Corynebacterium genus microorganism having improved L-arginine productivity, and method for producing L-arginine using same

Abstract

The present invention relates to a microorganism belonging to the genus Corynebacterium having improved L-arginine producibility and a method for producing L-arginine using the same, the microorganism belonging to the genus Corynebacterium having improved L-arginine production yield compared to a parent strain by introducing an argO gene encoding an arginine export protein, and being capable of efficiently producing L-arginine.

Description

Technical Field

[0001] The present invention relates to a Corynebacterium microorganism having improved L-arginine production ability and a method for producing L-arginine using the same. Background Art

[0002] Arginine is known to be contained in plants and the like in a free state. Arginine is a non-essential amino acid, but it is a semi-essential amino acid that must be supplied to growing children, under stress, trauma, cancer and other special conditions. It is widely used as a component of amino acid enhancers, pharmaceuticals, foods and the like. When used in medicine, it is used as a liver function enhancer, brain function enhancer, male infertility treatment agent, comprehensive amino acid preparation, etc. When used in food, it is used as a fish cake additive, health drink additive, and salt substitute for patients with hypertension.

[0003] Arginine can be produced by using wild-type strains obtained in nature or mutants modified in a way that improves its arginine production capacity. In recent years, in order to improve the production efficiency of arginine, gene recombination technology has been applied to microorganisms such as Escherichia coli and Corynebacterium. The biosynthesis of L-arginine in microorganisms is synthesized from L-glutamate as a starting material, through N-acetylglutamate, N-acetylglutamyl-P, N-acetylglutamate 5-semialdehyde, N-acetylornithine, L-ornithine, L-citrulline, and argininosuccinate. In this step-by-step synthesis process, various proteins such as enzymes, transcription factors, and transporters are involved. Therefore, attempts have been made to increase the production of L-arginine by inducing mutations in various genes involved in the biosynthesis pathway of L-arginine or regulatory sequences such as promoters that regulate their expression using genetic recombination techniques.

[0004] According to the literature [Bellmann et al., Microbiology 2001; 147: 1765-74], lysine exporter is not only involved in lysine, but also in the discharge of arginine, citrulline, and ornithine. When the expression of the lysE gene encoding the lysine exporter is enhanced, the arginine discharge capacity is enhanced. However, due to the enhancement of the lysE gene, the discharge of byproducts other than arginine is also increased, and in particular, the production increase of ornithine, which is equivalent to a byproduct, has a negative impact on the production of arginine.

[0005] Therefore, in order to develop a recombinant strain or a mutant strain having reduced production of by-products and excellent L-arginine-producing ability, a lot of research is still required.

[0006] Prior art literature

[0007] Patent Literature

[0008] Korean Patent No. 10-2269637 Summary of the invention

[0009] An object of the present invention is to provide a Corynebacterium microorganism having an improved L-arginine production ability.

[0010] Another object of the present invention is to provide a method for producing L-arginine using the above-mentioned microorganism.

[0011] One embodiment of the present invention provides a Corynebacterium microorganism having enhanced activity of an arginine exporter and improved L-arginine production capacity.

[0012] The "arginine exporter" used in the present invention regulates the intracellular arginine level and plays a role in balancing arginine and lysine. The arginine exporter in the present invention can be a polypeptide encoded by the argO gene and having arginine exporter activity or arginine excretion ability, but is not limited thereto.

[0013] The nucleic acid sequence and protein sequence information of the above-mentioned arginine exporter can be obtained through known sequence databases (eg, GenBank, UniProt).

[0014] The "activity enhancement" used in the present invention refers to the new introduction or increase in the expression of genes encoding target enzymes, transcription factors, transporters and other proteins, thereby increasing the expression level compared to the wild-type strain or the strain before transformation. Such activity enhancement includes the following situations: the activity of the protein itself is increased compared to the activity of the protein originally possessed by the microorganism through nucleotide substitution, insertion, deletion or a combination thereof of the encoding gene, and the overall protein activity level in the cell is higher than that of the wild-type strain or the strain before transformation through increased expression or increased translation of the gene encoding it, and also includes their combination.

[0015] According to a specific example of the present invention, the increase in the activity of the arginine exporter can be achieved by introducing a gene encoding the arginine exporter.

[0016] According to one embodiment of the present invention, the arginine exporter or the gene encoding the arginine exporter may be derived from a microorganism of the genus Escherichia.

[0017] Examples of the Escherichia microorganism include, but are not limited to, Escherichia coli, Escherichia albertii, Escherichia blattae, Escherichia fergusonii, Escherichia hermannii, and Escherichia vulneris.

[0018] For example, the above-mentioned arginine exporter or the gene encoding the same may be derived from Escherichia coli.

[0019] According to a specific example of the present invention, the arginine exporter may comprise the amino acid sequence of SEQ ID NO:1.

[0020] According to a specific example of the present invention, the gene encoding the above-mentioned arginine exporter may include the base sequence of SEQ ID NO:2.

[0021] The amino acid sequence or base sequence of the arginine exporter according to the present invention can be composed of a sequence having 70% or more, 80% or more, 90% or more, 98% or more, 99% or more, or 99.9% or more homology with the amino acid sequence of SEQ ID NO: 1 or the base sequence of SEQ ID NO: 2, or must contain it and be able to maintain the original function.

[0022] "Productivity is improved" used in the present invention means that the productivity of L-arginine increases compared to the parent strain. The above-mentioned parent strain refers to the wild type or mutant that becomes the object of variation, including the object that directly becomes the object of variation or the object transformed by the vector of recombinant. In the present invention, the parent strain can be a wild-type coryneform bacteria microorganism or a coryneform bacteria microorganism or strain mutated by the wild type.

[0023] The above-mentioned Corynebacterium microorganism may be any known microorganism in the art, for example, Corynebacterium glutamicum, Corynebacterium crudilactis, Corynebacterium deserti, Corynebacterium callunae, Corynebacterium suranareeae, Corynebacterium lubricantis, Corynebacterium doosanense, Corynebacterium efficiens, Corynebacterium uterequi, Corynebacterium stationis, Corynebacterium pacaense, Corynebacterium spp. singulare), Corynebacterium humireducens, Corynebacterium marinum, Corynebacterium halotolerans, Corynebacterium spheniscorum, Corynebacterium freiburgense, Corynebacterium striatum, Corynebacterium canis, Corynebacterium ammoniagenes, Corynebacterium renale, Corynebacterium pollutisoli, Corynebacterium imitans, Corynebacterium caspium, Corynebacterium testudinoris, Corynebacterium pseudocorynebacterium pseudopelargi) or Corynebacterium flavescens, but is not limited thereto.

[0024] According to a specific example of the present invention, the Corynebacterium microorganism may be Corynebacterium glutamicum.

[0025] According to the present invention, the Corynebacterium microorganism having improved L-arginine productivity can improve L-arginine productivity because the activity of the arginine exporter is enhanced, thereby increasing the amount of arginine excreted from the cell. Specifically, the Corynebacterium microorganism having improved L-arginine productivity shows increased L-arginine productivity compared to the parent strain, and in particular, the L-arginine production amount can be increased by at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 100% compared to the parent strain, or can be increased by 1.1 times, 1.5 times, 2 times, 2.5 times, 3 times, 3.5 times, 4 times, 4.5 times, 5 times, 5.5 times, 6 times, 6.5 times, 7 times, 7.5 times, 8 times, 8.5 times, 9 times, 9.5 times or 10 times, but is not limited thereto.

[0026] The Corynebacterium microorganism according to one embodiment of the present invention can be realized by a recombinant vector comprising a gene encoding an arginine exporter protein of a parent strain.

[0027] "Carrier (vector)" used in the present invention refers to all types of nucleic acid sequence transport structures used as a means for delivering and expressing target genes to host cells. Unless otherwise specified, the above-mentioned carrier can refer to inserting the nucleic acid sequence carried into the host cell gene for expression and / or independently expressing. Such a carrier includes an essential regulatory sequence operably connected so that the gene insert can be expressed, and "operably connected (operably linked)" refers to that the target gene and its regulatory sequence are functionally combined with each other and connected in a manner that can carry out gene expression, and "regulatory sequence" includes a promoter sequence for implementing transcription, an arbitrary operator sequence for regulating transcription, a sequence encoding a suitable mRNA ribosome binding site, and a sequence regulating the termination of transcription and translation.

[0028] The vector used in the present invention is not particularly limited as long as it can replicate in the host cell, and any vector known in the art can be used. As an example of the above-mentioned vector, a plasmid, a cosmid, a virus and a phage in a natural state or a recombinant state can be cited. For example, as a phage vector or a cosmid vector, there are pWE15, M13, λMBL3, λMBL4, λIXII, λASHII, λAPII, λt10, λt11, Charon4A, Charon21A, etc., and as a plasmid vector, there are pBR system, pUC system, pBluescriptII system, pGEM system, pTZ system, pCL system and pET system, etc., but it is not limited thereto.

[0029] The above-mentioned vector can be constructed as a vector for cloning or a vector for expression. The vector for expression can use a conventional vector for expressing foreign genes or proteins in plants, animals or microorganisms in the field, and can be constructed by various methods known in the field.

[0030] The "recombinant vector" used in the present invention can be constructed with prokaryotic cells or eukaryotic cells as hosts, can be replicated independently of the genome of the host cell, or can be sutured to the genome itself. The above-mentioned host cells are capable of replicating the vector and may include a replication origin as a specific base sequence for starting replication. For example, when the vector used is an expression vector and a prokaryotic cell is used as a host, it usually contains a strong promoter (e.g., pLλ promoter, CMV promoter, trp promoter, lac promoter, tac promoter, T7 promoter) that can enable transcription, a ribosome binding site for translation initiation, and a transcription / translation termination sequence. When eukaryotic cells are used as hosts, the replication origins initiated in the eukaryotic cells contained in the vector include f1 replication origin, SV40 replication origin, pMB1 replication origin, adenovirus replication origin, AAV replication origin, and BBV replication origin, but are not limited thereto. In addition, promoters derived from the genome of mammalian cells (e.g., metallothionein promoter) or promoters derived from mammalian viruses (e.g., adenovirus late promoter, vaccinia virus 7.5K promoter, SV40 promoter, cytomegalovirus promoter, tk promoter of HSV) can be used, and usually have a polyadenylation sequence as a transcription termination sequence.

[0031] The above-mentioned recombinant vector may include a selection marker, which is used to screen transformants (host cells) transformed by the vector. In the culture medium treated with the above-mentioned selection marker, only cells expressing the selection marker can survive, so transformed cells can be screened. As representative examples, the above-mentioned selection markers include ampicillin, kanamycin, streptomycin, chloramphenicol, etc., but are not limited thereto.

[0032] By inserting the above-mentioned recombinant vector into a host cell, a transformant can be prepared, and the above-mentioned transformant can be obtained by introducing the recombinant vector into a suitable host cell. The host cell is a cell that can stably and continuously clone or express the above-mentioned expression vector, and any host cell known in the art can also be used.

[0033] When prokaryotic cells are transformed to produce recombinant microorganisms, Escherichia coli strains such as E. coli DH5α, E. coli JM109, E. coli BL21, E. coli RR1, E. coli LE392, E. coli B, E. coli X1776, E. coli W3110, and E. coli XL1-Blue; Corynebacterium strains; Bacillus strains such as Bacillus subtilis and Bacillus thuringiensis; various enteric bacteria and strains such as Salmonella typhimurium, Serratia marcescens, and Pseudomonas can be used as host cells, but are not limited to these.

[0034] When eukaryotic cells are transformed to produce recombinant microorganisms, yeast (e.g., Saccharomyces cerevisiae), insect cells, plant cells, and animal cells can be used as host cells, for example, Sp2 / 0, CHO K1, CHO DG44, PER.C6, W138, BHK, COS7, 293, HepG2, Huh7, 3T3, RIN, MDCK cell lines, etc., but are not limited to these.

[0035] As used in the present invention, “transformation” refers to the phenomenon of introducing foreign DNA into host cells to cause artificial genetic changes, and “transformant” refers to a host cell into which foreign DNA has been introduced and which stably maintains the expression of the target gene.

[0036] In the above transformation, a suitable vector introduction technique is selected according to the host cell, so that the target gene or the recombinant vector containing the same can be expressed in the host cell. For example, the vector introduction can be carried out by electroporation, heat shock, calcium phosphate (CaPO 4 ) precipitation, calcium chloride (CaCl 2 ) precipitation, microinjection, polyethylene glycol (PEG) method, DEAE-dextran method, cationic liposome method, lithium acetate-DMSO method, or a combination thereof, but is not limited thereto. As long as the transformed gene can be expressed in the host cell, it can be included, without being limited to being inserted into the chromosome of the host cell or being located outside the chromosome.

[0037] The transformant includes cells transfected, transformed or infected with the recombinant vector according to the present invention in vivo or in vitro, and can be used as the same term as a recombinant host cell, a recombinant cell or a recombinant microorganism.

[0038] The gene inserted into the recombinant vector for transformation of the present invention can be replaced into a host cell such as a microorganism of the genus Corynebacterium by homologous recombination and crossing.

[0039] According to a specific example of the present invention, the host cell may be a microorganism of the genus Corynebacterium, for example, the microorganism of the genus Corynebacterium may be Corynebacterium glutamicum.

[0040] Another aspect of the present invention provides a method for producing L-arginine, comprising the steps of: culturing the above-mentioned Corynebacterium microorganism in a culture medium; and recovering L-arginine from the above-mentioned microorganism or the culture medium in which the microorganism is cultured.

[0041] The above-mentioned culture can be carried out according to suitable culture medium and culture conditions known in the art, and those skilled in the art can easily adjust the culture medium and culture conditions for use. Specifically, the above-mentioned culture medium can be a liquid culture medium, but is not limited thereto. The culture method can include, for example, batch culture, continuous culture, fed-batch culture, or a combination thereof, but is not limited thereto.

[0042] According to a specific example of the present invention, the above-mentioned culture medium must meet the requirements of a specific strain in a suitable manner and can be appropriately modified by those skilled in the art. Regarding the culture medium of Corynebacterium strains, reference can be made to known literature (Manual of Methods for General Bacteriology. American Society for Bacteriology. Washington DC, USA, 1981), but it is not limited thereto.

[0043] According to a specific example of the present invention, the culture medium may contain various carbon sources, nitrogen sources and trace element components. As the carbon source that can be used, sugars and carbohydrates such as glucose, sucrose, lactose, fructose, maltose, starch, cellulose, etc. are included; oils and fats such as soybean oil, sunflower oil, castor oil, coconut oil, etc.; fatty acids such as palmitic acid, stearic acid, linoleic acid; alcohols such as glycerol and ethanol; organic acids such as acetic acid. These substances can be used alone or in the form of a mixture, but are not limited to this. As the nitrogen source that can be used, peptone, yeast extract, broth, malt extract, corn steep liquor, soybean meal and urea or inorganic compounds such as ammonium sulfate, ammonium chloride, ammonium phosphate, ammonium carbonate and ammonium nitrate can be included. Nitrogen sources can also be used alone or in the form of a mixture, but are not limited to this. As a source of phosphorus that can be used, potassium dihydrogen phosphate or dipotassium hydrogen phosphate or corresponding sodium-containing salts can be included, but are not limited to this. In addition, the culture medium may contain metal salts such as magnesium sulfate or ferrous sulfate required for growth, but is not limited to this. In addition, essential growth substances such as amino acids and vitamins may be included. In addition, precursors suitable for the culture medium may be used. The above-mentioned culture medium or individual components may be added to the culture solution in batches or continuously in a suitable manner during the culture process, but are not limited thereto.

[0044] According to a specific example of the present invention, during the culture process, compounds such as ammonium hydroxide, potassium hydroxide, ammonia, phosphoric acid and sulfuric acid can be added to the microbial culture solution in an appropriate manner to adjust the pH of the culture solution. In addition, during the culture process, defoamers such as fatty acid polyethylene glycol esters can be used to inhibit the generation of bubbles. Further, in order to maintain the aerobic state of the culture solution, oxygen or oxygen-containing gas (such as air) can be injected into the culture solution. The temperature of the culture solution can generally be 20°C to 45°C, for example, 25°C to 40°C. The culture time can continue until the desired production of useful substances is obtained, for example, it can be 10 to 160 hours.

[0045] According to a specific example of the present invention, in the step of recovering L-arginine from the cultured Corynebacterium microorganism and the culture medium for culturing the microorganism, the produced L-arginine can be collected or recovered from the culture medium according to the culture method and using a suitable method known in the art. For example, centrifugation, filtration, extraction, spraying, drying, evaporation, precipitation, crystallization, electrophoresis, fractional dissolution (e.g., ammonium sulfate precipitation), chromatography (e.g., ion exchange, affinity, hydrophobicity and size exclusion) and other methods can be used, but are not limited thereto.

[0046] According to a specific example of the present invention, in the step of recovering L-arginine, the culture medium may be centrifuged at a low speed to remove biomass, and the obtained supernatant may be separated by ion exchange chromatography.

[0047] According to a specific example of the present invention, the step of recovering L-arginine may include a process of purifying L-arginine.

[0048] The Corynebacterium microorganism according to the present invention can efficiently produce L-arginine by introducing a gene encoding an arginine exporter, thereby improving the L-arginine production yield compared to the parent strain. DETAILED DESCRIPTION

[0049] The present invention will be described in more detail below, but such description is provided for illustrative purposes only to help understand the present invention, and the scope of the present invention is not limited to such illustrative description.

[0050] Example 1. Preparation of strains into which argO gene is introduced

[0051] In order to introduce the argO gene encoding the arginine export protein derived from an Escherichia microorganism into an arginine-producing strain derived from Corynebacterium glutamicum, Corynebacterium glutamicum 14GR (KCCM13219P) and E. coli DH5a (HIT Competent cells) as L-arginine-producing strains were used. TM , Cat No.RH618).

[0052] The above-mentioned Corynebacterium glutamicum 14GR was prepared in a mixture of 1L of distilled water, 10.5g of 98% glucose, 1g of beef extract, 4g of yeast extract, 2g of polypeptone, 2g of NaCl and 40g of (NH 4 ) 2 SO 4 The cells were cultured in ARG-broth (pH 7.2) at 30°C.

[0053] The above-mentioned E. coli DH5a was cultured at 37°C in an LB medium composed of 1 L of distilled water, 10.0 g of tryptone, 10.0 g of NaCl, and 5.0 g of yeast extract.

[0054] The antibiotic kanamycin (kanamycin) used was a product of Sigma.

[0055] DNA sequencing analysis and gene synthesis were commissioned to MacroGene Co., Ltd.

[0056] 1-1. Recombinant vector

[0057] Made for destroying the lysE gene that participates in the discharge of lysine and arginine in Corynebacterium glutamicum 14GR and importing the recombinant vector of the argO gene of the arginine export protein that coding derives from Escherichia microorganism in its position. Codon optimized argO base sequence (SEQ ID NO:2) and the right arm base sequence for cloning are obtained by gene synthesis. Centered on the lysE gene on the Corynebacterium glutamicum genome, the argO gene that is connected with the left arm 655bp part and the right arm 636bp part and promoter is amplified with PCR, after being connected by overlapping PCR (overlap PCR) method, is cloned in pk19mobsacB (ATCC, 87098) carrier.In order to make above-mentioned plasmid, each gene fragment is amplified, the primer of following table 1 is used.

[0058]

Table 1

[0059]

[0060] PCR was performed using the above primers under the following conditions. Using a thermocycler (TP600, TAKARA BIO Inc., Japan), 1 pM of oligonucleotides and 10 ng of chromosomal DNA of Corynebacterium glutamicum ATCC 13032 were used as templates in a reaction solution to which 100 μM of each deoxynucleotide triphosphate (dATP, dCTP, dGTP, dTTP) was added, and 25 to 30 cycles were performed in the presence of 1 unit of pfu-X DNA polymerase mixture (Solgent). Regarding the PCR implementation conditions, it was performed under the conditions of (i) denaturation step: 94°C for 30 seconds, (ii) annealing step: 58°C for 30 seconds, and (iii) extension step: 72°C for 1 to 2 minutes (giving 2 minutes of polymerization time per 1 kb).

[0061] The gene fragments prepared as described above were cloned into the pk19mobsacB vector using self-assembly cloning. The vector was transformed into E. coli DH5a, smeared on LB-agar plates containing 50 μg / ml of kanamycin, and cultured at 37°C for 24 hours. The resulting colonies were isolated, and after confirming whether the insert was correctly present in the vector, the vector was isolated and used for the recombination of the Corynebacterium glutamicum strain.

[0062] As a common process in the above method, the amplification of the corresponding gene is amplified by the PCR method from the genomic DNA of Corynebacterium glutamicum ATCC13032, and according to the strategy, it is inserted into the pk19mobsacB vector by the self-assembled cloning method, and screened in E.coli DH5a. Chromosomal base substitution is to amplify the genes of each fragment separately, and the target DNA fragment is produced by stacked PCR. In gene manipulation, Ex Taq polymerase (Takara) and Pfu polymerase (Solgent) were used as PCR amplification enzymes, and various restriction enzymes and DNA modifying enzymes used NEB products, which were used according to the supplied buffer and protocol.

[0063] 1-2. Corynebacterium glutamicum mutant DS1

[0064] The DS1 strain was prepared as a mutant strain using the cloning vector. The cloning vector was prepared in such a way that the final concentration was 1 μg / μl or more, and the electroporation method (reference [Tauch et al., FEMS Microbiology letters 123 (1994) 343-347]) was used to induce recombination once on Corynebacterium glutamicum 14GR. At this time, the electroporated strain was spread on an agar medium containing 50 μg / μl of kanamycin, and after the colonies were isolated, PCR and base sequence analysis were used to confirm whether the induction position on the genome was correctly inserted. In order to induce recombination twice again in the strain isolated in this way, it was inoculated in a liquid culture medium, cultured overnight or more, and spread on an agar medium containing 10% sucrose to isolate colonies. After confirming whether the colonies finally isolated had resistance to kanamycin, base sequence analysis was used to confirm whether the argO gene was introduced into the strain without antibiotic resistance (see reference [Schafer et al., Gene 145 (1994) 69-73]). Finally, a Corynebacterium glutamicum mutant strain DS1 capable of producing L-arginine was prepared, into which the argO gene was introduced.

[0065] Experimental Example 1. Evaluation of L-arginine production capacity of strains into which argO gene was introduced

[0066] The L-arginine production abilities of the parent strain, Corynebacterium glutamicum 14GR, and the Corynebacterium glutamicum DA1 into which the argO gene was introduced prepared in Example 1 were evaluated.

[0067] Each strain was patched in a flask seed culture solid medium and cultured at 30°C for 24 hours. The cultured colonies were inoculated into a 10 ml flask titer medium and cultured at 32°C and 200 rpm for 30 hours. The components of the culture medium used here are shown in Table 2 below. After the culture was completed, the culture solution was diluted 50 times with distilled water, filtered with a 0.45 μm filter, and the L-arginine production was analyzed using a high performance liquid chromatograph (HPLC) (agilent technologies 1260infinity, agilent technologies) equipped with a column (Dionex IonPacTM CS12A) and an ultraviolet detector (195 mm), and the results are shown in Table 3 below.

[0068]

Table 2

[0069]

[0070]

Table 3

[0071] strain <![CDATA[OD 610 ]]> L-Arginine(%) Residual sugar (%) L-Citrulline (%) L-Lysine(%) 14GR 32.0 2.37 1.43 0.026 0.023 DA1 40.0 2.62 0.75 0.012 0.008

[0072] As shown in Table 3 above, it was confirmed that the argO gene-introduced Corynebacterium glutamicum DA1 had enhanced arginine excretion and an approximately 10% increase in L-arginine production compared to the parent strain Corynebacterium glutamicum 14GR, and had excellent bacterial growth and sugar consumption, and reduced production of by-products such as L-citrulline and L-lysine.

[0073] So far, the present invention has been studied with its preferred embodiments as the center. It will be appreciated by those skilled in the art that the present invention can be implemented in a modified form without departing from the essential characteristics of the present invention. Therefore, the disclosed embodiments should be considered from an illustrative perspective rather than a restrictive perspective. The scope of the present invention is shown in the claims rather than in the above description, and should be interpreted as all differences within the scope of equivalence thereto are included in the present invention.

[0074]

Collection Information

[0075] Name of depository institution: Korea Collection of Microorganisms (KCCM)

[0076] Accession number: KCCM13219P

[0077] Collection date: June 29, 2022

[0078] Classification and nomenclature of biological material: Corynebacterium glutamicum.

Claims

1. A Corynebacterium microorganism, wherein the activity of an arginine export protein is enhanced and the L-arginine production capacity is improved.

2. The Corynebacterium microorganism according to claim 1, wherein The increase in the activity of the arginine exporter is achieved by introducing a gene encoding the arginine exporter.

3. The Corynebacterium microorganism according to claim 1, wherein The arginine exporter is derived from a microorganism of the genus Escherichia.

4. The Corynebacterium microorganism according to claim 1, wherein The arginine exporter comprises the amino acid sequence of SEQ ID NO:

1.

5. The Corynebacterium microorganism according to claim 1, wherein The Corynebacterium microorganism is Corynebacterium glutamicum.

6. A method for producing L-arginine, comprising the steps of: a step of culturing the Corynebacterium microorganism according to claim 1 in a culture medium, and A step of recovering L-arginine from the microorganism or the culture medium in which the microorganism is cultured.

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