CN115895980B

CN115895980B - Pallor for producing L-carnosine and application thereof

Info

Publication number: CN115895980B
Application number: CN202211727024.8A
Authority: CN
Inventors: 周宁; 李帆; 乔莉苹; 于鲁孟; 郭学平; 马欣娟; 毛华
Original assignee: Bloomage Biotech Co Ltd
Current assignee: Bloomage Biotech Co Ltd
Priority date: 2022-12-30
Filing date: 2022-12-30
Publication date: 2024-03-29
Anticipated expiration: 2042-12-30
Also published as: CN115895980A

Abstract

The application discloses a kind of pallor bacillus with preservation number of CGMCC No.25590 and its application in synthesizing L-carnosine. The strain can directly utilize L-histidine and beta-alanine to synthesize carnosine, and the yield of the L-carnosine can be up to about 20mg/L. The pallidum directly utilizes the L-histidine and the beta-alanine to synthesize the L-carnosine, and has the advantages of low cost, no methanol generation in the L-carnosine synthesis process and simple subsequent separation and purification.

Description

Pallor for producing L-carnosine and application thereof

Technical Field

The application relates to the field of microbial fermentation engineering, in particular to a pallor bacillus for producing L-carnosine and application thereof.

Background

Carnosine was first discovered by russian sch Gulewitsch 1900. He isolated carnosine from the meat extract of Liebig, which was later confirmed to have the structure β -alanyl-L-histidine. Carnosine is widely present in cells of body tissues, and is expressed highest in muscle and central nervous system, and in vertebrate bones, carnosine is present in amounts of about 1.6 to 20.6 mu mol/g, in pork, beef and chicken, 4.8, 2.0 and 15.0mmol, respectively, and in human muscle, 2 to 20mmol. At present, the production of L-carnosine is mainly extracted from the muscle of mammals, such as beef, pork, etc. However, the carnosine extract in animal muscle also contains a mixture of iron, heme, and the like, which can generate superoxide anions, hydroperoxides and hydroxyl radicals, and affects the antioxidant effect of L-carnosine to different degrees.

With the intensive research of carnosine, scholars have studied the biological properties of carnosine. To date, many biological activities of carnosine have been demonstrated, such as physiological pH buffering, chelating metal ions, scavenging free radicals, antioxidant, preventing protein glycation, anti-aging, anti-inflammatory and anti-tumor ^[5] Etc.

At present, the production of L-carnosine mainly adopts a chemical synthesis method, and is mainly divided into two main categories: (1) The main route of the synthesis of beta-alanine is that the beta-alanine is condensed with the protected L-histidine after amino protection and carboxyl activation, and then the protecting group is removed to obtain the L-carnosine. (2) reaction without beta-alanine involvement: l-histidine forms peptide bonds with different β -alanine precursors before further conversion to carnosine. Because harmful substances such as ethyl cyanoacetate and the like are used in the reaction process, water pollution and toxic reaction are easy to cause.

Depending on the enzymes used and the type of enzymatic carnosine synthesis reaction, the biological enzymatic processes for preparing L-carnosine can be divided into two main classes: (1) Beta-aminopeptidase catalyzes beta-alanyl amide or beta-alanine ester to be condensed and converted with L-histidine to synthesize L-carnosine, and the method has the defects that a chemical method is needed to activate carboxyl of beta-alanine, an organic solvent is used in the activation process, environmental pollution is easy to bring, and the cost is high; (2) Compared with the reaction of synthesizing the L-carnosine by the catalysis of aminopeptidase, the method does not need to chemically activate and protect a substrate, has fewer steps and is more environment-friendly. However, the currently published carnosine hydrolase has few varieties, low catalytic activity, long reaction time and low product concentration.

Patent CN107217048A discloses a method for synthesizing L-carnosine by using beta-alanine methyl ester hydrochloride and L-histidine as substrates, the method needs to react beta-alanine with methanol to generate beta-alanine methyl ester hydrochloride, and the L-carnosine generated in the conversion is decomposed into beta-alanine and L-histidine by aminopeptidase, which results in complex reaction products, difficult separation and purification and low L-carnosine yield.

Disclosure of Invention

Based on the current situation, the application aims at providing the pallor bacillus for producing the L-carnosine and the application thereof. The pallium (Ochrobactrum sp.) HXQ-1 is preserved in China general microbiological culture Collection center (CGMCC) for 8 months and 26 days in 2022, the preservation address is China, north Chen West Lu No. 1, 3 of the Korean region of Beijing, the post code is 100101, and the preservation number is CGMCC No.25590.

Specifically, the application adopts the following technical scheme:

1. the pallidum is characterized in that the preservation number of the pallidum is CGMCCNO.25590:

2. the pale bacillus according to item 1, wherein the 16s rRNA gene sequence of the pale bacillus is shown in SEQ ID NO. 1.

3. A culture containing the pallor bacillus of item 1.

4. The application of the pallor bacillus in the preparation of L-carnosine.

5. The use of the culture of item 3 for the preparation of L-carnosine.

6. The use of the pallidum of item 1 or the culture of item 2 in the preparation of a carnosine hydrolase.

7. A method for producing carnosine, comprising:

l-carnosine is obtained using the pallor bacterium of claim 1 or 2 or the culture of claim 3.

8. The method according to item 7, characterized in that it comprises the steps of:

placing the pallidum thallus or the culture in a reaction liquid containing L-histidine and beta-alanine for catalytic reaction to obtain L-carnosine, wherein the pallidum thallus is obtained by placing the pallidum thallus in a fermentation medium for culture and centrifugation.

9. The method of item 7, wherein the fermentation medium comprises the following components:

glucose 5-20g/L, peptone 5-10g/L, yeast powder 5-10g/L, ammonium chloride 5-10g/L, potassium dihydrogen phosphate 3-8g/L, sodium chloride 5-10g/L, and magnesium sulfate 0.5-3g/L.

10. The method according to item 7, wherein the conditions of the culture in the fermentation medium are:

culturing at 25-32deg.C under shaking at 120-220rpm for 12-24 hr.

11. The method according to item 7, wherein the reaction solution contains 5 to 30g/L of the pale bacteria, 2 to 10g/L of L-histidine and 1 to 8g/L of beta-alanine.

12. The method according to claim 7, wherein the pH of the reaction solution is 6.5 to 9.0, and preferably the buffer used in the reaction solution is Tris-HCl buffer;

further preferably, the conditions of the catalytic reaction are:

reacting for 12-36 h under the conditions of 25-37 ℃ and 100-200rpm oscillation.

13. The method according to item 7, wherein the culturing of the pallidum in the fermentation medium comprises the steps of:

activating the pallidum and inoculating the pallidum into a seed culture medium to obtain seed liquid;

inoculating the seed solution into a fermentation medium for culturing.

The pallidum is activated before being inoculated in a seed culture medium.

The application screens a strain of pallidum from fish intestinal tracts, and the strain can directly utilize L-histidine and beta-alanine to synthesize carnosine, and the carnosine yield is about 20mg/L. The method can directly utilize L-histidine and beta-alanine to synthesize the carnosine, and has the advantages of low cost, no methanol generation in the L-carnosine synthesis process and simple subsequent separation and purification.

Drawings

FIG. 1 is a liquid chromatogram of an L-carnosine standard;

FIG. 2 is a liquid chromatogram of a sample.

Detailed Description

Unless defined otherwise, technical and scientific terms used in this specification have the same meaning as commonly understood by one of ordinary skill in the art. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, the materials and methods are described herein below. In case of conflict, the present specification, including definitions therein, will control and materials, methods, and examples, will control and be in no way limiting. The present application is further illustrated below in conjunction with specific examples, but is not intended to limit the scope of the present application.

The experimental methods used below are conventional methods if no special requirements are imposed.

Materials, reagents and the like used in the following are commercially available unless otherwise specified.

The application provides an Ochrobactrum sp.) HXQ-1 which has been preserved in China general microbiological culture Collection center (CGMCC) for 8 months and 26 days in 2022, wherein the preservation address is China, north Chen West Lu No. 1, 3 of the Korean region of Beijing, and the postal code is 100101 and the preservation number is CGMCC No.25590.

The 16s rRNA gene sequence of the pallidum is shown in the following SEQ ID NO: 1:

GGCTCAGAACGAACGCTGGCGGCAGGCTTAACACATGCAAGTCGAGCGCCCCGCAAGGGGAGCGGCAGACGGGTGAGTAACGCGTGGGAACGTACCTTTTGCTACGGAATAACTCAGGGAAACTTGTGCTAATACCGTATGTGCCCTTCGGGGGAAAGATTTATCGGCAAAGGATCGGCCCGCGTTGGATTAGCTAGTTGGTGAGGTAAAGGCTCACCAAGGCGACGATCCATAGCTGGTCTGAGAGGATGATCAGCCACACTGGGACTGAGACACGGCCCAGACTCCTACGGGAGGCAGCAGTGGGGAATATTGGACAATGGGCGCAAGCCTGATCCAGCCATGCCGCGTGAGTGATGAAGGCCCTAGGGTTGTAAAGCTCTTTCACCGGTGAAGATAATGACGGTAACCGGAGAAGAAGCCCCGGCTAACTTCGTGCCAGCAGCCGCGGTAATACGAAGGGGGCTAGCGTTGTTCGGATTTACTGGGCGTAAAGCGCACGTAGGCGGACTTTTAAGTCAGGGGTGAAATCCCGGGGCTCAACCCCGGAACTGCCTTTGATACTGGAAGTCTTGAGTATGGTAGAGGTGAGTGGAATTCCGAGTGTAGAGGTGAAATTCGTAGATATTCGGAGGAACACCAGTGGCGAAGGCGGCTCACTGGACCATTACTGACGCTGAGGTGCGAAAGCGTGGGGAGCAAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGAATGTTAGCCGTTGGGGAGTTTACTCTTCGGTGGCGCAGCTAACGCATTAAACATTCCGCCTGGGGAGTACGGTCGCAAGATTAAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAACGCGCAGAACCTTACCAGCCCTTGACATACCGGTCGCGGACACAGAGATGTGTCTTTCAGTTCGGCTGGACCGGATACAGGTGCTGCATGGCTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTCGCCCTTAGTTGCCAGCATTTAGTTGGGCACTCTAAGGGGACTGCCGGTGATAAGCCGAGAGGAAGGTGGGGATGACGTCAAGTCCTCATGGCCCTTACGGGCTGGGCTACACACGTGCTACAATGGTGGTGACAGTGGGCAGCGAGCACGCGAGTGTGAGCTAATCTCCAAAAGCCATCTCAGTTCGGATTGCACTCTGCAACTCGAGTGCATGAAGTTGGAATCGCTAGTAATCGCGGATCAGCATGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCATGGGAGTTGGTTTTACCCGAAGGCGCTGTGCTAACCGCAAGGAGGCAGGCGACCACGGTAGGGTCAGCGACTGGGGTGAAGTCGT

the pallidum is obtained by screening by a transparent ring method.

Nutrient components which are poorly soluble and available to specific microorganisms penetrate into the solid medium, resulting in a cloudy, opaque medium background. L-carnosine is added into the solid culture medium, and the reaction is carried out by using a phthalaldehyde color development method to form blue color, which is used as a background of the screening culture medium. After the colonies to be screened are inoculated into the screening medium, colorless transparent circles are formed because they can decompose and utilize carnosine. The size of the transparent ring reflects the ability of the colony to utilize this substance. The transparent circle method is often used as a rapid method for preliminary screening of colonies.

In a specific embodiment, the screening method of the pallidum comprises the following steps:

after the sample is treated and diluted, the sample is coated in a screening culture medium containing L-carnosine for enrichment culture, and then colonies with different forms are picked up and cultured in a new screening culture medium containing L-carnosine.

And (3) verifying a transparent ring: after the bacteria in the newly screened plate grow out, reagents are sequentially added into the plate according to the reaction system shown in Table 1. After the reaction was completed, the plate was rinsed with tap water to remove the interference of the cells, and the strain with the transparent ring was observed and recorded.

TABLE 1

Wherein the sample can be beef and mutton, bean products, fish intestinal canal, seafood, etc.

In a specific embodiment, the reagents are added to the plate in sequence according to the reaction system of table 1, comprising the steps of:

0.2mL of 1% TCA standard solution and 8mL of 0.1mol/L NaOH solution are added for reaction for 5min,

then 0.8mL of 1% phthalic aldehyde solution is added for reaction for 1min,

0.8mL of 1mol/L HCl solution was added thereto, and the mixture was reacted at 20℃for 30 minutes.

In a specific embodiment, each 1L of the screening medium comprises: 3-10g of sodium chloride, 0.2-2g of peptone, 0.2-2g of yeast powder, 1-10g of L-carnosine, 20g of agar powder and the balance of water. The pH of the culture medium is 6.8-7.2.

The application also provides a culture containing the pallidum. In the present application, the culture of the pallidum means a substance obtained by culturing the pallidum of the present application, more specifically, a mixture of pallidum cells, a medium for culturing the pallidum, a substance produced by the cultured pallidum, and the like.

The application also provides application of the pallidum or the culture in preparation of L-carnosine.

The application also provides application of the pallidum or the culture in preparing carnosine hydrolase.

The present application also provides a method for producing carnosine, which obtains L-carnosine using the above-mentioned pallidum or a culture containing the above-mentioned pallidum.

Specifically, the method for producing carnosine comprises the following steps:

placing the pallidum into a fermentation culture medium for culturing and centrifuging to obtain pallidum thallus,

placing the pallidum thallus in a reaction liquid containing L-histidine and beta-alanine for catalytic reaction to obtain L-carnosine.

Wherein, the culturing of the pallidum in the fermentation medium can be directly culturing the pallidum in the fermentation medium, and the method also comprises the following steps:

inoculating the seed solution into a fermentation medium for culturing.

In a specific embodiment, the method of producing carnosine comprises the steps of:

inoculating the seed solution into a fermentation culture medium for culturing and centrifuging to obtain the pallidum thallus,

In a specific embodiment, the activation is performed in a slant medium at 25 to 32℃for 12 to 24 hours.

In a specific embodiment, the seed medium comprises the following components:

5-20g/L of glucose, 5-10g/L of peptone, 5-10g/L of yeast powder, 5-10g/L of ammonium chloride, 3-8g/L of monopotassium phosphate, 5-10g/L of sodium chloride, 0.5-3g/L of magnesium sulfate and the balance of water. The pH of the seed culture medium is 6.8-7.2.

In a specific embodiment, the conditions for culturing the pallidum in the seed medium are: shake culturing at 25-32 deg.c and 120-220rpm for 12-24 hr.

The seed liquid may be inoculated into the fermentation medium in any ratio. In a specific embodiment, the seed solution is inoculated into the fermentation medium in a volume ratio of 1% -10%.

In a specific embodiment, the fermentation medium comprises the following components:

In a specific embodiment, the conditions under which the pallidum is cultured in the fermentation medium are: shake culturing at 25-32 deg.c and 120-220rpm for 12-24 hr.

In a specific embodiment, the fermentation broth obtained after the completion of the fermentation culture is centrifuged at 6000 to 12000rpm, and the supernatant is removed to obtain the pallidum cells.

The obtained pallidum cells are used for catalyzing the reaction of L-histidine and beta-alanine to obtain the L-carnosine.

In a specific embodiment, the reaction solution contains 5-30g/L of the pallidum cells, 2-10g/L of L-histidine and 1-8g/L of beta-alanine. Wherein the mass of the pale bacteria is that the fermentation liquid is centrifuged at 6000-12000rpm, the supernatant is removed, and the obtained pale bacteria has a wet weight, and the content of the pale bacteria in the reaction liquid is the wet weight of the pale bacteria obtained under the condition unless otherwise specified.

In a specific embodiment, the pH of the reaction solution is from 6.5 to 9.0.

The reaction solution may be any buffer solution having a pH of 6.5 to 9.0.

In a specific embodiment, the reaction solution is Tris-HCl buffer with pH of 6.5-9.0.

In a specific embodiment, the conditions under which the catalytic reaction is carried out are:

placing the pallidum bacteria in a reaction solution containing L-histidine and beta-alanine for catalytic reaction to obtain L-carnosine, wherein the content of the pallidum bacteria in the reaction solution is 5-30g/L, the content of the L-histidine is 2-10g/L, the content of the beta-alanine is 1-8g/L,

wherein the seed culture medium comprises the following components:

5-20g/L of glucose, 5-10g/L of peptone, 5-10g/L of yeast powder, 5-10g/L of ammonium chloride, 3-8g/L of monopotassium phosphate, 5-10g/L of sodium chloride, 0.5-3g/L of magnesium sulfate and the balance of water. The pH of the seed culture medium is 6.8-7.2,

the fermentation medium comprises the following components:

inoculating the seed solution into a fermentation culture medium, shake culturing for 12-24 h at 25-32 ℃ and 120-220rpm, centrifuging to obtain pallidum thallus,

wherein the seed culture medium comprises the following components:

the fermentation medium comprises the following components:

placing the pallidum bacteria in a reaction solution containing L-histidine and beta-alanine and having a pH of 6.5-9.0, and reacting for 12-36 h at 25-37 ℃ under the condition of shaking at 100-200rpm to obtain L-carnosine, wherein the content of the pallidum bacteria in the reaction solution is 5-30g/L, the content of the L-histidine is 2-10g/L, the content of the beta-alanine is 1-8g/L,

wherein the seed culture medium comprises the following components:

the fermentation medium comprises the following components:

inoculating the screened pallidum strain into a test tube filled with 5mL of seed culture medium, and shake culturing at 30 ℃ for 1-3 days;

then transferring to a triangular flask filled with 50mL of fermentation medium, and culturing at 30 ℃ and 200rpm for 1-3 days;

20g of cells were collected by centrifugation at 8000rpm and resuspended in 1L of Tris-HCl buffer, pH=8.0, 5g of L-histidine and 2g of beta-alanine were added, and the reaction was catalyzed at 30℃and 200 rpm.

Wherein, seed culture medium: every 1L of the seed culture medium contains 10g of glucose, 5g of peptone, 10g of yeast powder, 5g of ammonium chloride, 3g of monopotassium phosphate, 5g of sodium chloride, 1.5g of magnesium sulfate and the balance of water, wherein the pH value is 6.8-7.2.

Fermentation medium: every 1L of the fermentation medium contains 10g of glucose, 5g of peptone, 10g of yeast powder, 5g of ammonium chloride, 3g of monopotassium phosphate, 5g of sodium chloride, 1.5g of magnesium sulfate and the balance of water, wherein the pH value is 6.8-7.2.

The L-carnosine obtained in the present application can be detected by High Performance Liquid Chromatography (HPLC).

The present application also provides a composition comprising 12mg/L or more of L-carnosine and the above-mentioned thallus of pallor bacillus.

In a specific embodiment, the composition comprises about 12-20mg/L of L-carnosine and the above-mentioned thallus of the pallidum bacteria.

In a specific embodiment, the composition comprises greater than about 20mg/L of L-carnosine and the above-described bacteria of the genus Ochrobactrum.

In a specific embodiment, the composition is used in the technical field of cosmetic or pharmaceutical products.

The present application also provides a cosmetic or pharmaceutical product comprising any of the above L-carnosine compositions.

Compared with the prior art, the pallidum can directly utilize histidine and alanine to synthesize carnosine, the histidine and the alanine have low price, no methanol is produced in the synthesis process, and the product separation is simple. Thereby avoiding the problems of the prior art that high-cost alanine methyl ester hydrochloride and the like are used as substrates to synthesize the carnosine and more methanol is generated, and providing a new choice for biologically synthesizing the carnosine.

The following examples of the present application are merely illustrative of specific embodiments for carrying out the present application and are not to be construed as limiting the present application. Any other changes, modifications, substitutions, combinations, and simplifications that do not depart from the spirit and principles of the present application are intended to be equivalent arrangements which are within the scope of the present application.

Examples

EXAMPLE 1 screening of the genus Xanthium

75 samples are collected from beef and mutton, bean products, fish intestinal tracts, seafood products and the like, the samples are respectively ground, 5g of the samples are added into 45mL of 0.9% sterilized normal saline, shaking culture is carried out for 30min at 30 ℃ and 200rpm, then dilution and coating are carried out on a screening culture medium plate, and culture is carried out for 1-3 days at 30 ℃ to obtain about 280 strains of the primary screening strain.

Wherein, the screening culture medium is: every 1L of screening culture medium contains 10g of sodium chloride, 1g of peptone, 0.5g of yeast powder, 5g of L-carnosine, 20g of agar powder and the balance of water, wherein the pH=7.0.

The strains with different colony forms are picked up by using a sterile toothpick or gun head and cultured on a new screening culture medium plate at 30 ℃ for 1-3 days.

After the bacteria in the newly screened plate grow out, reagents are sequentially added into the plate according to the reaction system shown in Table 1. Specifically, 0.2mL of 1% TCA standard solution and 8mL of 0.1mol/L NaOH solution are added for reaction for 5min; then 0.8mL of 1% o-phthalaldehyde solution is added for reaction for 1min; 0.8mL of 1mol/L HCl solution was added thereto, and the mixture was reacted at 20℃for 30 minutes.

After the reaction is completed, the flat plate is washed by tap water, the interference of thalli is removed, and the strain with the transparent ring is observed and recorded, which is a re-screened strain, about 20 strains.

Inoculating the re-screened strains with transparent circles into test tubes filled with 5mL of seed culture medium respectively, and shake culturing at 30 ℃ for 1-3 days; then transferring to a triangular flask filled with 50mL of fermentation medium, and culturing at 30 ℃ and 200rpm for 1-3 days; bacterial cells (0.2 g) were collected by centrifugation at 8000rpm and resuspended in 10mL of Tris-HCl buffer, pH=8.0, and catalytic reaction was carried out by adding L-histidine (0.05 g) and beta-alanine (0.02 g) at 30℃and 200 rpm.

And after the catalytic reaction is finished, derivatization is carried out by a pre-column derivatization method, and then the content of the L-carnosine in the reaction liquid is accurately measured by a high performance liquid chromatography.

Wherein the derivatization steps are as follows:

TABLE 1 derivatization reaction system (50 mL)

Adding the components in the table 1, mixing uniformly, carrying out water bath at 60 ℃ for 1h, cooling to room temperature after the reaction is finished, and fixing the volume of the 0.01mol/L potassium dihydrogen phosphate solution into a 50mL volumetric flask.

The specific conditions of the high performance liquid chromatography are as follows:

stationary phase: eclipse PLUS C18 (4.6X1250 mm);

mobile phase: fluidity a (0.05M sodium acetate, phosphoric acid adjusted ph=4.0): mobile phase B (acetonitrile) =70: 30;

elution mode: isocratic elution;

column temperature: 35 DEG C

Flow rate: 0.8mL/min;

sample injection amount: 20. Mu.L;

ultraviolet detection wavelength: 360nm.

The liquid chromatogram of the L-carnosine standard is shown in FIG. 1, wherein the L-carnosine concentration in the L-carnosine standard is 0.1g/L. The reaction liquid of the strain with the number of HXQ-1 and the L-carnosine standard product have the same peak positions, and the liquid chromatogram of the reaction liquid is shown in figure 2. . The yield of L-carnosine produced by 7 strains was calculated as shown in Table 2, and the strain numbered HXQ-1 produced the highest yield of L-carnosine, about 20mg/L.

TABLE 2L-carnosine yield of different isolates

Strain numbering	Carnosine yield (mg/L)
		HXQ-1	20
8311-2	10
		8311-3	8
8311-4	5
		8311-5	5
8311-6	7
		8311-7	12

And the strain provides a method for synthesizing the L-carnosine by utilizing the pallidum and using the histidine and the alanine for the first time, and the yield of the generated L-carnosine can reach about 20mg/L.

Example 2 identification of strains

The genome of the strain is sequenced and identified by Shanghai biological engineering (Shanghai) stock, and the strain is identified as the pallidum, and the 16s rRNA gene sequence of the strain is shown as SEQ ID NO. 1.

Claims

1. The method is characterized in that the pallor bacillus is pallor bacillus (Ochrobactrum sp.) HXQ-1, and the preservation number of the pallor bacillus in the China general microbiological culture collection center is CGMCC NO.25590.

2. A culture comprising the pallidum of claim 1.

3. The use of the pallor bacillus according to claim 1 for the preparation of L-carnosine.

4. Use of a culture according to claim 2 for the preparation of L-carnosine.

5. Use of the pallidum of claim 1 or the culture of claim 2 for the preparation of a carnosine hydrolase.

6. A method for producing carnosine, comprising obtaining L-carnosine using the pallidobacter of claim 1 or the culture of claim 2.

7. The method according to claim 6, characterized in that it comprises the steps of:

placing the pallidum thallus or the culture in a reaction liquid containing L-histidine and beta-alanine for catalytic reaction to obtain the L-carnosine.

8. The method according to claim 7, wherein the pallidum cells are obtained by culturing the pallidum cells in a fermentation medium and centrifuging the cells.