WO2001002543A1 - Procede de production d'acide l-amine - Google Patents

Procede de production d'acide l-amine Download PDF

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Publication number
WO2001002543A1
WO2001002543A1 PCT/JP2000/004343 JP0004343W WO0102543A1 WO 2001002543 A1 WO2001002543 A1 WO 2001002543A1 JP 0004343 W JP0004343 W JP 0004343W WO 0102543 A1 WO0102543 A1 WO 0102543A1
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Prior art keywords
lysine
producing
enolase
amino acid
acid
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PCT/JP2000/004343
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English (en)
Japanese (ja)
Inventor
Masakazu Sugimoto
Hisao Ito
Osamu Kurahashi
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Ajinomoto Co., Inc.
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Priority to AU57071/00A priority Critical patent/AU5707100A/en
Publication of WO2001002543A1 publication Critical patent/WO2001002543A1/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
    • C12P13/00—Preparation of nitrogen-containing organic compounds
    • C12P13/04—Alpha- or beta- amino acids
    • C12P13/14—Glutamic acid; Glutamine
    • 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
    • C12P13/00—Preparation of nitrogen-containing organic compounds
    • C12P13/04—Alpha- or beta- amino acids
    • C12P13/08—Lysine; Diaminopimelic acid; Threonine; Valine

Definitions

  • the present invention relates to a method for producing an L-amino acid by a fermentation method, and particularly to a method for producing L-lysine and L-glutamic acid.
  • L-lysine is widely used as a feed additive and the like
  • L-glutamic acid is widely used as a seasoning material and the like.
  • L-amino acids such as L-lysine and L-glutamic acid have been fermented using a coryneform bacterium belonging to the genus Brevipacterium or Corynebacterium, which has the ability to produce these L-amino acids. It is industrially produced by the law. For these coryneform bacteria, strains isolated from the natural world or artificial mutants of the strains are used in order to improve productivity.
  • diaminobi L-lysine producing ability can be obtained by successively amplifying or introducing the methacrylate dehydrogenase gene (ddh) (or the succinylase tetrahydrodipicolinate gene (dapD) and the succinyldiaminopimelate deacylase gene (dapE)). Is known to be improved (W0 95/16042).
  • ddh methacrylate dehydrogenase gene
  • dapD succinylase tetrahydrodipicolinate gene
  • dapE succinyldiaminopimelate deacylase gene
  • JP-A-63-214189 discloses that L-glutamic acid dehydrogenase gene, isocitrate dehydrogenase gene, aconitate hydratase gene, and citrate synthase gene are amplified or introduced. Techniques for increasing the ability to produce glutamate have been disclosed.
  • An object of the present invention is to provide a method for producing an L-amino acid such as L-lysine or L-glumic acid by a fermentation method which has been further improved, and a strain used therefor.
  • the present inventors have conducted intensive studies in order to solve the above-mentioned problems, and as a result, by introducing an enolase-encoding gene into coryneform bacteria to enhance enolase activity, L-lysine or L-glutamine
  • the present inventors have found that the production of minic acid can be increased, and have completed the present invention. That is, the present invention is as follows.
  • a coryneform bacterium having enhanced enolase activity in a cell and capable of producing L-amino acid (1) A coryneform bacterium having enhanced enolase activity in a cell and capable of producing L-amino acid.
  • L-amino acid is selected from L-lysine, L-glutamic acid, L-threonine, L-isoleucine, and L-serine.
  • the coryneform bacterium according to any of (1) to (4) is cultured in a medium, L-amino acids are produced and accumulated in the culture, and L-amino acids are collected from the culture.
  • a method for producing L-amino acid is produced.
  • the coryneform bacterium of the present invention is a coryneform bacterium having L-amino acid-producing ability and enhanced enolase activity in cells.
  • L-amino acids include L-lysine, L-glutamic acid, L-threonine, L-isoleucine, L-serine and the like. Of these, L-lysine and L-glutamic acid are preferred.
  • an embodiment of the present invention will be described mainly with respect to a coryneform bacterium having L-lysine-producing ability or L-glucamic acid-producing ability, but the present invention relates to a biosynthesis system specific to an L-amino acid of interest.
  • the coryneform bacterium referred to in the present invention is a group of microorganisms defined in Purges' Manual of Determinative Bacteriology, 8th edition, p. 599 (1974). Yes, aerobic, gram positive, It is a non-acid-fast, non-spore-forming bacillus, including bacteria that were previously classified into the genus Brevipacterium, but are now integrated as corynebacteria (Int. J. Syst. Bacterid., 41, 255 (1981)), and also includes bacteria of the genus Brevipacterium and Microbatterium, which are very closely related to the genus Corynepacterium. Examples of the strains of coryneform bacteria suitably used for producing L-lysine or L-glutamic acid include, for example, those shown below.
  • Corynebacterium thermoaminogenes AJ12340 (FERM BP-1539) To obtain these, for example, American Type Culture Collection, Address 12301 Parklawn Drive, Rockville, Maryland 20852, United States of America). That is, a registration number corresponding to each microorganism is assigned, and the microorganism can be ordered by referring to this registration number. The registration number corresponding to each microorganism is listed in the catalog of the American Type Culture Collection.
  • AJ12340 strain was deposited under the Budapest Treaty with the Ministry of International Trade and Industry, National Institute of Advanced Industrial Science and Technology, Institute of Biotechnology and Industrial Technology (zip code 305-8566, 1-3 1-3 Tsukuba-Higashi, Ibaraki, Japan). Have been.
  • mutants having L-amino acid-producing ability such as L-lysine or L-glucamic acid, derived from these strains can also be used in the present invention.
  • Such artificial mutants include the following. S- (2-aminoethyl) -cysteine (hereinafter abbreviated as "AEC") resistant mutant (for example, Brevibacterium 'Lactofamentum AJ11082 (NRRL B-11470), Japanese Patent Publication No. 56-1914, Japanese Patent Publication No. No. 56-1915, No. 57-14157, No. 57-14158, No. 57-30474, No. 58-10075, No. 59-4993, No. 61-35840, No.
  • AEC S- (2-aminoethyl) -cysteine
  • JP-A-53-86089 JP-A-55-9783, JP-A-55-9759, JP-A-56-32995, JP-A-56-39778, JP-B-53-43591, and JP-A-53-43591. No. 53-1833
  • L-lysine-producing mutants requiring inositol or acetic acid Japanese Patent Application Laid-Open Nos.
  • coryneform bacteria having L-threonine-producing ability include corynebacterium 'acetacidophila AJ12318 (FERM BP-1172) (see US Pat. No. 5,188,949) and L-isoleucine.
  • coryneform bacteria having the ability to produce are Brevibacterium flavum AJ12149 (FERM BP-759) (see US Pat. No. 4,656,135) and the like.
  • the ability to produce L-amino acids such as L-lysine means that when a coryneform bacterium is cultured in a medium, a significant amount of L-amino acid such as L-lysine is accumulated in the medium.
  • a recombinant DNA is prepared by ligating a gene fragment encoding the enolase to a vector that functions in the bacterium, preferably a multicopy vector. This may be introduced into a coryneform bacterium capable of producing L-lysine or L-glucamic acid for transformation.
  • the enolase activity is enhanced as a result of an increase in the copy number of the gene encoding the enolase in the cells of the transformed strain.
  • Enola is encoded by the eno gene in Escherichia coli.
  • enolase gene a gene of a coryneform bacterium or a gene derived from another organism such as a bacterium belonging to the genus Escherichia can be used.
  • the nucleotide sequence of the Escherichia coli eno gene has already been determined (Klein, M. et al., DNA Seq. 6, 315-355 (1996), Genbank / EMBL / DDBJ accetion No. X82400).
  • a primer prepared based on the base sequence for example, the primers shown in SEQ ID NOs: 1 and 2 in the Sequence Listing, a PCR method using polymerase chain reaction (PCR: polymerase chain reaction; White, TJ et al.) Eno gene can be obtained according to Trends Genet. 5, 185 (1989)).
  • Genes encoding enolase of other microorganisms such as coryneform bacteria can be obtained in a similar manner.
  • Chromosomal DNA is obtained from the DNA donor bacterium, for example, by the method of Saito and Miura (H. Saito and K. Miura Biochem. Biophys. Acta, 72, 619 (1963)) Pp. 97-98, Baifukan, 1992).
  • the gene encoding the enolase amplified by the PCR method is connected to a vector DNA capable of autonomously replicating in the cells of Escherichia coli and / or coryneform bacteria to prepare a recombinant DNA, which is then ligated to Escherichia coli. Introduced into E. coli cells Then, the operation later becomes difficult.
  • a vector capable of autonomous replication in Escherichia coli cells plasmid vectors are preferable, and those capable of autonomous replication in host cells are preferable.For example, pUC19, pUC18, pBR322, pHSG299, pHSG399 N pHS G398, RSF1010, etc. Is mentioned.
  • Examples of a vector capable of autonomous replication in a coryneform bacterium include PAM330 (see Japanese Patent Application Laid-Open No. 58-67699), pHM1519 (see Japanese Patent Application Laid-Open No. 58-77895), and the like.
  • a DNA fragment having the ability to enable autonomous replication of plasmid in coryneform bacteria is extracted from these vectors and inserted into the Escherichia coli vector, whereby autonomously expressed in both Escherichia coli and coryneform bacteria. It can be used as any shuttle vector that is not replicable.
  • Such shuttle vectors include: Microorganisms carrying the respective vectors and the accession numbers of the international depository organizations are shown in parentheses.
  • the vector is cut with a restriction enzyme that matches the end of the gene encoding the enolase. Ligation is usually performed using a ligase such as T4DNA ligase.
  • recombinant DNA can be obtained by transforming cells of a DNA-receiving bacterium into a protoplast or spheroblast that readily incorporates the recombinant DNA, as is known for Bacillus subtilis, actinomycetes and yeast.
  • Method for introduction into DNA recipient bacteria (Chang, S. and Choen, SN, Molec. Gen. Genet., 168, 111 (1979); Bibb, MJ, Ward, JM and Hopwood, 0.A., ature, 274, 398 (1978); Hinnen, A., Hicks, JBand Fink, GR, Proc. Natl. Acad. Sci. USA, 75 1929 (1978)).
  • the transformation method used in the examples of the present invention is the electric pulse method (see Japanese Patent Application Laid-Open No. 2-207791).
  • Enolase-encoding activity can also be enhanced by having multiple copies of the enolase-encoding gene on the chromosomal DNA of the host.
  • homologous recombination is performed by using a sequence that exists in multiple copies on the chromosome DNA as a target.
  • a sequence present in multiple copies on the chromosome DNA repetitive DNA and inverted repeats present at the end of a transposable element can be used.
  • Enolase activity is enhanced not only by the above-mentioned gene amplification but also by replacing the expression regulatory sequence such as the promoter of the gene encoding the enolase on chromosome DNA or plasmid with a strong one. Is also achieved (see Japanese Patent Application Laid-Open No. Hei 1-215280). For example, la. Promoters, trp promoters, trc promoters, ac promoters, PR promoters for lambda phage, PL promoters, etc. are known as strong promoters. The substitution with these promoters enhances the enolase activity by enhancing the expression of the gene encoding the enolase.
  • the coryneform bacterium of the present invention may have an enhanced enzymatic activity by enhancing an enzyme gene such as another amino acid synthesizing pathway or a glycolytic pathway in addition to the enolase activity.
  • an enzyme gene such as another amino acid synthesizing pathway or a glycolytic pathway in addition to the enolase activity.
  • genes that can be used for the production of L-lysine include aspartokinase subunit protein or / ?, in which synergistic feedback inhibition by L-lysine and L-threonine has been substantially eliminated.
  • genes that can be used for the production of L-glutamic acid include glutamate dehydrogenase (GDH, Japanese Patent Application Laid-Open No. Sho 61-268185), glutamate synthase, and glutamate. Synthase, isoquenate dehydrogenase (Japanese Patent Application Laid-open No. Sho 62-166890, Japanese Patent Application Laid-Open No. 63-214189), aconitate hydratase (Japanese Patent Application Laid-Open No. 62-294,086), citrate synthase, pyruvate carboxylase (Japanese Patent Laid-Open No. 60-87788, Japanese Patent Laid-open No.
  • an enzyme that catalyzes a reaction that diverges from a target L-amino acid biosynthetic pathway to produce a compound other than the same L-amino acid may be reduced or lacking.
  • homoserine dehydrogenase is an enzyme that catalyzes a reaction that produces a compound other than L-lysine by branching off from the L-lysine biosynthetic pathway (see W095 / 23864).
  • enzymes that catalyze the reaction that diverges from the biosynthetic pathway of L-glutamic acid to produce compounds other than L-glutamic acid include ketoglutarate dehydrogenase, isoquenate lyase, acetyl phosphate transferase, and acetate.
  • the biotin action inhibitor can be added to a medium containing an excess amount of biotin.
  • L-glutamic acid can be produced in the absence of E. coli (see W096 / 06180).
  • Examples of such coryneform bacteria include Brevipacterium. Lactofarmentum AJ13029 described in W096 / 06180. The AJ13029 strain was registered with the Institute of Biotechnology, Industrial Science and Technology (Postal Code 305-8566, Tsukuba 1-3-1, Ibaraki, Japan) on September 2, 1994 under the accession number FERM P-14501.
  • biotin-inhibiting substance by imparting a temperature-sensitive mutation to a biotin-inhibiting substance to a coryneform bacterium capable of producing L-lysine and L-glutamic acid, the biotin-inhibiting substance can be expressed in a medium containing an excessive amount of biotin. L-Lysine and L-glutamic acid can be produced simultaneously in the absence (see W096 / 06180). Examples of such a strain include Brevipacterium. Lactofermentum AJ12993 strain described in W096 / 06180.
  • the stock was issued to the Institute of Biotechnology, Institute of Industrial Technology (Postal Code 305-8566, Tsukuba 1-3-1-3, Ibaraki, Japan) on June 3, 1994, with the accession number FERM P- Deposited at 14348, transferred to the International Deposit under the Budapest Treaty on August 1, 1995, and given accession number FERM BP-5188.
  • the phrase “enhanced activity” of an enzyme generally means that the enzyme activity in a cell is higher than that of a wild-type strain, and is modified by a gene recombination technique or the like. When a strain with enhanced enzyme activity is obtained, it means that the enzyme activity in the cell is higher than that of the strain before modification.
  • reduced activity of an enzyme usually means that the enzyme activity in a cell is lower than that of a wild-type strain, and the enzyme activity has been reduced by modification by genetic recombination technology or the like. When a strain is obtained, it means that the enzyme activity in the cell is lower than that of the strain before modification.
  • a coryneform bacterium having enhanced enolase activity and capable of producing L-amino acid When a coryneform bacterium having enhanced enolase activity and capable of producing L-amino acid is cultured in a suitable medium, the L-amino acid accumulates in the medium. If it ’s clear, Enola If a coryneform bacterium having enhanced L-lysine activity and capable of producing L-lysine acid is cultured in a suitable medium, L-lysine accumulates in the medium. In addition, when a coryneform bacterium having enhanced enolase activity and capable of producing L-glutamic acid is cultured in a suitable medium, L-glutamic acid accumulates in the medium.
  • L-lysine and L-glutamic acid accumulate in the medium.
  • the L-lysine-producing bacterium may be cultured under L-glucaminic acid-producing conditions, or a L-lysine-producing coryneform A bacterium and a coryneform bacterium capable of producing L-glutamic acid may be mixedly cultured (Japanese Patent Application Laid-Open No. 5-3793).
  • the medium used for producing L-amino acids such as L-lysine or L-glutamic acid using the microorganism of the present invention contains a carbon source, a nitrogen source, inorganic ions, and other organic micronutrients as required. This is a normal medium.
  • Carbon sources include glucose, lactose, galactose, fructos, sucrose, molasses, carbohydrates such as starch hydrolysates, alcohols such as ethanol and inositol, acetic acid, fumaric acid, citric acid, An organic acid such as succinic acid can be used.
  • Nitrogen sources include inorganic ammonium salts such as ammonium sulfate, ammonium nitrate, ammonium chloride, ammonium phosphate, ammonium acetate, ammonia, peptone, meat extract, yeast extract, yeast extract, corn 'steep' liquor, soy hydrolyzate, etc.
  • Organic nitrogen, ammonia gas, aqueous ammonia, etc. can be used.
  • inorganic ions small amounts of potassium phosphate, magnesium sulfate, iron ions, manganese ions and the like are added.
  • organic trace nutrients it is desirable to include a required substance such as vitamin B1 or a yeast extract in an appropriate amount as necessary.
  • the culture is preferably carried out for 16 to 72 hours under aerobic conditions, such as shaking culture and aeration / agitation culture, at a culture temperature of 30 ° (up to 45 ° C, pH during culture). Is controlled to 5 to 9.
  • aerobic conditions such as shaking culture and aeration / agitation culture
  • a culture temperature of 30 ° (up to 45 ° C, pH during culture). Is controlled to 5 to 9.
  • an inorganic or organic acidic or alkaline substance, ammonia gas or the like can be used.
  • L-amino acid can be collected from the fermentation liquor in the same manner as in the normal L-amino acid production method. Can be done.
  • L-lysine can be collected usually by a combination of an ion exchange resin method, a precipitation method and other known methods.
  • the method of collecting L-glucamic acid may be a conventional method, for example, an ion exchange resin method, a crystallization method, or the like.
  • L-glutamic acid may be adsorbed and separated by an anion exchange resin, or may be neutralized and crystallized.
  • the nucleotide sequence of the Escherichia coli eno gene has already been determined (Klein, M. et al., DNA Seq. 6, 315-355 (1996), Genbank / EMBL / DDBJ accetion No. X82400). Based on the reported nucleotide sequence, the primers shown in SEQ ID NOs: 1 and 2 in the Sequence Listing were synthesized, and the pyruvate dehydrogenase gene was amplified by the PCR method using the chromosome DNA of Escherichia coli JM109 strain as type III.
  • SEQ ID NO: 1 corresponds to the sequence from the first to the 24th base of the base sequence of the eno gene described in Genbank / EMBL / DDBJ accetion No. X 82400
  • SEQ ID NO: 1 2 corresponds to the sequence from the 289th base to the 266th base.
  • the chromosome DNA of Escherichia coli JM109 strain was prepared by a conventional method (Biotechnological Experiments, edited by The Society of Biotechnology, Japan, pages 97-98, Baifukan, 1992).
  • PCR reaction standard reaction conditions described on page 185 of the PCR method forefront (edited by Takeo Sekiya et al., Kyoritsu Shuppan, 1989) are used.
  • the resulting PCR product was purified by a conventional method, ligated with Smal-cleaved plasmid pHC4 using a ligation kit (Takara Shuzo), and then combined with Escherichia coli JM109 (Takara Shuzo). Transformation was performed using L-culture medium containing 30 g / ml of chloram fenicol (10 g / L of Pak Tributone, The extract was applied to 5 g / L of extract, 5 g / L of NaCl, 15 g / L of agar, pH 7.2), cultured overnight, and the white colonies that appeared were picked and separated into single colonies to obtain a transformant. Plasmid was extracted from the obtained transformant to obtain a plasmid pHC4eno in which the eno gene was linked to a vector.
  • Escherichia coli which holds pHC 4, was named private number AJ12617, and on April 24, 1999, the Institute of Biotechnology and Industrial Technology, Institute of Industrial Science and Technology, Ministry of International Trade and Industry (zip code 305-8566, Ibaraki, Japan) Deposit No. FE RM P—122 1 15 at Tsukuba 1-3-1, Higashi-chome) and transferred to an international deposit based on the Budapest Convention on August 26, 1999, accession no. BP— 3 5 3 2 is awarded.
  • the enolase activity of the JM109 strain and the JM109 strain retaining pHC4eno was determined by the method of Bucher, T. et al. , Meth. Enzymol. 1, 427-435 (1955).
  • the JM109 strain carrying pHC4eno exhibited about 15 times the enolase activity of the JM109 strain not carrying pHC4en0, indicating that the eno gene was expressed. confirmed.
  • Brevibacterium lactofermentum AJ13029 was transformed with the plasmid pHC4eno by the electric pulse method (see Japanese Patent Application Laid-Open No. 2-207791) to obtain the resulting transformant.
  • cultivation for producing L-glucamic acid was performed as follows.
  • the cells of the AJ13029 / pHC4eno strain obtained by culturing on a CM2B plate medium containing 5 ⁇ g / ml chloramphenicol were added to L-cells having the following composition containing 5 ⁇ g / ml chloramphenicol.
  • the medium was inoculated into a glucanic acid-producing medium, shake-cultured at 31.5, and shaken until the sugar in the medium was consumed.
  • the obtained culture was inoculated into a medium having the same composition in an amount of 5%, and cultured at 37 ° C with shaking until the sugar in the medium was consumed.
  • a strain obtained by transforming a plasmid pHC4, which can be autonomously replicated with a previously obtained corynebacterium bacterium, into the corynebacterium bacterium AJ13029 as a control by the electric pulse method and culturing in the same manner as described above did.
  • the cells of the AJ11082 / pHC4eno strain obtained by culturing in a 0 ⁇ 12B plate medium containing 5 ⁇ / 1111 chloramphenicol were added to L-cells of the following composition containing 5 g / ml of chloramphenicol.
  • a lysine production medium was inoculated and cultured with shaking at 31.5 ° C until the sugar in the medium was consumed.
  • Corynepterium as control A strain obtained by transforming the strain AJ11082 with a plasmid pHC4 capable of autonomously replicating by a corynebacterium already obtained by the electric pulse method was cultured in the same manner as described above.
  • Brevi Pacterium 'Lactafamentum AJ11082 was established on January 31, 1981 at the Agricultural Research Culture Collection, America, Illinois, 6 1604 Peoria Northuniversity Street 18 1 5 (1815 N. University Street, Peoria, Illinois 61604 USA) and deposited under accession number NRRL B-11470.
  • Protein hydrolyzate (bean concentrate) 30 ml
  • Brevipacterium lactofermentum AJ12993 was transformed with plasmid pHC4eno by the electric pulse method (see Japanese Patent Application Laid-Open No. 2-207791) to obtain the obtained transformant.
  • culture for producing L-lysine and L-glucamic acid was performed as follows.
  • the cells of the AJ12993 / pHC4eno strain obtained by culturing in a CM2B plate medium containing 5 ⁇ g / ml chloramphenicol were added to the L-lysine production medium containing 5 / g / ml chloramphenicol. The cells were inoculated and cultured at 31.5 ° C.
  • the culture temperature was shifted to 34 ° C, and the culture was performed with shaking until the sugar in the medium was consumed.
  • a strain obtained by transforming a plasmid pHC4 capable of autonomously replicating with a previously obtained corynebacterium bacterium into an AJ12993 bacterium of the genus Corynebacterium by an electric pulse method was cultured in the same manner as described above.
  • L-anoic acid such as L-lysine or L-monoglutamic acid

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Abstract

Cette invention concerne un procédé de production d'acide L-aminé, tel que de l'acide de L-lysine ou L-glutamique, lequel procédé fait appel à une technique de fermentation améliorée par rapport aux techniques traditionnelles. Cette technique consiste à transférer un gène codant l'énolase dans une bactérie corynéforme capable de produire un acide L-aminé tel que de l'acide de L-lysine ou L-glutamique, ce qui augmente l'activité de l'énolase et accroît la production d'acide L-aminé. Cette invention concerne également des souches utilisées dans cette technique.
PCT/JP2000/004343 1999-07-02 2000-06-30 Procede de production d'acide l-amine WO2001002543A1 (fr)

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Application Number Priority Date Filing Date Title
AU57071/00A AU5707100A (en) 1999-07-02 2000-06-30 Process for producing l-amino acid

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JP11/189515 1999-07-02
JP18951599A JP2003180355A (ja) 1999-07-02 1999-07-02 Ｌ−アミノ酸の製造法

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Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1090998A1 (fr) * 1999-10-05 2001-04-11 Degussa-Hüls Aktiengesellschaft Séquences nucléotides codantes pour le gene eno
US6713289B2 (en) 1999-10-05 2004-03-30 Degussa Ag Nucleotide sequences which code for the eno gene
US7037690B2 (en) 2002-03-27 2006-05-02 Ajinomoto Co., Inc. Method for producing L-amino acid
US7217543B2 (en) 2002-11-20 2007-05-15 Ajinomoto Co., Inc. Method for producing L-amino acid using methylotroph

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Publication number Priority date Publication date Assignee Title
JP4504314B2 (ja) * 2003-11-11 2010-07-14 ヒゲタ醤油株式会社 新規ブレビバチルス・チョウシネンシス及び該微生物を宿主とするタンパク質の製造方法
JP4582573B2 (ja) * 2004-05-25 2010-11-17 味の素株式会社 ピルビン酸の製造方法及びｌ−バリンの製造方法
KR100892577B1 (ko) * 2004-08-10 2009-04-08 아지노모토 가부시키가이샤 유용한 대사산물을 생산하기 위한 포스포케톨라제의 용도

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EP1090998A1 (fr) * 1999-10-05 2001-04-11 Degussa-Hüls Aktiengesellschaft Séquences nucléotides codantes pour le gene eno
US6713289B2 (en) 1999-10-05 2004-03-30 Degussa Ag Nucleotide sequences which code for the eno gene
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US7432085B2 (en) 2002-03-27 2008-10-07 Ajinomoto Co., Inc. Method for producing L-amino acid
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