CN116555071A - 一株产蛋白酶的粪肠球菌及应用该粪肠球菌的饲料发酵方法 - Google Patents
一株产蛋白酶的粪肠球菌及应用该粪肠球菌的饲料发酵方法 Download PDFInfo
- Publication number
- CN116555071A CN116555071A CN202310036365.9A CN202310036365A CN116555071A CN 116555071 A CN116555071 A CN 116555071A CN 202310036365 A CN202310036365 A CN 202310036365A CN 116555071 A CN116555071 A CN 116555071A
- Authority
- CN
- China
- Prior art keywords
- fermentation
- enterococcus faecalis
- feed
- soybean meal
- substrate
- 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.)
- Pending
Links
- 238000000855 fermentation Methods 0.000 title claims abstract description 103
- 230000004151 fermentation Effects 0.000 title claims abstract description 102
- 241000194032 Enterococcus faecalis Species 0.000 title claims abstract description 47
- 229940032049 enterococcus faecalis Drugs 0.000 title claims abstract description 44
- 108091005804 Peptidases Proteins 0.000 title claims abstract description 21
- 239000004365 Protease Substances 0.000 title claims abstract description 21
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 title claims abstract description 21
- 238000000034 method Methods 0.000 title claims abstract description 13
- 235000019764 Soybean Meal Nutrition 0.000 claims abstract description 55
- 239000004455 soybean meal Substances 0.000 claims abstract description 55
- 238000004321 preservation Methods 0.000 claims abstract description 7
- 239000000758 substrate Substances 0.000 claims description 31
- 239000007788 liquid Substances 0.000 claims description 22
- 241000894006 Bacteria Species 0.000 claims description 19
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 13
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims description 12
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 claims description 12
- 238000002156 mixing Methods 0.000 claims description 12
- 238000012258 culturing Methods 0.000 claims description 6
- 238000010563 solid-state fermentation Methods 0.000 claims description 5
- 235000015099 wheat brans Nutrition 0.000 claims description 5
- 239000011159 matrix material Substances 0.000 claims description 4
- 239000000126 substance Substances 0.000 claims description 3
- 229930006000 Sucrose Natural products 0.000 claims 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 claims description 2
- 230000000694 effects Effects 0.000 abstract description 13
- 238000012216 screening Methods 0.000 abstract description 13
- 108091007433 antigens Proteins 0.000 abstract description 9
- 102000036639 antigens Human genes 0.000 abstract description 9
- 238000002360 preparation method Methods 0.000 abstract description 9
- 244000005700 microbiome Species 0.000 abstract description 7
- 230000000433 anti-nutritional effect Effects 0.000 abstract description 5
- 108090000765 processed proteins & peptides Proteins 0.000 abstract description 5
- 238000004519 manufacturing process Methods 0.000 abstract description 4
- 238000009629 microbiological culture Methods 0.000 abstract description 4
- 239000002028 Biomass Substances 0.000 abstract description 3
- 239000002207 metabolite Substances 0.000 abstract description 3
- 238000006243 chemical reaction Methods 0.000 abstract description 2
- 102000004196 processed proteins & peptides Human genes 0.000 abstract 1
- 230000013777 protein digestion Effects 0.000 abstract 1
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 18
- 244000046052 Phaseolus vulgaris Species 0.000 description 15
- 235000010627 Phaseolus vulgaris Nutrition 0.000 description 15
- 239000002609 medium Substances 0.000 description 11
- 241000282887 Suidae Species 0.000 description 10
- 239000001963 growth medium Substances 0.000 description 10
- 230000001580 bacterial effect Effects 0.000 description 9
- 239000004310 lactic acid Substances 0.000 description 9
- 235000014655 lactic acid Nutrition 0.000 description 9
- 206010012735 Diarrhoea Diseases 0.000 description 8
- 239000007787 solid Substances 0.000 description 7
- 239000000843 powder Substances 0.000 description 6
- 108090000623 proteins and genes Proteins 0.000 description 6
- 238000012360 testing method Methods 0.000 description 6
- 230000015556 catabolic process Effects 0.000 description 5
- 238000006731 degradation reaction Methods 0.000 description 5
- 235000021050 feed intake Nutrition 0.000 description 5
- 235000016709 nutrition Nutrition 0.000 description 5
- 235000018102 proteins Nutrition 0.000 description 5
- 102000004169 proteins and genes Human genes 0.000 description 5
- 239000000243 solution Substances 0.000 description 5
- 229920001817 Agar Polymers 0.000 description 4
- 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 4
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 4
- 239000008272 agar Substances 0.000 description 4
- 238000002474 experimental method Methods 0.000 description 4
- 210000001035 gastrointestinal tract Anatomy 0.000 description 4
- 239000008103 glucose Substances 0.000 description 4
- 108010083391 glycinin Proteins 0.000 description 4
- 239000000463 material Substances 0.000 description 4
- 230000002797 proteolythic effect Effects 0.000 description 4
- 239000000523 sample Substances 0.000 description 4
- 230000001954 sterilising effect Effects 0.000 description 4
- 108020004465 16S ribosomal RNA Proteins 0.000 description 3
- 241000193830 Bacillus <bacterium> Species 0.000 description 3
- 244000068988 Glycine max Species 0.000 description 3
- 235000010469 Glycine max Nutrition 0.000 description 3
- 108700037728 Glycine max beta-conglycinin Proteins 0.000 description 3
- 241000186660 Lactobacillus Species 0.000 description 3
- 241001465754 Metazoa Species 0.000 description 3
- 239000001888 Peptone Substances 0.000 description 3
- 108010080698 Peptones Proteins 0.000 description 3
- 240000008042 Zea mays Species 0.000 description 3
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 3
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 3
- 238000010564 aerobic fermentation Methods 0.000 description 3
- 150000001413 amino acids Chemical class 0.000 description 3
- 235000015278 beef Nutrition 0.000 description 3
- 235000005822 corn Nutrition 0.000 description 3
- 230000012010 growth Effects 0.000 description 3
- 230000003301 hydrolyzing effect Effects 0.000 description 3
- 230000000968 intestinal effect Effects 0.000 description 3
- 230000006799 invasive growth in response to glucose limitation Effects 0.000 description 3
- 229940039696 lactobacillus Drugs 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 230000035764 nutrition Effects 0.000 description 3
- 235000019319 peptone Nutrition 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 238000003756 stirring Methods 0.000 description 3
- 238000009631 Broth culture Methods 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 2
- 238000003794 Gram staining Methods 0.000 description 2
- IMQLKJBTEOYOSI-GPIVLXJGSA-N Inositol-hexakisphosphate Chemical compound OP(O)(=O)O[C@H]1[C@H](OP(O)(O)=O)[C@@H](OP(O)(O)=O)[C@H](OP(O)(O)=O)[C@H](OP(O)(O)=O)[C@@H]1OP(O)(O)=O IMQLKJBTEOYOSI-GPIVLXJGSA-N 0.000 description 2
- 108090001090 Lectins Proteins 0.000 description 2
- 102000004856 Lectins Human genes 0.000 description 2
- IMQLKJBTEOYOSI-UHFFFAOYSA-N Phytic acid Natural products OP(O)(=O)OC1C(OP(O)(O)=O)C(OP(O)(O)=O)C(OP(O)(O)=O)C(OP(O)(O)=O)C1OP(O)(O)=O IMQLKJBTEOYOSI-UHFFFAOYSA-N 0.000 description 2
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 101710162629 Trypsin inhibitor Proteins 0.000 description 2
- 229940122618 Trypsin inhibitor Drugs 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 230000002378 acidificating effect Effects 0.000 description 2
- 230000003078 antioxidant effect Effects 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 239000012895 dilution Substances 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
- 239000012153 distilled water Substances 0.000 description 2
- 239000000835 fiber Substances 0.000 description 2
- 230000037308 hair color Effects 0.000 description 2
- 230000007062 hydrolysis Effects 0.000 description 2
- 238000006460 hydrolysis reaction Methods 0.000 description 2
- 230000036039 immunity Effects 0.000 description 2
- 239000002054 inoculum Substances 0.000 description 2
- 239000002523 lectin Substances 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
- 239000011702 manganese sulphate Substances 0.000 description 2
- 235000007079 manganese sulphate Nutrition 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
- 230000000813 microbial effect Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 235000019629 palatability Nutrition 0.000 description 2
- 235000002949 phytic acid Nutrition 0.000 description 2
- 239000000467 phytic acid Substances 0.000 description 2
- 229940068041 phytic acid Drugs 0.000 description 2
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 description 2
- 229920000053 polysorbate 80 Polymers 0.000 description 2
- 244000144977 poultry Species 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 238000005070 sampling Methods 0.000 description 2
- 238000007789 sealing Methods 0.000 description 2
- 238000011218 seed culture Methods 0.000 description 2
- 235000020183 skimmed milk Nutrition 0.000 description 2
- 239000001632 sodium acetate Substances 0.000 description 2
- 235000017281 sodium acetate Nutrition 0.000 description 2
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 2
- 230000003068 static effect Effects 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 239000008399 tap water Substances 0.000 description 2
- 235000020679 tap water Nutrition 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
- 239000002753 trypsin inhibitor Substances 0.000 description 2
- 239000011782 vitamin Substances 0.000 description 2
- 235000013343 vitamin Nutrition 0.000 description 2
- 229940088594 vitamin Drugs 0.000 description 2
- 229930003231 vitamin Natural products 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- NLMDJJTUQPXZFG-UHFFFAOYSA-N 1,4,10,13-tetraoxa-7,16-diazacyclooctadecane Chemical compound C1COCCOCCNCCOCCOCCN1 NLMDJJTUQPXZFG-UHFFFAOYSA-N 0.000 description 1
- 102000006734 Beta-Globulins Human genes 0.000 description 1
- 108010087504 Beta-Globulins Proteins 0.000 description 1
- 108050009302 Claudin Proteins 0.000 description 1
- 235000019750 Crude protein Nutrition 0.000 description 1
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 1
- 239000004472 Lysine Substances 0.000 description 1
- 102000015728 Mucins Human genes 0.000 description 1
- 108010063954 Mucins Proteins 0.000 description 1
- 108010038807 Oligopeptides Proteins 0.000 description 1
- 102000015636 Oligopeptides Human genes 0.000 description 1
- 238000012408 PCR amplification Methods 0.000 description 1
- 241000235342 Saccharomycetes Species 0.000 description 1
- XUIMIQQOPSSXEZ-UHFFFAOYSA-N Silicon Chemical compound [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 238000005273 aeration Methods 0.000 description 1
- 238000000246 agarose gel electrophoresis Methods 0.000 description 1
- 235000021120 animal protein Nutrition 0.000 description 1
- 230000003110 anti-inflammatory effect Effects 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 235000006708 antioxidants Nutrition 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 239000011324 bead Substances 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 235000013339 cereals Nutrition 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 239000013530 defoamer Substances 0.000 description 1
- 230000000593 degrading effect Effects 0.000 description 1
- 238000013400 design of experiment Methods 0.000 description 1
- 239000012470 diluted sample Substances 0.000 description 1
- 230000002183 duodenal effect Effects 0.000 description 1
- 238000005265 energy consumption Methods 0.000 description 1
- 238000011049 filling Methods 0.000 description 1
- 239000000796 flavoring agent Substances 0.000 description 1
- 235000019634 flavors Nutrition 0.000 description 1
- 235000013312 flour Nutrition 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 210000003405 ileum Anatomy 0.000 description 1
- 230000036737 immune function Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- GOMNOOKGLZYEJT-UHFFFAOYSA-N isoflavone Chemical compound C=1OC2=CC=CC=C2C(=O)C=1C1=CC=CC=C1 GOMNOOKGLZYEJT-UHFFFAOYSA-N 0.000 description 1
- CJWQYWQDLBZGPD-UHFFFAOYSA-N isoflavone Natural products C1=C(OC)C(OC)=CC(OC)=C1C1=COC2=C(C=CC(C)(C)O3)C3=C(OC)C=C2C1=O CJWQYWQDLBZGPD-UHFFFAOYSA-N 0.000 description 1
- 235000008696 isoflavones Nutrition 0.000 description 1
- 210000001630 jejunum Anatomy 0.000 description 1
- 229920002521 macromolecule Polymers 0.000 description 1
- 235000012054 meals Nutrition 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 239000002068 microbial inoculum Substances 0.000 description 1
- 239000011707 mineral Substances 0.000 description 1
- 235000010755 mineral Nutrition 0.000 description 1
- 239000006872 mrs medium Substances 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- 230000014075 nitrogen utilization Effects 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 229920001542 oligosaccharide Polymers 0.000 description 1
- 150000002482 oligosaccharides Chemical class 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- 238000004806 packaging method and process Methods 0.000 description 1
- 239000012286 potassium permanganate Substances 0.000 description 1
- 239000006041 probiotic Substances 0.000 description 1
- 235000018291 probiotics Nutrition 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 238000007873 sieving Methods 0.000 description 1
- 229910052710 silicon Inorganic materials 0.000 description 1
- 239000010703 silicon Substances 0.000 description 1
- 210000000813 small intestine Anatomy 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 239000008223 sterile water Substances 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 210000002784 stomach Anatomy 0.000 description 1
- 230000002195 synergetic effect Effects 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
Classifications
-
- 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
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/12—Animal feeding-stuffs obtained by microbiological or biochemical processes by fermentation of natural products, e.g. of vegetable material, animal waste material or biomass
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
- A23K10/37—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms from waste material
-
- 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
- A23K50/30—Feeding-stuffs specially adapted for particular animals for swines
-
- 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
- A23K50/60—Feeding-stuffs specially adapted for particular animals for weanlings
-
- 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
- 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
- C12R2001/46—Streptococcus ; Enterococcus; Lactococcus
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
- Y02P60/80—Food processing, e.g. use of renewable energies or variable speed drives in handling, conveying or stacking
- Y02P60/87—Re-use of by-products of food processing for fodder production
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Polymers & Plastics (AREA)
- Zoology (AREA)
- Health & Medical Sciences (AREA)
- Biotechnology (AREA)
- Food Science & Technology (AREA)
- Animal Husbandry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Organic Chemistry (AREA)
- Genetics & Genomics (AREA)
- Wood Science & Technology (AREA)
- Biochemistry (AREA)
- Biomedical Technology (AREA)
- Microbiology (AREA)
- Physiology (AREA)
- General Health & Medical Sciences (AREA)
- Molecular Biology (AREA)
- General Engineering & Computer Science (AREA)
- Virology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Medicinal Chemistry (AREA)
- Birds (AREA)
- Sustainable Development (AREA)
- Botany (AREA)
- Mycology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
本发明公开一株产蛋白酶的粪肠球菌及应用该粪肠球菌的生物发酵饲料的制作方法,涉及功能性微生物筛选及发酵酶解豆粕技术领域。所述的粪肠球菌于2022年6月6日保藏于中国普通微生物菌种保藏管理中心(China General Microbiological Culture Collection Center,CGMCC),保藏编号为CGMCC NO.25007。本发明采用驯化筛选的方法进行功能性微生物的选育,得到了一株产蛋白酶能力强并且生物量好、代谢产物活性好的粪肠球菌。该菌株能用于豆粕发酵,能极大程度地降解大分子抗原蛋白,改善豆粕的抗营养特性,产生丰富的功能小肽,提高豆粕的蛋白质消化利用率,提高饲料的转化率,从而在无抗时代下发挥降本增效的价值。
Description
技术领域
本发明涉及微生物及发酵工程领域,具体涉及一株产蛋白酶的粪肠球菌及其应用。
背景技术
玉米、豆粕等粮食是我国主要的动物蛋白质饲料之一,特别是豆粕,其具有蛋白质含量高(43%-48%),氨基酸含量更为均衡(2.5%-2.8%)以及赖氨酸含量特别丰富等优点,可以均衡家禽和猪的营养。豆粕中抗营养因子主要包括胰蛋白酶抑制因子、大豆抗原蛋白、寡糖、植酸、凝集素等,抗营养因子可降低机体对氮的利用率,降低维生素的效价,造成肠道菌群失调,影响机体正常代谢,从而降低豆粕的饲用价值。目前,利用微生物发酵能充分发挥豆粕的营养价值,微生物发酵豆粕可使抗原蛋白大豆球蛋白和β-伴大豆球蛋白含量降低67%、62%,基本消除了豆粕中的胰蛋白酶抑制因子、植酸、凝集素。微生物发酵过程中产生大量益生菌、寡肽、乳酸、维生素、矿物质元素等物质,抑制肠道有害菌繁殖,降低中性洗涤纤维和酸性洗涤纤维含量,促进动物对营养物质的吸收,提高饲料利用率。同时微生物发酵豆粕显著提高大豆异黄酮含量,增强抗氧化活性,提高动物抗氧化能力,增强机体免疫。Feng等[1]用24.5%的发酵豆粕饲喂仔猪发现,发酵豆粕组可显著升高小肠各肠段的绒毛高度,增加十二指肠隐窝深度,增加仔猪空肠和回肠中黏蛋白和紧密连接蛋白基因的表达,说明发酵豆粕可以调节肠道相关基因的表达来增加肠道的免疫功能,减少断奶应激,提高经济价值。
乳酸菌是人和动物消化道内栖居的重要菌群,在维持肠道菌群平衡中起重要作用。用乳酸菌发酵豆粕会使豆粕中的蛋白质水解,增加游离氨基酸的释放,由此产生的发酵豆粕中总的游离氨基酸含量显著高于豆粕。同时采用乳酸菌发酵后的豆粕能改善饲料风味,提高发酵饲料适口性,从而有助于提高采食量。同时,乳酸菌发酵的豆粕中有着大量乳酸菌及其代谢产物,能够提高机体特异性免疫水平,有抗炎作用。本发明获得的粪肠球菌CGMCC NO.25007是经过梯度选育后的高产蛋白酶的乳酸菌,除具有上述乳酸菌发酵豆粕的优势以外,还可产生大量高活性的蛋白酶,对豆粕中大分子的抗营养因子的降解起到重要作用。本发明通过粪肠球菌所得的发酵豆粕对仔猪断奶应激具有明显的缓解作用,提高仔猪采食量和日增重。
目前发酵豆粕广泛用于无抗饲料的生产中,其主要生产工艺为菌酶协同发酵、芽孢杆菌好氧发酵以及芽孢杆菌好氧发酵与乳酸菌、酵母菌厌氧发酵相结合发酵工艺,工艺能耗高,好氧发酵对豆粕中的能量物质消耗大,生产损耗高。本发明通过选育得到一株高产蛋白酶乳酸菌,对豆粕中大分子蛋白降解效率高,本发明的菌株可以和酿酒酵母协同,在微氧环境下对豆粕进行有效发酵,通过简单混合,装袋密封,静态发酵3-7d,发酵产物含有丰富的有机酸、小肽。豆粕中的抗营养因子也能得到有效的降解,从而提高豆粕的营养价值。
发明内容
针对现有技术存在的不足,本发明要解决的技术问题是采用菌种驯化法选育高产蛋白酶活性的粪肠球菌,并用于豆粕发酵及豆粕资源的高值化利用。
为解决上述技术问题,本发明提供以下技术方案:
一方面,本发明提供一株产蛋白酶的粪肠球菌(Enterococcus faecalisstrain),于2022年6月6日保藏于中国微生物菌种保藏管理委员会普通微生物中心(CGMCC),保藏单位地址为北京市朝阳路1号院3号,保藏编号为CGMCC NO.25007,分类命名为:粪肠球菌Enterococcus faecalis。
另一方面,本发明提供了上述粪肠球菌在豆粕及含豆粕饲料发酵中的应用。
所述饲料发酵方法为:
S101.将粪肠球菌按2-5%的比例接种于发酵培养基,并培养获得发酵菌液;
S102.将发酵菌液与发酵基质按重量比(1.5-2):(4.5-5)混匀,得到混合发酵基质;
S103.向混合发酵基质中添加酿酒酵母菌,调整混合发酵基质的含水量为35%-45%,并发酵,获得目标发酵饲料。
进一步地,所述S101中,发酵菌液基于厌氧条件下培养。
进一步地,所述发酵菌液中活菌数为2.0*108-1.0*1010cfu/g。
进一步地,所述混合发酵基质中菌落总数为5.0*106-1.0*108cfu/g。
进一步地,所述发酵菌液与发酵基质重量比为1.6:4.5。
进一步地,所述发酵培养基为MRS肉汤培养基;所述发酵基质包括70-90重量份的豆粕、2-5重量份的糖和10-30重量份的麦麸,所述糖具体为含糖物质,可选为红糖、白糖、黑糖、黄糖。
进一步地,所述酿酒酵母添加终浓度为5*106-1.0*107cfu/g发酵基质。
进一步地,所述S103中,混合发酵基质的初始含水量为35%-40%,发酵方式为厌氧固态发酵。
本发明相对于现有技术,具有以下有益效果:
1、本发明实施例采用菌种驯化筛选的方法,获得一株产蛋白酶并且生物量好、代谢产物活性好的粪肠球菌(Enterococcus faecalis strain)EF-JF-10,可提高饲料中的生物量;
2、利用该菌株的产酶特性能够用于豆粕及含豆粕饲料中发酵,能有效降解大豆球蛋白和β-伴大豆球蛋白,提高豆粕的利用率,改善了豆粕的营养成分;
3、经实验证明,应用粪肠球菌的饲料可有效缓解仔猪的断奶应激,提高断奶仔猪采食量和日增重,对豆粕的高值化利用具有重要意义。
附图说明
图1为本发明实施例粪肠球菌EF-JF-10在MRS平板上的菌落形态;
图2为本发明实施例粪肠球菌菌落革兰氏染色镜检图;
图3为本发明实施例蛋白酶筛选培养基平板;
图3中:A为粪肠球菌EF-JF-10水解透明圈,B和C为重复样;D、E和F为对照菌株的水解透明圈;
图4为SDS-聚丙烯酰胺凝胶电泳测定豆粕中抗原蛋白的降解效果图
图4中:A、B均为重复样品,为豆粕原料(未发酵);C、D为重复样,为固体发酵豆粕中大分子抗原蛋白降解情况;E、F为两个重复样品,是将10%豆粕液体发酵后大分子抗原蛋白降解情况。
具体实施方式
下面结合具体实施例,对本发明作进一步详细的阐述,下述实施例不用于限制本发明,仅用于说明本发明。以下实施例中所使用的实验方法如无特殊说明,实施例中未注明具体条件的实验方法,通常按照常规条件,下述实施例中所使用的材料、试剂等,如无特殊说明,均可从商业途径得到。
本发明实施例的粪肠球菌(Enterococcus faecalis strain),于2022年6月6日保藏于中国微生物菌种保藏管理委员会普通微生物中心(CGMCC),保藏编号为CGMCCNO.25007,分类命名为:粪肠球菌Enterococcus faecalis。在实际使用时,发现本发明实施例的粪肠球菌能有效降解豆粕的大豆球蛋白(11S)和β-伴大豆球蛋白(7S),提高豆粕的适口性。
本发明实施例的粪肠球菌应用于发酵饲料的制备过程,具体为:将上述的粪肠球菌按2-5%的比例接种于发酵培养基,于37℃条件下培养48h-60h,得到发酵菌液;将发酵菌液与发酵基质按重量比(1.5-2):(4.5-5)混匀,得到混合发酵基质;向混合发酵基质中添加酿酒酵母菌,调整混合发酵基质的含水量为35%-45%,于37℃条件下发酵3d-7d。
优选地,所述的发酵菌液培养温度为37℃,培养条件为厌氧培养,培养时间48h-60h。
更优选地,所述的发酵菌液活菌数为2.0*108-1.0*1010cfu/g。
优选地,所述的混合发酵基质中菌落总数为1.0*107-1.0*108cfu/g。
优选地,所述的发酵菌液与发酵基质重量比为1.6:4.5。
优选地,所述的发酵培养基为MRS肉汤培养基。
优选地,所述的发酵基质包括80%豆粕、5%玉米和15%麦麸。
优选地,所述的酿酒酵母添加终浓度为25-30g/t。
优选地,所述的混合发酵基质的初始含水量为40%-45%,发酵温度为37℃,发酵方式为厌氧固态发酵,发酵时间为4d-5d。
现提供实施例1-4来具体说明本发明实施例的应用过程,。
实施例1蛋白酶粪肠球菌的筛选和驯化
1、实验材料
菌种源:农家无抗养殖家禽粪便
富集培养基:含粉碎豆粕(过80目筛)100g,葡萄糖20g,加水至1000mL,充分搅拌混匀,pH 6.0-6.5,在110℃下灭菌20min制得;
初筛培养基:含牛肉膏3g,蛋白胨10g,氯化钠5g,琼脂15g,脱脂牛奶15g,加水至1000mL,充分搅拌混匀,,培养基在110℃下灭菌15min;
MRS培养基(种子培养基):由10g蛋白胨、5g牛肉粉、4g酵母粉、2g葡萄糖、1ml吐温80、2g磷酸氢二钾、5g乙酸钠、2g柠檬酸三铵、0.2g硫酸镁、0.05g硫酸锰、1000ml蒸馏水组成。
斜面保藏培养基:由10g蛋白胨、5g牛肉粉、4g酵母粉、2g葡萄糖、1ml吐温80、2g磷酸氢二钾、5g乙酸钠、2g柠檬酸三铵、0.2g硫酸镁、0.05g硫酸锰、15g琼脂粉、1000ml蒸馏水组成。
基础发酵培养基:由豆粕40g,玉米粉30g,麸皮30g,KH2PO4 0.3g,Na2HPO4-12H2O4g,加水至1000mL,充分搅拌混匀,培养基在110℃下灭菌15min.
2、菌种筛选
富集过程:将采集的10g样品加入到装有玻璃珠的无菌水100mL/250mL的锥形瓶中,放在180rpm、37℃摇床培养30min,将样品混合均匀。取1mL混合液加入到装有100mL的500mL锥形瓶,37℃静态培养48h。
初筛过程:取富集后的菌液用十倍稀释法梯度稀释至不同倍数(10-1-10-8),取10-5、10-6、10-7和10-8五个稀释梯度,将稀释的样品各吸取100uL菌液涂布于蛋白酶初筛培养基平板上,每个梯度做三个平行,平板在37℃恒温厌氧培养24h后有明显的蛋白水解圈出现(见图3),用接种环挑取有明显水解圈的菌株划线挑取单菌落,用游标卡尺测量水解圈和菌落直径,选取水解圈直径与菌落直径比值(H/C)较大的菌株作为初筛的产蛋白酶菌株,将筛选的菌株保藏在试管斜面上,并在4℃冰箱保存。
复筛:将初筛中水解圈直径与菌落比值较大的5个单菌落接种到装有100mL种子培养基的250mL三角瓶中,在摇床转速为100rpm,温度为37℃条件下培养18h。将种子培养基按4%的接种量添加到装有200mL基础发酵培养基的500mL三角瓶中,在摇床转速100rpm,温度为37℃条件下发酵48h后,离心取上清,测定蛋白酶活力,筛选出蛋白酶活力最高的菌株。
3、菌种鉴定
采用菌落形态学观察及革兰氏染色:用接种环挑取产蛋白酶活力最高的菌株在初筛琼脂平板上划线,观察单菌落的形状、颜色和大小等特征,用革兰氏试剂盒(购买于上海生工)进行革兰氏染色,镜检。
16s rDNA测定:以固体平板上的单菌落为模板,采用细菌16s rDNA通用引物进行PCR扩增,引物由上海生工合成。将PCR扩增产物经琼脂糖凝胶电泳检测后送湖南擎科生物有限公司测序。
5、实验结果
本发明实施例1通过多次富集后筛选分离获得一株高产蛋白酶菌株,摇瓶发酵上清蛋白酶活力为72U/mL,其菌落形态图和在显微镜下菌体形态见图1、图2所示。将筛选到的此菌株接种于脱脂牛奶琼脂平板,具有明显的蛋白质水解圈如图3所示。该菌株16S rDNA序列通过在genbank数据库中进行序列比对(具体序列参见序列表),与粪肠球菌Enterococcus faecalis strain H4相似性最高,相似度99.86%,结合其生理生化特性确定本发明实施例1菌株为粪肠球菌,命名为:EF-JF-10,具体序列见下表。本发明实施例1所得粪肠球菌已经保藏至中国微生物菌种保藏中心(CGMCC),并获得保藏编号为CGMCCNO.25007。
实施例2乳酸菌液体发酵豆粕的制备
实施步骤:
S1 10%豆粕溶液的制备:称取100g粉碎(过80目筛)的豆粕粉,重悬于900g自来水中,加入2g葡萄糖,摇匀后110℃灭菌20min.
S2粪肠球菌种子液制备:取从粪肠球菌EF-JF-10斜面上挑取单菌落接种至灭菌MRS肉汤培养基中置于摇床培养16h,培养条件37℃、100r/min;
S3液态发酵豆粕制备:将粪肠球菌种子液种子液以4%的接种量接种到10%豆粕溶液中,37℃,100rpm摇床发酵48小时后检测小肽含量以及抗原蛋白降解情况。
实验结果:
豆粕经本发明的粪肠球菌液体发酵,发酵后的溶液中小肽含量丰富,占总蛋白的40%以上。豆粕中抗原蛋白主要由β-球蛋白(7S)和大豆球蛋白(11S)组成,7S蛋白是由三个亚基α(约67kD)、α’(约71kD)、β(约50kD)组成,11S蛋白是由酸性亚基(约40kDa)和碱性亚基(约20kDa)组成。采用SDS-聚丙烯酰胺凝胶电泳法测定发酵豆粕溶液中抗原蛋白的降解情况,结果显示发酵后,超过85%的7S和11S蛋白被降解成20000道尔顿以下的小肽分子。如图4中E和F所示。
实施例3发酵豆粕的制备
本发明实施例3的发酵豆粕是通过添加发酵菌剂对豆粕进行固态发酵,具体的发酵菌剂微生物包括粪肠球菌EF-JF-10和酿酒酵母菌(本发明实施例购自安琪酵母公司)。
S1:粪肠球菌EF-JF-10菌液制备:
(1)种子液制备:取纯化的粪肠球菌EF-JF-10斜面接种至灭菌MRS肉汤培养基中置于摇床培养16h,培养条件37℃、100rpm;
(2)液体发酵:发酵罐中加入MRS液体发酵培养基和0.05%有机硅消泡剂灭菌,将培养好的种子液按体积比2%的接种量接入发酵罐培养24h即可,发酵条件:罐压0.01Mpa、通气量0L/min、转速125rpm、温度37℃;
S2:豆粕固态发酵料制备
将S1的EF-JF-10粪肠球菌菌液与豆粕固态发酵基质按重量比(1.5-2):(4.5-5)混匀(固态发酵基质包括90%豆粕、2%红糖和8%麦麸),本发明实施例采用EF-JF-10粪肠球菌菌液与豆粕固态发酵基质按照1.5:5重量比混合,得到混合发酵基质;向混合发酵基质中添加酿酒酵母菌(20g/t),调整混合发酵基质的含水量为40%,将混合发酵基质装入带有呼吸阀的发酵袋中,每包10kg,排空空气,于37℃条件下密封发酵5d,发酵终止后取样检测水分,pH,乳酸菌活菌数,粗蛋白含量,酸溶蛋白含量以及总酸等指标。
对照组的为添加菌剂的豆粕样品。将干净无菌自来水与豆粕固态发酵基质按重量比(1.5-2):(4.5-5)混匀(固态发酵基质包括90%豆粕、2%红糖和8%麦麸),按照S2进行打包,发酵,取样检测。
实验结果如下表所示:
实施例4发酵豆粕对断奶仔猪的应用效果
1、实验设计
CK:基础日粮
T1:基础日粮+6%豆粕(未发酵)
T2:基础日粮+6%发酵豆粕
试验选定126头初始体重为5.66±0.52kg的断奶仔猪(日龄30d左右),按体重一致原则分为3组,每组6栏,每栏7头,试验为期9天。其营养配比和基础日粮饲喂量参照中华人民共和国农业行业标准猪饲养标准《NY/T65-2004》执行。
2、实验方法
试验开始前对猪舍及所有饲养用具进行清洗、熏蒸消毒(福尔马林/高锰酸钾为1:15),通风晾晒一周后进行使用。CK、T1和T2分栏饲养,猪群饲养在同栋舍内,地面平养。自由采食,自由饮水。温度控制在28±2℃之内,湿度控制在65±5%之间。保持舍内通风干燥,卫生清洁。
3、评定指标
生长性能包括:初重、末重、日增重、日采食、料肉比、腹泻率。
试验第l天和最后一天清晨空腹称重、计算耗料量、平均日增重(average dailygain,ADG)、平均日采食量(average daily feed intake,ADFI)及料重比(feed/gain,F/G)。
腹泻率=(腹泻只数×腹泻天数)/(试验只数×试验天数)×100%
外观指标:毛色评分。
表1皮毛外观评分标准
标准 | 评分 |
很好 | 5 |
较好 | 4 |
普通(一般) | 3 |
差 | 2 |
很差 | 1 |
4、实验结果
实验结果如下表所示,为不同原料的教槽料饲喂断奶仔猪的生长性能。
从表2可知,本发明实施例的饲料具有如下优点:
1、T2能有效提高断奶仔猪生长性能,使用本发明实施例的产品后能显著提高断奶仔猪日增重,降低料肉比,显著减少腹泻率;
2、采用本发明实施例饲喂断奶仔猪,日增重达285g左右,较CK提高了5.95%,较T1提高了5.17%,提升效果较为明显。
3、在防治腹泻率方面,饲喂本发明实施例的断奶仔猪腹泻率低,腹泻率1.87%,较CK降低63.55%,较T1降低54.61%。
4、本发明实施例T2组的料肉比较CK组,减低了4.03%,断奶仔猪的毛色也得到了改善,说明本发明实施例可以提高断奶仔猪日粮的适口性,对断奶应激具有明显的缓解效果。
综上所述,本发明的目的是利用粪肠球菌的产蛋白酶特性发酵降解豆粕中的抗营养因子,提高豆粕在断奶仔猪中的应用效果,添加的微生物仅为乳酸菌和酿酒酵母菌,无需添加芽孢杆菌,此外,本发明实施例发酵豆粕制备发酵方式为厌氧固态发酵,此方式发酵可以充分发挥粪肠球菌的特性,同时更好的保留发酵料的营养成分,区别于现有技术的好氧发酵。
最后应当说明的是,以上实施例仅用以说明本发明的技术方案而非对其限制,尽管参照上述实施例对本发明进行了详细的说明,所属领域的普通技术人员应当理解,技术人员阅读本申请说明书后依然可以对本发明的具体实施方式进行修改或者等同替换,但这些修改或变更均未脱离本发明申请待批权利要求保护范围之内。
Claims (10)
1.一株产蛋白酶的粪肠球菌,其特征在于,所述粪肠球菌的保藏编号为CGMCCNO.25007。
2.一种饲料发酵方法,所述饲料发酵方法中添加权利要求1所述的粪肠球菌。
3.根据权利要求2所述饲料发酵方法,其特征在于,所述饲料发酵方法为:
S101.将粪肠球菌按2-5%的比例接种于发酵培养基,并培养获得发酵菌液;
S102.将发酵菌液与发酵基质按重量比(1.5-2):(4.5-5)混匀,得到混合发酵基质;
S103.向混合发酵基质中添加酿酒酵母菌,调整混合发酵基质的含水量为35%-45%,并发酵3-7d,获得目标发酵饲料。
4.根据权利要求3所述饲料发酵方法,其特征在于,所述S101中,发酵菌液基于厌氧条件下培养。
5.根据权利要求4所述饲料发酵方法,其特征在于,所述发酵菌液中活菌数为2.0*108-1.0*1010cfu/g。
6.根据权利要求3所述饲料发酵方法,其特征在于,所述混合发酵基质中菌落总数为5.0*106-1.0*108cfu/g。
7.根据权利要求3所述饲料发酵方法,其特征在于,所述发酵菌液与发酵基质重量比为1.6:4.5。
8.根据权利要求3所述饲料发酵方法,其特征在于,所述发酵培养基为MRS肉汤培养基;所述发酵基质包括70-90重量份的豆粕、2-5重量份的糖和10-30重量份的麦麸,所述糖具体为含糖物质,可选为红糖、白糖、黑糖、黄糖。
9.根据权利要求3所述饲料发酵方法,其特征在于,所述酿酒酵母添加终浓度为5*106-1.0*107cfu/g发酵基质。
10.根据权利要求3所述饲料发酵方法,其特征在于,所述S103中,混合发酵基质的初始含水量为35%-40%,发酵方式为厌氧固态发酵。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202310036365.9A CN116555071A (zh) | 2023-01-09 | 2023-01-09 | 一株产蛋白酶的粪肠球菌及应用该粪肠球菌的饲料发酵方法 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202310036365.9A CN116555071A (zh) | 2023-01-09 | 2023-01-09 | 一株产蛋白酶的粪肠球菌及应用该粪肠球菌的饲料发酵方法 |
Publications (1)
Publication Number | Publication Date |
---|---|
CN116555071A true CN116555071A (zh) | 2023-08-08 |
Family
ID=87493541
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202310036365.9A Pending CN116555071A (zh) | 2023-01-09 | 2023-01-09 | 一株产蛋白酶的粪肠球菌及应用该粪肠球菌的饲料发酵方法 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN116555071A (zh) |
-
2023
- 2023-01-09 CN CN202310036365.9A patent/CN116555071A/zh active Pending
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN111363698B (zh) | 一种降低发酵饲料霉变及霉菌毒素危害的菌剂与应用 | |
CN106085881B (zh) | 一种黑曲霉菌株及其应用 | |
CN111436526A (zh) | 改善育肥猪生长性能菌酶协同发酵米糠粕制备方法及应用 | |
CN102334611A (zh) | 米糠基质纳豆芽孢杆菌与酵母菌复合菌剂的固态发酵方法 | |
CN107853478B (zh) | 一种降低牛甲烷排放的含酶中草药发酵饲料添加剂 | |
CN111700157A (zh) | 一种提高水产动物免疫力的益生菌饲料添加剂 | |
CN113736716B (zh) | 一株副干酪乳杆菌及黄贮复合生物发酵剂和黄贮方法 | |
CN116083262A (zh) | 有水产病原菌拮抗特性的植物乳杆菌菌株及其制剂的制备及应用 | |
CN109757603A (zh) | 一种发酵大豆饼粕制备猪饲料的方法 | |
CN103525724A (zh) | 一种棉粕微生物发酵剂及其制备方法 | |
CN112544787A (zh) | 菌酶协同发酵构树复合饲料的方法及其复合饲料 | |
CN114540232B (zh) | 有水产病原菌拮抗特性的鼠李糖乳杆菌及其制剂的制备及应用 | |
CN112980740B (zh) | 一种地衣芽孢杆菌及其应用 | |
CN114304379A (zh) | 一种含有复合微生物菌剂的发酵饲料的制备方法 | |
CN112961806B (zh) | 高产乳酸的凝结芽孢杆菌、生物发酵饲料及其制备方法和应用 | |
CN109423466B (zh) | 一种复合发酵菌剂及其应用 | |
CN106721278A (zh) | 微生物发酵保育期仔猪液态饲料及其制备方法与应用 | |
CN114190486B (zh) | 一种预防和治疗母猪便秘的微生物发酵饲料及母猪养殖方法 | |
CN114886008A (zh) | 一种生物发酵富硒饲料及其制备方法 | |
CN110140810B (zh) | 饲用发酵黄酒糟的制备方法及其发酵黄酒糟 | |
CN111642639B (zh) | 一种乳仔猪生物发酵饲料 | |
CN115927032A (zh) | 枯草芽孢杆菌、发酵剂及制备方法和它们的应用 | |
CN116555071A (zh) | 一株产蛋白酶的粪肠球菌及应用该粪肠球菌的饲料发酵方法 | |
KR101470995B1 (ko) | 프로피온산 생산능을 갖는 미생물 및 그를 포함하는 조사료 조성물 | |
CN107897505B (zh) | 发酵菌液、包含其的用于保育猪的产品及产品的制备方法和应用 |
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 |