CN115612634B - 一株分离的黑氏乳杆菌及其发酵的昆虫蛋白及应用 - Google Patents
一株分离的黑氏乳杆菌及其发酵的昆虫蛋白及应用 Download PDFInfo
- Publication number
- CN115612634B CN115612634B CN202210661357.9A CN202210661357A CN115612634B CN 115612634 B CN115612634 B CN 115612634B CN 202210661357 A CN202210661357 A CN 202210661357A CN 115612634 B CN115612634 B CN 115612634B
- Authority
- CN
- China
- Prior art keywords
- lactobacillus
- feed
- fermented
- niger
- strain
- 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.)
- Active
Links
- 241000186660 Lactobacillus Species 0.000 title claims abstract description 44
- 229940039696 lactobacillus Drugs 0.000 title claims abstract description 44
- 108010060231 Insect Proteins Proteins 0.000 title claims abstract description 29
- 241000894006 Bacteria Species 0.000 claims abstract description 16
- 241000238557 Decapoda Species 0.000 claims description 9
- 241001125889 Micropterus salmoides Species 0.000 claims description 7
- 241000193830 Bacillus <bacterium> Species 0.000 claims description 4
- 241001465754 Metazoa Species 0.000 claims description 4
- 238000002360 preparation method Methods 0.000 claims description 4
- 241000186781 Listeria Species 0.000 claims 1
- 241000607598 Vibrio Species 0.000 claims 1
- 238000000855 fermentation Methods 0.000 abstract description 20
- 230000000844 anti-bacterial effect Effects 0.000 abstract description 15
- 241000588724 Escherichia coli Species 0.000 abstract description 9
- 244000063299 Bacillus subtilis Species 0.000 abstract description 7
- 235000014469 Bacillus subtilis Nutrition 0.000 abstract description 7
- 241000191967 Staphylococcus aureus Species 0.000 abstract description 7
- 241000186779 Listeria monocytogenes Species 0.000 abstract description 5
- 241000589776 Pseudomonas putida Species 0.000 abstract description 5
- 241000607272 Vibrio parahaemolyticus Species 0.000 abstract description 5
- 238000009360 aquaculture Methods 0.000 abstract description 4
- 244000144974 aquaculture Species 0.000 abstract description 4
- 244000052616 bacterial pathogen Species 0.000 abstract description 4
- 150000007524 organic acids Chemical class 0.000 abstract description 4
- 239000002253 acid Substances 0.000 abstract description 3
- 238000003860 storage Methods 0.000 abstract description 2
- 230000007774 longterm Effects 0.000 abstract 1
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 22
- 230000004151 fermentation Effects 0.000 description 19
- 241000238631 Hexapoda Species 0.000 description 17
- 238000012360 testing method Methods 0.000 description 13
- 150000001413 amino acids Chemical class 0.000 description 11
- 239000004310 lactic acid Substances 0.000 description 11
- 235000014655 lactic acid Nutrition 0.000 description 11
- 108090000623 proteins and genes Proteins 0.000 description 10
- 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 9
- 241000111368 Lactobacillus hammesii Species 0.000 description 9
- 239000008103 glucose Substances 0.000 description 9
- 239000007788 liquid Substances 0.000 description 8
- 239000000047 product Substances 0.000 description 8
- 239000006228 supernatant Substances 0.000 description 8
- 235000018102 proteins Nutrition 0.000 description 7
- 102000004169 proteins and genes Human genes 0.000 description 7
- 241000709785 Hermetia illucens Species 0.000 description 6
- 230000029087 digestion Effects 0.000 description 6
- 238000004519 manufacturing process Methods 0.000 description 6
- 230000000694 effects Effects 0.000 description 5
- 239000001963 growth medium Substances 0.000 description 5
- 239000000126 substance Substances 0.000 description 5
- JVTAAEKCZFNVCJ-REOHCLBHSA-N L-lactic acid Chemical compound C[C@H](O)C(O)=O JVTAAEKCZFNVCJ-REOHCLBHSA-N 0.000 description 4
- 230000001580 bacterial effect Effects 0.000 description 4
- 150000001875 compounds Chemical class 0.000 description 4
- 230000005764 inhibitory process Effects 0.000 description 4
- 239000002609 medium Substances 0.000 description 4
- 229920001817 Agar Polymers 0.000 description 3
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 3
- 235000019733 Fish meal Nutrition 0.000 description 3
- 241000186605 Lactobacillus paracasei Species 0.000 description 3
- 239000008272 agar Substances 0.000 description 3
- 238000004458 analytical method Methods 0.000 description 3
- 230000009286 beneficial effect Effects 0.000 description 3
- 239000002131 composite material Substances 0.000 description 3
- 238000012258 culturing Methods 0.000 description 3
- 238000011161 development Methods 0.000 description 3
- 238000009792 diffusion process Methods 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 239000004467 fishmeal Substances 0.000 description 3
- 239000000796 flavoring agent Substances 0.000 description 3
- 235000019634 flavors Nutrition 0.000 description 3
- 244000005700 microbiome Species 0.000 description 3
- 238000004321 preservation Methods 0.000 description 3
- 238000012216 screening Methods 0.000 description 3
- 239000007787 solid Substances 0.000 description 3
- 230000004584 weight gain Effects 0.000 description 3
- 235000019786 weight gain Nutrition 0.000 description 3
- 239000007222 ypd medium Substances 0.000 description 3
- KSEBMYQBYZTDHS-HWKANZROSA-M (E)-Ferulic acid Natural products COC1=CC(\C=C\C([O-])=O)=CC=C1O KSEBMYQBYZTDHS-HWKANZROSA-M 0.000 description 2
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 2
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 2
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 2
- 239000004472 Lysine Substances 0.000 description 2
- 239000001888 Peptone Substances 0.000 description 2
- 108010080698 Peptones Proteins 0.000 description 2
- 241000235342 Saccharomycetes Species 0.000 description 2
- 241001052560 Thallis Species 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- 239000013543 active substance Substances 0.000 description 2
- 235000021120 animal protein Nutrition 0.000 description 2
- 235000015278 beef Nutrition 0.000 description 2
- 238000009395 breeding Methods 0.000 description 2
- 230000001488 breeding effect Effects 0.000 description 2
- 229940041514 candida albicans extract Drugs 0.000 description 2
- BOLQJTPHPSDZHR-UHFFFAOYSA-N dihydroferulic acid Chemical compound COC1=CC(CCC(O)=O)=CC=C1O BOLQJTPHPSDZHR-UHFFFAOYSA-N 0.000 description 2
- KSEBMYQBYZTDHS-HWKANZROSA-N ferulic acid Chemical compound COC1=CC(\C=C\C(O)=O)=CC=C1O KSEBMYQBYZTDHS-HWKANZROSA-N 0.000 description 2
- 229940114124 ferulic acid Drugs 0.000 description 2
- KSEBMYQBYZTDHS-UHFFFAOYSA-N ferulic acid Natural products COC1=CC(C=CC(O)=O)=CC=C1O KSEBMYQBYZTDHS-UHFFFAOYSA-N 0.000 description 2
- 235000001785 ferulic acid Nutrition 0.000 description 2
- 210000001035 gastrointestinal tract Anatomy 0.000 description 2
- 238000009630 liquid culture Methods 0.000 description 2
- 210000004185 liver Anatomy 0.000 description 2
- 239000011159 matrix material Substances 0.000 description 2
- 239000002207 metabolite Substances 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 230000008520 organization Effects 0.000 description 2
- 235000019319 peptone Nutrition 0.000 description 2
- 108090000765 processed proteins & peptides Proteins 0.000 description 2
- 238000012163 sequencing technique Methods 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 241000894007 species Species 0.000 description 2
- QURCVMIEKCOAJU-UHFFFAOYSA-N trans-isoferulic acid Natural products COC1=CC=C(C=CC(O)=O)C=C1O QURCVMIEKCOAJU-UHFFFAOYSA-N 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 239000012138 yeast extract Substances 0.000 description 2
- 108020004465 16S ribosomal RNA Proteins 0.000 description 1
- 241000251468 Actinopterygii Species 0.000 description 1
- 239000004475 Arginine Substances 0.000 description 1
- 108010062877 Bacteriocins Proteins 0.000 description 1
- 241000255789 Bombyx mori Species 0.000 description 1
- 241000222178 Candida tropicalis Species 0.000 description 1
- 108010082495 Dietary Plant Proteins Proteins 0.000 description 1
- 241000194032 Enterococcus faecalis Species 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- 244000068988 Glycine max Species 0.000 description 1
- 235000010469 Glycine max Nutrition 0.000 description 1
- FFEARJCKVFRZRR-BYPYZUCNSA-N L-methionine Chemical compound CSCC[C@H](N)C(O)=O FFEARJCKVFRZRR-BYPYZUCNSA-N 0.000 description 1
- 240000001046 Lactobacillus acidophilus Species 0.000 description 1
- 235000013956 Lactobacillus acidophilus Nutrition 0.000 description 1
- 244000199866 Lactobacillus casei Species 0.000 description 1
- 235000013958 Lactobacillus casei Nutrition 0.000 description 1
- 240000006024 Lactobacillus plantarum Species 0.000 description 1
- 235000013965 Lactobacillus plantarum Nutrition 0.000 description 1
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 description 1
- IOVCWXUNBOPUCH-UHFFFAOYSA-M Nitrite anion Chemical compound [O-]N=O IOVCWXUNBOPUCH-UHFFFAOYSA-M 0.000 description 1
- 241000191998 Pediococcus acidilactici Species 0.000 description 1
- 241000269799 Perca fluviatilis Species 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 description 1
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 241000254109 Tenebrio molitor Species 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 229910021529 ammonia Inorganic materials 0.000 description 1
- 239000003674 animal food additive Substances 0.000 description 1
- 230000008485 antagonism Effects 0.000 description 1
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 description 1
- 239000001569 carbon dioxide Substances 0.000 description 1
- 229910002092 carbon dioxide Inorganic materials 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 239000006071 cream Substances 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- 230000009615 deamination Effects 0.000 description 1
- 238000006481 deamination reaction Methods 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- YXVFQADLFFNVDS-UHFFFAOYSA-N diammonium citrate Chemical compound [NH4+].[NH4+].[O-]C(=O)CC(O)(C(=O)O)CC([O-])=O YXVFQADLFFNVDS-UHFFFAOYSA-N 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 229940032049 enterococcus faecalis Drugs 0.000 description 1
- 239000003797 essential amino acid Substances 0.000 description 1
- 235000020776 essential amino acid Nutrition 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- XJRPTMORGOIMMI-UHFFFAOYSA-N ethyl 2-amino-4-(trifluoromethyl)-1,3-thiazole-5-carboxylate Chemical compound CCOC(=O)C=1SC(N)=NC=1C(F)(F)F XJRPTMORGOIMMI-UHFFFAOYSA-N 0.000 description 1
- 210000001723 extracellular space Anatomy 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 210000003495 flagella Anatomy 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- 210000003736 gastrointestinal content Anatomy 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 229940039695 lactobacillus acidophilus Drugs 0.000 description 1
- 229940017800 lactobacillus casei Drugs 0.000 description 1
- 229940072205 lactobacillus plantarum Drugs 0.000 description 1
- 239000008101 lactose Substances 0.000 description 1
- 229920002521 macromolecule Polymers 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- 229930182817 methionine Natural products 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 238000000386 microscopy Methods 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 239000006872 mrs medium Substances 0.000 description 1
- 235000019629 palatability Nutrition 0.000 description 1
- 235000011837 pasties Nutrition 0.000 description 1
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 description 1
- 239000003910 polypeptide antibiotic agent Substances 0.000 description 1
- 229920000053 polysorbate 80 Polymers 0.000 description 1
- 235000015277 pork Nutrition 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 239000006041 probiotic Substances 0.000 description 1
- 230000000529 probiotic effect Effects 0.000 description 1
- 235000018291 probiotics Nutrition 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 229940081969 saccharomyces cerevisiae Drugs 0.000 description 1
- 235000021108 sauerkraut Nutrition 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000001632 sodium acetate Substances 0.000 description 1
- 235000017281 sodium acetate Nutrition 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 239000002699 waste material Substances 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
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
- A01N63/20—Bacteria; Substances produced thereby or obtained therefrom
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01P—BIOCIDAL, PEST REPELLANT, PEST ATTRACTANT OR PLANT GROWTH REGULATORY ACTIVITY OF CHEMICAL COMPOUNDS OR PREPARATIONS
- A01P1/00—Disinfectants; Antimicrobial compounds or mixtures thereof
-
- 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
- A23K10/20—Animal feeding-stuffs from material of animal origin
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/163—Sugars; Polysaccharides
-
- 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
- A23K50/80—Feeding-stuffs specially adapted for particular animals for aquatic animals, e.g. fish, crustaceans or molluscs
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L3/00—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs
- A23L3/34—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals
- A23L3/3454—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals in the form of liquids or solids
- A23L3/3463—Organic compounds; Microorganisms; Enzymes
- A23L3/3571—Microorganisms; Enzymes
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/40—Preparation of oxygen-containing organic compounds containing a carboxyl group including Peroxycarboxylic acids
- C12P7/56—Lactic acid
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/225—Lactobacillus
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Polymers & Plastics (AREA)
- Health & Medical Sciences (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Wood Science & Technology (AREA)
- Food Science & Technology (AREA)
- Organic Chemistry (AREA)
- Animal Husbandry (AREA)
- General Health & Medical Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Genetics & Genomics (AREA)
- Biomedical Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Chemical & Material Sciences (AREA)
- Biochemistry (AREA)
- Molecular Biology (AREA)
- General Engineering & Computer Science (AREA)
- Environmental Sciences (AREA)
- Plant Pathology (AREA)
- Virology (AREA)
- Pest Control & Pesticides (AREA)
- Physiology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Medicinal Chemistry (AREA)
- Sustainable Development (AREA)
- Nutrition Science (AREA)
- Agronomy & Crop Science (AREA)
- Birds (AREA)
- Marine Sciences & Fisheries (AREA)
- Dentistry (AREA)
- Insects & Arthropods (AREA)
- Fodder In General (AREA)
Abstract
本发明属于生物饲料技术领域，公开了一株分离的黑氏乳杆菌及其发酵的昆虫蛋白及应用，所述乳杆菌为Lactobacillus hammesiiTC38，保藏编号为：CCTCC NO：M2022723。该菌株具有较强的产酸能力，同时对大肠杆菌、金黄色葡萄球菌、单核增生李斯特菌、副溶血弧菌、枯草芽孢杆菌、恶臭假单胞菌等致病菌或腐败菌具有较强的抑菌活性。利用该菌株进行昆虫蛋白的厌氧发酵，产品富含有机酸，便于长期贮存，在水产养殖中具有较好的应用前景。
Description
技术领域
本发明属于生物饲料技术领域，具体涉及一株分离的黑氏乳杆菌及其发酵的昆虫蛋白及 应用。
背景技术
乳酸菌种类繁多，近年来不断有新种被报道。Lactobacillus hammesii最早在2005年被 Valcheva分离于法国酸面团中并被鉴定命名，在东北发酵酸菜中也被发现。有研究报道黑氏 乳杆菌和植物乳杆菌共发酵可释放结合的阿魏酸，并能将生成的游离阿魏酸转化为二氢阿魏 酸和挥发性代谢产物。
昆虫是地球上最大的生物类群，全世界现存昆虫可能超过1000万种，目前已定名的昆 虫约100万种，遍布所有的生态系统，具有种类多、周期短、繁殖快、易于饲养且可利用废 弃物进行大规模饲养等优点，其蛋白质含量高，可达干重的50％-82％，是一类巨大的生物 资源，被认为是目前最大且最具开发应用潜力的动物蛋白源。
昆虫蛋白是一种高质量的动物蛋白质资源，富含各种氨基酸，氨基酸种类和含量比例符 合联合国粮农组织/世界卫生组织(FAO/WHO)提出的氨基酸模式。昆虫中蛋白质含量与鱼 粉相当，优于猪肉、大豆以及牛肉。常见昆虫的蛋白质含量如黑水虻幼虫40％－44％，黄粉 虫47.70％－64.70％，蚕蛹60％左右，蝇蛆为40％-65％，并且其赖氨酸、蛋氨酸和必需氨基 酸总量分别是鱼粉的2.6、2.7倍和2.3倍，氨基酸质量接近鱼粉。另外，昆虫氨基酸的消 化利用率非常高，可达70％-98.93％，接近或超过肉、鱼的消化率，高于植物性蛋白的消化 利用率。
在昆虫蛋白饲料的应用中发现，不同加工方式的生产成本和应用效果有差异，热处理会 增加生产成本，降低活性物质含量，而冻虫或鲜虫不利于保存。因此，对昆虫蛋白饲料产品 的应用开发还需不断延伸和拓展，如对昆虫蛋白进行酶解、发酵，将大分子物质转化成小肽、 氨基酸等小分子物质，以利于水产动物消化利用，是一种高效创新的产品形式。
其中，在发酵昆虫蛋白饲料中应用较多的微生物包括芽孢杆菌、酵母菌、乳酸菌等。这 些菌株的联合应用能将昆虫蛋白质迅速降解，并在发酵过程中产生有机酸、细菌素、过氧化 氢等物质，并具有广泛的抗菌作用。有机酸的产生还能够有效延长昆虫蛋白饲料的保质期并 改善其口感。基于这些特点，添加复合微生物菌株对昆虫饲料进行发酵，在发酵过程中可产 生乳酸、高级醇和酯类等香味物质，改善饲料的风味，同时使其具有良好的诱食性且发酵后 昆虫饲料中的抗菌肽等活性物质仍具有较高的抗菌活性。可极大的的改善其保质期短、风味 较差等问题。
但上述复合微生物菌剂的主要问题是上述复合菌株组合没有抑制腐败细菌的能力，同时 芽孢杆菌容易过度发酵将蛋白质脱氨基释放出氨气产生异味，酵母菌容易产气影响产品储 存，因此筛选适宜发酵昆虫蛋白并具备抑菌和益生性能的乳酸菌对产品开发具有重要意义。
发明内容
本发明的目的是提供一株具有抑菌特性且适用于昆虫发酵的菌株，该菌株为Lactobacillus hammesiiTC38，保藏编号为：CCTCC NO：M2022723。
本发明的另一个目的在于提供了Lactobacillus hammesiiTC38在制备发酵昆虫蛋白饲料 中的应用。
本发明的最后一个目的在于提供了发酵昆虫蛋白饲料在水产养殖中的应用。
为了达到上述目的，本发明采取以下技术措施：
申请人自鲈鱼肠道中分离得到一株乳酸菌，经16S rDNA基因序列鉴定为Lactobacillus hammesii，该菌株已于2022年5月26日送至中国典型培养物保藏中心保藏，分类命名： Lactobacillus hammesiiTC38，保藏编号为：CCTCC NO：M2022723，地址：中国武汉武汉 大学。
所述Lactobacillus hammesiiTC38为革兰氏阳性菌，杆状，不产芽孢。在MRS固体培养 基上菌落较小、圆形、表面光滑湿润、边缘整齐、淡乳白色，菌落直径约2mm。菌株经简单 染色后在显微镜下观察，形状呈杆状，单个、成对生或短链状，无鞭毛。利用葡萄糖同型乳 酸发酵产L-乳酸；最适生长温度为37℃。
本发明提供的Lactobacillus hammesiiTC38发酵液具有同时对大肠杆菌、金黄色葡萄球 菌、单核增生李斯特菌、副溶血弧菌、枯草芽孢杆菌、恶臭假单胞菌等致病菌或腐败菌具有 较强的抑菌活性。
一种发酵昆虫蛋白饲料，采用新鲜昆虫匀浆，添加葡萄糖，经LactobacillushammesiiTC38 发酵而成。
以上所述的发酵昆虫蛋白饲料，优选的，将新鲜蝇蛆匀浆，加入2％葡萄糖(质量比)， 接种5％Lactobacillus hammesii种子液(质量比)，在发酵罐中37℃发酵48小时，pH为3.8， 乳酸含量2％(质量比)。
当用于水产养殖时，优选动物为河蟹和加州鲈。
与现有技术相比，本发明具有以下优点：
1.本发明的Lactobacillus hammesiiTC38对同时对大肠杆菌、金黄色葡萄球菌、单核增生 李斯特菌、枯草芽孢杆菌、恶臭假单胞菌等致病菌或腐败菌具有较强的抑菌活性和产酸性能， 能延长发酵昆虫蛋白饲料的保质期。
2.本发明的发酵昆虫蛋白饲料在水产养殖中的应用效果良好，在河蟹和加州鲈养殖中可 显著提高产量。
3.本发明的Lactobacillus hammesiiTC38为同型乳酸发酵菌株，能利用糖类发酵产生乳 酸，同时不产生二氧化碳，也不会导致蛋白质脱氨基释放出氨气。
具体实施方式
本发明所述技术方案，如未特别说明，均为本领域的常规方案；所述试剂或材料，如未 特别说明，均来源于商业渠道。
实施例1：
黑氏乳杆菌Lactobacillus hammesiiTC38菌株的筛选和鉴定：
培养基：
BCP培养基:蛋白胨5g；酵母膏3g；乳糖5g；琼脂20g；0.5％溴甲酚紫10ml；蒸馏 水1000ml；pH值6.8～7.0。
MRS液体培养基：葡萄糖20g/L；蛋白胨10g/L；酵母提取物5g/L；牛肉浸粉10g/L；柠檬酸二铵2g/L；醋酸钠5g/L；吐温80 1g/L；磷酸氢二钾2g/L。
固体培养基在液体培养基的基础上添加1.5％的琼脂，所有培养基115℃灭菌20min。
取鲈鱼肠道内容物10g置于100ml生理盐水中，150rpm振荡15min后取1ml培养液梯度稀释，涂布BCP平板，37℃培养48h后挑选菌落周边变黄的单菌落划线分离培养，共 挑选89个单菌落。将得到的单菌落转接到MRS液体培养基中，37℃培养48h，MRS斜面 保藏，同时用琼脂点扩散交叉拮抗实验初筛得到10株对大肠杆菌K88和金黄色葡萄球菌抑 菌作用较好的菌株。之后对这10株菌进行培养，分别进行牛津杯定量扩散抑制大肠杆菌K88 试验，通过抑菌圈大小比较抑菌效果，得到一株发酵液抑制大肠杆菌K88效果较好的乳酸 菌TC38。
菌株TC38生化鉴定：用结晶紫对菌株F153进行简单染色，并通过光学显微镜进行观 察，该菌株为革兰氏阳性杆菌，无芽孢。在MRS固体培养基上长成圆形、表面光滑湿润、边缘整齐、淡乳白色的较小菌落，直径约2mm。该菌株利用葡萄糖同型乳酸发酵产L-乳酸；最适生长温度为37℃。
菌株的16SrRNA基因测序分析：挑取TC38单菌落用通用引物进行扩增，对扩增产物进行16SrRNA基因测序分析。结果表明该序列与Lactobacillus hammesii相似性最高，达100％， 结合菌落形态可确定该菌株为Lactobacillus hammesii，命名为LactobacillushammesiiTC38。 该菌株已于2022年5月26日送至中国典型培养物保藏中心保藏，分类命名：Lactobacillus hammesiiTC38，保藏编号为：CCTCC NO：M2022723，地址：中国武汉武汉大学。
实施例2：
Lactobacillus hammesiiTC38的抑菌实验：
取在MRS液体培养基中培养24h的Lactobacillus hammesiiTC38发酵液50ml，10000 rpm离心10min后分离菌体，菌体重悬2次后加10ml水进行超声破碎，分别用发酵液上清 和破碎后的菌液进行牛津杯定量扩散抑菌试验(牛津杯外径7.5mm)。用乳酸调节MRS培养基的pH值使其与乳酸菌上清液pH值相同，作为对照(CK)。抑菌对象为大肠杆菌(Escherichia Coli)K88、金黄色葡萄球菌(Staphylococcus aureus)ATCC 27217、单增李斯 特菌(Listeria monocytogenes)、副溶血弧菌(Vibrio parahaemolyticus)、枯草芽孢杆菌(Bacillus subtilis)、恶臭假单胞菌(Pseudomonas putida)。
表1：Lactobacillus hammesiiTC38的抑菌试验结果
通过比较表1中上清液、菌体破碎液和CK对各指示菌的抑菌圈直径大小，可知Lactobacillus hammesiiTC38的发酵液上清和菌体破碎菌液均对大肠杆菌、金黄色葡萄球菌、 副溶血弧菌、单增李斯特菌及枯草芽孢杆菌都有抑菌作用，且发酵液上清的抑菌效果明显高 于细胞破碎液的抑菌效果；仅发酵液上清对乳酸片球菌有较明显抑制作用；发酵液上清和破 碎菌液均对干酪乳杆菌、粪肠球菌、嗜酸乳杆菌、酿酒酵母和热带假丝酵母抑菌能力较弱。
上述结果表明Lactobacillus hammesiiTC38分泌到胞外的代谢产物中存在抑菌活性较强 的物质，故菌体破碎液抑菌圈直径大于上清液抑菌圈直径。
实施例3：
蝇蛆为基质生产发酵昆虫蛋白饲料的方法，方法如下：
实验组：
1)取新鲜蝇蛆匀浆，加入2％葡萄糖(质量比)。
2)添加5％(质量比)Lactobacillus hammesiiTC38的YPD培养基发酵液。
3)在发酵罐中，在37℃条件下恒温培养48h。
4)检测有效活菌数量为Lactobacillus hammesii65×108CFU/g，pH3.8，乳酸含量2.1％(质 量比)，无杂菌。
对照组：
1)取新鲜蝇蛆匀浆，加入2％葡萄糖(质量比)。
2)添加5％(质量比)Lactobacillus paracaseiF204(CCTCC NO：M2020386，公开号： CN111944730B)的YPD培养基发酵液。
3)在发酵罐中，在37℃条件下恒温培养48h。
4)检测有效活菌数量为Lactobacillus paracasei12×108CFU/g，pH4.5，乳酸含量1.2％ (质量比)，杂菌含量较高。
实施例4：
黑水虻幼虫为基质生产发酵昆虫蛋白饲料的方法，方法如下：
实验组：
1)取新鲜黑水虻幼虫匀浆，加入2％葡萄糖(质量比)。
2)添加5％(质量比)Lactobacillus hammesiiTC38的YPD培养基发酵液。
3)在发酵罐中，在37℃条件下恒温培养48h。
4)检测有效活菌数量为Lactobacillus hammesii72×108CFU/g，pH3.9，乳酸含量1.95％ (质量比)，无杂菌。。
对照组：
1)取新鲜黑水虻幼虫匀浆，加入2％葡萄糖(质量比)。
2)添加5％(质量比)Lactobacillus paracaseiF204的YPD培养基发酵液。
3)在发酵罐中，在37℃条件下恒温培养48h。
4)检测有效活菌数量为Lactobacillus paracaseiF204 9.5×108CFU/g，pH4.6，乳酸含量 1.0％(质量比)，杂菌含量较高。
实施例5：
将实施例3、4中的发酵昆虫蛋白饲料进行理化指标分析，结果如表2所示。发酵昆虫 蛋白饲料为膏状，带有比较浓的酸香味。实验组产品中Lactobacillus hammesii菌数达到62 亿/g，大肠杆菌和其他致病菌未检出，说明在发酵过程中乳酸菌益生菌占据优势地位，抑制 了其他有害微生物的生长繁殖。而对照组Lactobacillus paracasei菌数较低，有较多大肠杆 菌检出，乳酸含量相对较低同时pH较高。亚硝酸盐的检测结果表明四组乳酸菌昆虫蛋白培 养物符合国家饲料卫生标准的要求，可以用作饲料原料或者饲料添加剂使用。
表2发酵昆虫蛋白饲料的理化指标
实施例6：
发酵昆虫蛋白饲料在加州鲈中的应用
将实施例3的产品应用于加州鲈(50g)饲喂试验，试验组采取15％的拌料比(海大集 团48蛋白鲈鱼配合饲料)，对照组直接饲喂鲈鱼配合饲料，试验周期22天(湖南省岳阳市君山区采桑湖镇二分厂；试验时间2019年8月21日-9月11日，其中8月29日开始拌料， 拌料14天，共20桶(20kg/桶)；试用塘面积18亩，加州鲈数量11万)。结果表明，Lactobacillushammesii发酵蝇蛆昆虫饲料对加州鲈的生长有明显的促进作用，且饲料的效率明显高于对照塘，可能是因为发酵后改善了饲料的风味，提高了饲料的适口性，且发酵后产生的小肽、氨基酸等更利于加州鲈消化吸收。实验塘中加州鲈的肝脏明显较对照组更健康，说明发酵饲 料的使用可减轻其肝脏的负担。
增重率(WGR)＝(末重-初重)/初重×100％
特定生长率(SGR)＝(ln末重-ln初重)/养殖时间×100％
饲料系数(FCR)＝饲料消耗量/增重量×100％
饲料效率＝1/饲料系数(FCR)×100％
表3加州鲈试验塘和对照塘各项指标对比
实施例7：
发酵昆虫蛋白饲料在河蟹中的应用
使用实施例4发酵黑水虻昆虫饲料对河蟹(公蟹70g，母蟹50g)进行饲喂试验，设置试验塘和对照塘，试验塘添加比例15％(海大集团36蛋白河蟹配合饲料)，加水量为饲料重量的1/5，将饲料混匀后静置半小时开始投喂，对照塘直接饲喂河蟹配合饲料，试验周期23天(孝感汉川市湾潭乡；试验时间2019年8月15日-9月7日，共投喂35桶(20kg/桶)， 试用塘面积35亩，螃蟹数量45000)。结果表明，饲喂Lactobacillus hammesii发酵黑水虻蛋 白饲料的试验塘中河蟹的增重率、特定生长率均高于Lactobacillus paracasei组和对照塘，具有明显的促生长作用，且饲料的利用率也显著高于对照塘，说明发酵后产生的小分子蛋白、 氨基酸等更利于河蟹的消化吸收。且发酵后产生的乳酸等有机酸能够明显改善河蟹肠道的健 康，以及产生的精氨酸、赖氨酸等氨基酸可以显著提高蟹膏的积累。
表4河蟹试验塘和对照塘各项指标对比
Claims (4)
1. 一株分离的黑氏乳杆菌（Lactobacillus hammesii）TC38，所述黑氏乳杆菌的保藏编号为：CCTCC NO：M2022723。
2.权利要求1所述的黑氏乳杆菌在制备发酵昆虫蛋白饲料中的应用。
3.权利要求1所述的黑氏乳杆菌在制备水产动物饲料中的应用，所述的水产动物为：河蟹和加州鲈。
4. 权利要求1所述的黑氏乳杆菌在制备细菌抑菌剂中的应用，所述的细菌为：大肠杆菌（Escherichia Coli）、金黄色葡萄球菌（Staphylococcus aureus）、单增李斯特菌（Listeria monocytogenes）、副溶血弧菌（Vibrio parahaemolyticus）、枯草芽孢杆菌（Bacillus subtilis）或恶臭假单胞菌（Pseudomonas putida）。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210661357.9A CN115612634B (zh) | 2022-06-13 | 2022-06-13 | 一株分离的黑氏乳杆菌及其发酵的昆虫蛋白及应用 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210661357.9A CN115612634B (zh) | 2022-06-13 | 2022-06-13 | 一株分离的黑氏乳杆菌及其发酵的昆虫蛋白及应用 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN115612634A CN115612634A (zh) | 2023-01-17 |
| CN115612634B true CN115612634B (zh) | 2023-12-08 |
Family
ID=84856975
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210661357.9A Active CN115612634B (zh) | 2022-06-13 | 2022-06-13 | 一株分离的黑氏乳杆菌及其发酵的昆虫蛋白及应用 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN115612634B (zh) |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20120022291A (ko) * | 2010-09-01 | 2012-03-12 | 한국과학기술원 | 흑운모와 유용미생물을 이용한 발효물을 함유하는 생육촉진용 가축사료 첨가제 |
| CN110741073A (zh) * | 2017-06-09 | 2020-01-31 | 曾根农场股份公司 | 新乳酸菌及其用途 |
-
2022
- 2022-06-13 CN CN202210661357.9A patent/CN115612634B/zh active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20120022291A (ko) * | 2010-09-01 | 2012-03-12 | 한국과학기술원 | 흑운모와 유용미생물을 이용한 발효물을 함유하는 생육촉진용 가축사료 첨가제 |
| CN110741073A (zh) * | 2017-06-09 | 2020-01-31 | 曾根农场股份公司 | 新乳酸菌及其用途 |
Non-Patent Citations (2)
| Title |
|---|
| Lactobacillus hammesii sp. nov., isolated from French sourdough;Rosica Valcheva等;International Journal of Systematic and Evolutionary Microbiology;第55卷(第2期);第763-767页，参见全文 * |
| 传统发面面肥中乳酸菌的分离与鉴定;尹雪等;食品工业科技;第38卷(第14期);第141-145页，参见全文 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN115612634A (zh) | 2023-01-17 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN109251874B (zh) | 一种益生菌制剂及其制备方法和应用 | |
| CN113317413B (zh) | 一种基于黑水虻的水产用发酵饲料加工方法及应用 | |
| CN109897803B (zh) | 一种水产益生菌剂及其制备方法和应用 | |
| CN103820363A (zh) | 一种屎肠球菌菌粉的制备与应用 | |
| WO2023159960A1 (zh) | 一株益生植物乳杆菌及其在低盐发酵肉食品制备中的应用 | |
| KR101349692B1 (ko) | 알코올 내성 유산균 페디오코커스 애시디락티시 및 그 응용 | |
| CN105132321B (zh) | 一种屎肠球菌及其高密度固态发酵的培养基及方法 | |
| CN112980735B (zh) | 丁酸梭菌、菌剂及它们的应用以及菌剂的制备方法 | |
| CN115287202B (zh) | 一株马克斯克鲁维酵母菌及其在制备功能性饲料方面的应用 | |
| CN111593010B (zh) | 一株植物乳杆菌及应用该菌株生产发酵饲料的方法 | |
| CN115074290B (zh) | 一种联产苯乳酸和γ-氨基丁酸的干酪乳杆菌及其应用 | |
| CN111534459A (zh) | 一株高产淀粉酶的发酵乳杆菌及其在制备发酵饲料中的应用 | |
| 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 | |
| CN110804571B (zh) | 一种复合乳酸菌制剂及在制备成饲料添加剂中的应用 | |
| KR100541379B1 (ko) | 사료 첨가물 및 그 제조방법 | |
| 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 | |
| CN111676153B (zh) | 一株植物乳杆菌及其在水产发酵饲料中的应用 | |
| KR20000073915A (ko) | 가축용 복합 미생물 제제 | |
| CN105802871B (zh) | 乳酸菌群及其在秸秆饲料制备中的应用 | |
| CN115612634B (zh) | 一株分离的黑氏乳杆菌及其发酵的昆虫蛋白及应用 | |
| CN114015626A (zh) | 一种具有抑菌功能的类植物乳杆菌及其复合制剂及应用 | |
| CN115572691A (zh) | 一种含植物乳杆菌的水产发酵剂及其制备方法和应用 | |
| RU2658977C1 (ru) | Способ получения белковой кормовой добавки | |
| Kannan et al. | Single cell protein (scp) production from Marine Lactobacillus spp. | |
| CN103966145A (zh) | 一株乳酸乳杆菌及其在发酵产抑菌多肽中的应用 |
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 | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |