CN113502243A - Lactobacillus plantarum GBW-LP001 capable of highly producing lactic acid and antibacterial agent alternative thereof and application - Google Patents
Lactobacillus plantarum GBW-LP001 capable of highly producing lactic acid and antibacterial agent alternative thereof and application Download PDFInfo
- Publication number
- CN113502243A CN113502243A CN202110733301.5A CN202110733301A CN113502243A CN 113502243 A CN113502243 A CN 113502243A CN 202110733301 A CN202110733301 A CN 202110733301A CN 113502243 A CN113502243 A CN 113502243A
- Authority
- CN
- China
- Prior art keywords
- lactobacillus plantarum
- gbw
- antibacterial agent
- lactic acid
- bacillus coagulans
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 240000006024 Lactobacillus plantarum Species 0.000 title claims abstract description 105
- 235000013965 Lactobacillus plantarum Nutrition 0.000 title claims abstract description 105
- 229940072205 lactobacillus plantarum Drugs 0.000 title claims abstract description 105
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 title claims abstract description 34
- 239000003242 anti bacterial agent Substances 0.000 title claims abstract description 33
- 239000004310 lactic acid Substances 0.000 title claims abstract description 17
- 235000014655 lactic acid Nutrition 0.000 title claims abstract description 17
- 238000004321 preservation Methods 0.000 claims abstract description 18
- 241000193749 Bacillus coagulans Species 0.000 claims abstract description 15
- 241000193171 Clostridium butyricum Species 0.000 claims abstract description 15
- 229940054340 bacillus coagulans Drugs 0.000 claims abstract description 15
- 244000144977 poultry Species 0.000 claims abstract description 12
- 230000000694 effects Effects 0.000 claims abstract description 9
- 108010015776 Glucose oxidase Proteins 0.000 claims abstract description 8
- 239000004366 Glucose oxidase Substances 0.000 claims abstract description 8
- 229940116332 glucose oxidase Drugs 0.000 claims abstract description 8
- 235000019420 glucose oxidase Nutrition 0.000 claims abstract description 8
- 239000002773 nucleotide Substances 0.000 claims abstract description 8
- 125000003729 nucleotide group Chemical group 0.000 claims abstract description 8
- 108020004465 16S ribosomal RNA Proteins 0.000 claims abstract description 5
- 230000001580 bacterial effect Effects 0.000 claims description 24
- 239000000843 powder Substances 0.000 claims description 22
- 241000894006 Bacteria Species 0.000 claims description 20
- 238000004519 manufacturing process Methods 0.000 claims description 8
- 238000002360 preparation method Methods 0.000 claims description 6
- 239000003674 animal food additive Substances 0.000 claims description 5
- 238000009629 microbiological culture Methods 0.000 claims description 5
- 230000012010 growth Effects 0.000 claims description 4
- 102000004190 Enzymes Human genes 0.000 claims description 3
- 108090000790 Enzymes Proteins 0.000 claims description 3
- 229940088598 enzyme Drugs 0.000 claims description 3
- 210000003495 flagella Anatomy 0.000 claims description 3
- 239000007921 spray Substances 0.000 claims description 3
- 239000002068 microbial inoculum Substances 0.000 claims 1
- 230000001488 breeding effect Effects 0.000 abstract description 24
- 238000009395 breeding Methods 0.000 abstract description 22
- 241001465754 Metazoa Species 0.000 abstract description 5
- 229940088710 antibiotic agent Drugs 0.000 abstract description 5
- 230000008901 benefit Effects 0.000 abstract description 5
- 239000000463 material Substances 0.000 abstract description 5
- 241000186660 Lactobacillus Species 0.000 abstract description 3
- 229940039696 lactobacillus Drugs 0.000 abstract description 2
- 241000287828 Gallus gallus Species 0.000 description 28
- 238000012360 testing method Methods 0.000 description 19
- 108020004414 DNA Proteins 0.000 description 14
- 235000013372 meat Nutrition 0.000 description 10
- 239000006041 probiotic Substances 0.000 description 10
- 235000018291 probiotics Nutrition 0.000 description 10
- 238000005507 spraying Methods 0.000 description 10
- 239000002253 acid Substances 0.000 description 9
- 235000013330 chicken meat Nutrition 0.000 description 9
- 235000013594 poultry meat Nutrition 0.000 description 9
- 239000003833 bile salt Substances 0.000 description 8
- 239000000243 solution Substances 0.000 description 8
- 238000006467 substitution reaction Methods 0.000 description 6
- 230000004083 survival effect Effects 0.000 description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 6
- 241000272525 Anas platyrhynchos Species 0.000 description 5
- 210000004051 gastric juice Anatomy 0.000 description 5
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 4
- 230000003115 biocidal effect Effects 0.000 description 4
- 239000003651 drinking water Substances 0.000 description 4
- 235000020188 drinking water Nutrition 0.000 description 4
- 239000001963 growth medium Substances 0.000 description 4
- 230000000968 intestinal effect Effects 0.000 description 4
- 230000000529 probiotic effect Effects 0.000 description 4
- 238000003307 slaughter Methods 0.000 description 4
- 238000003860 storage Methods 0.000 description 4
- 241000272517 Anseriformes Species 0.000 description 3
- 239000000654 additive Substances 0.000 description 3
- 239000000872 buffer Substances 0.000 description 3
- 239000007853 buffer solution Substances 0.000 description 3
- 239000003814 drug Substances 0.000 description 3
- 210000003746 feather Anatomy 0.000 description 3
- 239000012634 fragment Substances 0.000 description 3
- 230000014759 maintenance of location Effects 0.000 description 3
- 238000000034 method Methods 0.000 description 3
- 108090000623 proteins and genes Proteins 0.000 description 3
- 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
- 239000001888 Peptone Substances 0.000 description 2
- 108010080698 Peptones Proteins 0.000 description 2
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 2
- 230000000996 additive effect Effects 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 235000015278 beef Nutrition 0.000 description 2
- 229940041514 candida albicans extract Drugs 0.000 description 2
- 235000012730 carminic acid Nutrition 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- 230000007547 defect Effects 0.000 description 2
- 238000013461 design Methods 0.000 description 2
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 description 2
- 229910000396 dipotassium phosphate Inorganic materials 0.000 description 2
- 235000019797 dipotassium phosphate Nutrition 0.000 description 2
- 238000013401 experimental design Methods 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 239000000284 extract Substances 0.000 description 2
- 235000021050 feed intake Nutrition 0.000 description 2
- 238000000855 fermentation Methods 0.000 description 2
- 230000004151 fermentation Effects 0.000 description 2
- 239000008103 glucose Substances 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 238000009630 liquid culture Methods 0.000 description 2
- 244000144972 livestock Species 0.000 description 2
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 2
- 235000019341 magnesium sulphate Nutrition 0.000 description 2
- 229940099596 manganese sulfate Drugs 0.000 description 2
- 235000007079 manganese sulphate Nutrition 0.000 description 2
- 239000011702 manganese sulphate Substances 0.000 description 2
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
- 235000019319 peptone Nutrition 0.000 description 2
- 101150080777 pheS gene Proteins 0.000 description 2
- 229920000136 polysorbate Polymers 0.000 description 2
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 description 2
- 101150054232 pyrG gene Proteins 0.000 description 2
- 101150037064 rpoA gene Proteins 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 239000001632 sodium acetate Substances 0.000 description 2
- 235000017281 sodium acetate Nutrition 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 239000007858 starting material Substances 0.000 description 2
- YWYZEGXAUVWDED-UHFFFAOYSA-N triammonium citrate Chemical compound [NH4+].[NH4+].[NH4+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O YWYZEGXAUVWDED-UHFFFAOYSA-N 0.000 description 2
- 239000001393 triammonium citrate Substances 0.000 description 2
- 235000011046 triammonium citrate Nutrition 0.000 description 2
- 101150003576 uvrC gene Proteins 0.000 description 2
- 238000005303 weighing Methods 0.000 description 2
- 239000012138 yeast extract Substances 0.000 description 2
- 229920001817 Agar Polymers 0.000 description 1
- 241000271566 Aves Species 0.000 description 1
- 108010018956 CTP synthetase Proteins 0.000 description 1
- 108010078777 Colistin Proteins 0.000 description 1
- 101100311938 Dictyostelium discoideum phesA gene Proteins 0.000 description 1
- 101100423325 Dictyostelium discoideum phesB gene Proteins 0.000 description 1
- 101100038183 Dictyostelium discoideum polr2a gene Proteins 0.000 description 1
- 102000004533 Endonucleases Human genes 0.000 description 1
- 108010042407 Endonucleases Proteins 0.000 description 1
- 108091028043 Nucleic acid sequence Proteins 0.000 description 1
- 238000012408 PCR amplification Methods 0.000 description 1
- 102000057297 Pepsin A Human genes 0.000 description 1
- 108090000284 Pepsin A Proteins 0.000 description 1
- 102100035215 Phenylalanine-tRNA ligase alpha subunit Human genes 0.000 description 1
- 101710147128 Phenylalanine-tRNA ligase alpha subunit Proteins 0.000 description 1
- 101710202161 Probable phenylalanine-tRNA ligase alpha subunit Proteins 0.000 description 1
- 108010032060 RNA polymerase alpha subunit Proteins 0.000 description 1
- QAOWNCQODCNURD-UHFFFAOYSA-L Sulfate Chemical compound [O-]S([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 description 1
- VLSOAXRVHARBEQ-UHFFFAOYSA-N [4-fluoro-2-(hydroxymethyl)phenyl]methanol Chemical compound OCC1=CC=C(F)C=C1CO VLSOAXRVHARBEQ-UHFFFAOYSA-N 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 239000003124 biologic agent Substances 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 235000012000 cholesterol Nutrition 0.000 description 1
- OEYIOHPDSNJKLS-UHFFFAOYSA-N choline Chemical compound C[N+](C)(C)CCO OEYIOHPDSNJKLS-UHFFFAOYSA-N 0.000 description 1
- 229960001231 choline Drugs 0.000 description 1
- 239000011436 cob Substances 0.000 description 1
- 229960001127 colistin sulfate Drugs 0.000 description 1
- 108091036078 conserved sequence Proteins 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000035622 drinking Effects 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 238000009313 farming Methods 0.000 description 1
- 210000003608 fece Anatomy 0.000 description 1
- 235000021001 fermented dairy product Nutrition 0.000 description 1
- 235000021107 fermented food Nutrition 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 210000001035 gastrointestinal tract Anatomy 0.000 description 1
- 230000036512 infertility Effects 0.000 description 1
- 230000006799 invasive growth in response to glucose limitation Effects 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 230000000877 morphologic effect Effects 0.000 description 1
- ZESIAEVDVPWEKB-ORCFLVBFSA-N n-[(2s)-4-amino-1-[[(2s,3r)-1-[[(2s)-4-amino-1-oxo-1-[[(3s,6s,9s,12s,15r,18s,21s)-6,9,18-tris(2-aminoethyl)-3-[(1r)-1-hydroxyethyl]-12,15-bis(2-methylpropyl)-2,5,8,11,14,17,20-heptaoxo-1,4,7,10,13,16,19-heptazacyclotricos-21-yl]amino]butan-2-yl]amino]-3-h Chemical compound OS(O)(=O)=O.OS(O)(=O)=O.CC(C)CCCCC(=O)N[C@@H](CCN)C(=O)N[C@H]([C@@H](C)O)CN[C@@H](CCN)C(=O)N[C@H]1CCNC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](CCN)NC(=O)[C@H](CCN)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](CC(C)C)NC(=O)[C@H](CCN)NC1=O.CCC(C)CCCCC(=O)N[C@@H](CCN)C(=O)N[C@H]([C@@H](C)O)CN[C@@H](CCN)C(=O)N[C@H]1CCNC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](CCN)NC(=O)[C@H](CCN)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](CC(C)C)NC(=O)[C@H](CCN)NC1=O ZESIAEVDVPWEKB-ORCFLVBFSA-N 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 238000009304 pastoral farming Methods 0.000 description 1
- 229940111202 pepsin Drugs 0.000 description 1
- 239000000049 pigment Substances 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 210000002345 respiratory system Anatomy 0.000 description 1
- 101150047139 rpo1N gene Proteins 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000004460 silage Substances 0.000 description 1
- 235000020183 skimmed milk Nutrition 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
Images
Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/16—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
- A23K10/18—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions of live microorganisms
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/70—Feeding-stuffs specially adapted for particular animals for birds
- A23K50/75—Feeding-stuffs specially adapted for particular animals for birds for poultry
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
- A61K35/741—Probiotics
- A61K35/742—Spore-forming bacteria, e.g. Bacillus coagulans, Bacillus subtilis, clostridium or Lactobacillus sporogenes
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
- A61K35/741—Probiotics
- A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
- A61K35/747—Lactobacilli, e.g. L. acidophilus or L. brevis
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/43—Enzymes; Proenzymes; Derivatives thereof
- A61K38/44—Oxidoreductases (1)
- A61K38/443—Oxidoreductases (1) acting on CH-OH groups as donors, e.g. glucose oxidase, lactate dehydrogenase (1.1)
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/007—Pulmonary tract; Aromatherapy
- A61K9/0073—Sprays or powders for inhalation; Aerolised or nebulised preparations generated by other means than thermal energy
- A61K9/0078—Sprays or powders for inhalation; Aerolised or nebulised preparations generated by other means than thermal energy for inhalation via a nebulizer such as a jet nebulizer, ultrasonic nebulizer, e.g. in the form of aqueous drug solutions or dispersions
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/10—Dispersions; Emulsions
- A61K9/12—Aerosols; Foams
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y101/00—Oxidoreductases acting on the CH-OH group of donors (1.1)
- C12Y101/03—Oxidoreductases acting on the CH-OH group of donors (1.1) with a oxygen as acceptor (1.1.3)
- C12Y101/03004—Glucose oxidase (1.1.3.4)
-
- 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
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Microbiology (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Zoology (AREA)
- Veterinary Medicine (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Organic Chemistry (AREA)
- Epidemiology (AREA)
- Mycology (AREA)
- Polymers & Plastics (AREA)
- Biochemistry (AREA)
- Biotechnology (AREA)
- Dispersion Chemistry (AREA)
- Molecular Biology (AREA)
- Wood Science & Technology (AREA)
- Genetics & Genomics (AREA)
- Food Science & Technology (AREA)
- Birds (AREA)
- General Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Animal Husbandry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Virology (AREA)
- Otolaryngology (AREA)
- Communicable Diseases (AREA)
- Oncology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Pulmonology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Gastroenterology & Hepatology (AREA)
- Immunology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
Abstract
The invention provides lactobacillus plantarum GBW-LP001 capable of highly producing lactic acid, and a substituted antibacterial agent and application thereof. The classification of the Lactobacillus plantarum GBW-LP001 is named as Lactobacillus plantarumLactobacillus plantarumThe preservation number is CGMCC No.22078, the nucleotide sequence of 16S rRNA is shown in SEQ ID No.1, and the lactic acid yield is more than 18 after 24h of culture.1g/L, and has good use effect when being used as an opening material for young poultry animals. The invention also utilizes the lactobacillus plantarum GBW-LP001, the bacillus coagulans E21, the clostridium butyricum GBW-N1 and the glucose oxidase to jointly prepare a novel antibacterial agent, can be used for breeding poultry animals, can completely replace antibiotics to obtain a good breeding effect, and increases the breeding benefit.
Description
Technical Field
The invention belongs to the technical field of microorganisms, and relates to lactobacillus plantarum GBW-LP001 capable of highly producing lactic acid, a substituted antibacterial agent and application thereof.
Background
The production of commercial feed containing the growth-promoting drug feed additive is gradually stopped (short for 'forbidden resistance'). Probiotics, especially lactic acid bacteria, have become new anti-bacterial products and gradually highlight their unique advantages.
Lactobacillus plantarum is one of the lactobacilli, and is widely found in intestinal tracts of humans and animals, silage, fermented foods and dairy products. Usually, lactobacillus plantarum is found only in the late fermentation period, mainly because lactobacillus plantarum has high acid resistance and can endure various adverse environments. At present, lactobacillus plantarum is used as a probiotic for human, is used for reducing cholesterol and blood fat and losing weight, but is rarely used for breeding livestock and poultry animals.
At present, the lactobacillus plantarum is used for drinking water feeding in the breeding field of meat poultry. However, in the process of transporting the chicks from the poultry farm to the farm, a blank period of probiotics exists, and the blank period is the beginning and the key period of establishing intestinal flora from sterility to the intestinal flora of the chicks, so that the problem of how to increase the use of the probiotics after the chicks are shelved is very valuable to research, and most of microorganisms contacted with the chicks after the chicks are shelved are probiotics. Moreover, the stability of the lactobacillus plantarum is always a difficult problem, and the lactobacillus plantarum is short in storage time at normal temperature. Therefore, the stability of the lactobacillus plantarum is the first to solve, and the application range of the strain in the animal husbandry industry is expanded secondarily.
Disclosure of Invention
In order to overcome the defects and defects in the prior art, the invention provides a lactobacillus plantarum GBW-LP001 capable of highly producing lactic acid, the lactobacillus plantarum GBW-LP001 has excellent stability and stress resistance, a substitute antibacterial agent is prepared by using the lactobacillus plantarum GBW-LP001 and other strains, and the agent can be used for breeding livestock and poultry with remarkable effect.
In order to achieve the purpose of the invention, the invention is realized by the following technical scheme:
the invention provides Lactobacillus plantarum GBW-LP001 capable of highly producing lactic acid, which is classified and named as Lactobacillus plantarum and is preserved in China general microbiological culture Collection center (CGMCC) with the preservation number of CGMCC No. 22078.
Further, the nucleotide sequence of 16S rRNA of Lactobacillus plantarum GBW-LP001 is shown as SEQ ID No. 1.
Further, the nucleotide sequence of pheS of lactobacillus plantarum GBW-LP001 is shown as SEQ ID No. 2; the nucleotide sequence of pyrG is shown as SEQ ID No. 3; the nucleotide sequence of uvrC is shown in SEQ ID No. 4; the nucleotide sequence of rpoA is shown in SEQ ID No. 5.
Furthermore, the bacterial colony of the lactobacillus plantarum GBW-LP001 is in a very regular circle shape, is milky white, has the diameter of 2-3mm, has a smooth surface and neat edges, and is convex and opaque in the middle; the thallus is in a short rod shape, two ends are blunt and round, and the thallus is single, paired or chain-shaped, and has no flagellum and no spore.
Furthermore, the lactic acid yield of the lactobacillus plantarum GBW-LP001 after 24 hours of culture is more than or equal to 18.1 g/L.
The invention also provides an antibacterial agent containing the lactobacillus plantarum GBW-LP001, and the antibacterial agent also comprises bacillus coagulans, clostridium butyricum and glucose oxidase.
Further, the bacillus coagulans adopts bacillus coagulans E21 with the preservation number of CGMCC No.13044, and the clostridium butyricum adopts clostridium butyricum GBW-N1 with the preservation number of CGMCC No. 14499.
Furthermore, the viable bacteria content of the lactobacillus plantarum GBW-LP001 is 50 hundred million CFU/g, the viable bacteria content of the bacillus coagulans E21 is 50 hundred million CFU/g, the viable bacteria content of the clostridium butyricum GBW-N1 is 10 hundred million CFU/g, and the enzyme activity of the glucose oxidase is 1000U/g.
The invention also provides application of the lactobacillus plantarum GBW-LP001 or the antibacterial agent in preparation of an opening material or a spray for breeding poultries.
Further, when in use, the lactobacillus plantarum GBW-LP001 bacterial powder is used in the following amount: adding 50g of lactobacillus plantarum GBW-LP001 bacterial powder into 10000 young birds.
Further, the chicks are chicks.
The invention also provides the application of the lactobacillus plantarum GBW-LP001 or the antibacterial agent in preparing feed additives or drinking water additives for improving the growth performance of poultry.
Furthermore, when the antibacterial agent is used as a feed additive, the dosage of the antibacterial agent is 500g per ton of feed per day according to the feed intake of the poultry all day.
Further, when the antibacterial agent is used as a drinking water additive, the dosage of the antibacterial agent is 250g/m3And (4) adding water.
Compared with the prior art, the invention has the following advantages and technical effects:
1. the lactobacillus plantarum GBW-LP001 with high lactic acid yield is extracted from intestinal sections of healthy white feather broilers, has good stability and stress resistance, is higher than those of similar products in the market, has acid resistance, bile salt resistance and acid yield obviously better than those of similar products in the market, is safe and reliable, can be used as an opening material for chicks, and can obviously improve the survival rate and the marketing weight of the broilers through experimental verification, thereby improving the breeding benefit of the broilers.
2. The invention utilizes lactobacillus plantarum GBW-LP001, bacillus coagulans E21, clostridium butyricum GBW-N1 and glucose oxidase to prepare a novel alternative antibacterial agent, and experiments prove that the alternative antibacterial agent can completely replace antibiotics to be used in daily breeding of broilers and meat ducks, so that the growth performance of the broilers and the meat ducks is improved, the survival rate is improved, the feed-meat ratio is reduced, the slaughter weight is increased, and the breeding benefit is improved.
Drawings
FIG. 1 is the colony morphology of Lactobacillus plantarum GBW-LP001 on MRS plates.
FIG. 2 shows morphological characteristics of Lactobacillus plantarum GBW-LP001 under a microscope.
FIG. 3 shows the use of Lactobacillus plantarum GBW-LP001 as starter feed in a breeder's chicken house using the breeding data.
FIG. 4 is field 'substitution antibiotic' test data of the substitution antibiotic agent in the later stage of broiler chicken breeding; wherein A is the stool form of the test group, B is the stool form of the control group 1, C is the stool form of the control group 2, and D is the daily gain data of the broiler chickens.
Detailed Description
The technical solution of the present invention will be further described in detail with reference to the following specific examples.
In the following examples, unless otherwise specified, the experimental methods used were all conventional methods, and materials, reagents and the like used were all available from biological or chemical reagents companies.
The formulations of the media required in the following examples are as follows:
1. MRS solid medium: 10.0g/L of peptone, 5.0g/L of beef extract powder, 4.0g/L of yeast extract powder, 20.0g/L of glucose, 2.0g/L of dipotassium phosphate, 2.0g/L of triammonium citrate, 5.0g/L of sodium acetate, 0.2g/L of magnesium sulfate, 0.05g/L of manganese sulfate, 15.0g/L of agar, 801.0 g/L of tween, and the pH value of 6.5 +/-0.2 (25 ℃).
2. MRS liquid medium: 10.0g/L of peptone, 5.0g/L of beef extract powder, 4.0g/L of yeast extract powder, 20.0g/L of glucose, 2.0g/L of dipotassium phosphate, 2.0g/L of triammonium citrate, 5.0g/L of sodium acetate, 0.2g/L of magnesium sulfate, 0.05g/L of manganese sulfate, 801.0 g/L of tween, pH value of 6.5 +/-0.2 (25 ℃), and 0.5g/L of L-cysteine hydrochloride.
The above culture medium is sterilized at 116 deg.C for 30min before use, and then stored at room temperature.
Example 1: screening, separation and molecular identification of lactobacillus plantarum GBW-LP001
1. Screening and purifying of lactobacillus plantarum GBW-LP001 strain
Sampling different intestinal sections of healthy white feather broilers, separating and purifying the samples on an MRS liquid culture medium and an MRS solid culture medium for multiple times to obtain a lactobacillus strain with good acid productivity, and naming the single colony as GBW-LP 001.
As shown in figure 1, the bacterial colony of the bacterial strain GBW-LP001 on an MRS plate is in a very regular circle shape, is milky white, has the diameter of 2-3mm, has a smooth surface and neat edges, and is convex in the middle and opaque; the thallus under the microscope is in a short rod shape, two ends are blunt and round, and the thallus is single, paired or chain-shaped, and has no flagellum and no spore.
2. Identification of Lactobacillus plantarum GBW-LP001
(1)16S rRNA gene identification: the DNA of the strain GBW-LP001 is used as a template, 16S rRNA universal primers 27F and 1492R are used for amplification, amplified fragments are sent to Shanghai workers for sequence determination, the obtained nucleotide sequence is shown as SEQ ID No.1, the obtained sequence is compared by BLAST, and the result shows that the homology of the GBW-LP001 and a Lactobacillus plantarum sequence in GenBank is 99.93 percent, so that the strain is preliminarily determined to be Lactobacillus plantarum.
(2) Other conserved genes were further identified: the method comprises the steps of designing proper primers (the primer sequences are shown in table 1) by utilizing 4 conserved genes of pheS gene (phenylalanyl-tRNA synthetase alpha subunit), pyrG gene (CTP synthetase), uvrC gene (excision endonuclease subunit) and rpoA gene (RNA polymerase alpha subunit), carrying out PCR amplification, sending amplified fragments to Shanghai to carry out sequence determination, and respectively obtaining nucleotide sequences shown as SEQ ID No.2-SEQ ID No.5, wherein the sequence shows that the homology comparison similarities of the amplified fragments with Lactobacillus plantarum sequences in GenBank are respectively 99.86%, 99.41%, 100% and 99.88% through BLAST comparison. Therefore, it was finally confirmed that the strain was Lactobacillus plantarum.
Table 1: conserved gene name and primer
(3) Strain preservation
And (3) performing strain preservation on the screened strain GBW-LP001, wherein the preservation unit of the lactobacillus plantarum GBW-LP001 is as follows: china general microbiological culture Collection center (CGMCC); address: western road No.1, north west city of township, beijing, institute of microbiology, china academy of sciences; the preservation date is as follows: 26/03/2021; the preservation number of the Lactobacillus plantarum is CGMCC No. 22078.
Example 2: stability and stress resistance analysis of Lactobacillus plantarum GBW-LP001
1. Stability analysis of Lactobacillus plantarum GBW-LP001
Fermenting 1000 hundred million CFU/g pure bacterial powder of lactobacillus plantarum GBW-LP001 in a fermentation tank at 37 deg.C for about 12h, wherein the carrier is skimmed milk powder, and lyophilizing. The same line 1 in the market is lactobacillus plantarum produced by a certain Shanghai company, and the bacterial count is 2000 hundred million CFU/g; the market peer 2 is Lactobacillus plantarum produced by a certain company in Jiangsu, and the bacterial count is 1000 hundred million CFU/g; the market peer 3 is Lactobacillus plantarum produced by Taiwan, the bacterial load is 1000 hundred million CFU/g; three market samples were available commercially.
Sealing different lactobacillus plantarum pure bacteria powder, and placing the sealed lactobacillus plantarum pure bacteria powder at the temperature of 20+1 ℃ to track the number of viable bacteria. The results are shown in table 2, the bacterial load of lactobacillus plantarum GBW-LP001 after 1 month of storage is 83.9% of the original bacterial load, and the bacterial load after 2 months is 48.6% of the original bacterial load, although the number of viable bacteria is reduced during storage, the bacterial loads are all higher than those of market isors 1, 2 and 3, which indicates that the stability of lactobacillus plantarum GBW-LP001 after 1 month and 2 months of storage is better than that of the existing lactobacillus plantarum products.
Table 2: stability of Lactobacillus plantarum GBW-LP001
2. Acid resistance of Lactobacillus plantarum GBW-LP001
Firstly, preparing artificial gastric juice with pH value of 3.0, containing 1% pepsin for standby. Weighing different lactobacillus plantarum powder, adding the powder into artificial gastric juice according to the mass-volume ratio of 1: 100, and oscillating the powder at the constant temperature of 37 ℃ and 200rpm for 1 h; meanwhile, blank control without artificial gastric juice is made, and the bacteria content is respectively determined. The bacteria retention rate is the bacterial quantity after the artificial gastric juice treatment/blank bacterial quantity multiplied by 100 percent. As shown in table 3, the bacteria retention rates of GBW-LP001 in artificial gastric juice at pH 3.0 for 1h and 2h were 96.6% and 72.4%, respectively, which are higher than 3 market peers, indicating that lactobacillus plantarum GBW-LP001 has better acid resistance than the existing lactobacillus plantarum products.
Table 3: stress resistance of Lactobacillus plantarum GBW-LP001 under acidic conditions
3. Bile salt resistance of Lactobacillus plantarum GBW-LP001
First, PBS buffer solution with pH of 6.8 is prepared and sterilized for 30min at 121 ℃ for standby. Then, 0.3% of pig bile salt was added to the PBS buffer to prepare a PBS buffer containing 0.3% of bile salt. Weighing different lactobacillus plantarum powder, adding the powder into PBS buffer solution containing 0.3% bile salt according to the mass-volume ratio of 1: 100, and oscillating at the constant temperature of 37 ℃ and 200rpm for 4 hours; meanwhile, the blank control without the bile salt solution is used for measuring the bacteria content. And (5) calculating the survival rate of the bacteria by taking the number of bacteria measured in the blank as a control.
The results are shown in Table 4, the bacterial retention rate of GBW-LP001 in 0.3% bile salt after being treated for 4h at 37 ℃ is 28.9%, which is obviously higher than that of 3 market lines, and the results show that the Lactobacillus plantarum GBW-LP001 has certain choline resistance and the effect is superior to that of the existing Lactobacillus plantarum product.
Table 4: stress resistance of Lactobacillus plantarum GBW-LP001 under bile salt condition
4. Acid productivity of Lactobacillus plantarum GBW-LP001
Respectively selecting single colonies of different lactobacillus plantarum, inoculating the single colonies into an MRS liquid culture medium, culturing at 37 ℃ for 24 hours, taking a proper amount of culture solution to detect the pH value, and detecting the lactic acid yield by using a lactic acid test kit (Nanjing constructed biology). As shown in Table 5, the pH of the bacterial liquid of Lactobacillus plantarum GBW-LP001 was 3.74, the lactic acid production amount was 18.1g/L, the pH was lower than 3 market peers, and the lactic acid production amounts were all higher than 3 market peers, indicating that Lactobacillus plantarum GBW-LP001 had more excellent acid production performance.
Table 5: acid production performance of Lactobacillus plantarum GBW-LP001
Example 3: application case of lactobacillus plantarum GBW-LP001 in culture site
Lactobacillus plantarum GBW-LP001 used as chick starter feed in chicken house
Test site: fuxi (Weihai) farming-grazing development Co Ltd
Test protocol:
1. test materials: lactobacillus plantarum GBW-LP001 with the strain content of 2000 hundred million CFU/g; edible carmine pigment: procurement by Tianjin food company; PBS buffer: the Qingdao root research and development center provides; spraying equipment: is provided by a Fox breeding poultry factory.
2. Application protocol
2.1 preparation of probiotic spray solution
Taking out Lactobacillus plantarum GBW-LP001 bacterial powder from a refrigerator, adding 50g of bacterial powder into 10000 chicks according to the using amount of the bacterial powder, dissolving the bacterial powder into 2100mL of PBS buffer solution, adding 1g of carmine, and uniformly mixing. Adding the prepared probiotic spraying solution into spraying equipment.
2.2, grouping
300 ten thousand chicks in the test group use the lactobacillus plantarum GBW-LP001 bacterial powder; the control group 300 ten thousand chicks were not used.
2.3 spraying
And adjusting a spraying device, spraying the chicks for three times, stopping for 5s, and simultaneously controlling the speed of the chick conveying belt to enable the chicks in each paper box to stay for 5s under the spraying device.
3. Effects of use
After spraying, the probiotic spraying solution forms fog in the spraying equipment and can enter the bodies of the chickens through a respiratory system, meanwhile, red small water drops are formed on the feathers of the chickens, the chickens can peck the water drops mutually, and the probiotics further enter the bodies of the chickens.
The farming and pasturing company adopts Cobb ground flat breeding, and after the chicks are hatched and sprayed, the comprehensive data of 600 ten thousand broilers which are out of the farm are counted after two months, wherein the weight of the farm is the average weight of each chicken, the input-output ratio is the ratio of input/output, the input means the cost of each chicken for increasing probiotics, and the output means the yield increase rate of each chicken after the same number of chicks are out of the farm.
The production performance data of broiler breeding after chicks are bred is shown in fig. 3, and it can be seen that the broiler survival rate of a test group is improved by 0.91% compared with that of a control group, the slaughter weight is increased by 79g, the feed-weight ratio is reduced by 0.02%, the input-output ratio is 1: 33, which shows that after lactobacillus plantarum GBW-LP001 is used, the growth performance of the broiler is obviously improved, and the broiler breeding effect is very considerable because the broiler breeding performance data is safe and reliable for the chicks.
Second, the use of 'replacing antibiotics' in the later stage of the cultivation of the meat poultry
A novel antibacterial agent is prepared by utilizing lactobacillus plantarum GBW-LP001, bacillus coagulans E21, clostridium butyricum GBW-N1 and glucose oxidase, wherein the viable bacteria content of the lactobacillus plantarum GBW-LP001 is 50 hundred million CFU/g, the viable bacteria content of the bacillus coagulans E21 is 50 hundred million CFU/g (the bacillus coagulans preservation number is CGMCC No.13044), the viable bacteria content of the clostridium butyricum GBW-N1 is 10 hundred million CFU/g (the clostridium butyricum preservation number is CGMCC No.14499), and the enzyme activity of the glucose oxidase is 1000U/g.
The collection unit of the bacillus coagulans E21: china general microbiological culture Collection center (CGMCC); address: western road No.1, north west city of township, beijing, institute of microbiology, china academy of sciences; the preservation date is as follows: 2016, 9, 26 months; the preservation number of the Bacillus coagulans is CGMCC NO. 13044.
The collection of Clostridium butyricum GBW-N1: china general microbiological culture Collection center (CGMCC); address: western road No.1, north west city of township, beijing, institute of microbiology, china academy of sciences; the preservation date is as follows: year 2017, month 08, day 07; the preservation number of the Clostridium butyricum is CGMCC No. 14499.
Substitution resistance test 1: feed additive for later stage of broiler chicken breeding
The experimental design is shown in table 6, the control group 1 (antibiotic group) starts to add 20% colistin sulfate according to the conventional addition amount at the age of 30 days, the control group 2 starts to add the compound traditional Chinese medicine preparation according to the conventional addition amount at the age of 30 days, and the experimental group starts to add the antibacterial agent at the age of 30 days according to the feed intake of 500 g/ton per day.
Table 6: on-site 'substitution resistance' test design in later stage of broiler chicken breeding
The test results are shown in fig. 4: compared with the control group 1, the test group has better forming degree and no water excrement phenomenon in the excrement form of the broiler chicken; control 2 was dark brown in color relative to the feces of the other two groups, with a small amount of visible feed particles. In the aspect of daily gain, the daily gain of the broiler chickens in the test group is obviously higher than that of the broiler chickens in the control group, which shows that the antibacterial agent can completely replace the antibiotic product, and the using effect is obviously better than that of the antibiotic and the compound traditional Chinese medicine preparation.
Substitution resistance test 2: water drinking additive for later stage of meat duck breeding
The experimental design is shown in Table 7, and the control group was started to add 20% at 28 days of ageColistin sulfate, the test group started to add the alternative antibacterial agent to drinking water at the age of 28 days, and the addition amount of the alternative antibacterial agent is 250g/m3And (3) water.
Table 7: on-site 'resistance replacing' test design for meat duck breeding later stage
The results of the meat duck breeding test are shown in table 8: compared with a control group, the survival rate of the meat duck in the test group is improved by 2.59 percent, the feed-meat ratio is reduced by 0.025 percent, and the slaughter weight is increased by 0.14 jin, which shows that the alternative antibacterial agent taking the lactobacillus plantarum GBW-LP001 as the main component can completely replace the use of antibiotics, and meanwhile, positive results are obtained in the production performance of the meat duck.
Table 8: on-site 'substitute resistance' test breeding data in later breeding period of meat ducks
The total data of the survival rate, the feed-meat ratio and the slaughter weight are calculated according to a weighted average method.
The above examples are only intended to illustrate the technical solution of the present invention, but not to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, it will be apparent to those skilled in the art that various changes may be made and equivalents may be substituted for elements thereof; and such modifications or substitutions do not depart from the spirit and scope of the corresponding technical solutions.
Sequence listing
<110> Qingdao Shangde Biotechnology Co., Ltd
<120> lactobacillus plantarum GBW-LP001 capable of highly producing lactic acid and antibacterial agent and application thereof
<160> 13
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1466
<212> DNA
<213> Lactobacillus plantarum (Lactobacillus plantarum)
<400> 1
ggcgggcggg gtgctataca tgcagtcgaa cgaactctgg tattgattgg tgcttgcatc 60
atgatttaca tttgagtgag tggcgaactg gtgagtaaca cgtgggaaac ctgcccagaa 120
gcgggggata acacctggaa acagatgcta ataccgcata acaacttgga ccgcatggtc 180
cgagtttgaa agatggcttc ggctatcact tttggatggt cccgcggcgt attagctaga 240
tggtggggta acggctcacc atggcaatga tacgtagccg acctgagagg gtaatcggcc 300
acattgggac tgagacacgg cccaaactcc tacgggaggc agcagtaggg aatcttccac 360
aatggacgaa agtctgatgg agcaacgccg cgtgagtgaa gaagggtttc ggctcgtaaa 420
actctgttgt taaagaagaa catatctgag agtaactgtt caggtattga cggtatttaa 480
ccagaaagcc acggctaact acgtgccagc agccgcggta atacgtaggt ggcaagcgtt 540
gtccggattt attgggcgta aagcgagcgc aggcggtttt ttaagtctga tgtgaaagcc 600
ttcggctcaa ccgaagaagt gcatcggaaa ctgggaaact tgagtgcaga agaggacagt 660
ggaactccat gtgtagcggt gaaatgcgta gatatatgga agaacaccag tggcgaaggc 720
ggctgtctgg tctgtaactg acgctgaggc tcgaaagtat gggtagcaaa caggattaga 780
taccctggta gtccataccg taaacgatga atgctaagtg ttggagggtt tccgcccttc 840
agtgctgcag ctaacgcatt aagcattccg cctggggagt acggccgcaa ggctgaaact 900
caaaggaatt gacgggggcc cgcacaagcg gtggagcatg tggtttaatt cgaagctacg 960
cgaagaacct taccaggtct tgacatacta tgcaaatcta agagattaga cgttcccttc 1020
ggggacatgg atacaggtgg tgcatggttg tcgtcagctc gtgtcgtgag atgttgggtt 1080
aagtcccgca acgagcgcaa cccttattat cagttgccag cattaagttg ggcactctgg 1140
tgagactgcc ggtgacaaac cggaggaagg tggggatgac gtcaaatcat catgcccctt 1200
atgacctggg ctacacacgt gctacaatgg atggtacaac gagttgcgaa ctcgcgagag 1260
taagctaatc tcttaaagcc attctcagtt cggattgtag gctgcaactc gcctacatga 1320
agtcggaatc gctagtaatc gcggatcagc atgccgcggt gaatacgttc ccgggccttg 1380
tacacaccgc ccgtcacacc atgagagttt gtaacaccca aagtcggtgg ggtaaccttt 1440
taggaaccag ccgctaaggt gaccgt 1466
<210> 2
<211> 697
<212> DNA
<213> Lactobacillus plantarum (Lactobacillus plantarum)
<400> 2
cggaacaagc agccgttaat gagcaattgg ctgccgaaaa actggacgtg acgttaccgg 60
gtcgggaagt tccacaaggt cagcctcacg tgattaccca gattattact gaattggaag 120
atctatttat gggaatgggc tatcaaattg ttgatggtga tgaagttgaa gaagattact 180
acaactttga acggttgaac ttaccgaagg accatcccgc ccgtgacatg caagacacgt 240
tctatattac caaagacgtg ctactacgca cgcagacgtc tgctgatcag ccgcggtcac 300
ttgaaaatca cgatttttct aaaggaccgc tgaaggtctt gtcacctggc cgcgtttatc 360
ggcgtgatac ggatgatgca acccattccc atcaatttca tcaaattgaa gggttagtcg 420
tggacaagca tattacgatg gctgatttga agggcacctt aattctggtt gccaagactt 480
tgtttggcga tcaattcgat gttcggctac ggccaagctt ctttccattc acggaaccat 540
ccgtagaagc tgatgtaact tgctttaatt gcaatggcaa gggctgtgca atctgtaagc 600
aaacgggttg gatcgaagta ctgggtgccg gcatggttca cccccacgtg ttagaaatgt 660
ctggcattga tccagaagaa tatggtggtt ttgcttt 697
<210> 3
<211> 673
<212> DNA
<213> Lactobacillus plantarum (Lactobacillus plantarum)
<400> 3
gagttgattt taagttccga caatgttttc tatatccata ccacattgat cccatattta 60
cgggcagctg gtgaaatgaa gacgaagcca acgcaacatt ctgttaagga attgcggagt 120
tatgggattc agccgaacat gttagttgtc cggactgaac aaccaattac gcgggaaatg 180
cggaataaga ttgcgtcctt ctgtgacgtg gaaccagaag cagtcattga atccttagac 240
gttaagacga tttattcaat tccgttgaat gttcagaaac aaaacatgga ccaaatcgtc 300
cttgaccatt ttgatgtaca ggcacctaag gccgacatga gtgaatggat tgacttagaa 360
catcatgttc agaacttatc acggaccatc aagattgcgc tagtcggaaa atacgtcgct 420
ttacaggatg cttatatctc agtgacggaa gcattgaagc atgctggcta tacggatgat 480
gccgacattg atttgaagaa gatttctgct gaagatgtta cgccagaaaa tgtcgaagaa 540
ctactcggcg atgctgacgg aatcttagtt cctggtggct ttggtgatcg gggaattgaa 600
ggtaagatta cggcaatcaa gtatgcccgt gaaaacgacg tgccattctt aggtatctgc 660
ttggggaatg caa 673
<210> 4
<211> 770
<212> DNA
<213> Lactobacillus plantarum (Lactobacillus plantarum)
<400> 4
gatccatttt ttaatgggta aggccaagaa cttgaaaaat cgggtgcgtt catatttcaa 60
aagcagccat actggtaaga ccgcgcggtt agtatcagag attgcggact ttgagtttat 120
tgtgacctca acggataagg aagccttcct gctagaaatt acgttaattc aaaaacatca 180
accgtacttt aatattaaat taaaaaaggg aaccggttat ccatacatta agattaccaa 240
tgaacgagat ccccagatcc tgatcgtcag tgatgttcgt aaagacggtg gctactactt 300
tgggccatat cccaacgttt acgcggctca ggaaacggtc aattttattc agaaggtcta 360
tccgttgcgg cgttgccatg gtttccagaa gcggccgtgc ctctattatc atatgggcca 420
atgcttggga gcttgcttta aaacggtgcc ggtggcagaa tatgacgctc agatcaagcg 480
gattaaatcc tttttgaatg gtcacgttga aacggtcaag aagcagttaa ctaaacgaat 540
ggatcaagcg gccgctgacc tagaatttga acgagccgct gaattacgtg atcaactgaa 600
ttatattgaa atgacggttg aaaagcagaa aattatttca aacgataata cgccccgcga 660
cctgtttaat ttctatcttg ataaaggctg gctttcgatt caggtcttct ttattcggca 720
agcgcggttg atgaagcgcg agaaacgctt gttccgatta aatggttcaa 770
<210> 5
<211> 819
<212> DNA
<213> Lactobacillus plantarum (Lactobacillus plantarum)
<400> 5
ttatgattga aatttgaaaa acctaacatt cataaaattg atgaaaacga caactatggt 60
aagtttgttg tagaaccgct tgaacgcggt tatggtacaa ctttagggaa ttcacttcgt 120
cggattcttc tttcttcttt acctggcgct gctgttacta gtattcaaat tgatggtgtt 180
cttcatgaat tttcaacgat tgagggcgta acggaagacg ttacagcaat tatcttgaat 240
gttaagaaga ttgcacttaa gttggaatca gacgaaacca agacgttgga aatcgacgtt 300
aagggtcctg ctaacgttac tgccagtgat atcattggcg atgcggacgt agaagtcttg 360
aatccagact taccaatttg taccgtagca gacggggcac acttccatat gcgtatgacc 420
gcaaatactg gtcgtggtta tgtttccgct gaggataaca aacatcgtga agatgacatg 480
ccaattggcg ttttagctgt tgattcattg tattctccaa tcgaacgtgt caactaccaa 540
gttgaaaaca cgcgggttgg tcaacgtgat gatttcgata agttaacctt agacgtttgg 600
acaaatggtt caatcactcc aagtgaagcc attagtctat cagccaaaat cctgactgat 660
cacctttcaa tcttcgtaaa tctcactgat gaagctaaaa acactgatgt gatggtcgag 720
aaggaagaaa cgcataagga aaagatgtta gaaatgacga ttgaagagtt agatctctcc 780
gtccgttcat acaattgctt gaagcgtgca ggtactcac 819
<210> 6
<211> 18
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 6
ccgtgaagaa ctggaaca 18
<210> 7
<211> 18
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 7
cctaacccaa aggcaaaa 18
<210> 8
<211> 20
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 8
agtgatttag gttccgacaa 20
<210> 9
<211> 18
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 9
tgcattccca agcagata 18
<210> 10
<211> 21
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 10
gatcatttat gtgggtaagg c 21
<210> 11
<211> 20
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 11
tgacactact gggaacaagc 20
<210> 12
<211> 20
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 12
atgatygart ttgaaaaacc 20
<210> 13
<211> 20
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 13
achgtrttra tdccdgcrcg 20
Claims (10)
1. A lactobacillus plantarum GBW-LP001 capable of highly producing lactic acid is characterized in that the lactobacillus plantarum GBW-LP001 is named as lactobacillus plantarum in a classification modeLactobacillus plantarumThe microbial inoculum is preserved in the China general microbiological culture Collection center with the preservation number of CGMCC No. 22078.
2. Lactobacillus plantarum GBW-LP001 according to claim 1, characterized in that the nucleotide sequence of the 16S rRNA of Lactobacillus plantarum GBW-LP001 is shown in SEQ ID No. 1.
3. Lactobacillus plantarum GBW-LP001 according to claim 1, wherein the colonies of Lactobacillus plantarum GBW-LP001 are very regular circles, milky white, 2-3mm in diameter, smooth in surface, regular in edges, convex in the middle, opaque; the thallus is in a short rod shape, two ends are blunt and round, and the thallus is single, paired or chain-shaped, and has no flagellum and no spore.
4. Lactobacillus plantarum GBW-LP001 according to claim 1, wherein the lactic acid production of Lactobacillus plantarum GBW-LP001 after 24h cultivation is 18.1g/L or more.
5. An antibacterial agent comprising lactobacillus plantarum GBW-LP001 according to any one of claims 1 to 4, further comprising bacillus coagulans, clostridium butyricum and glucose oxidase.
6. The alternative antibacterial agent according to claim 5, wherein the Bacillus coagulans is Bacillus coagulans E21 with the preservation number of CGMCC No.13044, and the Clostridium butyricum is Clostridium butyricum GBW-N1 with the preservation number of CGMCC No. 14499.
7. The alternative antibacterial agent according to claim 6, wherein the viable bacteria content of Lactobacillus plantarum GBW-LP001 is 50 hundred million CFU/g, the viable bacteria content of Bacillus coagulans E21 is 50 hundred million CFU/g, the viable bacteria content of Clostridium butyricum GBW-N1 is 10 hundred million CFU/g, and the enzyme activity of glucose oxidase is 1000U/g.
8. Use of lactobacillus plantarum GBW-LP001 according to any one of claims 1-4 or the alternative antibacterial agent according to any one of claims 5-7 for the preparation of a mouth-opening or spray for hatchling.
9. The use according to claim 8, wherein the powder of Lactobacillus plantarum GBW-LP001 is used in an amount: adding 50g of lactobacillus plantarum GBW-LP001 bacterial powder into 10000 young birds.
10. Use of lactobacillus plantarum GBW-LP001 according to any one of claims 1 to 4 or a antibacterial agent according to any one of claims 5 to 7 for the preparation of a feed additive for improving the growth performance of poultry.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110733301.5A CN113502243B (en) | 2021-06-29 | 2021-06-29 | Lactobacillus plantarum GBW-LP001 capable of highly producing lactic acid and antibacterial agent alternative thereof and application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110733301.5A CN113502243B (en) | 2021-06-29 | 2021-06-29 | Lactobacillus plantarum GBW-LP001 capable of highly producing lactic acid and antibacterial agent alternative thereof and application |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN113502243A true CN113502243A (en) | 2021-10-15 |
| CN113502243B CN113502243B (en) | 2022-04-08 |
Family
ID=78009656
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110733301.5A Active CN113502243B (en) | 2021-06-29 | 2021-06-29 | Lactobacillus plantarum GBW-LP001 capable of highly producing lactic acid and antibacterial agent alternative thereof and application |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113502243B (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115261287A (en) * | 2022-09-13 | 2022-11-01 | 青岛根源生物技术集团有限公司 | Composite lactobacillus microbial inoculum and application thereof |
| CN115843921A (en) * | 2023-02-14 | 2023-03-28 | 成都铁骑力士饲料有限公司 | Biological pretreatment method for reducing mildew of wet residue in feed application |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106906154A (en) * | 2015-12-22 | 2017-06-30 | 北京大北农科技集团股份有限公司 | One lactobacillus plantarum strain and its feed addictive and feed |
| US20170333496A1 (en) * | 2016-05-20 | 2017-11-23 | Church & Dwight Co., Inc. | Lactobacillus and bacillus based direct fed microbial treatment for poultry and method of use |
| CN107868763A (en) * | 2017-11-21 | 2018-04-03 | 青岛根源生物技术集团有限公司 | One plant is improved and attacks the bacillus coagulans E21 of the anti-oxidation stress ability of livestock and poultry animal and its application after poison |
| CN112608861A (en) * | 2020-12-21 | 2021-04-06 | 青岛尚德生物技术有限公司 | Composite preparation containing clostridium butyricum and pediococcus acidilactici as well as preparation method and application of composite preparation |
-
2021
- 2021-06-29 CN CN202110733301.5A patent/CN113502243B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106906154A (en) * | 2015-12-22 | 2017-06-30 | 北京大北农科技集团股份有限公司 | One lactobacillus plantarum strain and its feed addictive and feed |
| US20170333496A1 (en) * | 2016-05-20 | 2017-11-23 | Church & Dwight Co., Inc. | Lactobacillus and bacillus based direct fed microbial treatment for poultry and method of use |
| CN107868763A (en) * | 2017-11-21 | 2018-04-03 | 青岛根源生物技术集团有限公司 | One plant is improved and attacks the bacillus coagulans E21 of the anti-oxidation stress ability of livestock and poultry animal and its application after poison |
| CN112608861A (en) * | 2020-12-21 | 2021-04-06 | 青岛尚德生物技术有限公司 | Composite preparation containing clostridium butyricum and pediococcus acidilactici as well as preparation method and application of composite preparation |
Non-Patent Citations (1)
| Title |
|---|
| 宋小燕等: "植物乳杆菌对大恒肉鸡生长性能、屠宰性能以及血清免疫和抗氧化功能的影响", 《动物营养学报》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115261287A (en) * | 2022-09-13 | 2022-11-01 | 青岛根源生物技术集团有限公司 | Composite lactobacillus microbial inoculum and application thereof |
| CN115843921A (en) * | 2023-02-14 | 2023-03-28 | 成都铁骑力士饲料有限公司 | Biological pretreatment method for reducing mildew of wet residue in feed application |
| CN115843921B (en) * | 2023-02-14 | 2023-07-04 | 成都铁骑力士饲料有限公司 | Biological pretreatment method for reducing mildew of wet residues in feed application |
Also Published As
| Publication number | Publication date |
|---|---|
| CN113502243B (en) | 2022-04-08 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106282072B (en) | Compound lactobacillus microecological preparation and preparation method and application thereof | |
| CN106906154B (en) | Lactobacillus plantarum strain, feed additive and feed thereof | |
| CN109161509B (en) | Bacterial strain capable of preventing and treating bovine and sheep diarrhea | |
| CN107164269A (en) | A kind of lactobacillus paracasei, preparation and its application in pig feed | |
| CN109749957B (en) | Preparation and application of lactobacillus gasseri preparation with aquatic pathogenic bacteria antagonistic property | |
| CN108753650B (en) | Enterococcus faecium and composite microecological preparation prepared from enterococcus faecium | |
| CN107312732B (en) | Probiotic feed additive | |
| CN104293696A (en) | Enterococcus faecalis HEW-A131 and application thereof | |
| CN108641979B (en) | Enterococcus faecium, high-density fermentation culture method thereof and microecological preparation prepared from enterococcus faecium | |
| CN113502243B (en) | Lactobacillus plantarum GBW-LP001 capable of highly producing lactic acid and antibacterial agent alternative thereof and application | |
| CN102559539A (en) | Lactobacillus acidophilus and application thereof and feed additive thereof and premix compound thereof | |
| CN111733117B (en) | Bacillus marinus for producing antibacterial peptide and fermentation method and application thereof | |
| CN103060222A (en) | Bacillus subtilis B27 with probiotic effect and application thereof | |
| CN102517227B (en) | Enterococcus faecalis and applications and feed additive and leavening agent thereof | |
| CN108522884B (en) | Application of composite microecological preparation in preventing fat deposition of broiler chickens | |
| CN103266074B (en) | B.subtilis spores strain and application thereof | |
| CN104263683B (en) | Bacillus pumilus 315 and its application with prebiotic effect | |
| CN107828703A (en) | Space lactobacillus reuteri Fullarton 9 35 and application | |
| CN107974424A (en) | Space lactobacillus reuteri Fullarton-9-25 and application | |
| CN116064324B (en) | Lactobacillus rhamnosus, culture method thereof and application thereof in preventing and treating diarrhea and enteritis | |
| CN117801981A (en) | Screening and application of probiotics lactic acid bacteria for promoting growth of broiler chickens | |
| CN106811429A (en) | The application of one bacillus subtilis strain and its feed addictive and feed | |
| CN113604387B (en) | Salt-tolerant and high-temperature-resistant lactobacillus reuteri and application thereof in prevention and treatment of pathogenic bacteria in livestock and poultry aquaculture | |
| CN111728081B (en) | Composite bacteria fermentation liquor for feed additive and preparation method thereof | |
| CN113999787B (en) | Clostridium butyricum, microbial inoculum, preparation method and application thereof, and feed |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |