprising genes which encode L-arabinose isomerase, L-ribulokinase, L-ribulose 5-phosphate 4-epimerase.
13. The vector of claim 11, comprising genes which encode transaldolase and transketolase.
14. The microorganism of claim 2, which contains the vector of claim 11.
15. The microorganism of claim 3, wherein the microorganism is Zymomonas mobilis.
16. The microorganism of claim 6, containing exogenous genes which encode L-arabinose isomerase, L-ribulokinase, and L-ribulose 5-phosphate 4-epimerase, expressed under the control of the Z. mobilis glyceraldehyde-3-phosphate dehydrogenase promoter, and exogenous genes which encode transaldolase and transketolase, expressed under the control of the Z.mobilis enolase promoter.
17. The microorganism of claim 14, wherein said genes are coordinately expressed.
18. A process for producing ethanol comprising, providing a feedstock containing pentose sugar, adding a microorganism of the genus Zymomonas to the feedstock wherein the microorganism contains at least one exogenous gene which imparts pentose sugar to ethanol fermentation capability to the microorganism and wherein said microorganism without said gene is incapable of fermenting said pentose sugar to ethanol, allowing the microorganism to ferment the pentose sugar in the feedstock to ethanol, and separating the ethanol.
19. The process of claim 18, wherein said exogenous gene encodes transaldolase or transketolase.
20. The process of claim 18, wherein said pentose sugar is arabinose.
21. The process of claim 20, wherein said exogenous gene encodes an enzyme selected from the group consisting of: L-arabinose isomerase, L-ribulokinase and L-ribulose 5-phosphate 4-epimerase, transaldolase and transketolase.
22. The process of claim 18, wherein said pentose sugar is xylose.
23. The process of claim 22, wherein said exogenous gene encodes an enzyme selected from the group consisting of: xylose isomerase, xylulokinase, transaldolase and transketolase.
24. The process of claim 18, wherein said exogenous gene is expressed under the control of one or more promoters located on said gene, whereby said gene is expressed in said microorganism to confer upon said microorganism an ability to ferment pentose sugar directly to ethanol.
25. The process of claim 18, wherein the exogenous gene is integrated into the host genome.
26. The process of claim 18, wherein the exogenous gene is located on a vector.
27. The process of claim 18, wherein the microorganism is Zymomonas mobilis.
28. A process for producing ethanol comprising, providing a mixed sugar feedstock, adding one or more microorganism containing an exogenous gene to the feedstock, allowing fermentation to ethanol of sugar contained in the feedstock to occur, and separating the ethanol, wherein the microorganism is selected from the group consisting of: Zymomonas mobilis containing exogenous genes from E. coli which encode L-arabinose isomerase, L-ribulokinase, and L-ribulose 5-phosphate 4-epimerase, and transaldolase and transketolase, and Zymomonas mobilis containing exogenous genes from E. coli which encode xylose isomerase and xylolukinase, and transaldolase and transketolase.
29. The process of claim 11 wherein the microorganism is Zymomonas mobilis containing exogenous genes encoding L-arabinose isomerase, L-ribulokinase, L-ribulose 5-phosphate 4-epimerase, transaldolase and transketolase, and the mixed sugar feedstock contains arabinose and glucose.
30. The process of claim 28 wherein the microorganism is Zymomonas mobilis containing exogenous genes encoding xylose isomerase, xylulokinase, transaldolase and transketolase and the mixed sugar feedstock contains xylose and glucose.
31. The process of claim 28 wherein the microorganism is Zymomonas mobilis containing exogenous genes encoding L-arabinose isomerase, L-ribulokinase, L-ribulose 5-phosphate 4-epimerase, transaldolase and transketolase, and Zymomonas mobilis containing exogenous genes encoding xylose isomerase, xylulokinase, transaldolase and transketolase and the mixed sugar feedstock contains xylose, arabinose and glucose.
32. The process of claim 29, further comprising the addition of hydrolytic enzymes such as cellulase enzyme complex and the mixed sugar feedstock also contains cellulose.
33. The process of claim 30, further comprising the addition of hydrolytic enzymes such as cellulase enzyme complex and the mixed sugar feedstock also contains cellulose.
34. The process of claim 14, further comprising the addition of hydrolytic enzymes such as cellulase enzyme complex and the mixed sugar feedstock also contains cellulose.
Applications Claiming Priority (4)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US08/421,996 US5726053A (en) | 1994-04-15 | 1995-04-14 | Recombinant Zymomonas for pentose fermentation |
| US08/422,424 | 1995-04-14 | ||
| US08/422,424 US5712133A (en) | 1994-04-15 | 1995-04-14 | Pentose fermentation by recombinant zymomonas |
| US08/421,996 | 1995-04-14 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CA2173793A1 true CA2173793A1 (en) | 1996-10-15 |
| CA2173793C CA2173793C (en) | 2011-01-11 |
Family
ID=27025447
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA2173793A Expired - Lifetime CA2173793C (en) | 1995-04-14 | 1996-04-10 | Pentose fermentation by recombinant zymomonas |
Country Status (3)
| Country | Link |
|---|---|
| KR (1) | KR100455096B1 (en) |
| AU (1) | AU698662B2 (en) |
| CA (1) | CA2173793C (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| AU774414B2 (en) * | 1997-05-06 | 2004-06-24 | Midwest Research Institute | Single (zymomonas mobilis) strain for xylose and arabinose fermentation |
-
1996
- 1996-04-10 CA CA2173793A patent/CA2173793C/en not_active Expired - Lifetime
- 1996-04-11 AU AU50582/96A patent/AU698662B2/en not_active Expired
- 1996-04-13 KR KR1019960011100A patent/KR100455096B1/en not_active IP Right Cessation
Also Published As
| Publication number | Publication date |
|---|---|
| KR960037834A (en) | 1996-11-19 |
| KR100455096B1 (en) | 2005-01-13 |
| CA2173793C (en) | 2011-01-11 |
| AU5058296A (en) | 1996-10-24 |
| AU698662B2 (en) | 1998-11-05 |
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Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| EEER | Examination request | ||
| MKEX | Expiry |
Effective date: 20160411 |