CN103173373B - Pediococcus acidilactici with high bacteriostatic activity and application thereof - Google Patents
Pediococcus acidilactici with high bacteriostatic activity and application thereof Download PDFInfo
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- CN103173373B CN103173373B CN201210240226.XA CN201210240226A CN103173373B CN 103173373 B CN103173373 B CN 103173373B CN 201210240226 A CN201210240226 A CN 201210240226A CN 103173373 B CN103173373 B CN 103173373B
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- 241000191998 Pediococcus acidilactici Species 0.000 title claims abstract description 38
- 230000003385 bacteriostatic effect Effects 0.000 title claims description 19
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- 239000007788 liquid Substances 0.000 abstract description 9
- 239000000843 powder Substances 0.000 abstract description 8
- 238000004519 manufacturing process Methods 0.000 abstract description 7
- 230000000844 anti-bacterial effect Effects 0.000 abstract description 5
- 238000001228 spectrum Methods 0.000 abstract description 5
- 238000002360 preparation method Methods 0.000 abstract description 4
- 238000000034 method Methods 0.000 abstract description 3
- 241000186660 Lactobacillus Species 0.000 abstract description 2
- 229940039696 lactobacillus Drugs 0.000 abstract description 2
- 241000192125 Firmicutes Species 0.000 abstract 1
- 239000003795 chemical substances by application Substances 0.000 abstract 1
- 238000000855 fermentation Methods 0.000 abstract 1
- 230000004151 fermentation Effects 0.000 abstract 1
- 230000000813 microbial effect Effects 0.000 abstract 1
- 238000009777 vacuum freeze-drying Methods 0.000 abstract 1
- 239000006041 probiotic Substances 0.000 description 11
- 235000018291 probiotics Nutrition 0.000 description 11
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 10
- 230000001580 bacterial effect Effects 0.000 description 8
- 241001465754 Metazoa Species 0.000 description 6
- 235000015097 nutrients Nutrition 0.000 description 6
- 239000003814 drug Substances 0.000 description 5
- 235000014655 lactic acid Nutrition 0.000 description 5
- 239000002609 medium Substances 0.000 description 5
- 238000004321 preservation Methods 0.000 description 5
- FRXSZNDVFUDTIR-UHFFFAOYSA-N 6-methoxy-1,2,3,4-tetrahydroquinoline Chemical compound N1CCCC2=CC(OC)=CC=C21 FRXSZNDVFUDTIR-UHFFFAOYSA-N 0.000 description 4
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 4
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 4
- 239000000654 additive Substances 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 244000144972 livestock Species 0.000 description 4
- 244000144977 poultry Species 0.000 description 4
- 239000000047 product Substances 0.000 description 4
- 239000006228 supernatant Substances 0.000 description 4
- 229960005486 vaccine Drugs 0.000 description 4
- 206010012735 Diarrhoea Diseases 0.000 description 3
- 239000003242 anti bacterial agent Substances 0.000 description 3
- 229940088710 antibiotic agent Drugs 0.000 description 3
- 230000000968 intestinal effect Effects 0.000 description 3
- 229920001817 Agar Polymers 0.000 description 2
- FBPFZTCFMRRESA-FSIIMWSLSA-N D-Glucitol Natural products OC[C@H](O)[C@H](O)[C@@H](O)[C@H](O)CO FBPFZTCFMRRESA-FSIIMWSLSA-N 0.000 description 2
- FBPFZTCFMRRESA-JGWLITMVSA-N D-glucitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-JGWLITMVSA-N 0.000 description 2
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 2
- 241000191938 Micrococcus luteus Species 0.000 description 2
- 229910019142 PO4 Inorganic materials 0.000 description 2
- 102000057297 Pepsin A Human genes 0.000 description 2
- 108090000284 Pepsin A Proteins 0.000 description 2
- 239000001888 Peptone Substances 0.000 description 2
- 108010080698 Peptones Proteins 0.000 description 2
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 2
- 229930006000 Sucrose Natural products 0.000 description 2
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 2
- DPDMMXDBJGCCQC-UHFFFAOYSA-N [Na].[Cl] Chemical compound [Na].[Cl] DPDMMXDBJGCCQC-UHFFFAOYSA-N 0.000 description 2
- 239000008272 agar Substances 0.000 description 2
- 230000000843 anti-fungal effect Effects 0.000 description 2
- 229940121375 antifungal agent Drugs 0.000 description 2
- 229940041514 candida albicans extract Drugs 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 239000003640 drug residue Substances 0.000 description 2
- 238000004108 freeze drying Methods 0.000 description 2
- 239000008103 glucose Substances 0.000 description 2
- 235000011187 glycerol Nutrition 0.000 description 2
- 230000036541 health Effects 0.000 description 2
- 238000011081 inoculation Methods 0.000 description 2
- 239000002054 inoculum Substances 0.000 description 2
- 235000019341 magnesium sulphate Nutrition 0.000 description 2
- 239000012533 medium component Substances 0.000 description 2
- 244000000010 microbial pathogen Species 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 239000006916 nutrient agar Substances 0.000 description 2
- 229940111202 pepsin Drugs 0.000 description 2
- 235000019319 peptone Nutrition 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 2
- 239000010452 phosphate Substances 0.000 description 2
- 229910052700 potassium Inorganic materials 0.000 description 2
- 239000011591 potassium Substances 0.000 description 2
- 238000011218 seed culture Methods 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 229960002920 sorbitol Drugs 0.000 description 2
- 239000005720 sucrose Substances 0.000 description 2
- 230000001988 toxicity Effects 0.000 description 2
- 231100000419 toxicity Toxicity 0.000 description 2
- 239000012138 yeast extract Substances 0.000 description 2
- DHPRQBPJLMKORJ-XRNKAMNCSA-N (4s,4as,5as,6s,12ar)-7-chloro-4-(dimethylamino)-1,6,10,11,12a-pentahydroxy-6-methyl-3,12-dioxo-4,4a,5,5a-tetrahydrotetracene-2-carboxamide Chemical compound C1=CC(Cl)=C2[C@](O)(C)[C@H]3C[C@H]4[C@H](N(C)C)C(=O)C(C(N)=O)=C(O)[C@@]4(O)C(=O)C3=C(O)C2=C1O DHPRQBPJLMKORJ-XRNKAMNCSA-N 0.000 description 1
- 108020004465 16S ribosomal RNA Proteins 0.000 description 1
- 102000035101 Aspartic proteases Human genes 0.000 description 1
- 108091005502 Aspartic proteases Proteins 0.000 description 1
- 206010059866 Drug resistance Diseases 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 241000607142 Salmonella Species 0.000 description 1
- 241000191967 Staphylococcus aureus Species 0.000 description 1
- 206010042566 Superinfection Diseases 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000000996 additive effect Effects 0.000 description 1
- 230000003042 antagnostic effect Effects 0.000 description 1
- 230000000845 anti-microbial effect Effects 0.000 description 1
- 244000052616 bacterial pathogen Species 0.000 description 1
- 239000002131 composite material Substances 0.000 description 1
- 235000020247 cow milk Nutrition 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000001627 detrimental effect Effects 0.000 description 1
- 230000001079 digestive effect Effects 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 235000012631 food intake Nutrition 0.000 description 1
- 230000001408 fungistatic effect Effects 0.000 description 1
- 230000002496 gastric effect Effects 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 230000036737 immune function Effects 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 230000003053 immunization Effects 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 239000004310 lactic acid Substances 0.000 description 1
- RWVUEZAROXKXRT-VQLSFVLHSA-N maduramicin Chemical compound O1[C@@H](C)[C@H](OC)[C@@H](OC)C[C@H]1O[C@@H]1[C@H]([C@@]2(C)O[C@H](CC2)[C@@]2(C)O[C@]3(O[C@@H]([C@H](C)[C@@H](O)C3)[C@@H](C)[C@H]3[C@@H]([C@@H](OC)[C@H](C)[C@@](O)(CC(O)=O)O3)OC)CC2)O[C@@H]([C@@H]2[C@H](C[C@@H](C)[C@@](C)(O)O2)C)C1 RWVUEZAROXKXRT-VQLSFVLHSA-N 0.000 description 1
- 229950006915 maduramicin Drugs 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- 238000009629 microbiological culture Methods 0.000 description 1
- 230000008520 organization Effects 0.000 description 1
- 230000000529 probiotic effect Effects 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 230000000452 restraining effect Effects 0.000 description 1
- 238000005728 strengthening Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
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- Fodder In General (AREA)
Abstract
The invention relates to a Pediococcus acidilactici vscc-1 CGMCC No.6055 with broad-spectrum high antibacterial activity and a powdery microecological preparation prepared by utilizing the Pediococcus acidilactici through high-density liquid submerged fermentation, centrifugal enrichment and vacuum freeze drying. The lactobacillus has wide antibacterial spectrum, and has good antibacterial effect on gram-positive bacteria and gram-negative bacteria; has high antibacterial activity. The microbial ecological agent prepared by the bacteria has simple process, short production period and low cost, and the viable bacteria contained in each gram of dry powder is about trillion cfu.
Description
Technical field
The present invention relates to a kind of pediococcus acidilactici, particularly relate to pediococcus acidilactici and the application of a plant height bacteriostatic activity.
Background introduction
Abuse of antibiotics in current feed and livestock and poultry cultivation, chemical synthetic drug, hormone cause series of problems and consequence, comprise: animal products drug residue is serious, directly be detrimental to health, even produce " three cause ", and the animal products quality of China is declined, lack the market competitiveness and trust; Bacterial drug resistance strengthens, Resistant strain increases, and result medication is invalid, threatens human security etc.World Health Organization expert thinks, the various resistance germs that livestock and poultry produce, and can pass to people by food, and produces the disease being difficult to cure, and harm humans is healthy.Meanwhile, the microbiotic that medicine, the Maduramicin etc. such as duomycin easily remain and toxicity is larger and a large amount of uses of synthesising bacteria anti-reflecting medicine, cause drug residue to increase, Resistant strain spreads, and animal flora imbalance, the immunizing power of animal is suppressed, secondary superinfection.And pediococcus acidilactici, produce acid, regulate gastrointestinal bacterial flora, maintain intestinal microecology balance.Have antagonistic action to pathogenic micro-organism in animal body, contestable ground suppresses pathogenic micro-organism, strengthens the immunologic function of animal body, produces useful meta-bolites, activates the activity of aspartic protease, participates in the metabolism of body, prevents objectionable impurities from producing.
In view of the problems existed in above-mentioned feed and livestock and poultry cultivation and prolongable space, China starts " feed safety engineering " and have disabled a collection of toxicity and residual larger medicine.Accelerating exploitation and the production of novel fodder additive, is particularly the production of the fodder additives of substitute antibiotics.Therefore, probiotics becomes the novel focus of industrialized development as a green fodder additives, is one of main products of substitute antibiotics.
As autochthonous flora in digestive tube, milk-acid bacteria has regulating intestinal canal colony balance, suppresses the effects such as harmful bacteria growth, strengthening immunity.Probiotics adopts milk-acid bacteria or milk-acid bacteria in the composite form of other probiotic bacteriums mostly on the market, in raising livestock and poultry food consumption, prevents there is remarkable effect in diarrhoea etc.But milk-acid bacteria also exists following defects as probiotics product: the viable count of dry powder of lactic acid bacteria is relatively low, and the viable bacteria survival time is short, the while of has a broad antifungal spectrum, the bacterial classification of good antimicrobial effect is few, and production cost is relatively high.
Summary of the invention
The object of this invention is to provide pediococcus acidilactici (Pediococcus acidilactici) the vscc-1CGMCC No.6055 of a strain wide spectrum high bacteriostatic activity.
The pediococcus acidilactici obtaining a strain wide spectrum high bacteriostatic activity is separated from cow's milk, pediococcus acidilactici (Pediococcus acidilactici) vscc-1 is accredited as through 16S rDNA, and in Song China Microbial Culture Preservation Commission common micro-organisms center preservation April 26 in 2012, be numbered CGMCC No.6055, preservation address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City institute of microbiology of Chinese Academy of Sciences postcode: 100101 phones: 86-10-64807596.
1, the pediococcus acidilactici of a plant height bacteriostatic activity and application, is characterized in that: this bacterium is pediococcus acidilactici (Pediococcus acidilactici) vscc-1CGMCC No.6055, has stronger bacteriostatic activity.
2, the pediococcus acidilactici of a plant height bacteriostatic activity according to claim 1 and application, is characterized in that described scope of restraining fungi activity refers to all have bacteriostatic action to intestinal bacteria, Salmonellas, micrococcus luteus, streptococcus aureus.
The bacteriostatic activity of the pediococcus acidilactici of 3, a described plant height bacteriostatic activity refers to that the antibacterial circle diameter of indicator reaches 20mm-30mm.
4, containing a probiotics for the pediococcus acidilactici of high bacteriostatic activity, it is characterized in that described probiotics is take pediococcus acidilactici as main raw material, and its concrete production technique is as follows:
1) bacterial classification is selected: bacterial classification used is pediococcus acidilactici (Pediococcus acidilactici) vscc-1 CGMCCNo.6055;
2) slant culture: pediococcus acidilactici (Pediococcus acidilactici) the vscc-1CGMCC No.6055 of-80 DEG C of glycerine pipes preservation is inoculated in solid slant culture base, cultivates 24h for 30 DEG C;
3) seed culture: by the strain inoculation of slant culture in liquid nutrient medium, 30 DEG C of quiescent culture 8h;
4) fermentor cultivation: primary seed solution is inoculated in fermentor cultivation liquid by 1% inoculum size, 30 DEG C, 150-250rpm ferments 10-20h;
Above-mentioned medium component is: sucrose 2-8g/L, Pepsin peptone 10-18g/L, yeast extract paste 2-10g/L, potassium primary phosphate 3-14g/L, sodium-chlor 1-10g/L, bitter salt 0.1-1.5g/L, and pH is adjusted to 7.0-7.35; Slant medium separately adds 1.5% agar powder.
5) thalline enrichment: fermented liquid is passed in tubular-bowl centrifuge, 16000-21000rpm collected by centrifugation thalline.
6) lyophilize: after being mixed with isopyknic 10%D-Sorbitol Solution USP by the bacterium mud after centrifugal, be placed in freeze-drying in freeze drier.
7) aftertreatment of freeze-dried vaccine powder: take glucose as carrier, mix with freeze-dried vaccine powder, makes mixed probiotics viable count reach 0.8 trillion cfu/g.
5, a kind of pediococcus acidilactici containing high bacteriostatic activity is as follows as the consumption in fodder additives at probiotics:
For normally planting fowl, addition be feed heavy 0.2%;
For diarrhoea kind of a fowl, addition be feed heavy 0.1%;
For normal piglets, addition be feed heavy 0.2%;
For diarrhoea piglet, addition be feed heavy 0.4%.
The present invention has the following advantages: described bacterial classification has a broad antifungal spectrum, bacteriostatic activity are high; Described probiotics viable count is the highest in commercially available at present Strains of lactobacillus; Production technique is simple, and the cycle is short, and production cost is low.
Accompanying drawing explanation
Fig. 1 be pediococcus acidilactici (Pediococcus acidilactici) vscc-1CGMCC No.6055 to the fungistatic effect of micrococcus luteus, be two identical experiment dosage groups in its figure; Test dose group is the supernatant liquor of the nutrient solution of pediococcus acidilactici (Pediococcus acidilactici) vscc-1CGMCC No.6055.
Embodiment
Embodiment one: the high bacteriostatic activity of pediococcus acidilactici (Pediococcus acidilactici) vscc-1CGMCC No.6055
1. the preparation of nutrient solution supernatant
The pediococcus acidilactici of slant culture is inoculated in liquid nutrient medium, 30 DEG C of quiescent culture 16h, gets the centrifugal 1min of 1mL bacterium liquid 12000rpm, get supernatant liquor.
2. bacteriostatic activity detects dull and stereotyped preparation
The LB nutrient agar dissolved completely by 15mL is poured in culture dish, after it solidifies, vertically put Oxford cup; Separately get a LB nutrient agar dissolved completely, be cooled to add when 55 DEG C indicator mixing until it, get the substratum that 10mL is mixed with indicator and pour in the culture dish being placed with Oxford cup, after solidifying completely, extract Oxford cup, make detect dull and stereotyped.
3. the mensuration of bacteriostatic activity
Getting 200 μ L nutrient solution supernatants adds in the hole detecting flat board, puts into 37 DEG C of incubators and cultivates 20h, measure inhibition zone size 20mm-30mm after 4 DEG C of placement 24h.
Embodiment two: high viable count probiotics preparation technology
1) bacterial classification is selected: bacterial classification used is pediococcus acidilactici (Pediococcus acidilactici) vscc-1CGMCCNo.6055.
2) slant culture: pediococcus acidilactici (Pediococcus acidilactici) the vscc-1CGMCC No.6055 of-80 DEG C of glycerine pipes preservation is inoculated in solid slant culture base, cultivates 24-30h for 30 DEG C.
3) seed culture: by the strain inoculation of slant culture in liquid nutrient medium, 30 DEG C of quiescent culture 8h.
4) fermentor cultivation: be inoculated in fermentor cultivation liquid by 1% inoculum size by seed liquor, 30 DEG C, 100rpm ferments 20h.Above-mentioned medium component is (g/L): sucrose 4, Pepsin peptone 12, yeast extract paste 4, potassium primary phosphate 5, sodium-chlor 2, and bitter salt 0.2, pH is adjusted to 7.0; Slant medium separately adds 1.5% agar powder
5) thalline enrichment: fermented liquid is passed in tubular-bowl centrifuge, 20000rpm collected by centrifugation thalline.
6) lyophilize: after being mixed with isopyknic 10%D-Sorbitol Solution USP by the bacterium mud after centrifugal, be placed in freeze-drying in freeze drier.
7) aftertreatment of freeze-dried vaccine powder: take glucose as carrier, mix with freeze-dried vaccine powder, makes mixed probiotics viable count reach 1,000,000,000,000 cfu/g.
Claims (1)
1. the pediococcus acidilactici of a plant height bacteriostatic activity, is characterized in that: this bacterium be pediococcus acidilactici (
pediococcus acidilactici) vscc-1 CGMCC No.6055, there is stronger bacteriostatic activity.
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| CN201210240226.XA CN103173373B (en) | 2012-07-12 | 2012-07-12 | Pediococcus acidilactici with high bacteriostatic activity and application thereof |
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| CN103173373A CN103173373A (en) | 2013-06-26 |
| CN103173373B true CN103173373B (en) | 2015-05-20 |
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Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103614325B (en) * | 2013-11-26 | 2015-07-08 | 江南大学 | Pediococcus pentosaceus and application thereof |
| CN103627637A (en) * | 2013-11-29 | 2014-03-12 | 天津大学 | Preparation method for pediococcus acidilactici strain freeze-drying preparation |
| LT6270B (en) * | 2014-09-24 | 2016-05-10 | Uab "Baltijos Biotechnologijos" | Probiotic fermented feed additives |
| CN106721260A (en) * | 2016-11-30 | 2017-05-31 | 江西正邦科技股份有限公司 | A kind of preparation and its application of anti diar rhea type piglet specific complex probiotics |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101050437A (en) * | 2007-03-21 | 2007-10-10 | 华东理工大学 | Method for producing bacterial strain of Pediococcus acidilactici, and bacterin of Pediococcus acidilactici |
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- 2012-07-12 CN CN201210240226.XA patent/CN103173373B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101050437A (en) * | 2007-03-21 | 2007-10-10 | 华东理工大学 | Method for producing bacterial strain of Pediococcus acidilactici, and bacterin of Pediococcus acidilactici |
Non-Patent Citations (2)
| Title |
|---|
| 乳酸片球菌细菌素的活性及特性的研究;韩雪 等;《食品研究与开发》;20061231;第27卷(第4期);第20页第2节 * |
| 从酸白菜中分离出一株产细菌素的乳酸片球菌;周志江 等;《食品科学》;20061231;第27卷(第4期);89-92 * |
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