215 results about "Corynebacterium" patented technology

The invention discloses a method for improving growth of corynebacterium glutamicum under high-temperature stress, and aims to overcome the problems of abnormal growth and metabolism, even death and the like caused by temperature rise of existing microorganisms in an industrial fermentation production process, adaptive laboratory evolution is adopted, corynebacterium glutamicum ATCC 13032 is used as an original strain, and the corynebacterium glutamicum ATCC 13032 is used as an original strain. And carrying out heat stress subculture for 42.9 days, and separating out the strains HES1 and HES2 with improved high temperature resistance. The whole genome is re-sequenced to obtain mutant elements cspB and gltA which enable the strain to generate high-temperature adaptive change. And carrying out high-temperature test on the abrupt change element. In specific implementation, the two mutants are respectively overexpressed, and in a test, compared with a control strain WT, the 42-DEG C high-temperature tolerance is respectively improved by about 14.4% and 7%. Therefore, the beneficial mutant obtained through adaptive laboratory evolution lays a good foundation for application of corynebacterium glutamicum in industrial high-temperature conditions.

The invention discloses corynebacterium glutamicum for producing N-acetylglucosamine and application of the corynebacterium glutamicum, and belongs to the technical field of synthetic biology. Specifically, the corynebacterium glutamicum overexpresses a GNA1 gene from a brewer's yeast S288C controlled by a tacM promoter, a glmS gene from corynebacterium glutamicum S9114 and a yqab gene from escherichia coli K12 MG1655, and simultaneously has five gene defect types of nagA, nagB, ldhA, msccg and msccg2 in a genome and two gene single base mutation types of odhA and glnA, and the codon of the glnA for coding the 405th amino acid is mutated from TAC to TTC. By blocking excretion of glutamic acid of corynebacterium glutamicum and enhancing conversion of glutamic acid to glutamine, the supply capacity of a precursor of corynebacterium glutamicum is enhanced, and an optimal way for synthesizing N-acetylglucosamine is obtained, so that efficient production of N-acetylglucosamine in corynebacterium glutamicum is realized, and the yield of N-acetylglucosamine in corynebacterium glutamicum is increased. The method has important economic value and social significance.

The present invention relates to an enzyme catalyst for preparing a dicarboxylic acid and a method for preparing a dicarboxylic acid using the same. The enzyme catalyst is prepared by a method comprising the following steps: (1) carrying out fermentation culture on enzyme-producing microorganisms in a culture medium to obtain a crude enzyme solution, and (2) immobilizing the crude enzyme solution on a molecular sieve to obtain the enzyme catalyst, wherein the enzyme producing microorganisms are selected from candida tropicalis, candida sake, pichia pastoris, saccharomyces cerevisiae, lactobacillus brevis, corynebacterium glutamicum, bacillus subtilis, bacillus cereus, escherichia coli and pseudomonas aeruginosa. The method comprises the following steps: in a PBS buffer solution, mixing triglyceride with the enzyme catalyst, and carrying out catalytic reaction under stirring at 20-50 DEG C; and then carrying out hydrolysis reaction at the temperature of 250-300 DEG C under the pressure of 30-100 bar. According to the method, the enzyme catalyst and the molecular sieve are immobilized, so that other oxidants or catalysts are not required to be added while the dicarboxylic acid with higher yield is obtained.

The invention discloses a method for producing 5-aminolevulinic acid by inhibiting expression of a global transcriptional regulatory factor gene mcbR in corynebacterium glutamicum. The method comprises the following steps: (1) constructing a pCRISPRi-mid-McbR plasmid and a pCRISPRi-high-McbR plasmid; (2) respectively introducing the plasmids into corynebacterium glutamicum Cgh3 through electrotransfection, verifying by using a bacterial colony PCR (Polymerase Chain Reaction) mode to obtain an expression strain Cgh3-m-McbR or Cgh3-h-McbR for inhibiting a global transcriptional regulation factor gene mcbR in the corynebacterium glutamicum, and fermenting to obtain 5-aminolevulinic acid; experiments prove that compared with a control strain Cgh3-pXi, the strain for inhibiting the expression of the global transcriptional regulation factor gene mcbR in corynebacterium glutamicum has the advantage that the yield of 5-aminolevulinic acid is increased by 1.27 times or more after fermentation.

The invention belongs to the technical field of biological medicine, and particularly relates to application of corynebacterium casei in preparation of medicine for preventing and treating lung injury. By constructing a radon inhalation-induced lung injury mouse model, after exposure at different radon levels, the weight of a mouse is reduced, the lung coefficient is increased, the tidal volume of the lung, the ventilation volume per minute and the maximum inspiration flow are reduced, the respiratory function is inhibited, the alveolar cavity of the mouse is abnormally expanded, the pulmonary septum is thickened and fractured, and more fibrin exudation is accompanied; the traditional Chinese medicine composition can be used for effectively inhibiting pulmonary fibrosis and lung injury due to obvious inflammatory infiltration beside bronchial tubes, extravasated blood in small blood vessels and lung tissue collagen deposition, and can be used for gavage of corynebacterium casei to effectively inhibit the above phenomena. Therefore, the corynebacterium casei can be used for preventing and treating lung injury, especially lung injury caused by radon exposure.

The embodiment of the invention discloses a recombinant III-type humanized collagen as well as a preparation method and application thereof. The amino acid sequence of the recombinant III-type humanized collagen is shown as SEQ ID NO.1, SEQ ID NO.2 or SEQ ID NO.3. The recombinant III-type humanized collagen has the advantages that the recombinant III-type humanized collagen can be used for preparing the recombinant III-type humanized collagen; the recombinant III-type humanized collagen provided by the invention has the activity of promoting cell adhesion, the amino acid sequence of the recombinant III-type humanized collagen is selected from the amino acid sequence of natural collagen, and the recombinant III-type humanized collagen does not generate immune response when being applied to a human body. The characteristic that corynebacterium glutamicum does not secrete endotoxin is utilized, the preparation method is simple, the product can meet the requirements of food, cosmetics, medical instruments and drugs without complex separation, and reference is provided for low-cost industrial production.

The invention discloses a corynebacterium glutamicum engineering strain produced by L-glutamine as well as a construction method and application of the corynebacterium glutamicum engineering strain. The corynebacterium glutamicum engineering strain is prepared by integrating a glutamine synthase encoding gene glnA which is sourced from saccharomyces cerevisiae and has a tyrosine residue mutation at the 99th site on the surface and an sRNA-MicC-EcHfq regulation module of a targeted adenylyltransferase gene glnE into a vector plasmid; the method comprises the following steps: constructing a recombinant plasmid for removing adenosine acylation of glutamine synthase; and transferring the recombinant plasmid into corynebacterium glutamicum of which the odhA gene expression is controlled by a growth stage specific promoter Pcg2705, so as to obtain a corynebacterium glutamicum engineering strain for producing L-glutamine. According to the invention, through multi-gene synergistic modification and space-time metabolic flux optimization, the problem of'growth-synthesis' tradeoff in the prior art is solved, the fermentation efficiency is improved, the fermentation process is simple, and the application prospect is good.

The invention discloses a corynebacterium glutamicum whole genome random mutation tool and application thereof, and belongs to the technical field of genetic engineering and biological mutagenesis. DsDNA is unwound in the transcription process based on DNA helicase, DNA adenine deaminase and the DNA helicase are fused, and whole-genome random mutation of corynebacterium glutamicum is achieved. The whole genome random mutation method disclosed by the invention is applied to mutagenesis of an original strain of Corynebacterium glutamicum ATCC 13032, and a mutant strain of which the biomass is 1.95 times of that of WT (OD600 is 2.2) and 1.99 times of that of WT / pXMJ19 (OD600 is 2.16) when being cultured under the condition that the pH value is 5.5 is obtained. The corynebacterium glutamicum whole genome random mutation method disclosed by the invention has important guiding significance for screening other mutation strains with enhanced robustness.

The invention discloses a CgGlmS mutant and application of the CgGlmS mutant in synthesis of N-acetylglucosamine (GlcNAc) from corynebacterium glutamicum, and belongs to the technical field of biology. According to the invention, a bottleneck enzyme CgGlmS in a synthetic route of N-acetylglucosamine is subjected to deep rational analysis and semi-rational transformation. Through systematic research on the structure and function of an original CgGlmS enzyme from corynebacterium glutamicum S9114, the efficient CgGlmS mutant A253M is successfully screened out, and the enzyme activity, stability and catalytic efficiency of the mutant are obviously improved compared with those of a wild type. The mutant CgGlmS is overexpressed in corynebacterium glutamicum S9114 by using plasmids, so that the yield of N-acetylglucosamine in corynebacterium glutamicum is increased. The CgGlmS enzyme is directionally modified, so that the catalytic efficiency of the CgGlmS enzyme is improved, accumulation of N-acetylglucosamine in corynebacterium glutamicum is further promoted, and powerful support is provided for efficient production of the corynebacterium glutamicum.

The invention discloses a construction method of recombinant corynebacterium glutamicum for fermentation synthesis of poly beta-(1, 6)-N-acetylglucosamine polysaccharide, and belongs to the technical field of bioengineering. The recombinant corynebacterium glutamicum capable of preparing the PNAG through fermentation is constructed by using synthetic biology and genetic engineering technologies for the first time, cheap carbon sources and nitrogen sources can be utilized, the recombinant corynebacterium glutamicum is suitable for high-density fed-batch fermentation, and the generation cost of the PNAG is reduced; the yield after 42 hours of fermentation can reach 6.82 g / L, and the method has industrial application potential.

A mixture includes a nickel-containing ore and / or concentrate thereof and an extraction composition including a fermentation broth obtained from an Aspergillus species (spp.), Acetobacter spp., Bacillus spp., Propionibacterium spp., Corynebacterium spp., Rhizopus spp., Clostridium spp., Fusobacterium spp., Pseudomonas spp., Bifidobacterium spp., Saccharomyces spp., Enterobacter spp., Escherichia spp., among others. The extraction composition is configured to selectively solubilize one or more components, such as a magnesium component from the nickel-containing ore and / or concentrate thereof. A method for extracting a metal from a nickel-containing ore and / or concentrate includes selectively solubilizing a magnesium component from the nickel-containing ore and / or concentrate thereof into an extraction composition including the fermentation broth. The method can include extracting a magnesium component from a nickel-containing ore and / or concentrate thereof into an extraction composition, separating the nickel-containing ore and / or concentrate thereof from an extracted magnesium component and broth, separating the broth from the extracted magnesium component, and repeating the extracting. The extracted magnesium component has a purity in a range from 10% to 90%.

The invention relates to the technical field of wastewater purification, and discloses a treatment method of high-salt and high-ammonia-nitrogen wastewater. The method comprises the following steps: (1) carrying out nitration treatment on the high-salt and high-ammonia-nitrogen wastewater under an aerobic condition to obtain first effluent containing nitrate; (2) carrying out denitrification treatment on the first effluent under an anaerobic condition to obtain nitrogen and purified water; wherein the nitrification treatment and the denitrification treatment are independently carried out in the presence of a compound microbial agent; the compound microbial agent at least comprises the following components: Thioalkalivibrio versutus D301, Thauera sp. Strain SND5 (Single Nucleotide Polymorphism 5), and Rhodococcus corynebacterium bacteria (Rhodococcus corynebacterium bacteria), and is characterized in that the compound microbial agent at least comprises the following components of: Thioalkalivibrio versutus D301, Thauera sp. Strain SND5 (Single Nucleotide Polymorphism 5) and Rhodococcus corynebacterium bacteria (Rhodococcus corynebacterium bacteria (Rhodococcus corynebacterium bacteria). The method provided by the invention can effectively remove ammonia nitrogen contained in the high-salt and high-ammonia-nitrogen wastewater, so that the concentration of the ammonia nitrogen in the purified water is lower than 8mg / L, and the emission requirement is met. The method also has the characteristics of extremely small sludge discharge amount, quick start and stable purification effect.

The invention discloses a screening and expression platform construction method for non-essential gene loci of corynebacterium glutamicum, and belongs to the technical field of biology. The center sites CGLRS02940, CGLRS03085, CGLRS08555, CGLRS09020 and CGLRS13560 of the corynebacterium glutamicum identified by the invention have the remarkable advantages that when gene knockout or knockin operation is carried out, the sites do not interfere with the normal physiological metabolism process of strains, and stable expression of exogenous genes can be guaranteed; meanwhile, the gene editing efficiency is relatively high, and an editing target with high universality and high reliability is provided for genetic engineering modification of corynebacterium glutamicum.

The present invention belongs to the field of biomedicine technology, and in particular to the application of Corynebacterium casei in the preparation of medicines for preventing and treating lung injury. The present invention constructs a mouse model of lung injury caused by radon inhalation and finds that after exposure to different radon levels, the mice lose weight, increase the lung coefficient, reduce the lung tidal volume, minute ventilation and maximum inspiratory flow, and their respiratory function is inhibited. The alveolar cavity of the mice is abnormally expanded, the lung septum is thickened and fractured, accompanied by more fibrin exudation, obvious inflammatory infiltration appears near the bronchus, congestion occurs in small blood vessels, and collagen deposition in lung tissue can lead to the occurrence of pulmonary fibrosis, causing lung injury. Oral administration of Corynebacterium casei can effectively inhibit the above-mentioned phenomenon. Therefore, Corynebacterium casei can be used for the prevention and treatment of lung injury, especially lung injury caused by radon exposure.

This invention relates to the fields of microbial genetic engineering and fermentation biomanufacturing technology, specifically disclosing a recombinant Corynebacterium glutamicum with high 5-aminolevulinic acid (ALA) production and its applications. Addressing the problems of existing strains such as crude enzyme activity regulation, inhibited cell growth, the need for external inhibitors, and low yield, this invention uses Corynebacterium glutamicum as a host and removes the gdhA gene without scarring to reduce precursor competitive metabolism. Site-directed mutations of the endogenous hemB gene (S288A, D128E, R231K) are performed to moderately reduce dehydratase activity. A high-expression plasmid pXMJ19-hemA containing the hemA gene from Rhodopsinus capsulatum is constructed and introduced into the modified strain to obtain a recombinant strain with optimized triple metabolism. This strain reduces product degradation without the need for external dehydratase inhibitors, does not affect cell growth, and achieves a yield up to 4.30 times higher than the control after 42 hours of fermentation, with ALA accumulation exceeding 8.9 g / L. The strain exhibits stable characteristics, the process is simple, and it has good industrial application value.

The invention belongs to the technical field of biochemical engineering, and provides a method for improving the high-temperature tolerance of corynebacterium glutamicum and the production capacity of L-tryptophan. The invention provides a novel high-temperature tolerance related gene target osrR of corynebacterium glutamicum, biomass accumulation of corynebacterium glutamicum under a high-temperature condition is remarkably improved by regulating and controlling the expression level of the novel high-temperature tolerance related gene target osrR, and the fermentation yield of L-tryptophan of recombinant corynebacterium glutamicum under the high-temperature condition is improved. Specifically, by reducing the expression level of the recombinant corynebacterium glutamicum, the biomass accumulation of the recombinant corynebacterium glutamicum under the condition of 40 DEG C and the yield of the L-tryptophan under the condition of 33.5 DEG C are remarkably improved, and support is provided for high-temperature fermentation production of the L-tryptophan. Wide application prospects are realized.

An exemplary embodiment of the present disclosure provides an allulose epimerase expression cassette including a polynucleotide encoding allulose epimerase and a promoter which is operably linked thereto and regulates the expression of the allulose epimerase in a Corynebacterium genus strain. The promoter consists of a base sequence represented by SEQ ID NO: 1, a base sequence represented by SEQ ID NO: 9, a base sequence represented by SEQ ID NO: 10, or a base sequence represented by SEQ ID NO: 11. According to the exemplary embodiment of the present disclosure, the allulose epimerase expression cassette is hardly operated under a normal environment of culturing a recombinant Corynebacterium genus strain and does not induce growth inhibition of the recombinant Corynebacterium genus strain. Therefore, the recombinant Corynebacterium genus strain transformed with the allulose epimerase expression cassette may grow and proliferate normally until a logarithmic phase and stably and highly express allulose epimerase after the logarithmic phase.

The application discloses a construction method and application of a genetically engineered corynebacterium glutamicum for synthesizing a bacteriochlorophyllide amino acid shinorine, and belongs to the field of genetic engineering and fermentation technology. The genetically engineered corynebacterium glutamicum is constructed by taking corynebacterium glutamicum ATCC 13032 as a host strain, knocking out a competitive pathway gene, integrating a key enzyme gene in multiple copies on a genome, knocking out an acid production pathway gene and a serine dehydrogenase gene, and expressing a gene cluster for synthesizing shinorine on a plasmid, so that the synthesis yield of shinorine in a microbial cell is improved. The strain can produce shinorine in a flask, and 1.12 g / L of shinorine can be accumulated in 72 hours. In fed-batch fermentation in a 50 L fermenter, the strain can be cultured to 8.6 g / L.

The invention relates to Corynebacterium glutamicum with high indigo yield as well as a construction method and application thereof, and belongs to the technical field of gene recombination fermentation. The recombinant corynebacterium glutamicum can be used for overexpression of the following genes: a D-glucose-6-phosphate dehydrogenase gene zwf, a II type deoxyarabinose type heptulose phosphate synthase gene aroG and a flavin-containing monooxygenase gene CcFMO. The recombinant corynebacterium glutamicum can weaken the following genes: a pyruvate kinase gene pyk and a tryptophan synthase gene trpB. The corynebacterium glutamicum is used for overexpressing a zwf gene, an aroG gene and a CcFMO gene, weakening a pyk gene and a trpB gene and enhancing the expression quantity of an indole synthetic pathway related enzyme gene, so that the yield of the corynebacterium glutamicum at a shake flask stage reaches 619mg / L in a fermentation production process of indigo under the condition of not additionally adding tryptophan. The invention provides the recombinant corynebacterium glutamicum with high indigo yield, which is low in production cost, good in stability and high in synthesis efficiency, and is suitable for practical popularization and application.

To provide: an allulose epimerase expression cassette capable of highly expressing an allulose enzyme in a stationary phase after a logarithmic phase of a Corynebacterium host microorganism without inducing growth inhibition of the Corynebacterium genus host microorganism because of a low expression level of the allulose enzyme in an early stage of culture; and uses thereof.SOLUTION: An exemplary embodiment of the present disclosure provides an allulose epimerase expression cassette including a polynucleotide encoding allulose epimerase and a promoter which is operably linked thereto and regulates the expression of the allulose epimerase in a Corynebacterium genus strain. The promoter consists of a specific base sequence.SELECTED DRAWING: Figure 3

The invention relates to the technical field of biology, and provides a bacterial strain capable of accelerating and promoting germination of tulipa seeds and application of the bacterial strain, the bacterial strain is obtained by being separated from soil of tulipa germinating seedlings, the bacterial strain is identified as aspergillus niger through ITS sequence sequencing after being purified, the taxonomic name is Aspergillus niger, and the preservation number of the bacterial strain is CCTCC NO: M 2025709. Through an associated germination test of the strain and tulipa plant seeds, the result shows that the strain can obviously accelerate the germination of the tulipa seeds and improve the germination rate of the tulipa seeds.

The invention discloses genetically engineered corynebacterium glutamicum and an application of the genetically engineered corynebacterium glutamicum in preparation of ectoine, and the genetically engineered corynebacterium glutamicum contains an L-diaminobutyric acid transaminase gene ectB, an L-diaminobutyric acid acetyltransferase gene ectA and an ectoine synthetase gene ectC, the invention relates to an aspartic acid transferase gene aspB, an aspartic acid kinase gene lysC and an aspartic acid semialdehyde dehydrogenase gene asd, wherein the nucleotide sequences of the gene aspB, the gene lysC and the gene asd are respectively as shown in SEQ ID NO.4, SEQ ID NO.5 and SEQ ID NO.6. The invention further discloses a preparation method of the aspartic acid transferase gene aspB, the aspartic acid kinase gene lysC and the aspartic acid semialdehyde dehydrogenase gene asd. After the recombinant corynebacterium glutamicum constructed by the invention is fermented for 60 hours, the yield of ectoine can reach 50g / L. The genetically engineered corynebacterium glutamicum disclosed by the invention has important practical significance on industrial production and large-scale application of ectoine.

This invention discloses a method for the efficient synthesis of gastrodin using recombinant Corynebacterium glutamicum, belonging to the fields of genetic engineering, fermentation engineering, and synthetic biology. This invention uses Corynebacterium glutamicum as the host bacterium and knocks out… pobA The gene was expressed, and the engineered strain WN01 / pB6 was constructed using the expression plasmid pEC-XK99E to express cladoid-pyruvate lyase, carboxylic acid reductase, and glycosyltransferase, achieving heterologous synthesis of gastrodin. Further overexpression was then performed. aroG Genes, overexpression of protein-coding genes that activate carboxylic acid reductase sfp The engineered strain WN05 / pB6 was able to synthesize 7.37 g / L of gastrodin at the shake flask level, and the gastrodin yield reached 17.77 g / L at 72 h of batch fermentation in a 5-L fermenter, with a yield of 0.247 g / L / h. This provides a brand-new biomanufacturing platform for the industrial production of gastrodin.

The present invention provides recombinant Corynebacterium glutamicum genetically engineered to introduce the N-acetylmannosamine (ManNAc) synthesis pathway, and genetically engineered to express an exogenous N-acetylneuraminic acid synthetase, an exogenous CMP-N-acetylneuraminic acid synthetase, an exogenous lactose transporter, and an exogenous sialyltransferase. The invention also provides a method for producing sialic acid lactose by using the recombinant corynebacterium glutamicum.

The present invention relates to a novel promoter derived from Corynebacterium and to a use of a variant thereof in the production of a protein or a rare sugar. In the present invention, a sod gene promoter derived from Corynebacterium ammoniagenes, which enables regulation of gene expression and exhibits significantly stronger activity compared to conventionally used promoters, was developed, promoter variants having various strengths were identified from the developed promoter, and by using the identified improved promoters, two enzymes capable of converting a common sugar into a rare sugar were highly expressed in a single vector system, thereby enabling production of a rare sugar via a one-pot reaction. Accordingly, the promoters can be utilized for the production of a target protein or a rare sugar.

The invention relates to the technical field of tag peptides, and particularly discloses a tag peptide combined on the surface of corynebacterium, and the amino acid sequence of the tag peptide is shown as SEQ ID NO: 1 or SEQ ID NO: 2. The tag peptide provided by the invention enriches the types of tag peptides bound to the surfaces of current bacteria, and corynebacterium bacteria bound with the tag peptide have wider adaptability to pH and can be compatible with the optimal activity conditions of more foreign proteins; the molecular weight of the tag peptide is only 3.5 kDa, so that the interference to the structure or function of an exogenous protein can be remarkably reduced; in addition, the tag peptide can be fused at the N end or the C end of the target protein, the binding efficiency is not affected, simultaneous display of multiple proteins is supported, and a basis is provided for multi-enzyme cascade reaction and multivalent vaccine development.

The application discloses an expression vector of a biosynthetic metabolic pathway gene of 2-azacyclohexanone and a synthesis method of 2-azacyclohexanone. The expression vector comprises a first promoter and, sequentially inserted downstream of the first promoter, davA-davB genes or davB-davA genes, a Pb promoter or a mutant thereof, an act gene or an orf26 gene or a caiC gene, a ChnR gene or a mutant thereof. The expression vector is further transferred into glutamic acid corynebacterium with high lysine yield, and is subjected to fermentation culture to efficiently synthesize 2-azacyclohexanone. The application synthesizes 2-azacyclohexanone in glutamic acid corynebacterium for the first time, and utilizes an optimized 2-azacyclohexanone molecular probe system to start the expression of key genes in the 2-azacyclohexanone biosynthetic metabolic pathway for the first time, so that high yield of 2-azacyclohexanone is realized. The synthesis method reduces the production cost required by using lysine as a synthesis raw material, eliminates a rate-limiting step, and greatly improves the biosynthesis yield of 2-azacyclohexanone.

As one aspect, the present disclosure provides a bacterium of the genus Corynebacterium capable of producing isopropanol with high productivity. As another aspect, the present disclosure provides a method which enables the production of isopropanol with high productivity while suppressing the production of by-products.　In one aspect, the present disclosure relates to a bacterium of the genus Corynebacterium capable of producing isopropanol, the bacterium being produced by introducing into a bacterium of the genus Corynebacterium a specific gene (A) that encodes an enzyme having an activity of producing acetoacetic acid from acetoacetyl-CoA and / or a specific gene (B) that encodes an enzyme having an activity of producing acetone from acetoacetic acid in a manner that enables expression thereof. In another aspect, the present disclosure relates to a method for producing isopropanol, the method comprising: allowing a genetically modified bacterium of the genus Corynebacterium capable of producing isopropanol to carry out a fermentation reaction in a fermentation tank to produce a fermentation product containing isopropanol; continuously or intermittently withdrawing a portion of a reaction solution containing the fermentation product from the fermentation tank during the fermentation reaction; separating isopropanol from the withdrawn reaction solution by pervaporation; and supplying to the fermentation tank the reaction solution from which isopropanol has been separated.

Provided are a novel gluconate repressor variant, a polynucleotide comprising the variant of the present disclosure, a microorganism of the genus Corynebacterium comprising the variant or polynucleotide of the present disclosure, and a method of producing L-arginine using the microorganism of the present disclosure.

Provided are a protein variant, a microorganism of the genus of Corynebacterium comprising the protein variant and a method of producing L-lysine using the microorganism in the present disclosure.