CN116970512A - Lactobacillus plantarum, and culture method and application thereof - Google Patents
Lactobacillus plantarum, and culture method and application thereof Download PDFInfo
- Publication number
- CN116970512A CN116970512A CN202310526238.7A CN202310526238A CN116970512A CN 116970512 A CN116970512 A CN 116970512A CN 202310526238 A CN202310526238 A CN 202310526238A CN 116970512 A CN116970512 A CN 116970512A
- Authority
- CN
- China
- Prior art keywords
- lactobacillus plantarum
- liquid
- culturing
- culture
- bacteria
- 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
- 240000006024 Lactobacillus plantarum Species 0.000 title claims abstract description 55
- 235000013965 Lactobacillus plantarum Nutrition 0.000 title claims abstract description 55
- 229940072205 lactobacillus plantarum Drugs 0.000 title claims abstract description 55
- 238000012136 culture method Methods 0.000 title description 8
- 239000006041 probiotic Substances 0.000 claims abstract description 48
- 235000018291 probiotics Nutrition 0.000 claims abstract description 48
- 238000000855 fermentation Methods 0.000 claims abstract description 38
- 230000004151 fermentation Effects 0.000 claims abstract description 38
- 241000894006 Bacteria Species 0.000 claims abstract description 31
- 239000007788 liquid Substances 0.000 claims abstract description 29
- 230000000529 probiotic effect Effects 0.000 claims abstract description 20
- 238000000034 method Methods 0.000 claims abstract description 19
- 238000012258 culturing Methods 0.000 claims abstract description 18
- 238000002360 preparation method Methods 0.000 claims abstract description 14
- 238000005516 engineering process Methods 0.000 claims abstract description 12
- 238000004321 preservation Methods 0.000 claims abstract description 8
- 238000009629 microbiological culture Methods 0.000 claims abstract description 4
- 230000001580 bacterial effect Effects 0.000 claims description 13
- 239000000843 powder Substances 0.000 claims description 13
- 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 claims description 12
- 239000008103 glucose Substances 0.000 claims description 12
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 claims description 12
- 230000000813 microbial effect Effects 0.000 claims description 11
- 238000005406 washing Methods 0.000 claims description 8
- 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 claims description 7
- 239000008101 lactose Substances 0.000 claims description 7
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 claims description 6
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 claims description 6
- 235000015278 beef Nutrition 0.000 claims description 6
- 235000019253 formic acid Nutrition 0.000 claims description 6
- 238000004108 freeze drying Methods 0.000 claims description 6
- PPASLZSBLFJQEF-RXSVEWSESA-M sodium-L-ascorbate Chemical compound [Na+].OC[C@H](O)[C@H]1OC(=O)C(O)=C1[O-] PPASLZSBLFJQEF-RXSVEWSESA-M 0.000 claims description 6
- 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 claims description 6
- 239000001393 triammonium citrate Substances 0.000 claims description 6
- 235000011046 triammonium citrate Nutrition 0.000 claims description 6
- 239000012137 tryptone Substances 0.000 claims description 6
- 229940041514 candida albicans extract Drugs 0.000 claims description 5
- 239000003795 chemical substances by application Substances 0.000 claims description 5
- 229920000136 polysorbate Polymers 0.000 claims description 5
- 239000001632 sodium acetate Substances 0.000 claims description 5
- 235000017281 sodium acetate Nutrition 0.000 claims description 5
- PPASLZSBLFJQEF-RKJRWTFHSA-M sodium ascorbate Substances [Na+].OC[C@@H](O)[C@H]1OC(=O)C(O)=C1[O-] PPASLZSBLFJQEF-RKJRWTFHSA-M 0.000 claims description 5
- 235000010378 sodium ascorbate Nutrition 0.000 claims description 5
- 229960005055 sodium ascorbate Drugs 0.000 claims description 5
- 229960002901 sodium glycerophosphate Drugs 0.000 claims description 5
- REULQIKBNNDNDX-UHFFFAOYSA-M sodium;2,3-dihydroxypropyl hydrogen phosphate Chemical compound [Na+].OCC(O)COP(O)([O-])=O REULQIKBNNDNDX-UHFFFAOYSA-M 0.000 claims description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 5
- 239000012138 yeast extract Substances 0.000 claims description 5
- 239000001888 Peptone Substances 0.000 claims description 4
- 108010080698 Peptones Proteins 0.000 claims description 4
- 235000019319 peptone Nutrition 0.000 claims description 4
- 239000008223 sterile water Substances 0.000 claims description 4
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 claims description 3
- 239000000203 mixture Substances 0.000 claims description 3
- 229910052760 oxygen Inorganic materials 0.000 claims description 3
- 239000001301 oxygen Substances 0.000 claims description 3
- 244000068988 Glycine max Species 0.000 claims description 2
- 235000010469 Glycine max Nutrition 0.000 claims description 2
- 239000004480 active ingredient Substances 0.000 claims description 2
- AIUDWMLXCFRVDR-UHFFFAOYSA-N dimethyl 2-(3-ethyl-3-methylpentyl)propanedioate Chemical compound CCC(C)(CC)CCC(C(=O)OC)C(=O)OC AIUDWMLXCFRVDR-UHFFFAOYSA-N 0.000 claims description 2
- 229940013688 formic acid Drugs 0.000 claims description 2
- 235000001727 glucose Nutrition 0.000 claims description 2
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 claims 1
- 238000009472 formulation Methods 0.000 claims 1
- 230000000968 intestinal effect Effects 0.000 abstract description 6
- 238000012216 screening Methods 0.000 abstract description 6
- 239000002028 Biomass Substances 0.000 abstract description 5
- 206010028980 Neoplasm Diseases 0.000 abstract description 4
- 201000011510 cancer Diseases 0.000 abstract description 4
- 239000000284 extract Substances 0.000 abstract description 2
- 230000002906 microbiologic effect Effects 0.000 abstract description 2
- 239000001963 growth medium Substances 0.000 description 12
- 239000002609 medium Substances 0.000 description 12
- 239000000243 solution Substances 0.000 description 12
- 244000052616 bacterial pathogen Species 0.000 description 7
- 238000002474 experimental method Methods 0.000 description 7
- 239000010410 layer Substances 0.000 description 7
- 238000012360 testing method Methods 0.000 description 7
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 6
- 239000002253 acid Substances 0.000 description 6
- 239000003833 bile salt Substances 0.000 description 5
- 230000005764 inhibitory process Effects 0.000 description 5
- 238000009630 liquid culture Methods 0.000 description 5
- 230000004083 survival effect Effects 0.000 description 5
- 238000010586 diagram Methods 0.000 description 4
- 238000005259 measurement Methods 0.000 description 4
- 238000002156 mixing Methods 0.000 description 4
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 230000021164 cell adhesion Effects 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 230000003647 oxidation Effects 0.000 description 3
- 238000007254 oxidation reaction Methods 0.000 description 3
- 230000001105 regulatory effect Effects 0.000 description 3
- 239000006228 supernatant Substances 0.000 description 3
- 239000000725 suspension Substances 0.000 description 3
- 229920001817 Agar Polymers 0.000 description 2
- GUBGYTABKSRVRQ-XLOQQCSPSA-N Alpha-Lactose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@H](O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-XLOQQCSPSA-N 0.000 description 2
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 2
- 238000003794 Gram staining Methods 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- XUJNEKJLAYXESH-REOHCLBHSA-N L-Cysteine Chemical compound SC[C@H](N)C(O)=O XUJNEKJLAYXESH-REOHCLBHSA-N 0.000 description 2
- 239000008272 agar Substances 0.000 description 2
- 239000002775 capsule Substances 0.000 description 2
- 238000007865 diluting Methods 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- 238000011156 evaluation Methods 0.000 description 2
- 210000003608 fece Anatomy 0.000 description 2
- 238000002955 isolation Methods 0.000 description 2
- -1 mgSO2 Chemical compound 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 239000000523 sample Substances 0.000 description 2
- 238000012163 sequencing technique Methods 0.000 description 2
- HDTRYLNUVZCQOY-UHFFFAOYSA-N α-D-glucopyranosyl-α-D-glucopyranoside Natural products OC1C(O)C(O)C(CO)OC1OC1C(O)C(O)C(O)C(CO)O1 HDTRYLNUVZCQOY-UHFFFAOYSA-N 0.000 description 1
- 108020004465 16S ribosomal RNA Proteins 0.000 description 1
- 241000222122 Candida albicans Species 0.000 description 1
- 238000007400 DNA extraction Methods 0.000 description 1
- 244000017020 Ipomoea batatas Species 0.000 description 1
- 235000002678 Ipomoea batatas Nutrition 0.000 description 1
- 239000004201 L-cysteine Substances 0.000 description 1
- 235000013878 L-cysteine Nutrition 0.000 description 1
- 239000012880 LB liquid culture medium Substances 0.000 description 1
- 241000186660 Lactobacillus Species 0.000 description 1
- 108010019160 Pancreatin Proteins 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 241001354013 Salmonella enterica subsp. enterica serovar Enteritidis Species 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- 241001052560 Thallis Species 0.000 description 1
- HDTRYLNUVZCQOY-WSWWMNSNSA-N Trehalose Natural products O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-WSWWMNSNSA-N 0.000 description 1
- 239000005862 Whey Substances 0.000 description 1
- 102000007544 Whey Proteins Human genes 0.000 description 1
- 108010046377 Whey Proteins Proteins 0.000 description 1
- QMIDDQFZXOSYIL-UHFFFAOYSA-K [Na+].[Na+].[Na+].[O-]P([O-])([O-])=O.OCC(O)CO Chemical compound [Na+].[Na+].[Na+].[O-]P([O-])([O-])=O.OCC(O)CO QMIDDQFZXOSYIL-UHFFFAOYSA-K 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 239000002671 adjuvant Substances 0.000 description 1
- 239000003513 alkali Substances 0.000 description 1
- HDTRYLNUVZCQOY-LIZSDCNHSA-N alpha,alpha-trehalose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-LIZSDCNHSA-N 0.000 description 1
- 230000008485 antagonism Effects 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 229940088710 antibiotic agent Drugs 0.000 description 1
- 239000011324 bead Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 238000007664 blowing Methods 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 239000006285 cell suspension Substances 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 229940099352 cholate Drugs 0.000 description 1
- BHQCQFFYRZLCQQ-OELDTZBJSA-N cholic acid Chemical compound C([C@H]1C[C@H]2O)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(O)=O)C)[C@@]2(C)[C@@H](O)C1 BHQCQFFYRZLCQQ-OELDTZBJSA-N 0.000 description 1
- 239000012228 culture supernatant Substances 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- HHEAADYXPMHMCT-UHFFFAOYSA-N dpph Chemical compound [O-][N+](=O)C1=CC([N+](=O)[O-])=CC([N+]([O-])=O)=C1[N]N(C=1C=CC=CC=1)C1=CC=CC=C1 HHEAADYXPMHMCT-UHFFFAOYSA-N 0.000 description 1
- 230000005284 excitation Effects 0.000 description 1
- 230000002550 fecal effect Effects 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 230000007760 free radical scavenging Effects 0.000 description 1
- 210000004211 gastric acid Anatomy 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 238000000227 grinding Methods 0.000 description 1
- TUJKJAMUKRIRHC-UHFFFAOYSA-N hydroxyl Chemical compound [OH] TUJKJAMUKRIRHC-UHFFFAOYSA-N 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 229940039696 lactobacillus Drugs 0.000 description 1
- 210000002429 large intestine Anatomy 0.000 description 1
- 238000002386 leaching Methods 0.000 description 1
- 239000010871 livestock manure Substances 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 239000011812 mixed powder Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 229940055695 pancreatin Drugs 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 230000001603 reducing effect Effects 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 230000002000 scavenging effect Effects 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000002356 single layer Substances 0.000 description 1
- 235000020183 skimmed milk Nutrition 0.000 description 1
- 210000000813 small intestine Anatomy 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 239000013589 supplement Substances 0.000 description 1
- 230000001502 supplementing effect Effects 0.000 description 1
- 239000002699 waste material Substances 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
- 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
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
-
- 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
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
- C12N11/10—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a carbohydrate
-
- 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
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/225—Lactobacillus
- C12R2001/25—Lactobacillus plantarum
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Biomedical Technology (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- Tropical Medicine & Parasitology (AREA)
- Virology (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Mycology (AREA)
- Nutrition Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses lactobacillus plantarum, a method for culturing lactobacillus plantarum and application thereof, wherein the lactobacillus plantarum sieve is selected from intestinal bacteria of healthy and longevity volunteers in Yuan-village in Fuchun town source county in Shangxi province. The lactobacillus plantarum is lactobacillus plantarum MH-301, the preservation number is CGMCC No.23397, the preservation time is 2021, 09 and 13 days, and the lactobacillus plantarum is preserved in the China general microbiological culture Collection center with the following addresses: beijing, chaoyang area, north Chen Xili No. 1, 3, china academy of sciences, microbiological institute. The invention extracts the probiotics from healthy, cancer-free and long-life population for the first time, and by means of the original high-density liquid fermentation technology, the number of the live bacteria of the probiotic preparation is obviously increased, and compared with the number of the live bacteria of the probiotic obtained by traditional screening, the biomass of the bacteria is improved by at least 2 orders of magnitude.
Description
Technical Field
The invention relates to the technical field of microorganisms, in particular to lactobacillus plantarum, a lactobacillus plantarum culture method and application thereof.
Background
Intestinal flora has evolved for millions of years in combination with human hosts and their ancestors, and has made a significant contribution to the human body in providing nutrition, protecting against pathogens, preventing diseases, etc. At present, intestinal flora of healthy, cancer-free and longevity people especially attracts attention of vast researchers.
Studies indicate that intestinal flora is of greater value in the case of the century old, as it is structurally distinct from the young healthy population.
At present, the biomass of the viable count of probiotics extracted from healthy, cancer-free and longevity population sources is low.
Disclosure of Invention
Based on the above, the invention aims to provide lactobacillus plantarum, a lactobacillus plantarum culture method and application thereof, and aims to solve the problem that the biomass of the viable bacteria number of the intestinal flora in the prior art is low in the background art.
The embodiment of the invention is realized as follows:
lactobacillus plantarum MH-301 is preserved in China general microbiological culture Collection center with a preservation number of CGMCC No.23397.
A method for culturing lactobacillus plantarum MH-301 as described above, comprising:
preparing a high-density fermentation medium, and inoculating activated lactobacillus plantarum MH-301 into the high-density fermentation medium for fermentation culture;
the viable count of the lactobacillus plantarum MH-301 is improved through temperature control and oxygen control and fed-batch fermentation technology, so that high-density bacterial liquid is obtained.
Further, the culture method of the lactobacillus plantarum MH-301 comprises the following steps of:
tryptone, dadaSoy peptone, glucose, beef powder, yeast extract powder, lactose, KHPO 2 ·7H 2 O, sodium glycerophosphate, sodium ascorbate, formic acid, tween and MgSO 2 Sodium acetate, tri-ammonium citrate and MnSO 2 。
Further, the culture method of lactobacillus plantarum MH-301 comprises the following steps of:
0.5-1% tryptone, 0.4-1% soybean peptone, 0.5-15% glucose, 0.3-0.5% beef powder, 0.3-0.5% yeast extract powder, 0.2-0.5% lactose, 0.1-0.2% KHPO 2 7H2O, 1-1.5% sodium glycerophosphate, the balance sodium ascorbate, formic acid, tween, mgSO2, sodium acetate, triammonium citrate and MnSO 2 。
Another object of the present invention is to provide a clinical microbial preparation comprising the above lactobacillus plantarum MH-301 as an active ingredient.
Another object of the present invention is to provide a method for preparing a microbial clinical agent for preparing the above microbial clinical agent, the method comprising:
fermenting and culturing the lactobacillus plantarum MH-301 by a high-density fermentation technology to ensure that the viable bacteria amount in a fermentation liquid reaches at least 1.5X109 CFU/mL;
and freeze-drying by adopting a freeze-drying technology to prepare an active microbial agent with an inner layer embedded layer, and then embedding an outer layer to obtain the microbial clinical preparation.
It is another object of the present invention to provide an edible probiotic, which is obtained by solidifying the lactobacillus plantarum MH-301 described above.
Another object of the present invention is to provide a method for preparing an edible probiotic, for preparing the edible probiotic, the method comprising:
adding the cultured mature lactobacillus plantarum MH-301 seeds into an immobilized carrier solution suitable for growth and propagation of probiotics according to a proportion;
dripping a curing liquid into the immobilized carrier solution; or adding the solidifying liquid into an immobilization carrier to solidify the solidifying liquid;
washing the solidified immobilization carrier containing lactobacillus plantarum MH-301 with sterile water, and then placing the immobilization carrier at a preset temperature for closed culture for a preset time.
Further, the preparation method of the edible probiotics is characterized in that the preset temperature is 10-45 ℃ and the preset time is 8-72 hours.
Compared with the prior art: the invention extracts the probiotics from healthy, cancer-free and longevity crowd for the first time, and obviously increases the viable count of the probiotic preparation (not less than 1.0x10) by means of the high-density liquid fermentation technology 9 CFU/mL), and the number of viable probiotics obtained by traditional screening (1.0x10) 6 CFU/mL～1.0×10 7 CFU/mL) at least 2 orders of magnitude higher cell biomass.
In addition, the invention has at least the following beneficial effects:
1. inoculating activated probiotics strains into a high-density fermentation medium for fermentation culture, and improving the viable count of probiotics by temperature control, oxygen control, fed-batch fermentation technology and the like, so as to obtain high-density bacterial liquid. The waste fermentation liquor is sterilized and diluted with the original fermentation liquor to be used, and the probiotics can be cultured in high density, so that the secondary utilization of the fermentation culture medium is realized;
2. fermenting and culturing intestinal probiotics by high-density fermentation technology to make the viable bacteria amount in fermentation liquid reach at least 1.5X10 9 CFU/mL. Freeze drying to obtain active microbial agent with inner layer embedding layer. And then embedding the outer layer. Effectively increases the stress resistance of the probiotics, prolongs the storage life activity of the probiotics, protects the probiotics from being eroded by gastric acid, ensures the probiotics to safely reach the small intestine and the large intestine and then target the field planting, and plays the biological effect of the probiotics. The probiotics embedded by the multilayer embedding technology have much better stress resistance, stability and survival rate than the probiotics which are not embedded in general.
3. The cultured mature probiotic seeds are added into an immobilized carrier solution containing suitable probiotics for growth and propagation, and then the immobilized carrier solution of the probiotic seeds is dripped into a solidifying liquid, or the solidifying liquid is added into the immobilized carrier to solidify the probiotic seeds. Washing the solidified immobilized carrier containing probiotics with sterile water, and sealing and culturing at 10-45 deg.c for 8-72 hr. The immobilized probiotics produced by the technology do not need to be separated and freeze-dried, can be directly taken as probiotics, and can also be put into other foods; can be stored in a sealed way for a long time at normal temperature, and the bacterial count can still grow continuously.
Drawings
FIG. 1 is a schematic diagram showing the effect of different fermentation pH on viable bacteria count in a method for culturing Lactobacillus plantarum according to an embodiment of the present invention;
FIG. 2 is a schematic diagram showing the effect of different bile salt concentrations on viable bacteria count in a Lactobacillus plantarum culture method according to an embodiment of the present invention;
FIG. 3 is a schematic diagram showing the measurement of the oxidation resistance of an isolated strain in a method for culturing Lactobacillus plantarum according to an embodiment of the present invention;
fig. 4 is a schematic diagram showing the experimental results of the pathogenic bacteria inhibition capability in the lactobacillus plantarum culturing method according to an embodiment of the present invention.
The invention will be further described in the following detailed description in conjunction with the above-described figures.
Detailed Description
The present invention will be described more fully hereinafter in order to facilitate an understanding of the present invention. This invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The terminology used herein in the description of the invention is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. The term "and/or" as used herein includes any and all combinations of one or more of the associated listed items.
Example 1
Acquisition of Lactobacillus plantarum MH-301
1) Isolation of strains
Collecting first manure sample of healthy and long-life volunteers in Dai-Yuan county, fu-Chun-Yuan-Chun-Yuan-village, shang-Yuan-xi province (no antibiotics, non-vegetarian) in the morning, and immediately transferring into a sealed anaerobic plastic operation bag (85% N) 2 ，5％ H 2 ，10％CO 2 ) After one hour of standing, 10g fecal samples were weighed into a centrifuge tube with 10 mL of sterilized glycerol inside, the glycerol content of the samples being 10%, and the centrifuge tube orifice was sealed with a sealer (the above was done in an anaerobic bag). The samples were stored in a 4 ℃ incubator and sent rapidly to the laboratory for further processing.
Aseptically collecting feces 0.5-1.0 g, adding into test tube, adding aseptic glass beads, shaking thoroughly, mixing, diluting at 10 times, and culturing and counting on commercial MRS agar medium at proper concentration. After the colonies grow out, the medium-sized yellow, convex, slightly white, moist and clean-edged colonies with circular shape and diameter of 3.0 mm +/-1 mm are selected as the medium-sized colonies in the plate. And meanwhile, carrying out gram staining on the bacterial colony, wherein the bacterial colony with positive gram staining is lactobacillus.
2) Authentication and preservation
Single-cell genomic DNA was isolated using a DNA extraction kit and 16S rRNA sequencing was performed. The sequencing result is to remove the carrier sequence, and the target sequence can be obtained. And (3) running BLAST program on NCBI, carrying out homology search on cloned target genes in Gene Bank, searching for a 16SrRNA sequence with higher homology, taking sequences with similarity more than 99% as references, and identifying. Finally, lactobacillus plantarum (Lactobacillus plantarum) is obtained through screening and is preserved, the lactobacillus plantarum is named as lactobacillus plantarum MH-301, the preservation number is CGMCC No.23397, the preservation time is 2021, 09 and 13 days, and the lactobacillus plantarum is preserved in the China general microbiological culture Collection center with the preservation address of: beijing, chaoyang area, north Chen Xili No. 1, 3, china academy of sciences, microbiological institute.
Example two
Acid resistance experiment, cholate resistance experiment, determination of oxidation resistance of isolated strain, pathogenic bacteria inhibition capability experiment and isolated strain cell adhesion capability evaluation
Acid resistance experiment flow:
the pH value of MRS liquid culture medium is regulated to 2.0, 3.0, 4.0, 5.0, 6.0 and 7.0 by using 1mol/L hydrochloric acid, the probiotic bacteria liquid cultured by 18 h is respectively connected into the conventional liquid culture medium and the liquid culture medium with the pH value of 2.0, 3.0, 4.0, 5.0, 6.0 and 7.0 according to the proportion of 1 percent, the growth condition of the probiotic bacteria cultured by 3 h is detected, the bacteria liquid is spotted on a flat plate culture medium by adopting a method of double dilution, the viable bacteria number is calculated, each count is repeatedly averaged three times, and the survival rate of each candidate strain is calculated according to a formula. Survival (%) = number of viable bacteria in different pH medium fermentation broth of candidate strain/number of viable bacteria in conventional medium fermentation broth of candidate strain x 100%.
As shown in FIG. 1, the test result shows that the cultured lactobacillus plantarum has good acid resistance and can well cope with the acidic condition in reality.
Bile salt resistance experimental procedure:
the strain with good acid resistance after acid resistance experiment screening is taken to cultivate 18 h, bacterial liquid is inoculated into a conventional liquid culture medium and a liquid culture medium respectively containing 0.1%,0.3% and 0.5% of ox gall salt according to the proportion of 1%, the growth condition of probiotics for culturing 18 h is detected, the probiotics liquid is spotted on a flat plate culture medium by adopting a method of double dilution, the viable count is calculated, the average value is obtained by repeating each count for three times, and the survival rate of the strain is calculated by taking the conventional liquid fermentation liquid of each candidate strain as a control. Survival rate (%) = number of viable bacteria in culture broth of candidate strain with different bile salt concentration/number of viable bacteria in conventional culture broth of candidate strain x 100%.
As shown in figure 2, the test result shows that the cultured lactobacillus plantarum has good bile salt resistance and can well cope with the condition of the concentration of the bile salt in reality.
Measurement of the oxidation resistance of the isolates:
will be divided intoThe bacteria obtained after isolation identification are cultured in corresponding (without L-cysteine) liquid medium for 1-2 d until growth reaches 10 10 And centrifuging to obtain supernatant when CFU/mL is about, and storing at 4 ℃ for later use. For DPPH free radical scavenging ability, scavenging superoxide free radical (O) 2- ) Ability, ability to scavenge hydroxyl radical (HO.) and ability to scavenge Fe 2+ Measurement of chelating ability and reducing Activity. Wherein, the test result is shown in figure 3,
experiment of pathogenic bacteria inhibition ability:
8 common food-borne pathogenic bacteria for testing bacterial antagonism are obtained by acid dough separation and screening, and compriseE.coliO157、S.aureus、S.hemolytic-β、S.castellani、C.albicans、S.enteritidis、S.typhimuriumActivating 7 pathogenic bacteria by using an LB plate culture medium, and performing expansion culture by using an LB liquid culture medium; regulating thallus concentration of 8 strains of pathogenic bacteria to 10 8 About CFU/mL, 200 mu L of bacterial liquid is respectively coated on LB solid culture medium (5% agar), and 3 plates are coated on each pathogenic bacteria strain; after the bacterial liquid is solidified, a sterile oxford cup is placed in each plate, 200 mu L of the original culture supernatant of the probiotics is added into the oxford cup, and the bacteria liquid is cultured at 37 ℃ for 6-8 h. And observing the shape of the inhibition zone, photographing, and simultaneously measuring the diameter of the inhibition zone by using a vernier caliper.
As shown in FIG. 4, the test results show that the cultured Lactobacillus plantarum has good bacteriostasis and can well cope with the bacterial environment in reality.
Cell adhesion ability evaluation of isolates:
digesting the cultured HT29 cells, diluting the cells with DMEM solution to a concentration of 5X 10 5 cell/mL from which 1 mL cell suspension was aspirated in 12 well cell culture plates, 5% CO 2 Cells were grown to a monolayer in an incubator at 37 ℃. Washing three times with sterilized PBS solution, then adding 1 mL of cFDA-SE labeled probiotic suspension, standing for culturing, and washing three times with sterilized solution to remove non-adhering probiotics. Adding 0.7. 0.7 mL pancreatin into each well, stopping digestion after the cells completely fall off from the bottom of the culture plate, adding 0.3 mL DMEM culture solution, collecting bacterial suspension in the well, and measuring probiotics by using a fluorescence spectrophotometerFluorescence intensity of the bacterial suspension. Fluorescent detection conditions: excitation wavelength 492 nm, emission wavelength 517 nm, slit width 2.5 nm. The adhesion rate is calculated as follows: adhesion (%) = adhesion probiotic fluorescence intensity/initial probiotic fluorescence intensity x 100%. The inventor carries out cell adhesion rate measurement of lactobacillus plantarum MH-301, and the adhesion rate reaches 50% +/-14%.
Example III
Lactobacillus plantarum MH-301 high-density culture
The embodiment provides a lactobacillus plantarum MH-301 high-density culture method, which specifically comprises the following steps:
1) Preparation of high Density fermentation Medium
The composition of the high-density medium is: 5-10g/L tryptone, 4-10g/L soyase peptone, 5-15 g/L glucose, 3-5 g/L beef powder, 3-5 g/L yeast extract powder, 2-5 g/L, KHPO lactose 2 ·7H 2 O1-2 g/L, sodium glycerophosphate 10-15 g/L, sodium ascorbate 0.1-0.5 g/L, formic acid 0.1-0.3 g/L, tween 0.5-2.0 g/L, mgSO 2 0.58-0.6 g/L, sodium acetate 1-10 g/L, tri-ammonium citrate 1.0-2.0 g/L and MnSO 2 0.28-0.3 g/L. The raw materials are fixed to 1000mL by distilled water, the pH is regulated to 6.8+/-0.3, and after uniform stirring and dissolution, the raw materials are sterilized for 15-20min under the pressure of 0.08-0.10 MPa, and then cooled.
2) Activated inoculation
Inoculating activated lactobacillus plantarum MH-301 into a high-density fermentation medium for culturing.
3) Fermentation
The culture conditions of the test tube suitable for the thalli in the culture medium are obtained by researching the conditions of temperature, initial pH, shaking table rotating speed and the like: the initial pH is 6.5, the temperature is 36.2 ℃, and the fermentation is kept at rest.
Feeding in batches:
the pH probe of the fermentation tank can monitor the pH change of the fermentation liquid in real time, so that a constant-current alkali supplementing method is adopted, and 15% NaOH solution is dripped, so that the pH of the fermentation liquid is kept at 6.5+/-0.1. The glucose is added according to the actual fermentation condition when the glucose concentration is reduced to 5 g.L -1 The 1 st sugar supplement is started, and 10% sterilized glucose 170 mL should be added for the first time to increase sugar concentration7 g·L -1 . When the glucose concentration is reduced to 5 g.L -1 Glucose is repeatedly added when the concentration of glucose is about 5-7g.L -1 Within the range.
The number of viable bacteria cultured in the high-density fermentation medium prepared by the composition of the medium under different parameters was measured, respectively, as shown in the following table one:
list one
| Example 1 | Example 2 | |
| Tryptone | 8g | 9.2g |
| Soy peptone | 7.5g | 6.2g |
| Glucose | 12.4g | 14.3g |
| Beef powder | 3.9g | 4.2g |
| Yeast leaching powder | 4.2g | 4.8g |
| Lactose and lactose | 3.7g | 2.9g |
| KHPO2·7H2O | 1.4g | 1.45g |
| Glycerol sodium phosphate | 12.8g | 11.3g |
| Ascorbic acid sodium salt | 0.4g | 0.4g |
| Formic acid | 0.2g | 0.1g |
| Tween-type oil | 1.0g | 1.3g |
| MgSO2 | 0.6g | 0.58g |
| Acetic acid sodium salt | 2.5g | 5.5g |
| Triammonium citrate | 1.5g | 1.3g |
| MnSO2 | 0.28-0.3 g/L | 0.28-0.3 g/L |
| Number of viable bacteria | 1.5×109 cfu·mL-1 | 2.1×109 cfu·mL-1 |
As apparent from the first table, the number of viable bacteria of the probiotic preparation is remarkably increased (not less than 1.0X10) 9 CFU/mL), and the number of viable probiotics obtained by traditional screening (1.0x10) 6 CFU/mL～1.0×10 7 CFU/mL) at least 2 orders of magnitude higher cell biomass.
Example IV
The embodiment provides a microbial clinical preparation and a preparation method thereof, and the preparation method specifically comprises the following steps:
1) Enlarged culture of flora
Fermenting Lactobacillus plantarum MH-301 by high density fermentation (such as high density culture in example 1) to obtain a viable count of at least 1.5X10 9 CFU/mL。
2) Freeze drying
Fermenting Shan Zhuyi bacteria with MRS culture medium 36-h-48 h before lyophilizing, and counting viable bacteria to 1.5X10 9 CFU/mL, 3500 r/min centrifugation for 15 min after reaching the corresponding dose, discarding the supernatant, washing with physiological saline, centrifuging and discarding the supernatant. Blowing and mixing with equal volume of skimmed milk, storing at-80deg.C, lyophilizing with a lyophilizing machine after 48 and h, taking out after about 48 and h, grinding, adding adjuvants such as sweet potato powder, mixing, weighing 1 and g mixed powder, counting viable bacteria, calculating dosage according to viable bacteria count, adding porous starch, whey powder and trehalose, mixing to form an embedding layer, embedding, making capsule, randomly selecting several capsules, counting viable bacteria, and storing at low temperature.
Example five
The embodiment provides an edible probiotic and a preparation method thereof, and the preparation method specifically comprises the following steps:
1) Curing
Adding the cultured mature lactobacillus plantarum MH-301 seeds into an immobilized carrier solution containing the bacteria suitable for growth and propagation in proportion, and dripping a curing solution into the immobilized carrier solution; or adding the solidifying liquid into the immobilized carrier to solidify.
2) Solidifying, washing and culturing
Washing the solidified immobilization carrier containing lactobacillus plantarum MH-301 with sterile water, and then placing the immobilization carrier in a sealed culture at 10-45 ℃ for 8-72h.
The foregoing examples illustrate only a few embodiments of the invention and are described in detail herein without thereby limiting the scope of the invention. It should be noted that it will be apparent to those skilled in the art that several variations and modifications can be made without departing from the spirit of the invention, which are all within the scope of the invention. Accordingly, the scope of protection of the present invention is to be determined by the appended claims.
Claims (9)
1. Lactobacillus plantarum MH-301 is preserved in China general microbiological culture Collection center with a preservation number of CGMCC No.23397.
2. A method for culturing lactobacillus plantarum MH-301, for culturing lactobacillus plantarum MH-301 as claimed in claim 1, comprising:
preparing a high-density fermentation medium, and inoculating activated lactobacillus plantarum MH-301 into the high-density fermentation medium for fermentation culture;
the viable count of the lactobacillus plantarum MH-301 is improved through temperature control and oxygen control and fed-batch fermentation technology, so that high-density bacterial liquid is obtained.
3. The method for culturing lactobacillus plantarum MH-301 according to claim 2, wherein the high density fermentation medium comprises:
tryptone, soy peptone, glucose, beef powder, yeast extract, lactose, KHPO 2 ·7H 2 O, sodium glycerophosphate, sodium ascorbate, formic acid, tween and MgSO 2 Sodium acetate, tri-ammonium citrate and MnSO 2 。
4. A method of culturing lactobacillus plantarum MH-301 according to claim 3, characterized in that the high density fermentation medium comprises, in mass percent:
0.5-1% tryptone, 0.4-1% soybean peptone, 0.5-15% glucose, 0.3-0.5% beef powder, 0.3-0.5% yeast extract powder, 0.2-0.5% lactose, 0.1-0.2% KHPO 2 ·7H 2 O, 1-1.5% sodium glycerophosphate, and the balance of sodium ascorbate, formic acid, tween and MgSO 2 Sodium acetate, tri-ammonium citrate and MnSO 2 。
5. A clinical microbial preparation comprising lactobacillus plantarum MH-301 as an active ingredient according to claim 1.
6. A method for preparing a microbial clinical formulation according to claim 5, comprising:
fermenting and culturing Lactobacillus plantarum MH-301 by high density fermentation technology to make the viable bacteria amount in fermentation liquid reach at least 1.5X10 9 CFU/mL；
And freeze-drying by adopting a freeze-drying technology to prepare an active microbial agent with an inner layer embedded layer, and then embedding an outer layer to obtain the microbial clinical preparation.
7. An edible probiotic bacterium, wherein the edible probiotic bacterium is obtained by solidifying lactobacillus plantarum MH-301 according to claim 1.
8. A method of preparing an edible probiotic, for use in preparing an edible probiotic as claimed in claim 7, the method comprising:
adding the cultured mature lactobacillus plantarum MH-301 seeds into an immobilized carrier solution suitable for growth and propagation of probiotics according to a proportion;
dripping a curing liquid into the immobilized carrier solution; or adding the solidifying liquid into an immobilization carrier to solidify the solidifying liquid;
washing the solidified immobilization carrier containing lactobacillus plantarum MH-301 with sterile water, and then placing the immobilization carrier at a preset temperature for closed culture for a preset time.
9. The method of claim 8, wherein the predetermined temperature is 10 ℃ to 45 ℃ and the predetermined time is 8 hours to 72 hours.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310526238.7A CN116970512A (en) | 2023-05-11 | 2023-05-11 | Lactobacillus plantarum, and culture method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310526238.7A CN116970512A (en) | 2023-05-11 | 2023-05-11 | Lactobacillus plantarum, and culture method and application thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN116970512A true CN116970512A (en) | 2023-10-31 |
Family
ID=88473848
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202310526238.7A Pending CN116970512A (en) | 2023-05-11 | 2023-05-11 | Lactobacillus plantarum, and culture method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN116970512A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116694525A (en) * | 2023-06-14 | 2023-09-05 | 南昌大学 | Lactobacillus plantarum and application thereof in preparing bacteria agent for inhibiting or relieving or treating oral mucositis |
-
2023
- 2023-05-11 CN CN202310526238.7A patent/CN116970512A/en active Pending
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116694525A (en) * | 2023-06-14 | 2023-09-05 | 南昌大学 | Lactobacillus plantarum and application thereof in preparing bacteria agent for inhibiting or relieving or treating oral mucositis |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105132318A (en) | Lactobacillus plantarum grx16 and application thereof | |
| CN109929773B (en) | Bifidobacterium capable of being used for selenium-rich culture and active protein and application thereof | |
| CN112143680B (en) | Lactobacillus paracasei ZJUIDS05 with antioxidant effect and application thereof | |
| CN110564638A (en) | Lactobacillus reuteri with probiotic characteristics and application thereof | |
| CN108004155A (en) | Lactobacillus plantarum pc-26 bacterial strains and its application | |
| CN112812999B (en) | Lactobacillus plantarum SLB01 with inhibition effect on enterobacter cloacae and derivative product and application thereof | |
| CN110577912A (en) | lactobacillus gasseri and application thereof in preparing fermented milk | |
| CN114958658B (en) | Lactobacillus rhamnosus A21149 for inhibiting helicobacter pylori and application thereof | |
| CN114480214A (en) | Lactobacillus paracasei separated from Tibet Aliyak milk keloid and application thereof | |
| CN116970512A (en) | Lactobacillus plantarum, and culture method and application thereof | |
| CN108546663B (en) | Porcine lactobacillus crispatus and application thereof | |
| CN104877940B (en) | One plant of streptococcus thermophilus | |
| CN115918909B (en) | Application of bacillus suis in preparation of antioxidant and immunity-improving products | |
| CN109576165B (en) | Saccharomyces bayanus and application thereof | |
| CN112608865A (en) | Lactobacillus rhamnosus with function of reducing triglyceride and application thereof | |
| CN114806944B (en) | Lactobacillus plantarum LP11, fermentation broth thereof, and preparation method and application thereof | |
| CN113088468B (en) | Lactobacillus casei Ma. GLRGJ1 and application thereof | |
| CN113046276B (en) | Breast milk source lactobacillus rhamnosus and application thereof | |
| CN114085789A (en) | Pediococcus pentosaceus MA.WTPQJ01 and application thereof | |
| CN114317366A (en) | Bacterial strain and application thereof | |
| CN114262680A (en) | Bacterial strain and application thereof | |
| CN109161501B (en) | Feeding bacillus licheniformis and application thereof | |
| CN113215067A (en) | VBNC (viable but non-viable) state lactobacillus brevis CSHRR5-3 strain and application thereof | |
| CN113508907A (en) | Application of heat-resistant lactobacillus fermentum in preparation of defecation promoting food or medicine | |
| CN109971667A (en) | One plant of porcine lactobacillus plantarum and application |
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 |