126 results about "Coryneform bacterium" patented technology

L-Arginine is produced by culturing a coryneform bacterium in which an arginine repressor involved in L-arginine biosynthesis is deleted by disrupting a gene coding for the repressor, and which has L-arginine producing ability in a medium to produce and accumulate L-arginine in the medium, and collecting the L-arginine from the medium.

A method of producing coryneform bacteria having improved amino acid or nucleic acid productivity comprising the steps of introducing a mutation in a promoter sequence of amino acid- or nucleic acid-biosynthesizing genes on a chromosome of a coryneform bacterium to make it close to a consensus sequence, or introducing a change in a promoter sequence of amino acid- or nucleic acid-biosynthesizing genes on a chromosome of a coryneform bacterium by gene recombination to make it close to a consensus sequence, to obtain mutants of the coryneform amino acid- or nucleic acid-producing microorganism, culturing the mutants and selecting a mutant capable of producing the intended amino acid or nucleic acid in a large amount. This method allows the construction of a mutant capable of enriching or controlling the expression of an intended gene without using a plasmid and to promote production of amino acids in a high yield by recombination or mutation.

A coryneform bacterium that is modified by using a yggB gene so that L-glutamic acid-producing ability is enhanced as compared to a non-modified strains is cultured in a medium to cause accumulation of L-glutamic acid in the medium or bacterial cells, and L-glutamic acid is collected from the medium or cells.

A plasmid which is able to be isolated from Corynebacterium thermoaminogenes, and which comprises a gene coding for a Rep protein having the amino acid sequence shown in SEQ ID NO: 4 or an amino acid sequence having homology of 90% or more to the amino acid sequence shown in SEQ ID NO: 4. and has a size of about 4.4 kb or about 6 kb, or a derivative thereof.

The invention relates to coryneform bacteria which have, in addition to at least one copy, present at the natural site (locus), of an open reading frame (ORF), gene or allele which codes for the synthesis of a protein or an RNA, in each case a second, optionally third or fourth copy of this open reading frame (ORF), gene or allele at in each case a second, optionally third or fourth site in a form integrated into the chromosome and processes for the preparation of chemical compounds by fermentation of these bacteria.

L-glutamic acid is produced by culturing in a medium a microorganism belonging to enterobacteria and having L-glutamic acid productivity, into which a citrate synthase gene derived from a coryneform bacterium is introduced to produce and accumulate L-glutamic acid in the medium and collecting the L-glutamic acid from the medium.

The invention relates to coryneform bacteria which, instead of the singular copy of an open reading frame (ORF), gene or allele naturally present at the particular desired site (locus), have at least two copies of the open reading frame (ORF), gene or allele in question, preferably in tandem arrangement, and optionally at least a third copy of the open reading frame (ORF), gene or allele in question at a further gene site, and processes for the preparation of chemical compounds by fermentation of these bacteria.

The present invention discloses a method for producing a target substance using a coryneform bacterium comprising culturing a coryneform bacterium having an ability to produce the target substance in a medium, resulting in accumulation of the target substance in the medium or cells of the bacterium, and collecting the target substance from the medium or the cells of the bacterium. Also disclosed is a coryneform bacterium which is introduced with a methanol dehydrogenase gene and which has enhanced activities of hexulose phosphate synthase and phosphohexuloisomerase, and to which an ability to utilize methanol is imparted or which has enhanced ability to utilize methanol, and the medium contains methanol as a carbon source.

The object of the present invention is to provide a method of producing a heterologous protein by making a coryneform bacterium to produce and efficiently extracellularly secrete (secreto-production) an industrially useful heterologous protein. According to the present invention, a genetic construct is used where a gene sequence encoding an intended protein which is ligated to the downstream of a sequence encoding the signal peptide derived from a coryneform bacterium, the gene construct is introduced into a mutant coryneform bacterium which has a capacity of secreting the heterologous protein at least 2-fold higher than the wild type Corynebacterium glutamicum ATCC 13869, the mutant coryneform bacterium is cultured and the extracellularly released heterologous protein is recovered.

A method of producing coryneform bacteria having an improved amino acid- or nucleic acid-productivity comprises the steps of introducing a mutation in a promoter sequence of amino acid- or nucleic acid-biosynthesizing genes on a chromosome of a coryneform bacterium to make it close to a consensus sequence or introducing a change in a promoter sequence of amino acid- or nucleic acid-biosynthesizing genes on a chromosome of a coryneform bacterium by gene recombination to make it close to a consensus sequence, to obtain mutants of the coryneform amino acid d- or nucleic acid-producing microorganism, culturing the mutants and select a mutant capable of producing the intended amino acid or nucleic acid in a large amount. This method can construct a mutant capable of suitably enriching or controling the expression of an intended gene without using a plasmid and also capable of producing amino acids in a high yield, by the recombination or mutation.

A coryneform bacterium which has an L-glutamic acid-producing ability and grows at least at the same growth rate as a non-mutated strain or a wild-type strain and has intracellular α-ketoglutarate dehydrogenase activity which is less than half that of the non-mutated or wild-type strain, and is obtained by introducing a mutation into a coding region or an expression control region of the chromosomal odhA gene encoding the E1o subunit of the α-ketoglutarate dehydrogenase complex.

Coryneform bacteria are described that have an ability to produce L-amino acids and are modified so that acetyl-CoA hydrolase activity is decreased. The bacteria are used to produce L-amino acids generated by a biosynthetic pathway in which pyruvic acid is an intermediate, such as L-glutamic acid, L-arginine, L-glutamine, L-proline, L-alanine, L-valine, and L-lysine.

Succinic acid is produced by using a coryneform bacterium which has succinic acid-producing ability and has been modified so that an activity of pyruvate oxidase is decreased.

Non-amino organic acids such as succinic acid, malic acid and fumaric acid are produced by reacting bacterial cells or treated bacterial cells of a coryneform bacterium with an organic raw material in an aqueous medium containing magnesium carbonate and / or magnesium hydroxide, and a certain range of concentration of a monovalent cation, while maintaining the pH within a certain range without increasing the volume of the aqueous medium.

L-cysteine is produced by culturing a coryneform bacterium in which intracellular serine acetyltransferase activity is increased by transformation with a gene coding for serine acetyltransferase, such modification of an expression control sequence of a gene coding for serine acetyltransferase that expression of the gene in a cell should be enhanced and for forth, preferably in which L-cysteine decomposition system is further suppressed, in a medium to produce and accumulate L-cysteine in culture and collecting the L-cysteine from the culture.

The invention relates to mutants of coryneform bacteria in which genes have been enhanced by the use of a mutated promoter region, and to processes for the production of amino acids using bacteria according to the invention.

The invention relates to a process for the production of L-amino acids, in which the following steps are carried out: a) fermentation of the coryneform bacteria producing the desired L-amino acid, in which at least the mqo gene is attenuated, b) concentration of the desired L-amino acid in the medium or in the cells of the bacteria, and c) isolation of the L-amino acid, and, optionally, bacteria are used in which further genes of the biosynthesis pathway of the desired L-amino acid are additionally enhanced, or bacteria are used in which at least some of the metabolic pathways reducing formation of the desired L-amino acid are excluded.

The invention belongs to the technical field of bioengineering, and discloses coryneform bacteria capable of producing lysine at high yield as well as a construction method and an application thereof.The coryneform bacterium capable of producing lysine at high yield has L-lysine production capacity, and the activity of at least one of promoters of encoding genes of key enzymes of an intracellularL-lysine synthesis pathway and / or a TCA circulating back-supplement pathway of the coryneform bacterium is enhanced. According to the coryneform bacterium capable of producing lysine at high yield, the sequence of the promoter of the coding gene of the key enzyme in the lysine biosynthesis pathway or the TCA circulating back-supplement pathway is modified, compared with an endogenous promoter, the L-lysine promoter has higher activity, so that the initial transcription efficiency is enhanced, the lysine production level is further improved, and the production capacity of an L-lysine strain isimproved. The experiments show that the coryneform bacteria provided by the invention significantly improve the yield and conversion rate of L-lysine, lay a foundation for industrial production of L-lysine, and have a wide industrial application prospect.

A plasmid isolatable from Corynebacterium thermoaminogenes, which comprises a gene coding for a Rep protein having the amino acid sequence shown in SEQ ID NO: 8 or an amino acid sequence having homology of 90% or more to the amino acid sequence shown in SEQ ID NO: 8, and has a size of about 4.4 kb or about 6 kb, or a derivative thereof.

L-Arginine is produced by culturing a coryneform bacterium in which an arginine repressor involved in L-arginine biosynthesis is deleted by disrupting a gene coding for the repressor, and which has L-arginine producing ability in a medium to produce and accumulate L-arginine in the medium, and collecting the L-arginine from the medium.

Disclosed is a method for producing 1,5-pentanediamine through fermentation using a microorganism, said microorganism having LDC gene in the chromosome and showing regulated secondary production of L-lysine. In the method for producing 1,5-pentanediamine whereby 1,5-pentanediamine is produced by a coryneform bacterium having a gene encoding lysine decarboxylase in the chromosome, said coryneform bacterium continuously maintains a lysine decarboxylase activity of 50 mU / mg protein or greater during the culture.

A method for producing a target substance is provided. A target substance is produced by culturing a coryneform bacterium, which is able to produce a target substance, and which also has an improved ability to assimilate xylose as a result of the introduction of a mutation into a coding region and / or an expression control region of the NCgl2954 gene on the chromosome of the bacterium. The culture is conducted in a medium containing xylose, and the target substance can be collected from the medium.

This transformant is obtained by introducing, to a host coryneform bacterium, at least one of the DNAs (a), (b), and (c): (a) a DNA, or an analog thereof, obtained by introducing, to a DNA that encodes an acetohydroxy acid synthase derived from Corynebacterium glutamicum, a mutation that mutates the No. 156 glycine in the amino acid sequence encoded by this DNA to glutamic acid (G156E); (b) a DNA, or an analog thereof, obtained by introducing, to a DNA that encodes an acetohydroxy acid isomeroreductase derived from Corynebacterium glutamicum, mutations that mutate the No. 34 serine in the amino acid sequence encoded by this DNA to glycine (S34G), mutate the No. 48 leucine to glutamic acid (L48E), and mutate the No. 49 arginine to phenylalanine (R49F); and (c) a DNA, or an analog thereof, that encodes a leucine dehydrogenase derived from Lysinibacillus sphaericus.

The present invention provides an aerobic coryneform bacterium transformant in which a lactate dehydrogenase gene is disrupted, and a pyruvate carboxylase gene is recombined so as to be highly expressed by a genetic engineering method. The aerobic coryneform bacterium transformant of the present invention can produce dicarboxylic acids from saccharides at a high production rate.

Provided is a transformant having phenol-producing capability, wherein the gene encoding an enzyme having chorismate-pyruvate lyase activity and the gene encoding an enzyme having 4-hydroxybenzoate decarboxylase activity have been introduced to a host Coryneform bacterium. Also provided is a method for producing a phenol, said method comprising: a step for reacting, under reduction conditions, the transformant in a reaction solution containing saccharides; and a step for recovering the phenol in the reaction solution.

A method for producing an L-amino acid of glutamate family such as L-glutamic acid is provided. An L-amino acid of glutamate family is produced by culturing a coryneform bacterium having an ability for producing an L-amino acid of glutamate family, which has been modified so that the activity of an alpha-ketoglutaric acid (alpha-KG) uptake carrier is increased, in a medium, and collecting the L-amino acid of glutamate family from the medium.

A method for producing an L-amino acid is provided. An L-amino acid is produced by culturing a coryneform bacterium having an L-amino acid-producing ability, which has been modified so that the activity of a phosphate transporter is increased, in a medium, and collecting the L-amino acid from the medium.

L-Glutamine is produced by culturing a coryneform bacterium which has L-glutamine producing ability and has been modified so that its intracellular glutamine synthetase activity should be enhanced, preferably which has been further modified so that its intracellular glutamate dehydrogenase activity should be enhanced, in a medium to produce and accumulate L-glutamine in the medium and collecting the L-glutamine.

A transformant obtainable by introducing one or more of the following DNAs (a), (b), and (c) into a coryneform bacterium as a host.(a) A DNA which encodes acetohydroxy acid synthase derived from Corynebacterium glutamicum and which has a mutation changing the glycine at position 156 to glutamic acid (G156E) in an amino acid sequence encoded by the DNA, or an analog thereof.(b) A DNA which encodes acetohydroxy acid isomeroreductase derived from Corynebacterium glutamicum and which has mutations changing the serine at position 34 to glycine (S34G), the leucine at position 48 to glutamic acid (L48E), and the arginine at position 49 to phenylalanine (R49F) in an amino acid sequence encoded by the DNA, or an analog thereof.(c) A DNA which encodes leucine dehydrogenase derived from Lysinibacillus sphaericus, or an analog thereof.