CN108611299A - One plant of lactobacillus plantarum for producing bacteriostatic peptide and its application - Google Patents
One plant of lactobacillus plantarum for producing bacteriostatic peptide and its application Download PDFInfo
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- CN108611299A CN108611299A CN201810447104.5A CN201810447104A CN108611299A CN 108611299 A CN108611299 A CN 108611299A CN 201810447104 A CN201810447104 A CN 201810447104A CN 108611299 A CN108611299 A CN 108611299A
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- lactobacillus plantarum
- bacteriostatic
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- acid bacteria
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- 235000013965 Lactobacillus plantarum Nutrition 0.000 title claims abstract description 54
- 229940072205 lactobacillus plantarum Drugs 0.000 title claims abstract description 53
- 230000003385 bacteriostatic effect Effects 0.000 title claims abstract description 48
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 39
- 230000001580 bacterial effect Effects 0.000 claims abstract description 24
- 235000013305 food Nutrition 0.000 claims abstract description 24
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- 239000002253 acid Substances 0.000 claims abstract description 11
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- 241000894006 Bacteria Species 0.000 claims description 53
- 239000004310 lactic acid Substances 0.000 claims description 33
- 235000014655 lactic acid Nutrition 0.000 claims description 33
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- RPAJSBKBKSSMLJ-DFWYDOINSA-N (2s)-2-aminopentanedioic acid;hydrochloride Chemical class Cl.OC(=O)[C@@H](N)CCC(O)=O RPAJSBKBKSSMLJ-DFWYDOINSA-N 0.000 claims 1
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- FFRBMBIXVSCUFS-UHFFFAOYSA-N 2,4-dinitro-1-naphthol Chemical compound C1=CC=C2C(O)=C([N+]([O-])=O)C=C([N+]([O-])=O)C2=C1 FFRBMBIXVSCUFS-UHFFFAOYSA-N 0.000 description 1
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- 239000010985 leather Substances 0.000 description 1
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- 244000039328 opportunistic pathogen Species 0.000 description 1
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- 235000019834 papain Nutrition 0.000 description 1
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- 230000001717 pathogenic effect Effects 0.000 description 1
- 229940111202 pepsin Drugs 0.000 description 1
- 235000021110 pickles Nutrition 0.000 description 1
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- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
- 235000020795 whole food diet Nutrition 0.000 description 1
Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L3/00—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs
- A23L3/34—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals
- A23L3/3454—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals in the form of liquids or solids
- A23L3/3463—Organic compounds; Microorganisms; Enzymes
- A23L3/3571—Microorganisms; Enzymes
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/02—Separating microorganisms from their culture media
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/169—Plantarum
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/225—Lactobacillus
- C12R2001/25—Lactobacillus plantarum
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- Biotechnology (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Wood Science & Technology (AREA)
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- Tropical Medicine & Parasitology (AREA)
- Biomedical Technology (AREA)
- Virology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nutrition Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Food Preservation Except Freezing, Refrigeration, And Drying (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses the lactobacillus plantarums of one plant of production bacteriostatic peptide, the bacterial strain is lactobacillus plantarum (Lactobacillus plantarum) YLY 1, it is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on March 8th, 2018, and deposit number is CGMCC NO.15429.The invention also discloses applications of the lactobacillus plantarum YLY 1 of above-mentioned production bacteriostatic peptide in terms of inhibiting liquor-saturated spiral shell spoilage organisms.The lactobacillus plantarum YLY 1 of the present invention has good tolerance, stablizes in acid condition, and the bacteriostatic peptide of generation has good fungistatic effect to mud snail spoilage organisms, has broad application prospects in food antiseptic.
Description
Technical field
The present invention relates to a lactobacillus plantarum more particularly to one plant production bacteriostatic peptide lactobacillus plantarum and its inhibiting
Application in terms of liquor-saturated spiral shell spoilage organisms, belongs to food microorganisms technical field.
Technical background
Currently, a large amount of food preservative has been applied in food service industry, with prevent food spoilage, to extend food goods
The frame phase.Wherein, artificial synthesized chemical preservative due at low cost, easy to use and anti-corrosion effect it is good, in China's food row
Industry occupies leading position, but these artificial preservatives in use can have a negative impact to the mouthfeel of food, flavor,
And it can be absorbed as food is absorbed by body together, certain toxic action is caused to body.And animal, plant and microorganism
The natural antiseptic agent in source attracts the sight of numerous scholars because having the characteristics that safe and free of toxic and side effects, antibiotic property is strong, wherein
Microbial source such as lactic acid bacteria preservative are derived from a wealth of sources, are not limited by raw material, by scholar's bigger since producing strains type more
Concern.Therefore, it seeks the microbial source wholefood antisepsis antistaling agent that safe and non-toxic, antimicrobial spectrum is wide, antibiotic property is strong and has become food
Grind high hot spot in product field.
Lactic acid bacteria is the dominant bacteria in humans and animals normal intestinal flora, has many lifes healthy and beneficial to humans and animals
Function is managed, is commonly applied in fermented food, unexpected flavor and special quality structure is provided for food；It is prior
It is that the antibacterial materials such as organic acid, hydrogen peroxide and lactein can be generated in metabolic process, wherein lactein is in food
It plays an important role in anti-corrosive fresh-keeping.In view of the probiotic of lactic acid bacteria, lactic acid bacteria, which is one kind generally acknowledged at present, has potential exploitation
The producing strains of the microbial source natural biological food preservative of value.
Lactic acid bacteria bacteriostatic peptide is the protein with inhibition other microorganisms growth that lactic acid bacteria generates in metabolic process
Or polypeptides matter.It can effectively inhibit the growth of pathogenic bacteria and spoilage organisms in food.The bacteriostatic peptide that lactic acid bacteria generates has
Following feature so that it becomes natural food-preservative development object.(1) bacterial growth breeding is fast, with short production cycle, is suitble to
Industrialized production；(2) there is preferable stability, be suitble to food processing processing, it is easy to use；It (3) can be by the albumen in human body
Enzyme is degraded, and does not accumulate, will not have an impact to normal intestinal flora in vivo；(4) drug resistance is not will produce, food disease will not be induced
Resistance of the microorganisms such as opportunistic pathogen to treatment antibiotic.
There are many report that lactic acid bacteria detaches from dairy products, pickles, the enteron aisle of humans and animals, excrement, soil etc. at present, so
And the report of separating lactic acid bacterium is also less from the aquatic animals such as fish, shellfish, the animal of long-term life in water, since its is special
Environment, the microorganism in enteron aisle, which must develop, oneself unique metabolic pathway, and it is unique simultaneously to generate many structure novels, effect
It is different from the metabolite of Lu Yuan microorganisms.
Invention content
Technical problem to be solved by the invention is to provide one plant production bacteriostatic peptide lactobacillus plantarum YLY-1, the bacterial strain from
It is screened in perch enteron aisle, the bacteriostatic peptide for generation of fermenting can effectively inhibit the growth of liquor-saturated spiral shell spoilage organisms, have safe nothing
Poison, antibiotic property be strong, antibacterial general wide feature.
The lactobacillus plantarum YLY-1 that the present invention also technical problems to be solved are to provide above-mentioned production bacteriostatic peptide is inhibiting liquor-saturated spiral shell
Application in terms of spoilage organisms.
In order to solve the above technical problems, the technical solution adopted in the present invention is：
The present invention is with the lactic acid of the separation screening from the perch enteron aisle that sea route non-staple food central market on Nanjing acquires
Bacterium is to grind high object, therefrom filters out the lactic acid bacteria with the production bacteriostatic peptide for inhibiting food-borne spoilage organisms and pathogenic activity.
Strain isolation of the present invention from perch enteron aisle, through MRS inoculated and cultureds, isolate and purify after the strains of lactic acid bacteria strain that obtains,
It is named as lactobacillus plantarum (Lactobacillus plantarum) YLY-1, Classification And Nomenclature is lactobacillus plantarum
(Lactobacillus plantarum), is deposited in China Committee for Culture Collection of Microorganisms's common micro-organisms center, should
Microbial preservation number is CGMCC NO.15429, and the deposit date is on March 8th, 2018.
The biological property of the bacterial strain is as follows：Lactobacillus plantarum YLY-1 bacterium sources of the present invention are in perch enteron aisle, in MRS
In culture medium, 37 DEG C of grown under anaerobic conditions it is good, bacterium colony is at circle, diameter 0.8-1.5mm, and white colony is impermeable
Bright, the smooth moistening in surface, Gram's staining is positive, and cell shape is rod-short under micro- sem observation.
The present invention produces the lactic acid bacteria strains of bacteriostatic peptide, is sequenced through 16s rDNA, and using BLAST analytic approach, the present invention is planted
The Gene sequence comparison of the 16s rDNA complete sequences of object lactobacillus YLY-1 and the 16s rDNA of the lactobacillus plantarum of NCBI registrations,
Homology is 100%, obtains lactobacillus plantarum.
The screening technique of above-mentioned lactobacillus plantarum, includes the following steps：
Step 1, lactic acid bacteria isolates and purifies：It is screened from perch excrement with MRS culture mediums and isolates lactic acid bacteria strains, led to
Plate streak purified lactic acid bacteria strain is crossed, Gram's staining is carried out to bacterial strain after purification and catalase is tested, chooses leather
Blue Albert'stain Albert is the positive, the bacterial strain that catalase experiment is feminine gender；
Step 2, the screening of antibacterial peptide lactic acid bacteria is produced：The lactic acid bacteria strains of step 1 Economical Purification are inoculated into the training of MRS liquid
It supports in base, the stationary culture 24 hours under the conditions of 37 DEG C obtains bacterial strain fermentation liquor, and zymotic fluid is in 4 DEG C, 10000r/min conditions
Under, it centrifuges 10 minutes, retains supernatant, carry out bacteriostatic experiment with card punch punch method, sour effect and mistake are carried out in experimentation
The exclusion of hydrogen oxide effect；Finally filtering out still has the production bacteriostatic peptide breast of bacteriostatic activity after excluding acid effect and hydrogen peroxide effect
Sour bacterium.
Wherein, it in step 1, is screened from perch excrement and isolates lactic acid bacteria strains using plate dilution method, after dilution
Sample bacteria suspension be coated on the MRS solid plates of addition bromocresol purple, cultivate 48h under the conditions of 37 DEG C, choose culture medium
On tablet four rides are carried out at flaxen bacterium colony.
Applications of the lactobacillus plantarum YLY-1 of above-mentioned production bacteriostatic peptide in terms of inhibiting food spoilage bacterium.
Wherein, the food spoilage bacterium is Escherichia coli, bacillus subtilis and staphylococcus aureus.
Applications of the lactobacillus plantarum YLY-1 of above-mentioned production bacteriostatic peptide in terms of inhibiting liquor-saturated spiral shell spoilage organisms.
Wherein, the liquor-saturated spiral shell spoilage organisms is Leuconostoc mesenteroides.
Advantageous effect：The lactobacillus plantarum YLY-1 of the present invention has good tolerance, stablizes in acid condition, produces
Raw bacteriostatic peptide has good fungistatic effect to mud snail spoilage organisms, has broad application prospects in food antiseptic.
Description of the drawings
Fig. 1 is colonial morphologies of the lactobacillus plantarum YLY-1 of the present invention on MRS solid mediums；
Fig. 2 is the Gram's staining picture I of lactobacillus plantarum YLY-1 of the present invention；
Fig. 3 is the Gram's staining picture II of lactobacillus plantarum YLY-1 of the present invention；
The 16s of the 16s rDNA complete sequences that Fig. 4 is lactobacillus plantarum YLY-1 of the present invention and the lactobacillus plantarum of NCBI registrations
RDNA gene order comparison results；
Fig. 5 is fungistatic effects of the lactobacillus plantarum YLY-1 of the present invention to mud snail spoilage organisms.
Specific implementation mode
Technical scheme of the present invention is described further below in conjunction with the drawings and specific embodiments.
Embodiment 1：Lactic acid bacteria isolates and purifies
Perch excrement is taken out, 1g is weighed and is placed in the test tube for filling 9mL sterile salines, with turbine mixer mixing；
Aseptically, gradient dilution is carried out, is drawn from the sample bacteria suspension of tri- dilutions of 10-3,10-4,10-5 respectively
0.1mL dilutions are respectively coated on the MRS solid plates of addition bromocresol purple, and 48h is cultivated under the conditions of 37 DEG C, choose training
It supports and carries out four rides at flaxen bacterium colony on base tablet, 24~48h is cultivated under the conditions of being placed in 37 DEG C, repeat scribing line 2-3 times,
Obtain bacterial strain after purification；Gram's staining and catalase experiment are carried out to bacterial strain after purification, choosing Gram's staining is
Positive, catalase experiment is negative bacterial strain.
Embodiment 2：Produce the screening of antibacterial peptide lactic acid bacteria
(1) preparation of fermented supernatant fluid
In the inoculation for the Economical Purification that embodiment 1 is obtained to MRS fluid nutrient mediums, training is stood under the conditions of 37 DEG C
It supports 24 hours, obtains bacterial strain fermentation liquor, zymotic fluid centrifuges 10 minutes under the conditions of 4 DEG C, 10000r/min, retains supernatant.
(2) screening of biocidal property lactic acid bacteria
For the common spoilage organisms for causing food apoilage, Escherichia coli, bacillus subtilis, golden yellow Portugal are chosen
Grape coccus.Fermented supernatant fluid carries out bacteriostatic experiment by card punch punch method obtained by step (1), has filtered out bacteriostatic activity
Bacterial strain.Bacterial strain YLY-1 of the present invention concrete outcomes in biocidal property lactic acid bacteria screening process are shown in Table 1.
The In Vitro Bacteriostasis of 1 three kinds of indicator bacterias of lactobacillus plantarum YLY-1 fermented supernatant fluids pair of table tests primary dcreening operation result
(3) exclusion of sour inhibiting effect
Fermented supernatant fluid is likely to the inhibition of indicator bacteria in the work of lactic acid in metabolite, acetic acid and other organic acids
With as a result, to exclude this possibility, pH 1.0mol/L NaOH and the 1.0mol/L HCl of lactobacillus-fermented supernatant to be measured
5 are adjusted to, acid discharge experiment is carried out with card punch punch method, is retained in the bacterial strain for still having inhibition zone after excluding acid effect.
(4) exclusion of hydrogen peroxide effect
Its pH is adjusted to 7.0,20mg mistakes is added by the bacterial strain fermentation liquor for still having fungistatic effect after taking 2.0mL to exclude acid effect
Hydrogen oxide enzyme is jiggled to dissolving, is placed in 37 DEG C of heat preservation 2h, then pH is adjusted to identified control pH when exclusion acid acts on,
As a contrast with the acid discharge zymotic fluid without hydrogen peroxide enzymatic treatment, bacteriostatic experiment is carried out with card punch punch method.If bacterial strain is sent out
It is exactly the effect of bacteriostatic peptide caused by the bacterial strain that zymotic fluid still has inhibition zone, bacteriostasis after excluding acid, hydrogen peroxide effect.
(5) protease hydrolyzed is tested
It is pancreas still to have the bacterial strain fermentation liquor of bacteriostatic activity, adjusting pH value after taking 4 parts of 2.0mL to exclude acid, hydrogen peroxide effect
The optimum pH (be not 7.4,7.4,6.0 and 2.0) of protease, Proteinase K, papain and each self-applying of pepsin,
Each enzyme solution is added by the amount of final concentration 1.0mg/mL, original pH is recalled to after 4h is reacted under preference temperature, after measuring enzymatic treatment
With the bacteriostatic activity before processing.
The experimental results showed that lactobacillus plantarum YLY-1 of the present invention can generate bacteriostatic peptide and bacteriostasis is good, concrete outcome
It is shown in Table 2.
Table 2 is that the In Vitro Bacteriostasis of three kinds of indicator bacterias of lactobacillus plantarum YLY-1 fermented supernatant fluids pair tests secondary screening result
It can be seen from Table 2 that protease hydrolyzed is carried out to bacterial strain, three kinds of instructions of strain fermentation supernatant pair after enzymolysis
Bacterium loses bacteriostasis.
Embodiment 3：Produce the identification of bacteriostatic peptide lactic acid bacteria strains
Filter out one plant of production bacteriostatic peptide lactic acid bacteria strains YLY-1 is cultivated to exponential phase, its genome is extracted
DNA.Utilize primer (forward primer AGAGTTTGATCCTGGCTCAG；Reverse primer AAGGAGGTGATCCAGCCGCA) it amplifies
Its 16s rDNA segments are sent to Sangon Biotech (Shanghai) Co., Ltd. and are sequenced, and sequence is 1505bp (see sequence
Table, SEQ ID NO, 1).Pass through sequence alignment, it was demonstrated that be lactobacillus plantarum, be named as lactobacillus plantarum (Lactobacillus
plantarum)YLY-1。
Embodiment 4：Application of the lactobacillus plantarum YLY-1 bacteriostatic peptides to liquor-saturated spiral shell spoilage organisms
(1) preparation of bacteriostatic peptide semifinished product
Lactobacillus plantarum YLY-1 is inoculated into MRS fluid nutrient mediums, stationary culture 24 hours, obtains under the conditions of 37 DEG C
To bacterial strain fermentation liquor, zymotic fluid centrifuges 10 minutes under the conditions of 4 DEG C, 10000r/min, retains supernatant.By fermented supernatant fluid
It is frozen into powder, obtains powdered bacteriostatic peptide, is stored in 4 DEG C of refrigerators.
(2) fungistatic effect of the bacteriostatic peptide semifinished product to Leuconostoc mesenteroides in liquor-saturated spiral shell
With 10mL ddH2O back dissolving bacteriostatic peptide powder makees bacteriostatic agent, makees indicator bacteria with Leuconostoc mesenteroides, is beaten with card punch
Hole method carries out bacteriostatic test, measures antibacterial circle diameter.
The experimental results showed that the bacteriostatic peptide that lactobacillus plantarum YLY-1 of the invention is generated have to liquor-saturated spiral shell spoilage organisms it is good
Inhibiting effect, antibacterial circle diameter 17.36mm.Concrete outcome is as shown in Figure 5.
The bacteriostatic peptide that lactobacillus plantarum (Lactobacillus plantarum) YLY-1 of the present invention is generated is to gram sun
The growth of property and Gram-negative bacteria has good inhibiting effect, with good stability, and can be by multiple protein
Enzyme hydrolysis.The lactobacillus plantarum YLY-1 fermented supernatant fluids of the present invention, bacteriostatic peptide semifinished product can be obtained after freeze-dried；It should
The bacteriostatic peptide fungistatic effect that lactobacillus plantarum YLY-1 is generated is good, has wider antimicrobial spectrum, and safe and non-toxic, can be extensive
For the anti-corrosive fresh-keeping of food, to extend the shelf life of food.
The preservation title of lactobacillus plantarum strain of the present invention：YLY-1；
Preserving number：CGMCCNO.15429；
Preservation date：2018-3-8；
Preservation place：China Committee for Culture Collection of Microorganisms's common micro-organisms center；
Address：Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3 Institute of Microorganism, Academia Sinica.
Sequence table
<110>Nanjing Normal University
<120>One plant of lactobacillus plantarum for producing bacteriostatic peptide and its application
<160> 3
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1505
<212> DNA
<213>Lactobacillus plantarum (Lactobacillus plantarum)
<400> 1
aggcttagct ccttgttcga cttcacccta atcatctgtc ccaccttagg cggctggttc 60
ctaaaaggtt accccaccga ctttgggtgt tacaaactct catggtgtga cgggcggtgt 120
gtacaaggcc cgggaacgta ttcaccgcgg catgctgatc cgcgattact agcgattccg 180
acttcatgta ggcgagttgc agcctacaat ccgaactgag aatggcttta agagattagc 240
ttactctcgc gagttcgcaa ctcgttgtac catccattgt agcacgtgtg tagcccaggt 300
cataaggggc atgatgattt gacgtcatcc ccaccttcct ccggtttgtc accggcagtc 360
tcaccagagt gcccaactta atgctggcaa ctgataataa gggttgcgct cgttgcggga 420
cttaacccaa catctcacga cacgagctga cgacaaccat gcaccacctg tatccatgtc 480
cccgaaggga acgtctaatc tcttagattt gcatagtatg tcaagacctg gtaaggttct 540
tcgcgtagct tcgaattaaa ccacatgctc caccgcttgt gcgggccccc gtcaattcct 600
ttgagtttca gccttgcggc cgtactcccc aggcggaatg cttaatgcgt tagctgcagc 660
actgaagggc ggaaaccctc caacacttag cattcatcgt ttacggtatg gactaccagg 720
gtatctaatc ctgtttgcta cccatacttt cgagcctcag cgtcagttac agaccagaca 780
gccgccttcg ccactggtgt tcttccatat atctacgcat ttcaccgcta cacatggagt 840
tccactgtcc tcttctgcac tcaagtttcc cagtttccga tgcacttctt cggttgagcc 900
gaaggctttc acatcagact taaaaaaccg cctgcgctcg ctttacgccc aataaatccg 960
gacaacgctt gccacctacg tattaccgcg gctgctggca cgtagttagc cgtggctttc 1020
tggttaaata ccgtcaatac ctgaacagtt actctcagat atgttcttct ttaacaacag 1080
agttttacga gccgaaaccc ttcttcactc acgcggcgtt gctccatcag actttcgtcc 1140
attgtggaag attccctact gctgcctccc gtaggagttt gggccgtgtc tcagtcccaa 1200
tgtggccgat taccctctca ggtcggctac gtatcattgc catggtgagc cgttacccca 1260
ccatctagct aatacgccgc gggaccatcc aaaagtgata gccgaagcca tctttcaagc 1320
tcggaccatg cggtccaagt tgttatgcgg tattagcatc tgtttccagg tgttatcccc 1380
cgcttctggg caggtttccc acgtgttact caccagttcg ccactcactc aaatgtaaat 1440
catgatgcaa gcaccaatca ataccagagt tcgttcgact gcatgtatag cacccgccat 1500
gccaa 1505
<210> 2
<211> 20
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 2
agagtttgat cctggctcag 20
<210> 3
<211> 20
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 3
aaggaggtga tccagccgca 20
Claims (7)
1. the lactobacillus plantarum of one plant of production bacteriostatic peptide, it is characterised in that：The bacterial strain is lactobacillus plantarum (Lactobacillus
Plantarum) YLY-1 was preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms on March 8th, 2018
Center, deposit number are CGMCC NO.15429.
2. a kind of screening technique of the lactobacillus plantarum of production bacteriostatic peptide described in claim 1, which is characterized in that including walking as follows
Suddenly：
Step 1, lactic acid bacteria isolates and purifies：It is screened from perch excrement with MRS culture mediums and isolates lactic acid bacteria strains, by flat
Plate method of scoring purified lactic acid bacteria strain carries out Gram's staining to bacterial strain after purification and catalase is tested, chooses gram
Dyeing is the positive, the bacterial strain that catalase experiment is feminine gender；
Step 2, the screening of antibacterial peptide lactic acid bacteria is produced：The lactic acid bacteria strains of step 1 Economical Purification are inoculated into MRS fluid nutrient mediums
In, the stationary culture 24 hours under the conditions of 37 DEG C obtains bacterial strain fermentation liquor, zymotic fluid under the conditions of 4 DEG C, 10000r/min, from
The heart 10 minutes retains supernatant, carries out bacteriostatic experiment with card punch punch method, sour effect and hydrogen peroxide are carried out in experimentation
The exclusion of effect；Still there is the antibacterial peptide lactic acid bacteria of the production of bacteriostatic activity after finally filtering out the effect of exclusion acid and hydrogen peroxide effect.
3. the screening technique of the lactobacillus plantarum of production bacteriostatic peptide according to claim 2, it is characterised in that：In step 1, from
Screening isolates lactic acid bacteria strains and uses plate dilution method in perch excrement, and the sample bacteria suspension after dilution is coated on addition bromine
The MRS solid plate E of cresol-purple cultivate 48h under the conditions of 37 DEG C, choose culture medium flat plate E and carry out four at flaxen bacterium colony
Ride.
4. application of the lactobacillus plantarum of production bacteriostatic peptide described in claim 1 in terms of inhibiting food spoilage bacterium.
5. application of the lactobacillus plantarum of production bacteriostatic peptide described in claim 1 in terms of inhibiting liquor-saturated spiral shell spoilage organisms.
6. application of the lactobacillus plantarum of production bacteriostatic peptide according to claim 4 in terms of inhibiting food spoilage bacterium, special
Sign is：The food spoilage bacterium is Escherichia coli, bacillus subtilis and staphylococcus aureus.
7. application of the lactobacillus plantarum of production bacteriostatic peptide according to claim 5 in terms of inhibiting liquor-saturated spiral shell spoilage organisms, special
Sign is：The liquor-saturated spiral shell spoilage organisms is Leuconostoc mesenteroides.
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| CN110055188A (en) * | 2019-03-07 | 2019-07-26 | 南京师范大学 | One plant of Paenibacillus polymyxa XW4 for producing bacteriostatic peptide and its separation screening and application |
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| CN113061550A (en) * | 2021-04-01 | 2021-07-02 | 新疆农业科学院微生物应用研究所(中国新疆—亚美尼亚生物工程研究开发中心) | Lactobacillus new strain Z6 and application thereof in food |
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