CN113122475B

CN113122475B - Saccharopolyspora and application thereof in preparation of epsilon-polylysine

Info

Publication number: CN113122475B
Application number: CN202110429895.0A
Authority: CN
Inventors: 赵志军; 李沁雨; 史吉平; 陈枭嘉; 姜标
Original assignee: Shanghai Advanced Research Institute of CAS
Current assignee: Shanghai Advanced Research Institute of CAS
Priority date: 2021-04-21
Filing date: 2021-04-21
Publication date: 2022-07-22
Anticipated expiration: 2041-04-21
Also published as: CN113122475A

Abstract

The invention relates to the technical field of biology, in particular to saccharopolyspora and application thereof in preparation of epsilon-polylysine. The invention provides saccharopolyspora, the preservation number of which is CCTCC NO: m2020594. The saccharopolyspora provided by the invention has better substrate (sugar concentration) tolerance, has the characteristics of simple application and operation, good stability and repeatability and the like in the production of epsilon-polylysine, can realize the stable production of epsilon-polylysine, is beneficial to large-scale production, and has good industrialization potential.

Description

Saccharopolyspora and application thereof in preparation of epsilon-polylysine

Technical Field

The invention relates to the technical field of biology, in particular to saccharopolyspora and application thereof in preparation of epsilon-polylysine.

Background

Epsilon-polylysine (Epsilon-poly-L-lysine) is a novel biological preservative with high efficiency and broad spectrum. The epsilon-polylysine is a polymer formed by connecting alpha-carboxyl and epsilon-amino of L-lysine, generally contains 25-35 lysine residues, and a purified finished product is light yellow powder, is easy to dissolve in water, has strong hygroscopicity and is slightly bitter. Compared with other preservatives, the epsilon-polylysine has the characteristics of wide antibacterial spectrum, high antibacterial efficiency, good stability and high safety. The epsilon-polylysine has bacteriostatic effects on gram-positive bacteria, gram-negative bacteria, yeast and mould, and also has inhibitory effects on heat-resistant bacillus and some viruses; the addition amount of the food additive is small but the food additive is high-efficiency; can adapt to a wider pH range, can be sterilized along with food, and has high thermal stability; the application of epsilon-polylysine is very wide. In the food industry, it can also be used as an emulsifier, a dietetic agent; in the pharmaceutical industry, can be used as a drug carrier and a biological material with high water absorption. The common polylysine-producing strains are mostly concentrated in northern Lyocell, Streptomyces and the like, 1 saccharopolyspora wzj4 producing epsilon-polylysine is obtained by screening in the early stage of the experiment, the maximum fermentation yield of a fermentation tank is only 2.54g/L, and the epsilon-polylysine producing capability is limited.

Disclosure of Invention

In view of the above-mentioned drawbacks of the prior art, it is an object of the present invention to provide a saccharopolyspora and its use in the preparation of epsilon-polylysine, which solve the problems of the prior art.

In order to achieve the above objects and other related objects, the present invention provides a glycopolyspora, which has a collection number of CCTCC NO: m2020594.

In some embodiments of the invention, the 16S rDNA polynucleotide sequence of Saccharopolyspora comprises the sequence shown in SEQ ID No. 4.

In some embodiments of the invention, the glycopolyelectrolytes are gram positive bacteria;

and/or, the sugar multicellular carbon source utilizes a positive carbon source comprising one or more of fructose, glycerol and mannitol, and the sugar multicellular carbon source utilizes a negative carbon source comprising one or more of glucose, sucrose, lactose and beta-cyclodextrin;

and/or, the saccharopolyspora is starch hydrolysis positive;

and/or, the sugar multicellular bacteria do not produce pigment and/or hydrogen sulfide in the induction culture process.

In another aspect, the invention provides the use of the above saccharopolyspora for the preparation of epsilon-polylysine.

The invention also provides a preparation method of epsilon-polylysine, which is prepared from the saccharopolyspora.

In some embodiments of the present invention, the method for preparing epsilon-polylysine specifically comprises: inoculating the saccharopolysaccharose into a fermentation medium, and inducing the saccharopolysaccharose to produce epsilon-polylysine under appropriate conditions.

In some embodiments of the present invention, the conditions for inducing said saccharopolyspora to produce epsilon-polylysine under suitable conditions are: fermenting under the conditions of 25-35 ℃, pH 6.5-7 and DO value 3-5 mg/mL.

In some embodiments of the invention, the activated glycomulticellular bacteria are inoculated into a fermentation medium.

In some embodiments of the present invention, the method further comprises: purifying and separating the epsilon-polylysine.

The invention also provides epsilon-polylysine which is prepared by the saccharopolyspora or the preparation method of the epsilon-polylysine.

Drawings

FIG. 1 is a schematic representation of the phylogenetic tree of Saccharopolysporaerythraea TG01, a strain of Saccharopolyspora saccharolytica in example 1 of the present invention.

FIG. 2 is a schematic view showing the morphology of Saccharopolysporaerythraea TG01 on solid medium in example 1 of the present invention.

FIG. 3 is a schematic diagram showing the standard curve of ε -polylysine in the HPLC determination method in example 2 of the present invention.

FIG. 4 is a graph showing the fermentation yield of the saccharopolyspora erythraea TG01 in a 5L fermenter for 192 hours in accordance with example 3 of the present invention.

FIG. 5 is a schematic diagram showing the results of an experiment for inhibiting saccharopolyspora erythraea TG01 in example 3 of the present invention.

Detailed Description

In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail with reference to the following embodiments, and other advantages and effects of the present invention will be readily apparent to those skilled in the art from the disclosure of the present specification.

The inventor of the invention has surprisingly found a saccharopolysaccharomyces strain which has high yield of epsilon-polylysine under proper conditions, has stable antibacterial performance and is beneficial to realizing stable and cheap production of epsilon-polylysine through a great deal of practical research, and the invention is completed on the basis.

The invention provides a saccharopolyspora, the preservation number of which is CCTCC NO: m2020594. The saccharopolyspora provided by the application is obtained by removing surface soil from a collected soil sample, fully drying the soil sample, taking about 50g of soil with the distance of 5-15 cm from the surface, taking the soil as a separation source, and screening the separation source under the pressure condition of epsilon-polylysine and a composite antibiotic bacteriostatic agent (sodium naphthyridonate: nystatin: cycloheximide; 2: 1: 1).

In the saccharopolyspora provided by the invention, the polynucleotide sequence of 16S rDNA of the saccharopolyspora can comprise a sequence shown as SEQ ID NO.4, and the sequence length is 1511 bp. The 16S rRNA sequence determination method is adopted to know that the strain is similar to Saccharopolyspora saccharophagolyspora, and the 16S rDNA sequence analysis and identification show that the strains with the sequence similarity higher than 99 percent are all strains of the genus Saccharopolyspora, wherein the similarity with Saccharopolyspora saccharophagolyspora erythrae-D is 99.7 percent.

In the saccharopolyspora provided by the invention, the saccharopolyspora is usually a gram-positive bacterium, cells of a young culture of the saccharopolyspora are usually irregular rod-shaped, after the saccharopolyspora enters a stabilization period, thalli can be changed into spherical shapes, bacterial colonies are usually round powder, white or faint yellow, the bacterial colonies are usually dry and opaque, the thalli are usually divided into 2 or 3 petals with gaps in the middle, and the thalli are usually tightly connected with a culture medium and are not easy to pick up.

In the saccharopolyspora provided by the invention, the saccharopolyspora has certain selectivity on utilization of a carbon source. For example, carbon sources that are positive for carbon source utilization typically include a combination of one or more of fructose, glycerol, mannitol, and the like. As another example, carbon sources that are negative for carbon source utilization typically include a combination of one or more of glucose, sucrose, lactose, beta-cyclodextrin, and the like. Suitable methods for judging the carbon source utilization selectivity should be known to those skilled in the art. For example, the physiological and biochemical identification methods can be referred to in "Bergey's Manual of identification of bacteria," the foundation of Classification of Actinomycetes, "or" Manual of identification of Streptomyces ".

In the saccharopolyspora provided by the invention, the saccharopolyspora is usually positive to starch hydrolysis, and does not usually produce pigment and/or hydrogen sulfide in the induction culture process of the saccharopolyspora. Suitable methods for determining whether starch hydrolysis is positive, and whether pigments and/or hydrogen sulfide are produced during induction, will be known to those skilled in the art. For example, the relevant experimental methods can be referred to in "Bergey's Manual of identification of bacteria", in "Classification of Actinomycetes", or in "Manual of identification of Streptomyces".

In a second aspect, the invention provides the use of saccharopolyspora provided in the first aspect of the invention for the preparation of epsilon-polylysine.

The third aspect of the invention provides a preparation method of epsilon-polylysine, which is prepared from the saccharopolyspora provided by the first aspect of the invention. In general, the skilled person can select suitable methods for the preparation of epsilon-polylysine by the above saccharopolyspora, which may include, for example: inoculating saccharopolyspora into a fermentation medium, and inducing the saccharopolyspora to produce epsilon-polylysine under appropriate conditions.

In the preparation method of epsilon-polylysine provided by the invention, the saccharopolyspora is induced to produce epsilon-polylysine under proper conditions usually when thalli are in the early stage of logarithmic growth. The above-mentioned suitable conditions may generally include suitable conditions of temperature, pH, DO (dissolved oxygen) value, suitable inoculum amount, and the like. For example, the induction can be carried out at a temperature of 20 to 40 ℃, 20 to 22 ℃, 22 to 24 ℃, 24 to 25 ℃, 25 to 26 ℃, 26 to 27 ℃, 27 to 28 ℃, 28 to 29 ℃, 29 to 30 ℃, 30 to 31 ℃, 31 to 32 ℃, 32 to 33 ℃, 33 to 34 ℃, 34 to 35 ℃, 35 to 36 ℃, 36 to 38 ℃ or 38 to 40 ℃. For example, the induction may be carried out at a pH of 5 to 9, 5 to 5.5, 5.5 to 6, 6 to 6.5, 6.5 to 7, 7 to 7.5, 7.5 to 8, 8 to 8.5, or 8.5 to 9, and usually the pH of the medium may be suitably close to neutral, and if the pH is lowered during the fermentation (for example, when the pH is lowered to 4), a suitable base (for example, ammonia water or the like) may be added to maintain the pH of the fermentation system. For example, the induction can be carried out under conditions of DO value of 3 to 7mg/mL, 3 to 3.5mg/mL, 3.5 to 4mg/mL, 4 to 4.5mg/mL, 4.5 to 5mg/mL, 5 to 5.5mg/mL, 5.5 to 6mg/mL, 6 to 6.5mg/mL, 6.5 to 7mg/mL, and the induction condition is usually aerobic culture condition. For another example, the amount of inoculation may be 5-10%, 5-6%, 6-7%, 7-8%, 8-9%, or 9-10% of the volume of the fermentation medium.

The culture medium used in the method for producing epsilon-polylysine according to the present invention may be appropriately adjusted, as is known to those skilled in the art. For example, the fermentation medium may include glucose, yeast powder, (NH)₄)₂SO₄、 K₂HPO₄·3H₂O、KH₂PO₄And concentrated salt solution.

In the preparation method of epsilon-polylysine provided by the invention, the saccharopolyspora to be inoculated is usually activated saccharopolyspora, and can be usually included in a seed culture solution. Suitable methods for activating the bacterial cells will be known to those skilled in the art. For example, the saccharopolyspora may be inoculated into a seed medium and cultured under appropriate conditions. The culture conditions for activation of saccharopolyspora and/or the formulation of the seed medium used may be referred to the induction conditions in the production of epsilon-polylysine and the formulation of the fermentation medium used.

The preparation method of epsilon-polylysine provided by the invention can also comprise the following steps: purifying and separating the epsilon-polylysine. Suitable purification and separation methods should be known to those skilled in the art. For example, steps such as ultrafiltration, microfiltration, ion exchange, or activated carbon decolorization may be included.

The fourth aspect of the invention provides epsilon-polylysine, which is prepared by the saccharopolyspora provided by the first aspect of the invention or the preparation method of epsilon-polylysine provided by the third aspect of the invention. The prepared epsilon-polylysine can be dissolved in water, hydrochloric acid and the like, can be slightly dissolved in ethanol and the like, can be insoluble in organic solvents such as diethyl ether, ethyl acetate and the like, is positive to ninhydrin after being hydrolyzed by 6N HCl, and is detected by paper chromatography and thin layer chromatography after being hydrolyzed by 6N HCl, and the hydrolysate of the epsilon-polylysine is generally single amino acid lysine.

The saccharopolyspora provided by the invention has better substrate (sugar concentration) tolerance, has the characteristics of simple application and operation, good stability, good repeatability and the like in the production of epsilon-polylysine, can realize the stable production of epsilon-polylysine, is beneficial to large-scale production, and has good industrialization potential.

The invention of the present application is further illustrated by the following examples, which are not intended to limit the scope of the present application.

The culture media and components used in the examples are as follows:

primary screening of culture medium: 0.4% of yeast extract, 1% of malt extract, 0.4% of glucose, 1.5% of agar, 1mL of trace salt solution and pH 7.2. Sterilizing at 115 deg.C for 15 min.

Fermentation medium: glucose 5%, yeast powder 0.5%, (NH)₄)₂SO₄ 1％，K₂HPO₄·3H₂O 0.1048％，KH₂PO₄0.136% concentrated salt solution (MgSO)₄·7H₂O，FeSO₄·7H2O，ZnSO₄·7H₂O)100mL, pH 6.8. Glucose was sterilized separately.

And (3) preserving the culture medium: 1% of glucose, 0.1% of beef extract, 0.2% of peptone, 0.1% of yeast powder, 2% of agar and 7.5 of PH. Sterilizing at 115 deg.C for 15 min.

Trace salt solution: FeSO₄·7H₂O 0.1g，MnCl₂·4H₂O 0.1g，ZnSO₄·7H₂0.1g of O, and adding water to a constant volume of 1L.

Example 1

Screening and identifying program of saccharopolyspora erythraea TG 01.

1. Primary screening:

collecting a plurality of soil samples in suburbs of Shanghai, putting the soil samples into sterilized kraft paper bags, naturally drying the soil samples for 3-10 days at room temperature, grinding and sieving the soil samples, and drying the soil samples for 1 hour at 50 ℃. Adding 1g of the pretreated soil sample into 10mL of sterile water, shaking at 30 ℃ for 15min, standing for 30min, sucking supernatant liquid, diluting by a proper multiple, and coating the diluted supernatant liquid on an LB separation medium (primary screening medium) plate containing 2g/L of epsilon-polylysine. When the culture is carried out at 30 ℃ for the third day, the colonies generate obvious transparent circles, and the colonies with the obvious transparent circles are used for next re-screening.

2. Re-screening:

inoculating the actinomycetes obtained by primary screening to a fermentation culture medium of 30mL/250mL for fermentation for 96h, collecting the fermentation liquor, centrifuging, and detecting by using a bismuth potassium iodide reagent. And (3) hydrolyzing the supernatant strong acid obtained by centrifuging the fermentation liquor, and detecting by thin layer chromatography analysis to obtain a polylysine-producing strain, wherein the strain is named as TG 01.

3. Identification of the strain:

taking the bacterial liquid after shake flask culture of the strain to be identified, and extracting genome DNA by adopting a bacterial genome extraction kit. Amplification was performed using the universal primer 27F (5'-AGAGTTTGATCCTGGCTCAG-3') (SEQ ID NO.1)/1492R (5'-GGTTACCTTGTTACGACTT-3') (SEQ ID NO.2), and the 16S rDNA sequence obtained by sequencing was compared with the sequences of related species in the GenBank database, which revealed that TG01 was close to Saccharopolyspora Saccharopolyspora, whereas 16SrDNA sequence analysis identified that the strains more than 99% similar to the sequences were Saccharopolyspora strains in which the degree of similarity to Saccharopolyspora erythra-D was 99.7%, and thus the strain was finally named Saccharopolyspora Saccharopolyspora erythra TG 01.

The 16S rDNA sequence of the strain TG01 is shown in SEQ ID NO.3, the sequence length is 1511bp, the 16S rRNA sequence is submitted in NCBI, the homology sequence comparison analysis is carried out by software and GenBank, and MEGA 7 software is used for constructing a phylogenetic tree of the strain (figure 1). In view of the above information, the strain TG01 was identified as Saccharopolyspora erythraea TG 01.

The main biological characteristics of the strain TG01 are gram-positive bacteria, cells of a young culture are irregular rod-shaped, and after entering a stabilization period, the bacteria become spherical; the bacterial colony is round powder, white or light yellow, dry and opaque, has gaps in the middle to be divided into 2 and 3 petals, is tightly connected with the culture medium, and is not easy to lift. The morphology on solid medium is shown in FIG. 2.

AGAGTTTGATCCTGGCTCAGGACGAACGCTGGCGGCGTGCTTAACACATGCAAGTC GAACGCTGAAGCATCTTCGGGTGTGGATGAGTGGCGAACGGGTGAGTAACACGTGGGT AATCTGCCCTGCACTCTGGGATAAGCCCTGGAAACGGGGTCTAATACCGGATAGGACAT TCTGCCGCATGGTGGGGTGTGGAAAGTTCCGGCGGTGCAGGATGAGCCCGCGGCCTATC AGCTTGTTGGTGGGGTGATGGCCTACCAAGGCGACGACGGGTAGCCGGCCTGAGAGGG TGACCGGCCACACTGGGACTGAGACACGGCCCAGACTCCTACGGGAGGCAGCAGTGGG GAATCTTGCGCAATGGGCGAAAGCCTGACGCAGCAACGCCGCGTGGGGGATGACGGCC TTCGGGTTGTAAACCTCTTTCGACAGGGACGAAGCCTTCGGGTGACGGTACCTGTAGAA GAAGCACCGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGGTGCGAGCGTTGTC CGGATTTATTGGGCGTAAAGAGCTCGTAGGCGGTTTGTCGCGTCGGCCGTGAAAACCTG CAGCTTAACTGTGGGCGTGCGGTCGATACGGGCAGACTTGAGTTCGGCAGGGGAGACT GGAATTCCTGGTGTAGCGGTGAAATGCGCAGATATCAGGAGGAACACCGGTGGCGAAG GCGGGTCTCTGGGCCGATACTGACGCTGAGGAGCGAAAGCGTGGGGAGCGAACAGGAT TAGATACCCTGGTAGTCCACGCCGTAAACGTTGGGCGCTAGGTGTGGGGACCGGTTCCA CGGTTTCTGTGCCGTAGCTAACGCATTAAGCGCCCCGCCTGGGGAGTACGGCCGCAAGG CTAAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGCGGAGCATGTGGATTAATT CGATGCAACGCGAAGAACCTTACCTGGGTTTGACATGCACCAGATTGCCCCTGAGAGGG GGTTTCCCTTGTGGTTGGTGTACAGGTGGTGCATGGCTGTCGTCAGCTCGTGTCGTGAG ATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTGTCCTGTGTTGCCAGCACGTAATG GTGGGGACTCGCGGGAGACTGCCGGGGTCAACTCGGAGGAAGGTGGGGATGACGTCA AGTCATCATGCCCCTTATGCCCAGGGCTTCACACATGCTACAATGGCTGGTACAGAGGGT GGCGATACCGTGAGGTGGAGCGAATCCCTTAAAGCCGGTCTCAGTTCGGATCGGGGTCT GCAACTCGACCCCGTGAAGTCGGAGTCGCTAGTAATCGCAGATCAGCAGTGCTGCGGTG AATACGTTCCCGGGCCTTGTACACACCGCCCGTCACGTCATGAAAGTCGGTAACACCCG AAGCCCATGGCCTAACCCTGTTGGGGTGGAGTGGTCGAAGGTGGGACTGGCGATTGGG ACGAAGTCGTAACAAGGTAGCCGTACCGGAAGGTGCGGCTGGATCACCT(SEQ ID NO.3)

Example 2

Content determination method of epsilon-polylysine

And (3) measuring the content of epsilon-polylysine by adopting a high performance liquid chromatography method. Making an epsilon-polylysine standard curve:

weighing 1.2g of epsilon-polylysine standard substance, dissolving with deionized water, fixing the volume to 1L, and diluting 8 concentration gradients, namely 1.2g/L, 1.0g/L, 0.8g/L, 0.6g/L, 0.5g/L, 0.4g/L, 0.2g/L and 0. And establishing a standard curve by using peak areas of different concentration gradients epsilon-polylysine. The high performance liquid chromatography column is C18 column, the mobile phase is 8% acetonitrile, the flow rate is 0.4mL/min, the detection wavelength is 215nm, the injection volume is 20 μ L, and the temperature is 40 ℃. Collecting fermentation liquid, centrifuging at 12000r/min for 15min, diluting with proper times, preparing sample in standard curve range, and calculating the concentration of epsilon-polylysine in the fermentation liquid according to the standard curve (figure 3).

Example 3

Application of saccharopolyspora saccharophaga erythraea TG01 strain in fermentation production of epsilon-polylysine

Saccharopolyspora saccharopolyspora erythraea TG01 was inoculated into a seed medium (the same as fermentation medium) and cultured at 30 ℃. After the seed liquid is cultured for 24h, the seed liquid is inoculated into a 2.5L/5L fermentation tank according to the inoculation amount of 8 percent, the fermentation is carried out for 192h at 30 ℃ and pH6.8, fermentation broth obtained by fermentation culture is centrifuged for 15 minutes at 12000rpm, thalli are separated, clarified filtrate is obtained by filtration, then the pH is adjusted to 7.5, the yield of the epsilon-polylysine is measured by the measuring method given in the example 2, and the yield of the epsilon-polylysine is 7.19g/L after the saccharopolyspora erythraea TG01 is fermented for 192h finally (figure 4).

The antibacterial experiment is carried out by taking epsilon-polylysine hydrochloride produced by New Yinxing corporation as a reference substance. Four epsilon-polylysine concentrations of 0mg/L, 50mg/L, 150mg/L and 200mg/L are set as gradients to carry out inhibition experiments on the bacillus subtilis. The results show that the epsilon-polylysine extracted from the fermentation liquor of Saccharopolyspora erythraea TG01 has the bacteriostatic ability basically similar to that of the reference substance and the bacteriostatic property is stable (figure 5).

In conclusion, the present invention effectively overcomes various disadvantages of the prior art and has high industrial utilization value.

The foregoing embodiments are merely illustrative of the principles and utilities of the present invention and are not intended to limit the invention. Those skilled in the art can modify or change the above-described embodiments without departing from the spirit and scope of the present invention. Accordingly, it is intended that all equivalent modifications or changes which may be made by those skilled in the art without departing from the spirit and scope of the present invention as defined in the appended claims.

And (3) classification and naming: saccharopolyspora erythraea TG01

The preservation date is as follows: 2020.10.19

The preservation unit is as follows: china Center for Type Culture Collection (CCTCC)

The address of the depository: eight-path 299 Wuhan university school of Wuhan city, Wuhan province, Hubei province

The preservation number is: CCTCC NO: m2020594.

Sequence listing

<110> Shanghai higher research institute of Chinese academy of sciences

<120> saccharopolyspora and application thereof in preparation of epsilon-polylysine

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ggttaccttg ttacgactt 19

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agagtttgat cctggctcag gacgaacgct ggcggcgtgc ttaacacatg caagtcgaac 60

gctgaagcat cttcgggtgt ggatgagtgg cgaacgggtg agtaacacgt gggtaatctg 120

ccctgcactc tgggataagc cctggaaacg gggtctaata ccggatagga cattctgccg 180

catggtgggg tgtggaaagt tccggcggtg caggatgagc ccgcggccta tcagcttgtt 240

ggtggggtga tggcctacca aggcgacgac gggtagccgg cctgagaggg tgaccggcca 300

cactgggact gagacacggc ccagactcct acgggaggca gcagtgggga atcttgcgca 360

atgggcgaaa gcctgacgca gcaacgccgc gtgggggatg acggccttcg ggttgtaaac 420

ctctttcgac agggacgaag ccttcgggtg acggtacctg tagaagaagc accggctaac 480

tacgtgccag cagccgcggt aatacgtagg gtgcgagcgt tgtccggatt tattgggcgt 540

aaagagctcg taggcggttt gtcgcgtcgg ccgtgaaaac ctgcagctta actgtgggcg 600

tgcggtcgat acgggcagac ttgagttcgg caggggagac tggaattcct ggtgtagcgg 660

tgaaatgcgc agatatcagg aggaacaccg gtggcgaagg cgggtctctg ggccgatact 720

gacgctgagg agcgaaagcg tggggagcga acaggattag ataccctggt agtccacgcc 780

gtaaacgttg ggcgctaggt gtggggaccg gttccacggt ttctgtgccg tagctaacgc 840

attaagcgcc ccgcctgggg agtacggccg caaggctaaa actcaaagga attgacgggg 900

gcccgcacaa gcggcggagc atgtggatta attcgatgca acgcgaagaa ccttacctgg 960

gtttgacatg caccagattg cccctgagag ggggtttccc ttgtggttgg tgtacaggtg 1020

gtgcatggct gtcgtcagct cgtgtcgtga gatgttgggt taagtcccgc aacgagcgca 1080

acccttgtcc tgtgttgcca gcacgtaatg gtggggactc gcgggagact gccggggtca 1140

actcggagga aggtggggat gacgtcaagt catcatgccc cttatgccca gggcttcaca 1200

catgctacaa tggctggtac agagggtggc gataccgtga ggtggagcga atcccttaaa 1260

gccggtctca gttcggatcg gggtctgcaa ctcgaccccg tgaagtcgga gtcgctagta 1320

atcgcagatc agcagtgctg cggtgaatac gttcccgggc cttgtacaca ccgcccgtca 1380

cgtcatgaaa gtcggtaaca cccgaagccc atggcctaac cctgttgggg tggagtggtc 1440

gaaggtggga ctggcgattg ggacgaagtc gtaacaaggt agccgtaccg gaaggtgcgg 1500

ctggatcacc t 1511

Claims

1. A Saccharopolyspora (A. about., (B. about.), (A. about.), (B. about.), (A. about.), (B. about.), (A. about.), (B.) about., (B. about.), (C. about.), (B.) about.), (B. about.), (C.), (B. about.), (C. about.), (B.), (C. about.), (B.), (Saccharopolyspora erythraea) The saccharopolyspora is preserved in China center for type culture Collection with the preservation number of CCTCC NO: m2020594.

2. The glycomulticellular bacteria of claim 1 wherein the polynucleotide sequence of 16S rDNA of the glycomulticellular bacteria includes the sequence set forth in SEQ ID number 4.

3. Use of the saccharopolyspora as defined in any one of claims 1 to 2 for the preparation of epsilon-polylysine.

4. A process for producing epsilon-polylysine by fermentation with the saccharopolyspora as claimed in any one of claims 1 to 2.

5. The process for producing epsilon-polylysine according to claim 4, which comprises:

inoculating the saccharopolysaccharose into a fermentation medium, and inducing the saccharopolysaccharose to produce epsilon-polylysine under appropriate conditions.

6. The process for producing epsilon-polylysine according to claim 5, wherein the conditions for inducing said Saccharopolyspora to produce epsilon-polylysine under suitable conditions are as follows: fermenting at 25-35 deg.C, pH 6.5-7, and DO 3-5 mg/mL.

7. The process for producing epsilon-polylysine according to claim 5, wherein the activated saccharopolyspora is inoculated into a fermentation medium.

8. The process for the preparation of epsilon-polylysine of claim 5, further comprising: purifying and separating the epsilon-polylysine.