WO2010120000A1 - Procédé d'obtention d'éthanol ou d'un acide organique au moyen d'un micro-organisme procaryotique avec un gène faba surexprimé - Google Patents

Procédé d'obtention d'éthanol ou d'un acide organique au moyen d'un micro-organisme procaryotique avec un gène faba surexprimé Download PDF

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Publication number
WO2010120000A1
WO2010120000A1 PCT/KR2009/001936 KR2009001936W WO2010120000A1 WO 2010120000 A1 WO2010120000 A1 WO 2010120000A1 KR 2009001936 W KR2009001936 W KR 2009001936W WO 2010120000 A1 WO2010120000 A1 WO 2010120000A1
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WIPO (PCT)
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gene
faba
ethanol
organic acid
present
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PCT/KR2009/001936
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English (en)
Korean (ko)
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김철호
서정우
라련화
서필수
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한국생명공학연구원
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Priority to PCT/KR2009/001936 priority Critical patent/WO2010120000A1/fr
Publication of WO2010120000A1 publication Critical patent/WO2010120000A1/fr

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    • 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
    • C12P7/065—Ethanol, i.e. non-beverage with microorganisms other than yeasts
    • 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
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/88—Lyases (4.)
    • 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
    • C12P7/40—Preparation of oxygen-containing organic compounds containing a carboxyl group including Peroxycarboxylic acids
    • 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

  • the present invention relates to a method for producing ethanol or organic acid using an ethanol resistant strain, and more specifically, a recombinant prokaryotic nucleus overexpressed with ⁇ -hydroxydecanoyl thio ester dehydrase gene ( fabA). It relates to a method for producing ethanol or organic acid, characterized in that the use of microorganisms.
  • Saccharomyces cerevisiae a representative ethanol-producing strain, is resistant to ethanol when the unsaturated fatty acid generating gene is overexpressed or the fatty acid is added to the medium artificially to increase the amount of unsaturated fatty acid. It has been shown to improve ( Appl. Environ. Microbiol . 69: 1499-1503, 2003; Appl. Environ. Microbiol . 1996, 62: 4309, 1996).
  • ⁇ -Hydroxydecanoyl thio ester dehydrase (beta-hydroxyldecanoyl thio ester dehydrase, fabA) is a trans-beta-hydroxyl-Acid during intracellular fatty acid synthesis.
  • FabA FabA enzyme
  • delta 5-unsaturase derived from Bacillus subtilis ( ⁇ 5 desaturase, des) is known to have a function of introducing a double bond to the saturated fatty acids present in the cell membrane, the cell membrane when overexpressing the des gene in E. coli It has been reported to increase the contents of unsaturated citric acid 5-cis-palmitoleic acid (16: 1 ⁇ 5) and epedrenic acid (18: 2 ⁇ 5 ⁇ 11) ( J. Bacteriol ., 180: 2194, 1998).
  • the present inventors pay attention to the fact that when the microbial strain is exposed to alcohols, the ratio of unsaturated fatty acids in the cell membrane is increased by a biological defense mechanism, thereby artificially modifying the ratio of saturated fatty acids and unsaturated fatty acids in these cell membranes to obtain resistance to ethanol.
  • the study was conducted under the assumption that it could be.
  • the present inventors have made intensive efforts to develop prokaryotic microorganisms having high ethanol resistance.
  • the present inventors confirmed that recombinant E. coli overexpressing the ⁇ -hydroxydecanoyl thio ester dehydrase gene has resistance to high concentrations of ethanol and the present invention. To complete.
  • An object of the present invention is to provide a method for producing ethanol or organic acid using ethanol-resistant prokaryotic microorganisms that can grow in a high concentration of ethanol-containing medium.
  • the present invention comprises the steps of culturing a prokaryote microorganism overexpressing the ⁇ -hydroxydecanoyl thio ester dehydrase gene ( fabA) ; And it provides a method for producing ethanol or organic acid comprising the step of obtaining ethanol from the culture.
  • the present invention comprises the steps of culturing a prokaryotic microorganism overexpressing the ⁇ -hydroxydecanoyl thio ester dehydrase gene ( fabA) ; And it provides a method for producing an organic acid comprising the step of obtaining an organic acid from the culture.
  • Figure 1 shows the manufacturing process of a recombinant vector containing the fabA gene and the des gene according to the present invention.
  • Figure 2 shows the growth curve of the ethanol concentration in the medium of the recombinant strain according to the present invention.
  • the present invention provides a method for preparing a prokaryotic microorganism in which the ⁇ -hydroxydecanoyl thio ester dehydrase gene ( fabA) is overexpressed; And it relates to a method for producing ethanol or organic acid comprising the step of recovering ethanol or organic acid from the culture.
  • prokaryotic microorganisms overexpressed the ⁇ -hydroxydecanoyl thio ester dehydrase gene are ⁇ -hydroxydecanoyl thio ester dehydrase gene ( ⁇ -hydroxydecanoyl thio ester dehydrase gene: fabA) is or transformed with the inserted vector, ⁇ - hydroxy decanoyl thioester di Hydra dehydratase gene ( ⁇ -hydroxydecanoyl dehydrase gene thio ester: fabA) is inserted in a recombinant prokaryotic chromosome Microorganisms can be used.
  • Saccharomyces cerevisiae a representative ethanol-producing strain, is resistant to ethanol when an unsaturated fatty acid gene is overexpressed or an unsaturated fatty acid is added to the medium to artificially increase the amount of unsaturated fatty acid. It has been shown to improve ( Appl. Environ. Microbiol . 69: 1499-1503, 2003; Appl. Environ. Microbiol . 1996, 62: 4309, 1996).
  • E. coli overexpresses the ⁇ -hydroxydecanoyl thio ester dehydrase (FabA) gene to increase the content of saturated fatty acids and decrease the content of unsaturated fatty acids.
  • FAA ⁇ -hydroxydecanoyl thio ester dehydrase
  • Recombinant Escherichia coli E. coli / pBR-fabA was prepared and examined for ethanol resistance, interestingly it was confirmed that the resistance to ethanol is improved compared to the control. Therefore, in the present invention, the prokaryotic microorganism may be characterized as E. coli.
  • the method for recovering ethanol or organic acid from the culture medium of the variant may use a conventional separation technique, for example, distillation, electrodialysis, pervaporation, chromatography, solvent extraction, reaction extraction, etc. In order to separate substances of high purity, these may be used in combination.
  • the gene can be introduced into a variety of vectors, including plasmid vector, bacteriophage vector, cosmid vector, YAC (Yeast Artificial Chromosome) vector.
  • plasmid vectors include plasmid vector, bacteriophage vector, cosmid vector, YAC (Yeast Artificial Chromosome) vector.
  • YAC yeast Artificial Chromosome
  • Typical plasmid vectors that can be used for such purposes include (a) a replication initiation point that allows for efficient replication to include hundreds of plasmid vectors per host cell, and (b) host cells transformed with the plasmid vector. It has a structure comprising an antibiotic resistance gene and (c) a restriction enzyme cleavage site into which foreign DNA fragments can be inserted. Although no appropriate restriction enzyme cleavage site is present, the use of synthetic oligonucleotide adapters or linkers according to conventional methods can facilitate ligation of the vector and foreign DNA.
  • Various vectors including the T-vector used in the present invention, contain a multicloning site (MCS), which is a collection of several restriction enzyme cleavage sites, which are located in the lacZ gene. Since MCS does not destroy the reading frame of the lacZ gene, lacZ is expressed in a suitable host cell to synthesize ⁇ -galaccosidase enzyme having biological activity. When the host cell is cultured in a medium containing X-gal, a colorless chemical, the enzyme breaks down X-gal and creates an insoluble blue substance. Therefore, host cell colonies transformed with plasmid vectors without insertion of foreign DNA into MCS are blue.
  • MCS multicloning site
  • the vector can be used more usefully because it confirms whether the DNA desired to be integrated on the slide is normally cloned into the vector.
  • the vector should be transformed into the appropriate host cell.
  • the preferred host cell is a prokaryotic cell.
  • Suitable prokaryotic host cells include E. coli strain DH5a, E. coli strain JM101, E. coli K12 strain 294, E. coli strain W3110, E. coli strain X1776, E. coli XL-1Blue (Stratagene) and E. coli B And the like.
  • E. coli strains such as FMB101, NM522, NM538 and NM539 and other prokaryotic species and genera may also be used.
  • E. coli strains such as FMB101, NM522, NM538 and NM539 and other prokaryotic species and genera may also be used.
  • E. coli strains such as FMB101, NM522, NM538 and NM539 and other prokaryotic species and genera may also be used.
  • E. coli strains such as FMB101, NM522, NM538
  • Bacillus subtilis Bacillus subtilis with the same Bashile (bacilli), S. typhimurium (Salmonella typhimurium) or Serratia dry sense another enteric bacteria, and various Pseudomonas (Pseudomonas) in the strain, such as (Serratia marcescens) as a host cell Can be used.
  • Bacillus subtilis Bacillus subtilis with the same Bashile (bacilli), S. typhimurium (Salmonella typhimurium) or Serratia dry sense another enteric bacteria, and various Pseudomonas (Pseudomonas) in the strain, such as (Serratia marcescens) as a host cell Can be used.
  • Prokaryotic transformation can be readily accomplished using the calcium chloride method described in section 1.82 of Sambrook et al., Supra.
  • electroporation EMBO J. , 1: 841,1982
  • EMBO J. , 1: 841,1982 can also be used to transform these cells.
  • the method for inserting the gene on the chromosome of the host cell in the present invention can be used a commonly known genetic engineering method, for example retrovirus vector, adenovirus vector, adeno-associated virus vector, herpes simplex virus vector , Poxvirus vectors, lentiviral vectors or non-viral vectors.
  • retrovirus vector for example retrovirus vector, adenovirus vector, adeno-associated virus vector, herpes simplex virus vector , Poxvirus vectors, lentiviral vectors or non-viral vectors.
  • an expression vector known in the art may be used, and it is preferable to use a pET family vector (Novagen).
  • a pET family vector Novagen
  • histidine groups are bound to the ends of the expressed protein, and thus the protein can be effectively purified.
  • a general method known in the art may be used, and specifically, it may be separated by a chromatographic method using Ni-NTA His-binding resin (Novagen). .
  • expression “expression control sequence” in the present invention means a DNA sequence essential for the expression of a coding sequence operably linked in a particular host organism.
  • regulatory sequences include promoters for carrying out transcription, any operator sequence for regulating such transcription, sequences encoding suitable mRNA ribosomal binding sites, and sequences that control the termination of transcription and translation.
  • suitable control sequences for prokaryotes include promoters, optionally operator sequences, and ribosomal binding sites.
  • Eukaryotic cells include promoters, polyadenylation signals, and enhancers. The factor that most influences the amount of gene expression in the plasmid is the promoter.
  • an SR ⁇ promoter As the promoter for high expression, an SR ⁇ promoter, a promoter derived from a cytomegalovirus, and the like are preferably used.
  • any of a wide variety of expression control sequences can be used in the vector.
  • Useful expression control sequences include, for example, early and late promoters of SV40 or adenovirus, lac system, trp system, TAC or TRC system, T3 and T7 promoters, major operator and promoter regions of phage lambda, fd code protein Regulatory region of, promoter for 3-phosphoglycerate kinase or other glycolysis enzymes, promoters of the phosphatase such as Pho5, promoter of yeast alpha-crossing system and gene expression of prokaryotic or eukaryotic cells or viruses Other sequences known to modulate and various combinations thereof.
  • the T7 promoter can be usefully used to express the proteins of the invention in E. coli.
  • Nucleic acids are "operably linked” when placed in a functional relationship with other nucleic acid sequences. This may be genes and regulatory sequence (s) linked in such a way as to allow gene expression when appropriate molecules (eg, transcriptional activating proteins) bind to regulatory sequence (s).
  • DNA for a pre-sequence or secretion leader is operably linked to DNA for a polypeptide when expressed as a shear protein that participates in the secretion of the polypeptide;
  • a promoter or enhancer is operably linked to a coding sequence when it affects the transcription of the sequence;
  • the ribosomal binding site is operably linked to a coding sequence when it affects the transcription of the sequence;
  • the ribosomal binding site is operably linked to a coding sequence when positioned to facilitate translation.
  • "operably linked” means that the linked DNA sequence is in contact, and in the case of a secretory leader, is in contact and present within the reading frame.
  • enhancers do not need to touch. Linking of these sequences is performed by ligation (linking) at convenient restriction enzyme sites. If such sites do not exist, synthetic oligonucleotide adapters or linkers according to conventional methods are used.
  • expression vector generally refers to a fragment of DNA that is generally double stranded as a recombinant carrier into which fragments of heterologous DNA have been inserted.
  • heterologous DNA refers to heterologous DNA, which is DNA not naturally found in host cells.
  • the gene must be operably linked to transcriptional and translational expression control sequences that function in the selected expression host.
  • the expression control sequence and the gene of interest are included in one expression vector including the bacterial selection marker and the replication origin. If the host cell is a eukaryotic cell, the expression vector must further comprise an expression marker useful in the eukaryotic expression host.
  • the beta-hydroxyacyl- ACE dehydratase gene ( fabA ) was amplified by PCR.
  • SEQ ID NO: 2 5- GGATCC GCTAGC TCAGGCATTCTTCCGC-3 (fabA- BamHI - NheI -R)
  • the ⁇ 5 desaturase gene ( des ) was amplified by PCR, and the following DNA fragments were used as primer sets.
  • SEQ ID NO: 4 5- GGATCC CTCGAGTCAGGCATTCTTCCGC-3 ( des- BamHI -R)
  • the amplified fabA or des gene DNA fragments were cloned using pGEM T Easy vector (Promega, USA) to confirm the nucleotide sequence.
  • pGEM T Easy vector Promega, USA
  • the following primers were used to insert a lacZ promoter (PlacZ), a potent promoter upstream of each gene (FIG. 1).
  • SEQ ID NO: 6 5- GGATCC TCTAGA GCTGTTTCCTGTGT-3 (PlacZ-fabA- BamHI - XbaI -R)
  • SEQ ID NO: 8 5- GGATCC TCTAGA GCTGTTTCCTGTGT-3 (PlacZ-des- BamHI - XbaI -R)
  • the pBR-fabA and pBR-des plasmids were prepared by recloning the fabA or des genes linked to the lacZ promoter with pBR322 vectors, respectively, and at the same time, the pBR-fabA-des plasmid was prepared to express the fabA and des genes simultaneously.
  • the pBR-fabA, pBR-des and pBR-fabA-des plasmids were introduced into E. coli DH5 ⁇ by thermal shock, respectively, to prepare a recombinant strain, and a control strain was prepared by introducing an expression vector pBR322.
  • the recombinant strains were 10 ⁇ g / Cell proliferation was confirmed by culturing in LB medium containing ml tetracycline, 0.5 mM IPTG and ethanol at concentrations of 0%, 1%, 2% and 3%, respectively. Culture conditions were culture volume, 50 ml / 250 ml, culture temperature 37 ° C, suspension speed 120 rpm. Inoculation of 1 ml of the preculture strain grown at 600 nm to absorbance 2.0 to investigate the growth of the cells at 3 hour intervals.
  • the cell proliferation of the control at 2% ethanol concentration was sharply reduced, whereas the recombinant strain (pBR-fabA) overexpressing the fabA gene showed ethanol resistance and showed continuous cell growth. Meanwhile, the recombinant strain (pBR-des) overexpressed the des gene was more sensitive to ethanol than the control. Recombinant strain (pBR-fabA-des) overexpressing both fabA and des genes showed similar ethanol resistance as control. At 3% ethanol concentration, the cell proliferation of all the recombinant strains was severely inhibited, but the ethanol resistance pattern was maintained as it is.
  • the recombinant Escherichia coli (pBR-fabA) overexpressed the fabA gene was found to have a higher content of saturated fatty acid (SFA) and lower content of unsaturated fatty acid (UFA) compared to the control (pBR322).
  • SFA saturated fatty acid
  • UFA unsaturated fatty acid
  • fabA and des genes were overexpressed at the same time, the effect of each gene on fatty acid composition was found to be offset.

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Abstract

Procédé d'obtention d'éthanol ou d'un acide organique au moyen de souches résistant à l'éthanol et, plus particulièrement, procédé d'obtention d'éthanol ou d'un acide organique au moyen d'un micro-organisme procaryotique de recombinaison avec fabA (gène de β- hydroxydécanoyl thio ester déshydrase : fabA).Selon la présente invention, de l'éthanol fortement concentré et un acide organique peuvent être fabriqués dans la mesure où la croissance des souches résistantes n'est pas inhibée au cours de la fermentation, et ceci même lorsque l'éthanol fortement concentré produit par le micro-organisme s'accumule dans le milieu de culture.
PCT/KR2009/001936 2009-04-15 2009-04-15 Procédé d'obtention d'éthanol ou d'un acide organique au moyen d'un micro-organisme procaryotique avec un gène faba surexprimé WO2010120000A1 (fr)

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PCT/KR2009/001936 WO2010120000A1 (fr) 2009-04-15 2009-04-15 Procédé d'obtention d'éthanol ou d'un acide organique au moyen d'un micro-organisme procaryotique avec un gène faba surexprimé

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112143690A (zh) * 2019-06-28 2020-12-29 中国科学院青岛生物能源与过程研究所 一种酸耐受性提高的重组菌及其构建方法和应用

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20060104989A1 (en) * 2002-11-05 2006-05-18 Affinium Pharmaceuticals, Inc. Essential novel bacterial polypeptides
WO2008113041A2 (fr) * 2007-03-14 2008-09-18 Ls9, Inc. Procédé de production d'hydrocarbures de bas poids moléculaire à partir de ressources renouvelables

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20060104989A1 (en) * 2002-11-05 2006-05-18 Affinium Pharmaceuticals, Inc. Essential novel bacterial polypeptides
WO2008113041A2 (fr) * 2007-03-14 2008-09-18 Ls9, Inc. Procédé de production d'hydrocarbures de bas poids moléculaire à partir de ressources renouvelables

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
AKIHIKO KAWAGUCHI ET AL.: "Fatty acid synthetase from Brevibacterium ammoniagenes: Formation of monounsaturated fatty acids by a multienzyme complex.", PROC. NATL. ACAD. SCI., vol. 74, no. 8, 1977, pages 3180 - 3183 *
DAVID F. SILBERT ET AL.: "Fatty acid mutant ofE. coli lacking a beta-hydroxydecanoyl thioester dehydrase.", PROC. NATL. ACAD. SCI., vol. 58, no. 4., 1967, pages 1579 - 1586 *
MINSUN LEESONG ET AL.: "Structure of a dehydratase-isomerase from the bacterial pathway for biosynthesis of unsaturated fatty acids: two catalytic activities in one active site.", STRUCTURE., vol. 4, no. 3, 1996, pages 253 - 264 *
RICHARD J. HEATH ET AL.: "Roles of the FabA and FabZ beta-Hydroxyacyl-Acyl Carrier Protein Dehydratases in Escherichia coli Fatty Acid Biosynthesis.", THE JOURNAL OF BIOLOGICAL CHEMISTRY., vol. 271, no. 44, 1996, pages 27795 - 27801, XP055044776, DOI: doi:10.1074/jbc.271.44.27795 *

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112143690A (zh) * 2019-06-28 2020-12-29 中国科学院青岛生物能源与过程研究所 一种酸耐受性提高的重组菌及其构建方法和应用
CN112143690B (zh) * 2019-06-28 2024-01-12 中国科学院青岛生物能源与过程研究所 一种酸耐受性提高的重组菌及其构建方法和应用

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