WO2001002544A1 - Procede de production d'acide l-amine - Google Patents

Procede de production d'acide l-amine Download PDF

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Publication number
WO2001002544A1
WO2001002544A1 PCT/JP2000/004344 JP0004344W WO0102544A1 WO 2001002544 A1 WO2001002544 A1 WO 2001002544A1 JP 0004344 W JP0004344 W JP 0004344W WO 0102544 A1 WO0102544 A1 WO 0102544A1
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Prior art keywords
lysine
amino acid
producing
succinate dehydrogenase
acid
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PCT/JP2000/004344
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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 AU57072/00A priority Critical patent/AU5707200A/en
Publication of WO2001002544A1 publication Critical patent/WO2001002544A1/fr

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y103/00—Oxidoreductases acting on the CH-CH group of donors (1.3)
    • C12Y103/99—Oxidoreductases acting on the CH-CH group of donors (1.3) with other acceptors (1.3.99)
    • C12Y103/99001—Succinate dehydrogenase (1.3.99.1)
    • 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
    • C12N9/0004—Oxidoreductases (1.)
    • C12N9/001—Oxidoreductases (1.) acting on the CH-CH group of donors (1.3)
    • 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
    • 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

Definitions

  • the present invention relates to a method for producing an L-amino acid by a fermentation method, and more particularly to a method for producing L-lysine and L-glutamic acid.
  • L-lysine is widely used as a feed additive
  • L-glutamic acid is widely used as a seasoning material.
  • L-amino acids such as L-lysine and L-glutamic acid have been obtained by 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 fermentation. For these coryneform bacteria, strains isolated from the natural world or artificial mutants of the strains are used in order to improve productivity.
  • diaminobi Successive amplification or introduction of the mehydrogenate dehydrogenase gene (ddh) (or the tetrahydrodipicolinic acid succinylase gene (dapD) and the succinyldiaminobimelate deacylase gene (dapE)) may improve L-lysine production.
  • ddh mehydrogenate dehydrogenase gene
  • dapD tetrahydrodipicolinic acid succinylase gene
  • dapE succinyldiaminobimelate deacylase gene
  • JP-A-63-214189 discloses that a glutamate dehydrogenase gene, an isocitrate dehydrogenase gene, an aconitate hydrase gene, and a quinate synthase gene are amplified or introduced. Discloses a technique for increasing the ability to produce L-glutamic acid.
  • An object of the present invention is to provide a method for producing an L-amino acid such as L-lysine or L-glucaminic acid by a fermentation method which has been further improved than before, and a strain used therefor.
  • the present inventors have conducted intensive studies to solve the above problems, and as a result, introduced a gene encoding succinate dehydrogenase into a coryneform bacterium, thereby enhancing succinate dehydrogenase activity, thereby improving L-lysine or L-lysine.
  • the inventors have found that the production of glutamic acid can be increased, and have completed the present invention. That is, the present invention is as follows.
  • a coryneform bacterium having enhanced succinate dehydrogenase activity in cells and having L-amino acid producing ability (1) A coryneform bacterium having enhanced succinate dehydrogenase activity in cells and having L-amino acid producing ability.
  • 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. Method for producing L-amino acid.
  • the coryneform bacterium of the present invention is a coryneform bacterium having an ability to produce L-amino acid and having enhanced succinate dehydrogenase 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.
  • embodiments of the present invention will be described mainly with respect to a coryneform bacterium having L-lysine-producing ability or L-glutamate-producing ability.
  • the present invention provides a biosynthetic system specific to an L-amino acid of interest. The same applies to those that are located downstream of the drogenase.
  • the coryneform bacterium referred to in the present invention is a barges. Manual. Tibno, a group of microorganisms defined in Bergey's Manual of Determinative Bacteriology, 8th edition, p. 599 (1974), aerobic, gram-positive, nonacidic, and capable of sporulation. Bacteria that have not been classified into the genus Brevipacterium, but now include bacteria that have been integrated into the genus Corynebacterium (Int. J. Syst. Bacteriol., 41, 255 (1981)). Also, it includes bacteria of the genus Brevipacterium and Microbaterim, 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 samminoaminogenes AJ12340 (FERM BP-1539) To obtain them, for example, American Type Culture Collection, American Type Culture Collection, Address 12301 Park lawn 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 'Cultural Collection.
  • the AJ12340 strain has been deposited with the Ministry of International Trade and Industry at the National Institute of Bioscience and Human-Technology (Postal Code 305-856-6 1-3 1-3 Tsukuba-Higashi, Ibaraki, Japan) based on the Budapest Treaty.
  • mutants having L-amino acid-producing ability such as L-lysine or L-glutamic acid derived from these strains can also be used in the present invention.
  • Such artificial mutants include the following. S— (2-aminoethyl) -cystine (hereinafter abbreviated as “AEC”) resistant mutant (eg, 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 (2-aminoethyl) -cystine
  • coryneform bacteria having L-threonine-producing ability include: (See U.S. Pat. No. 5,188,949) is one example of a coryneform bacterium having the ability to produce L-isoleucine. Lium Flavam AJ12149 (FERM BP-759) (see US Pat. No. 4,656,135).
  • the ability to produce L-amino acid 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. Ability or ability to increase the content of amino acids such as L-lysine in cells.
  • a recombinant DNA is obtained by ligating a gene fragment encoding succinate dehydrogenase to a vector that functions in the bacterium, preferably a multicopy vector. May be prepared and introduced into a coryneform bacterium having the ability to produce L-lysine or L-glucaminic acid, followed by transformation.
  • succinate dehydrogenase activity is enhanced.
  • Succinate dehydrogenase is encoded by the sdh gene in Escherichia coli.
  • succinate dehydrogenase 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 sdh gene of Escherichia coli has already been determined (Genbank / EMBL / DDBJ accetion No. J01619 K00542 MU121)
  • primers prepared based on the nucleotide sequence for example, SEQ ID NO: 1 and Using the primers shown in 2 above, the PCR method (PCR: polymerase chain reaction; White, TJ et al; Trends Genet. 5, 185 (1989)) using the Escherichia coli chromosome DNA as type I was carried out.
  • the sdh gene can be obtained.
  • the gene encoding succinate dehydrogenase amplified by the PCR method is connected to a vector DNA capable of autonomous replication in cells of Escherichia coli and / or coryneform bacteria to prepare a recombinant DNA, which is then prepared. If introduced into Escherichia coli cells, subsequent operations will be slower. Plasmid vectors are preferred as vectors capable of autonomous replication in Escherichia coli cells, and those capable of autonomous replication in host cells are preferred.For example, pUC19, pUC18, pBR322, pHSG299, pHSG399, pHSG398, RSF1010 And the like.
  • Examples of vectors capable of autonomous replication in coryneform bacterium cells include PAM330 (see JP-A-58-67699), pHM1519 (see JP-A-58-77895), and the like.
  • a DNA fragment having the ability to enable autonomous replication of a plasmid in a coryneform bacterium is extracted from these vectors and inserted into the Escherichia coli vector, which results in autonomous expression in both the Escherichia coli and the coryneform bacteria. It can be used as a replicable shuttle vector.
  • the following are examples of such shuttle vectors. Microorganisms carrying the respective vectors and the accession numbers of the international depository organizations are shown in parentheses.
  • PAJ611 Escherichia Cori AJ11884 (FERM BP-138)
  • the vector To prepare recombinant DNA by ligating a gene encoding succinate dehydrogenase and a vector that functions in coryneform bacteria, the vector must be digested with a restriction enzyme that matches the end of the gene encoding succinate dehydrogenase. Disconnect. The ligation is usually performed using a ligase such as T4 DNA ligase.
  • Escherichia A method for increasing the permeability of DNA by treating recipient cells with calcium chloride, as reported for li-112 (Mandel, M. and Higa, A., J. Mol. Biol., 53 , 159 (1970)), and a method for preparing a recombinant cell from a cell at the growth stage and introducing DNA as described in Bacillus' subtilis (Duncan, CH, Wilson, GA and Young, FE, Gene, 1, 153 (1977)).
  • recombinant DNA can be obtained by transforming cells of a DNA-accepting bacterium into protoblasts or spherovlasts that readily incorporate the recombinant DNA, as is known for Bacillus subtilis, actinomycetes and yeast. Methods for introduction into DNA recipients (Chang, S. and Choen, SN, Molec. Gen. Genet., 168, 111 (1979); Bibb, MJ, Ward, JMand Hopwood, 0.A., Nature, 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).
  • Enhancement of the activity encoding succinate dehydrogenase can also be achieved by causing multiple copies of a gene encoding succinate dehydrogenase to be present on the chromosomal DNA of the host.
  • homologous recombination is performed using a sequence present on the chromosomal DNA in a multiple copy as a target. Sequences present in multiple copies on the chromosomal DNA include repetitive DNA and invert 'repeats at the ends of transposable elements.
  • a gene encoding succinate dehydrogenase can be mounted on a transposon and transferred, and multiple copies can be introduced into chromosomal DNA, as disclosed in JP-A-2-19985. is there. Either method increases the copy number of the gene encoding succinate dehydrogenase in the transformant, resulting in enhanced succinate dehydrogenase activity.
  • Enhancement of succinate dehydrogenase activity can be achieved not only by the above-described gene amplification but also by replacing an expression regulatory sequence such as a promoter of a gene encoding succinate dehydrogenase on chromosomal DNA or plasmid with a strong one.
  • an expression regulatory sequence such as a promoter of a gene encoding succinate dehydrogenase on chromosomal DNA or plasmid with a strong one.
  • lac promotion Yuichi, trp promoter, trc promoter, tac bromo, PR promoter and PL promoter of lambda phage are known as strong promoters. Substitution with these promoters enhances the expression of the gene encoding succinate dehydrogenase, thereby enhancing succinate dehydrogenase activity.
  • the coryneform bacterium of the present invention can enhance the enzymatic genes of other amino acid biosynthetic pathways or glycolytic pathways in addition to the succinate dehydrogenase activity, thereby enhancing the enzymatic activity.
  • genes that can be used for the production of L-lysine include aspartase kinase subunit protein and / or protein, in which synergistic feedback inhibition by L-lysine and L-threonine has been substantially released.
  • genes that can be used for the production of L-glutamic acid include glutamate dehydrogenase (GDH, Japanese Patent Application Laid-Open No. 61-268185), glutamate synthetase, glutamate synthetase, and isoquene.
  • Acid dehydrogenase JP-A-62-166890, JP-A-63-214189), aconitate hydrazine (JP-A-62-294086), quenic acid Synthase, pyruvate carboxylase (JP-A-60-87788, JP-A-62-55089), phosphoenolpyruvate carboxylase, phosphoenolpyruvate synthase, enolase, phosphoglyceromase, Phosphoglycerate kinase, glyceraldehyde 3-phosphate dehydrogenase, triosephosphate isomerase, fructosebisphosphate aldolase, phosphofruc Kinase (JP 63 - 1 02 692), there is a glucose phosphate Isomera Ichize like.
  • an enzyme that catalyzes a reaction that diverges from a target L-amino acid biosynthetic pathway to produce a compound other than the 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 (W09). 5/23864).
  • Enzymes that catalyze the reaction that diverges from the L-glutamic acid biosynthetic pathway to produce a compound other than L-glutamic acid include: ketoglutarate dehydrogenase, isoquenate lyase, acetyl phosphate transferase, and the like. There are acetate kinase, acetate hydroxysynthase, acetate lactate synthase, acetyl formate transferase, lactate dehydrogenase, glutamate decarboxylase, and monopyrophosphate dehydrogenase.
  • a temperature-sensitive mutation to a biotin-inhibitory substance such as a surfactant to a coryneform bacterium capable of producing L-glutamic acid
  • a biotin-inhibitory substance such as a surfactant
  • the biotin-inducing action in a medium containing an excessive amount of biotin can be achieved.
  • L-glutamic acid can be produced in the absence of inhibitors (see W096 / 06180).
  • Examples of such coryneform bacteria include Brevibacterium lactofermentum AJ13029 described in W096 / 06180.
  • the AJ13029 strain was registered on September 2, 1994, with the Institute of Biotechnology and Industrial Technology, National Institute of Advanced Industrial Science and Technology (Postal Code 305-8566, 1-3-1 Tsukuba East, Ibaraki Prefecture, Japan) under the accession number FERM P-14501. Deposited and transferred to an international deposit under the Budapest Treaty on August 1, 1995, and given accession number FERM BP-5189.
  • a temperature-sensitive mutation to a substance inhibiting the action of biotin to a coryneform bacterium capable of producing L-lysine and L-glutamic acid, the action of biotin in a medium containing an excessive amount of biotin is suppressed.
  • L-lysine and L-glutamic acid can be produced simultaneously in the absence of a substance (see W096 / 06180).
  • Such strains include Brevipacterium 'Lactofamentum AJ12993 strain described in W096 / 06180.
  • the stock was granted to the Institute of Biotechnology and Industrial Technology, Institute of Industrial Science and Technology (Postal Code 305-8566, 1-3-1, Higashi 1-3-chome, Tsukuba, Ibaraki, Japan) on June 3, 1994, with accession number FERM P-14348. And transferred to an 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. Also, “reduced activity” of an enzyme usually means that the enzyme activity in a cell is lower than that of a wild-type strain. However, when a strain whose enzyme activity is reduced by modification by genetic recombination technology or the like is obtained, it means that the enzyme activity in the cell is lower than that of the strain before modification.
  • the L-amino acid When a coryneform bacterium having enhanced succinate dehydrogenase activity and capable of producing L-amino acid is cultured in a suitable medium, the L-amino acid accumulates in the medium. For example, when a coryneform bacterium having enhanced succinate dehydrogenase activity and capable of producing L-lysine acid is cultured in a suitable medium, L-lysine accumulates in the medium. Further, when a coryneform bacterium having enhanced succinate dehydrogenase activity and capable of producing L-gluminic acid is cultured in a suitable medium, L-gluminic acid accumulates in the medium.
  • L-lysine and L-glucaminic acid accumulate in the medium.
  • the L-gine-producing bacterium may be cultured under L-glucamic acid production conditions, or L-lysine-producing ability
  • a coryneform bacterium having L-glutamic acid-producing ability may be mixedly cultured (JP-A-5-37993).
  • the medium used to produce L-amino acids such as L-lysine or L-glucamic acid using the microorganism of the present invention contains a carbon source, a nitrogen source, inorganic ions, and if necessary, other organic micronutrients. This is a normal medium.
  • Carbon sources include glucose, lactose, galactose, fructose, sucrose, molasses, carbohydrates such as starch hydrolysates, alcohols such as ethanol and inositol, acetic acid, fumaric acid, citric acid, Organic acids such as succinic acid can be used.
  • nitrogen source examples include inorganic ammonium salts such as ammonium sulfate, ammonium nitrate, ammonium chloride, ammonium phosphate, and ammonium acetate, ammonia, peptone, meat extract, yeast extract, yeast extract, corn steep liquor, soybean hydrolyzate, etc.
  • organic nitrogen, ammonia gas, ammonia water, and the like can be used.
  • Potassium phosphate, magnesium sulfate, iron ion, manga A small amount of ion or the like is 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 cultivation is preferably carried out for 16 to 72 hours under aerobic conditions such as shaking cultivation and aeration / agitation cultivation.
  • aerobic conditions such as shaking cultivation and aeration / agitation cultivation.
  • pH adjustment an inorganic or organic acidic or alkaline substance, ammonia gas or the like can be used.
  • the L-amino acid can be collected from the fermentation liquor in the same manner as in the usual method for producing an L-amino acid.
  • collection of L-lysine can be usually carried out by combining an ion exchange resin method, a precipitation method and other known methods.
  • the method for collecting L-glutamic acid may be a conventional method, such as an ion exchange resin method or a crystallization method.
  • L-glutamic acid may be adsorbed and separated by an anion exchange resin, or may be neutralized and crystallized.
  • the nucleotide sequence of the sdh gene of Escherichia coli has already been elucidated (Genbank / EMBL / DDBJ accetion No. J01619 K00542 M11121). Primers shown in SEQ ID NOS: 1 and 2 in the Sequence Listing were synthesized based on the reported nucleotide sequence, 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 extends from nucleotide 199 to nucleotide 215 of the nucleotide sequence of the sdh gene described in Genbank / EMBL / DDBJ accetion No. J 01619 K00542 M11121.
  • SEQ ID NO: 2 corresponds to the sequence from the 6309th to the 286th base.
  • the resulting PCR product is purified by a conventional method, and then ligated with Smal-cleaved plasmid pHC4 using a ligation kit (Takara Shuzo Co., Ltd.). Then, Escherichia coli KM JM109 competent cells (Takara Shuzo) Transformation is performed using L medium containing 30 g / ml of chloramphenicol (10 g / L of pact tributone, 5 g / L of pak toy extract, 5 g / L of NaCl, and 15 g / L of agar). , PH 7.2), and after culture, the white colonies that appeared were picked and separated into single colonies to obtain transformed strains. Plasmid was extracted from the obtained transformant to obtain a plasmid pHC4sdh in which the sdh gene was bound to the vector.
  • Escherichia coli harboring pHC4 was named private number AJ12617, and on April 24, 1991, the Ministry of International Trade and Industry, National Institute of Industrial Science and Technology, Institute of Biotechnology and Industrial Technology (Zip code 305-8566 Japan Deposit No. FE RM P—122 15 at Tsukuba 1-3-1 Higashi, Ibaraki Pref., Japan, and transferred to an international deposit based on the Budapest Agreement on August 26, 1999.
  • FERM BP—3532 is granted.
  • the succinate dehydrogenase activity of the JM109 strain and the JM109 strain carrying pHC4sdh was determined.
  • the sdh gene was expressed because the JM109 strain carrying pHC 4 sdh exhibited about 19 times the succinate dehydrogenase activity of the JM109 strain not carrying pHC4 sdh. .
  • Brevipacterium 'lactofermentum AJ13029 was transformed with plasmid pHC4sdh by the electric pulse method (see Japanese Patent Application Laid-Open No. 2-207791) to obtain the obtained transformant.
  • Using the resulting transformant AJ13029 / pHC4sdh to produce L-glutamic acid Cultivation was performed as follows.
  • the AJ13029 / pHC4sdh strain obtained by culturing on a CM2B plate medium containing chloramphenicol in L-glutamic acid producing medium containing 5 ⁇ g / ml chloramphenicol and having the following composition Then, the cells were shake-cultured at 31.5 ° C and shake-cultured 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 capable of autonomously replicating with a previously obtained Corynebacterium bacterium into an AJ13029 strain of Corynebacterium bacterium by the electric pulse method was cultured in the same manner as described above. did.
  • the cells of the AJ11082 / pHC4sdh strain obtained by culturing in ⁇ 2B plate medium containing chloramphenicol were inoculated into an L-lysine production medium having the following composition containing 5 / ml chloramphenicol, and Shaking culture was performed at ° C 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 by the electric pulse method was cultured in the corynebacterium bacterium AJ11082 strain in the same manner as described above.
  • Protein hydrolyzate (bean concentrate) 30 ml 50 g calcium carbonate
  • Brevipacterium lactofermentum AJ12993 was transformed with plasmid pHC4sdh 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.
  • AJ12993 / pHC4sdh strain cells obtained by culturing in a CM2B plate medium containing 5 ⁇ g / ml chloramphenicol were inoculated into the L-lysine production medium containing 5 ig / ml chloramphenicol. 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 belonging to the genus Corynebacterium by the electric pulse method was cultured in the same manner as described above.
  • L-anoic acid such as L-lysine or L-glutamic 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 la déhydrogénase de succinate 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 la déhydrogénase de succinate et accroît la production d'acide L-aminé. Cette invention concerne également des souches utilisées dans cette technique.
PCT/JP2000/004344 1999-07-02 2000-06-30 Procede de production d'acide l-amine WO2001002544A1 (fr)

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AU57072/00A AU5707200A (en) 1999-07-02 2000-06-30 Process for producing l-amino acid

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JP18951699A JP2003180348A (ja) 1999-07-02 1999-07-02 Ｌ−アミノ酸の製造法
JP11/189516 1999-07-02

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Cited By (2)

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Publication number Priority date Publication date Assignee Title
EP1106684A1 (fr) * 1999-12-10 2001-06-13 Degussa AG Séquences polynucleotidiques de Corynebacterium glutamicum codant pour des sous-unités de succinate déshydrogénase
EP1382686A1 (fr) * 2002-07-12 2004-01-21 Ajinomoto Co., Inc. Procédé pour la production fermentative d'une substance déesirée en utilisant une souche bactérienne qui manque le gen ArcA

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ATE463564T1 (de) * 2004-08-10 2010-04-15 Ajinomoto Kk Verwendung von phosphoketolase zur herstellung geeigneter metabolite
MX2012007718A (es) * 2009-12-30 2012-07-25 Metabolic Explorer Sa Aumento de la produccion de metionina mediante la sobreexpresion de succinato deshidrogenasa.

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EP0841395A1 (fr) * 1995-06-07 1998-05-13 Ajinomoto Co., Inc. Procede de production de l-lysine
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