WO2007084477A1 - Methods and compositions for ethanol producing cyanobacteria - Google Patents
Methods and compositions for ethanol producing cyanobacteria Download PDFInfo
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- WO2007084477A1 WO2007084477A1 PCT/US2007/001071 US2007001071W WO2007084477A1 WO 2007084477 A1 WO2007084477 A1 WO 2007084477A1 US 2007001071 W US2007001071 W US 2007001071W WO 2007084477 A1 WO2007084477 A1 WO 2007084477A1
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- cyanobacterium
- nucleic acid
- ethanol
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- host cell
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- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
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- C12N15/09—Recombinant DNA-technology
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- C12N9/0004—Oxidoreductases (1.)
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- 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
- C12P7/065—Ethanol, i.e. non-beverage with microorganisms other than yeasts
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- 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
Definitions
- Ethanol production from cyanobacteria using sunlight, CO 2 , and inorganic nutrients is an attractive pathway for obtaining a renewable fuel.
- the costs associated with plant growth/harvesting/processing are circumvented, reducing total input energy, and increasing net energy gain.
- the disclosed methods will directly utilize large quantities of CO 2 as a carbon source for fuel production and will thus help reduce this greenhouse gas from the atmosphere.
- homologous recombination refers to the process of recombination between two nucleic acid molecules based on nucleic acid sequence similarity.
- the term embraces both reciprocal and nonreciprocal recombination (also referred to as gene conversion).
- the recombination can be the result of equivalent or non- equivalent cross-over events. Equivalent crossing over occurs between two equivalent sequences or chromosome regions, whereas nonequivalent crossing over occurs between identical (or substantially identical) segments of nonequivalent sequences or chromosome regions. Unequal crossing over typically results in gene duplications and deletions.
- Watson et al. - Molecular Biology of the Gene pp 313-327, The Benjamin/Cummings Publishing Co. 4th ed. (1987).
- Heterologous polynucleotide sequences are considered "exogenous" because they are introduced to the host cell via transformation techniques.
- the heterologous polynucleotide can originate from a foreign source or from the same. source.
- Modification of the heterologous polynucleotide sequence may occur, e.g., by treating the polynucleotide with a restriction enzyme to generate a polynucleotide sequence that can be operably linked to a regulatory element. Modification can also occur by techniques such as site-directed mutagenesis.
- an "expression cassette” or “construct” refers to a series of polynucleotide elements that permit transcription of a gene in a host cell.
- the expression cassette includes a promoter and a heterologous or native polynucleotide sequence that is transcribed.
- Expression cassettes or constructs may also include, e.g., transcription termination signals, polyadenylation signals, and enhancer elements.
- a recombinant expression vector for transformation of a host cell and subsequent integration of the gene(s) of interest is prepared by first isolating the constituent polynucleotide sequences, as discussed herein.
- the gene(s) of interest are homologously integrated into the host cell genome.
- the genes are non-homologously integrated into the host cell genome.
- the gene(s) of interest are homologously integrated into the Synechocystis genome.
- the pMota vector integrates into the psbAII locus via double homologous recombination.
- One preferred method for obtaining specific polynucleotides combines the use of synthetic oligonucleotide primers with polymerase extension or ligation on a mRNA or DNA template.
- a method e.g., RT, PCR, or LCR, amplifies the desired nucleotide sequence (see U.S. Pat. Nos. 4,683,195 and 4,683,202). Restriction endonuclease sites can be incorporated into the primers.
- Amplified polynucleotides are purified and ligated to form an expression cassette. Alterations in the natural gene sequence can be introduced by techniques such as in vitro mutagenesis and PCR using primers that have been designed to incorporate appropriate mutations.
- the promoter is the nirA promoter, which is a 166 base pair Synechococcus sp. strain PCC 7942 promoter sequence described by Qi et al. (2005) in development of an inducible expression vector for Synechocystis sp. PCC 6803. [00421
- the promoter comprises the nucleic acid sequence shown in SEQ ID NO: 2,- which contains the psbAII promoter.
- the promoter comprises the nucleic acid sequence shown in SEQ ID NO: 5.
- the promoter comprises the Synechococcus nirA promoter sequence shown in SEQ ID NO: 6.
- any pdc gene capable of being expressed may be used in the present invention.
- the pdc gene is the Zymomonas mobilis pdc gene.
- the pdc gene is obtained from the Zymomonas mobilis plasmid pLOI295.
- the pdc gene comprises the nucleic acid sequence shown in SEQ ID NO: 3.
- the pdc gene is a nucleic acid sequence encoding the protein shown in SEQ ID NO: 7.
- the pdc gene is a nucleic acid encoding the pdc enzyme obtained from Zymobacter paimae.
- the adh gene is the Zymomonas mobilis adhll gene. In some embodiments, the adh gene is obtained from the Zymomonas mobilis plasmid pLOI295. In some embodiments, the adh gene comprises the nucleic acid sequence shown in SEQ ID NO: 4. In some embodiments, the pdc gene is a nucleic acid sequence encoding the amino acid sequence shown in SEQ ID NO: 9.
- the isolated polynucleotide sequence of choice e.g., the pdc/adh genes driven by the promoter sequence discussed above, is inserted into an "expression vector,” “cloning vector,” or “vector,” terms which usually refer to plasmids or other nucleic acid molecules that are able to replicate in a chosen host cell.
- Expression vectors can replicate autonomously, or they can replicate by being inserted into the genome of the host cell.
- [0049J After construction and isolation of the recombinant expression vector, it is used to transform a host cell for ethanol production.
- the particular procedure used to introduce the genetic material into the host cell for expression of a protein is not particularly critical. Any of the well known procedures for introducing foreign polynucleotide sequences into host cells can be used.
- the host cells can be transformed and screened sequentially via the protocol described by Williams (1988). This method exploits the natural transformability of the Synechocystis sp. PCC 6803 cyanobacteria, where transformation is possible via simple incubation of purified plasmid construct with exponentially growing cells.
- the host cell is incubated under conditions suitable for production of ethanol.
- the host cell will be grown in a photoautotrophic liquid culture in BG-I l media, with a lL/min air sparge rate and a pH setpoint of 8.5, controlled via sparging with CO 2 , and the temperature maintained at 30°C.
- BG-I l media with a lL/min air sparge rate and a pH setpoint of 8.5, controlled via sparging with CO 2 , and the temperature maintained at 30°C.
- Various media for growing cyanobacteria are known in the art. In some embodiments, Synechocystis sp.
- PCC 6803 is cultured on standard BG-1 1 media plates, with or without the addition of (final concentration): 5 mM glucose, 5% sucrose, and/or either 5 ⁇ g ml "1 , 25 ⁇ g ml "1 , or 50 ⁇ g ml "1 kanamycin. Plates containing Synechocystis sp. PCC 6803 were incubated at 30°C under ⁇ 100 microeinsteins m 2 s ⁇ '. All Synechocystis liquid cultures were grown in standard BG-1 1, with the addition of 50 ⁇ g ml "1 kanamycin when appropriate.
- Ethanol concentration levels may reach from about 5 mM to about 15 mM after about 2 to about 5 days of fermentation. In cases where the medium is changed, ethanol concentration levels may reach from about 25 to about 100 mM after 5 days of fermentation.
- the ethanol production level is about 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8 » 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9.0, 9.1, 9.2, 9.3, 9.4 , 9.5, 9.6, 9.7, 9.8, 9.9, 10.0,
- the pH setpoint is about 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9.0, 9.1, 9.2, 9.3, 9.4, or 9.5.
- the temperature during host cell growth may be from about 25°C to 35°C. In some embodiments, the temperature is about 25.0, 25.1, 25.2, 25.3,
- This Example illustrates the construction of a stable cyanobacteria line for production of ethanol.
- Each of the three reactions comprising the PCR assay shared a common upstream primer that lies outside of the psbAII gene loci, while each reaction is defined by the downstream primer that is specific for each of the three possible genetic constructs.
- the upstream primer used in all three reactions was: 5'- gTCAgTTCCAATCTgAACATCGA -3' (SEQ ID NO: 12), with the amplicon beginning 48 bp upstream of the psbAII start codon.
- the downstream primer for probing the WT psbAII gene 5'-
- E. coli was cultured on standard LB formulation in both liquid and solid media. Ampicillin and kanamycin were supplemented to the LB media plates at concentrations of 100 ⁇ g ml "1 and 50 ⁇ g ml "1 , respectively.
- a parallel batch culture system (six 10OmL bioreactors) was established to grow the ethanol-producing Synechocyst ⁇ s strains developed.
- Standard BG-1 1 liquid media was used for the all the experiments. Agitation was set at 400 rpm. Lighting intensity was 200 microeinsteins on the formost face of the bioreactors. Compressed air was sparged to provide CO 2 and remove the oxygen produced by Synechocystis.
- Semi batch operation mode was used to test the ethanol production. The total cell growth period was 20 days.
- This Example illustrates production of ethanol using autotrophic photobioreactor fermentation.
- a 7.5 L BIOFLO ® 110 bioreactor system (New Brunswick Scientific, Edison, NJ) was used for the experiments.
- the 7.5 L BIOFLO ® 110 bioreactor system includes controllers for temperature, pH and dissolved oxygen concentration (DO) adjustment.
- the Synechocystis cultivation process was monitored and controlled automatically by a Pentium ® II (233 MHz, Windows 98) computer equipped with an interface board PCI-MIO- 16E- 10 (National Instruments Corp., Austin, TX).
- the data acquisition program was written in LabVIEW7.1 (National Instruments Corp., Austin, TX).
- the data from the BIOFLO ® 110 bioreactor system, including pH, agitation, temperature and DO were acquired through the computer interface board.
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EP07718106A EP1979481B1 (en) | 2006-01-13 | 2007-01-16 | Methods and compositions for ethanol producing cyanobacteria |
CA2637031A CA2637031C (en) | 2006-01-13 | 2007-01-16 | Methods and compositions for ethanol producing cyanobacteria |
US12/160,770 US8372613B2 (en) | 2006-01-13 | 2007-01-16 | Methods and compositions for ethanol producing cyanobacteria |
DE602007012829T DE602007012829D1 (en) | 2006-01-13 | 2007-01-16 | METHOD AND COMPOSITIONS FOR ETHANOL-PRODUCING CYANOBACTERIA |
AT07718106T ATE500327T1 (en) | 2006-01-13 | 2007-01-16 | METHOD AND COMPOSITIONS FOR ETHANOL-PRODUCING CYANOBACTERIA |
JP2008550460A JP5306823B2 (en) | 2006-01-13 | 2007-01-16 | Methods and compositions for cyanobacteria producing ethanol |
AU2007207635A AU2007207635B2 (en) | 2006-01-13 | 2007-01-16 | Methods and compositions for ethanol producing cyanobacteria |
MX2008008970A MX2008008970A (en) | 2006-01-13 | 2007-01-16 | Methods and compositions for ethanol producing cyanobacteria. |
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WO2008055190A2 (en) | 2006-11-02 | 2008-05-08 | Algenol Biofuels Limited | Closed photobioreactor system for production of ethanol |
WO2009078712A2 (en) * | 2007-12-17 | 2009-06-25 | Universiteit Van Amsterdam | Light-driven co2 reduction to organic compounds to serve as fuels or as industrial half products by an autotroph containing a fermentative gene cassette |
WO2009105714A2 (en) * | 2008-02-22 | 2009-08-27 | James Weifu Lee | Designer oxyphotobacteria and greehouse distillation for photobiological ethanol phoduction from carbon dioxide and water |
WO2009111513A1 (en) | 2008-03-03 | 2009-09-11 | Joule Biotechnologies, Inc. | Engineered co2 fixing microorganisms producing carbon-based products of interest |
WO2010093365A1 (en) * | 2009-02-12 | 2010-08-19 | Helio Biotechnology Corporation | Methods and compositions for genetic engineering of cyanobacteria to produce ethanol |
US7785861B2 (en) | 2007-11-10 | 2010-08-31 | Joule Unlimited, Inc. | Hyperphotosynthetic organisms |
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US8323958B2 (en) | 2006-11-02 | 2012-12-04 | Algenol Biofuels Switzerland GmbH | Closed photobioreactor system for continued daily in situ production of ethanol from genetically enhanced photosynthetic organisms with means for separation and removal of ethanol |
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US20140154762A1 (en) * | 2011-06-24 | 2014-06-05 | Algenol Biofuels Inc. | Genetically Enhanced Cyanobacteria Lacking Functional Genes Conferring Biocide Resistance for the Production of Chemical Compounds |
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US8993303B2 (en) | 2011-02-24 | 2015-03-31 | South Dakota State University | Genetically engineered cyanobacteria |
US9163264B2 (en) | 2008-10-17 | 2015-10-20 | Joule Unlimited Technologies, Inc. | Ethanol production in microorganisms |
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Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
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Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US6306639B1 (en) | 1997-02-19 | 2001-10-23 | Enol Energy Inc. | Genetically modified cyanobacteria for the production of ethanol, the constructs and method thereof |
US20030087368A1 (en) * | 2001-05-04 | 2003-05-08 | University Of Florida | Cloning and sequencing of pyruvate decarboxylase (PDC) genes from bacteria and uses therefor |
Family Cites Families (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5000000A (en) * | 1988-08-31 | 1991-03-19 | University Of Florida | Ethanol production by Escherichia coli strains co-expressing Zymomonas PDC and ADH genes |
US5028539A (en) * | 1988-08-31 | 1991-07-02 | The University Of Florida | Ethanol production using engineered mutant E. coli |
US5270175A (en) * | 1991-07-12 | 1993-12-14 | Dna Plant Technology Corporation | Methods and compositions for producing metabolic products for algae |
US8753840B2 (en) | 2006-10-20 | 2014-06-17 | Arizona Board Of Regents On Behalf Of Arizona State University | Modified cyanobacteria |
-
2007
- 2007-01-16 WO PCT/US2007/001071 patent/WO2007084477A1/en active Application Filing
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Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US6306639B1 (en) | 1997-02-19 | 2001-10-23 | Enol Energy Inc. | Genetically modified cyanobacteria for the production of ethanol, the constructs and method thereof |
US6699696B2 (en) | 1997-02-19 | 2004-03-02 | Enol Energy Inc. | Genetically modified cyanobacteria for the production of ethanol, the constructs and method thereof |
US20030087368A1 (en) * | 2001-05-04 | 2003-05-08 | University Of Florida | Cloning and sequencing of pyruvate decarboxylase (PDC) genes from bacteria and uses therefor |
Non-Patent Citations (9)
Title |
---|
APPL. ENVIRON. MICROBIOL., vol. 68, no. 6, 2002, pages 2869 - 2876 * |
DATABASE GENBANK [online] 19 June 2002 (2002-06-19), XP003016238, accession no. NCBI Database accession no. (M15393) * |
DATABASE GENBANK [online] 26 April 1993 (1993-04-26), XP003016240, accession no. NCBI Database accession no. (M15394) * |
DATABASE GENBANK [online] 9 June 2002 (2002-06-09), XP003016239, accession no. NCBI Database accession no. (AAM49566) * |
DENG M.D. ET AL.: "Ethanol synthesis by genetic engineering in cyanobacteria", APPL. ENVIRON. MICROBIOL., vol. 65, no. 2, February 1999 (1999-02-01), pages 523 - 528, XP003016237 * |
DENG; COLEMAN: "Ethanol Synthesis by Genetic Engineering in Cyanobacteria", APPLIED AND ENVIRONMENTAL MICROBIOLOGY, vol. 65, no. 2, 1999, pages 523 - 428 |
J. BACTERIOL., vol. 169, no. 3, 1987, pages 949 - 954 * |
J. BACTERIOL., vol. 169, no. 6, 1987, pages 2591 - 2597 * |
LI R. ET AL.: "Enhancer activity of light-responsive regulatory elements in the untranslated leader regions of cyanobacterial psbA genes", PROC. NATL. ACAD. SCI. USA, vol. 90, no. 24, 15 December 1993 (1993-12-15), pages 11678 - 11682, XP003016236 * |
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DE602007012829D1 (en) | 2011-04-14 |
EP1979481B1 (en) | 2011-03-02 |
US20090155871A1 (en) | 2009-06-18 |
ATE500327T1 (en) | 2011-03-15 |
AU2007207635B2 (en) | 2013-01-17 |
JP5306823B2 (en) | 2013-10-02 |
ES2343776B1 (en) | 2011-06-24 |
CA2637031C (en) | 2015-12-08 |
EP1979481A4 (en) | 2009-03-25 |
EP1979481A1 (en) | 2008-10-15 |
MX2008008970A (en) | 2008-09-25 |
CN101395273A (en) | 2009-03-25 |
JP2009523424A (en) | 2009-06-25 |
ES2343776A1 (en) | 2010-08-09 |
KR20080113199A (en) | 2008-12-29 |
US8372613B2 (en) | 2013-02-12 |
CA2637031A1 (en) | 2007-07-26 |
AU2007207635A1 (en) | 2007-07-26 |
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