CN107788517A - A kind of method for strengthening ferment effect using lactobacillus bulgaricus - Google Patents
A kind of method for strengthening ferment effect using lactobacillus bulgaricus Download PDFInfo
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- CN107788517A CN107788517A CN201710972121.6A CN201710972121A CN107788517A CN 107788517 A CN107788517 A CN 107788517A CN 201710972121 A CN201710972121 A CN 201710972121A CN 107788517 A CN107788517 A CN 107788517A
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- Prior art keywords
- ferment
- lactobacillus
- gluconacetobacter
- saccharomycete
- bulgaricus
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Links
- 235000013960 Lactobacillus bulgaricus Nutrition 0.000 title claims abstract description 15
- 238000000034 method Methods 0.000 title claims abstract description 14
- 230000000694 effects Effects 0.000 title abstract description 13
- 229940004208 lactobacillus bulgaricus Drugs 0.000 title abstract description 4
- 238000005728 strengthening Methods 0.000 title abstract description 4
- 241000186672 Lactobacillus delbrueckii subsp. bulgaricus Species 0.000 title abstract 2
- 241000186660 Lactobacillus Species 0.000 claims abstract description 43
- 229940039696 lactobacillus Drugs 0.000 claims abstract description 43
- 241000032681 Gluconacetobacter Species 0.000 claims abstract description 26
- 239000000463 material Substances 0.000 claims abstract description 26
- 235000012055 fruits and vegetables Nutrition 0.000 claims abstract description 24
- 241000942810 Saccharomyces cerevisiae N85 Species 0.000 claims abstract description 16
- 238000002360 preparation method Methods 0.000 claims abstract description 5
- 241000894006 Bacteria Species 0.000 claims description 55
- 238000000855 fermentation Methods 0.000 claims description 24
- 230000004151 fermentation Effects 0.000 claims description 21
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims description 19
- 235000013399 edible fruits Nutrition 0.000 claims description 19
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 19
- 238000011081 inoculation Methods 0.000 claims description 18
- 239000002994 raw material Substances 0.000 claims description 18
- 238000003756 stirring Methods 0.000 claims description 18
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 claims description 17
- 235000013311 vegetables Nutrition 0.000 claims description 17
- 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 14
- 239000008103 glucose Substances 0.000 claims description 14
- 244000199885 Lactobacillus bulgaricus Species 0.000 claims description 13
- 241000235342 Saccharomycetes Species 0.000 claims description 13
- 244000157072 Hylocereus undatus Species 0.000 claims description 11
- 235000018481 Hylocereus undatus Nutrition 0.000 claims description 11
- 235000011430 Malus pumila Nutrition 0.000 claims description 11
- 235000015103 Malus silvestris Nutrition 0.000 claims description 11
- 244000068988 Glycine max Species 0.000 claims description 10
- 235000010469 Glycine max Nutrition 0.000 claims description 10
- 235000007688 Lycopersicon esculentum Nutrition 0.000 claims description 10
- 241000220225 Malus Species 0.000 claims description 10
- 240000003768 Solanum lycopersicum Species 0.000 claims description 10
- FRXSZNDVFUDTIR-UHFFFAOYSA-N 6-methoxy-1,2,3,4-tetrahydroquinoline Chemical compound N1CCCC2=CC(OC)=CC=C21 FRXSZNDVFUDTIR-UHFFFAOYSA-N 0.000 claims description 8
- 238000004321 preservation Methods 0.000 claims description 7
- SRBFZHDQGSBBOR-IOVATXLUSA-N D-xylopyranose Chemical compound O[C@@H]1COC(O)[C@H](O)[C@H]1O SRBFZHDQGSBBOR-IOVATXLUSA-N 0.000 claims description 6
- 241000186673 Lactobacillus delbrueckii Species 0.000 claims description 6
- 241000196324 Embryophyta Species 0.000 claims description 5
- 239000000796 flavoring agent Substances 0.000 claims description 5
- 235000019634 flavors Nutrition 0.000 claims description 5
- PYMYPHUHKUWMLA-UHFFFAOYSA-N arabinose Natural products OCC(O)C(O)C(O)C=O PYMYPHUHKUWMLA-UHFFFAOYSA-N 0.000 claims description 4
- SRBFZHDQGSBBOR-UHFFFAOYSA-N beta-D-Pyranose-Lyxose Natural products OC1COC(O)C(O)C1O SRBFZHDQGSBBOR-UHFFFAOYSA-N 0.000 claims description 4
- 239000000052 vinegar Substances 0.000 claims description 4
- 235000021419 vinegar Nutrition 0.000 claims description 4
- 125000000969 xylosyl group Chemical group C1([C@H](O)[C@@H](O)[C@H](O)CO1)* 0.000 claims 2
- 239000012141 concentrate Substances 0.000 claims 1
- 235000013305 food Nutrition 0.000 claims 1
- 230000000813 microbial effect Effects 0.000 claims 1
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 abstract description 27
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 abstract description 23
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 abstract description 11
- 239000004310 lactic acid Substances 0.000 abstract description 11
- 235000014655 lactic acid Nutrition 0.000 abstract description 11
- 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 abstract description 5
- 230000009286 beneficial effect Effects 0.000 abstract description 4
- PGMYKACGEOXYJE-UHFFFAOYSA-N pentyl acetate Chemical compound CCCCCOC(C)=O PGMYKACGEOXYJE-UHFFFAOYSA-N 0.000 abstract description 4
- 235000021107 fermented food Nutrition 0.000 abstract description 2
- 239000008369 fruit flavor Substances 0.000 abstract description 2
- 239000005418 vegetable material Substances 0.000 abstract description 2
- 239000000243 solution Substances 0.000 description 28
- 102000004190 Enzymes Human genes 0.000 description 21
- 108090000790 Enzymes Proteins 0.000 description 21
- 239000007788 liquid Substances 0.000 description 17
- 239000002609 medium Substances 0.000 description 13
- 239000006228 supernatant Substances 0.000 description 13
- 239000001963 growth medium Substances 0.000 description 10
- OUYCCCASQSFEME-QMMMGPOBSA-N L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-QMMMGPOBSA-N 0.000 description 8
- 238000002835 absorbance Methods 0.000 description 8
- OUYCCCASQSFEME-UHFFFAOYSA-N tyrosine Natural products OC(=O)C(N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-UHFFFAOYSA-N 0.000 description 8
- 238000002156 mixing Methods 0.000 description 7
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Natural products CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 6
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 6
- 238000004140 cleaning Methods 0.000 description 6
- 239000012530 fluid Substances 0.000 description 6
- 235000015097 nutrients Nutrition 0.000 description 6
- 239000002504 physiological saline solution Substances 0.000 description 6
- 239000011780 sodium chloride Substances 0.000 description 6
- 239000007787 solid Substances 0.000 description 6
- 108091005804 Peptidases Proteins 0.000 description 5
- 239000004365 Protease Substances 0.000 description 5
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 description 5
- 238000005259 measurement Methods 0.000 description 5
- 150000007524 organic acids Chemical class 0.000 description 5
- 230000003014 reinforcing effect Effects 0.000 description 5
- 239000000523 sample Substances 0.000 description 5
- 238000012360 testing method Methods 0.000 description 5
- 241000193830 Bacillus <bacterium> Species 0.000 description 4
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 description 4
- 235000019441 ethanol Nutrition 0.000 description 4
- 239000000047 product Substances 0.000 description 4
- 244000235858 Acetobacter xylinum Species 0.000 description 3
- 235000002837 Acetobacter xylinum Nutrition 0.000 description 3
- 230000000844 anti-bacterial effect Effects 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- -1 potassium ferricyanide Chemical compound 0.000 description 3
- 235000019991 rice wine Nutrition 0.000 description 3
- 244000063299 Bacillus subtilis Species 0.000 description 2
- 235000014469 Bacillus subtilis Nutrition 0.000 description 2
- 241000588724 Escherichia coli Species 0.000 description 2
- 241000282414 Homo sapiens Species 0.000 description 2
- 230000001580 bacterial effect Effects 0.000 description 2
- 210000004413 cardiac myocyte Anatomy 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 235000009508 confectionery Nutrition 0.000 description 2
- 235000013365 dairy product Nutrition 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 230000001605 fetal effect Effects 0.000 description 2
- 238000009413 insulation Methods 0.000 description 2
- 238000012269 metabolic engineering Methods 0.000 description 2
- 230000003647 oxidation Effects 0.000 description 2
- 238000007254 oxidation reaction Methods 0.000 description 2
- 239000006041 probiotic Substances 0.000 description 2
- 235000018291 probiotics Nutrition 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 229910000029 sodium carbonate Inorganic materials 0.000 description 2
- 238000007711 solidification Methods 0.000 description 2
- 230000008023 solidification Effects 0.000 description 2
- YNJBWRMUSHSURL-UHFFFAOYSA-N trichloroacetic acid Chemical compound OC(=O)C(Cl)(Cl)Cl YNJBWRMUSHSURL-UHFFFAOYSA-N 0.000 description 2
- 150000003673 urethanes Chemical class 0.000 description 2
- 229920001817 Agar Polymers 0.000 description 1
- 108010076119 Caseins Proteins 0.000 description 1
- 235000011511 Diospyros Nutrition 0.000 description 1
- 244000236655 Diospyros kaki Species 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 241000896292 Odontothrips loti Species 0.000 description 1
- 230000002292 Radical scavenging effect Effects 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 229940088710 antibiotic agent Drugs 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- 235000006708 antioxidants Nutrition 0.000 description 1
- 239000012620 biological material Substances 0.000 description 1
- 210000000481 breast Anatomy 0.000 description 1
- 239000005018 casein Substances 0.000 description 1
- 108010079058 casein hydrolysate Proteins 0.000 description 1
- BECPQYXYKAMYBN-UHFFFAOYSA-N casein, tech. Chemical compound NCCCCC(C(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(CC(C)C)N=C(O)C(CCC(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(C(C)O)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(COP(O)(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(N)CC1=CC=CC=C1 BECPQYXYKAMYBN-UHFFFAOYSA-N 0.000 description 1
- 235000021240 caseins Nutrition 0.000 description 1
- 235000013351 cheese Nutrition 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 239000013068 control sample Substances 0.000 description 1
- 230000001186 cumulative effect Effects 0.000 description 1
- 239000008367 deionised water Substances 0.000 description 1
- 229910021641 deionized water Inorganic materials 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- MGJZITXUQXWAKY-UHFFFAOYSA-N diphenyl-(2,4,6-trinitrophenyl)iminoazanium Chemical compound [O-][N+](=O)C1=CC([N+](=O)[O-])=CC([N+]([O-])=O)=C1N=[N+](C=1C=CC=CC=1)C1=CC=CC=C1 MGJZITXUQXWAKY-UHFFFAOYSA-N 0.000 description 1
- 125000005909 ethyl alcohol group Chemical group 0.000 description 1
- 239000000706 filtrate Substances 0.000 description 1
- 239000010200 folin Substances 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 238000002290 gas chromatography-mass spectrometry Methods 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- RBTARNINKXHZNM-UHFFFAOYSA-K iron trichloride Chemical compound Cl[Fe](Cl)Cl RBTARNINKXHZNM-UHFFFAOYSA-K 0.000 description 1
- 239000007791 liquid phase Substances 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 239000011707 mineral Substances 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 238000006396 nitration reaction Methods 0.000 description 1
- 235000005985 organic acids Nutrition 0.000 description 1
- 239000008363 phosphate buffer Substances 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 230000035484 reaction time Effects 0.000 description 1
- 239000012088 reference solution Substances 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 239000012488 sample solution Substances 0.000 description 1
- 229930000044 secondary metabolite Natural products 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 229910021642 ultra pure water Inorganic materials 0.000 description 1
- 239000012498 ultrapure water Substances 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 150000003722 vitamin derivatives Chemical class 0.000 description 1
- 239000003643 water by type Substances 0.000 description 1
- 235000014101 wine Nutrition 0.000 description 1
Classifications
-
- 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
-
- 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
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/123—Bulgaricus
-
- 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)
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Biotechnology (AREA)
- Genetics & Genomics (AREA)
- Food Science & Technology (AREA)
- Nutrition Science (AREA)
- Tropical Medicine & Parasitology (AREA)
- Biomedical Technology (AREA)
- Medicinal Chemistry (AREA)
- Microbiology (AREA)
- Polymers & Plastics (AREA)
- Virology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Mycology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Preparation Of Fruits And Vegetables (AREA)
- Storage Of Fruits Or Vegetables (AREA)
- General Preparation And Processing Of Foods (AREA)
Abstract
The invention discloses a kind of method for strengthening ferment effect using lactobacillus bulgaricus, belong to fermented food technical field.The present invention is by adding saccharomyces cerevisiae N85 into fruit and vegetable materials, gluconacetobacter Q1 and lactobacillus delbruockii subspecies bulgaricus B1, enhance the beneficial function of ferment, the ferment reducing power of preparation is set to improve 43.3% compared to control group, DPPH clearance rates have also brought up to 93.09% from 88.25%, lactic acid content improves 36.1%, and acetic acid content improves 35.2%.Prolease activity improves 11.9% compared to control group, adds the content of volatile materials, and ethyl acetate content improves 85.1%, and pentyl acetate content improves 89.1%, ferment is had special fruit flavor, and improves ferment bacteriostasis.
Description
Technical field
The present invention relates to a kind of method for strengthening ferment effect using lactobacillus bulgaricus, belongs to fermented food technology neck
Domain.
Background technology
Plant enzyme is using plants such as veterinary antibiotics as raw material, through multiple-microorganisms such as saccharomycete, acetic acid bacteria, lactobacillus
A kind of microorganism formulation containing abundant enzyme, vitamin, mineral matter and a variety of secondary metabolites that co-fermentation forms.Plant
Thing ferment is also referred to as ferment.Ferment is different from common enzyme preparation, due to promote to digest, relax bowel, it is anti-oxidant, antibacterial,
Suppress the multiple functions such as tumour, be a kind of health product with very high market value.Existing many natural plant enzymes have
Many restrictive factors, such as easy dye miscellaneous bacteria, mouthfeel or have a poor flavour, fermentation period length, it is difficult to product quality is controlled, by environment shadow
Sound is larger.Therefore, product oxidation resistance is improved, the taste and flavor of ferment is improved, strengthens the ferment preparation side of bacteriostasis
Method, it is more beneficial for the industrialized production of ferment.
The content of the invention
First purpose of the present invention is to provide method that is a kind of while improving pectase function factor and flavor, described
Method is that inoculation yeast bacterium, gluconacetobacter and lactic acid bacteria are fermented in using fruits and vegetables as the ferment of raw material.
In one embodiment of the invention, the saccharomyces cerevisiae is saccharomyces cerevisiae (Saccharomyces
Cerevisiae) N85, gluconacetobacter (Gluconacetobacter xylinus) Q1 and Lactobacillus delbrueckii are sub-
Kind (Lactobacillus delbrueckii subsp.bulgaricus strain) B1, B2.
Second purpose is to provide a kind of probiotics ferment, is inoculation yeast bacterium, glucose vinegar using vegetables and fruit as raw material
Bacillus and lactic acid bacteria co-fermentation are prepared.
In one embodiment of the invention, the probiotics ferment is the inoculation wine brewing using vegetables and fruit as raw material
Yeast, gluconacetobacter and Lactobacillus delbrueckii co-fermentation are prepared.
In one embodiment of the invention, the vegetables and fruit include dragon fruit, apple, tomato and soya bean,
In one embodiment of the invention, the saccharomyces cerevisiae is saccharomyces cerevisiae (Saccharomyces
Cerevisiae) N85, gluconacetobacter are xylose gluconacetobacter (Gluconacetobacter xylinus) Q1, lactobacillus
For lactobacillus delbruockii subspecies bulgaricus (Lactobacillus delbrueckii subsp.bulgaricus strain) B1
Or lactobacillus delbruockii subspecies bulgaricus (Lactobacillus delbrueckii subsp.bulgaricus strain)
B2。
In one embodiment of the invention, the saccharomyces cerevisiae N85 is industrialization bacterial strain, it has been disclosed that was inscribed in 2016
For《The research of metabolic engineering brewing yellow rice wine yeast low yield urethanes》Paper in.
In one embodiment of the invention, the gluconacetobacter Q1 is preserved in China on July 23rd, 2014
Type Tissue Collection, deposit number are CCTCC M 2014353.
In one embodiment of the invention, the lactobacillus delbruockii subspecies bulgaricus B1 is June 26 in 2017
It is preserved in China typical culture collection center day, deposit number is CCTCC M 2017370, and preservation address Wuhan, China is military
Chinese university.
In one embodiment of the invention, the lactobacillus delbruockii subspecies bulgaricus B2 is from Inner Mongol agricultural
Key lab of the dairy products biotechnology Ministry of Education of university lactic acid bacteria culturers resources bank (LABCC), numbering LABCC
IMAU20094 DBTE ML12-2-2。
In one embodiment of the invention, the ferment is prepared according to the following steps：(1) with dragon fruit, apple, western red
Persimmon and soya bean are raw material, add the glucose that material quality percentage is 15~20%, adding material quality percentage is
100% water, is stirred；(2) by volume, 1% saccharomycete and 2% acetic acid bacteria are inoculated with, is placed in 35~37 DEG C
Incubated, stirring in every 20~24 hours once, is fermented to the 3rd~4 day, the lactic acid bacteria of inoculation 3~5%, 28~30 DEG C of constant temperature
Closed culture, ferment and terminated fermentation to the 12nd~15 day.
Third object of the present invention is to provide a kind of preparation method of ferment effect, and methods described comprises the following steps：
(1) using dragon fruit, apple, tomato and soya bean as raw material, the glucose that material quality percentage is 15~20% is added, then add
Enter the water that material quality percentage is 100%, stir；(2) by volume, 1% saccharomycete and 2% vinegar are inoculated with
Sour bacterium, be placed in 35~37 DEG C it is incubated, every 20~24 hours stirring once, ferment to the 3rd~4 day, be inoculated with 5% lactic acid
Bacterium, 30 DEG C of constant-temperature enclosed cultures, ferment and terminated fermentation by the 15th day.
In one embodiment of the invention, the saccharomycete, acetic acid bacteria or the bacterial concentration of lactic acid bacteria are 1~9.9
×107CFU/mL。
In one embodiment of the invention, the saccharomycete is saccharomyces cerevisiae (Saccharomyces
cerevisiae)N85。
In one embodiment of the invention, the acetic acid bacteria is gluconacetobacter (Gluconacetobacter
xylinus)Q1。
In one embodiment of the invention, the lactic acid bacteria is lactobacillus delbruockii subspecies bulgaricus
(Lactobacillus delbrueckii subsp.bulgaricus strain) B1 or lactobacillus delbruockii subspecies bulgaricus
(Lactobacillus delbrueckii subsp.bulgaricus strain)B2。
In one embodiment of the invention, the step is specific as follows：(1) with dragon fruit, apple, tomato and Huang
Beans etc. are raw material, after being cleaned with hot water, are air-dried, and be cut into small pieces shape, several fruits and vegetables of the quality such as precise, and uniformly mixing, puts
Enter in round；(2) glucose that material quality percentage is 12~15% is added, adding fruit quality percentage is
100% water, is stirred；(3) saccharomycete and 2% acetic acid bacteria of addition 1%, 37 DEG C of incubated, holdings are placed in
One-way exhaust, and stirring in 24 hours is once；The concentration of bacterium solution is 107CFU/mL；(4) fermentation added 5% breast by the 4th day
Sour bacterium, 30 DEG C of constant-temperature enclosed cultures；The concentration of bacterium solution is 107CFU/mL；(5) fermentation terminated fermentation by the 15th day.
Beneficial effect：The present invention is by adding saccharomyces cerevisiae (Saccharomyces cerevisiae) into fruit and vegetable materials
N85, gluconacetobacter (Gluconacetobacter) Q1 and lactobacillus delbruockii subspecies bulgaricus (Lactobacillus
Delbrueckii subsp.bulgaricus strain) B1, the beneficial function of ferment is enhanced, reducing power compares control group
Improve 43.3%, DPPH clearance rates and also brought up to 93.09% from 88.25%, lactic acid content improves 36.1%, and acetic acid contains
Amount improves 35.2%.Prolease activity improves 11.9% compared to control group, adds the content of volatile materials, acetic acid
Ethyl ester content improves 85.1%, and pentyl acetate content improves 89.1%, ferment is had special fruit flavor, and improve
Ferment bacteriostasis, can effectively suppress bacillus subtilis, Escherichia coli and Human fetal cardiomyocytes.
Biomaterial preservation
One plant of lactobacillus delbruockii subspecies bulgaricus (Lactobacillus delbrueckii subsp.bulgaricus
Strain) B1 is preserved in China typical culture collection center on June 26th, 2017, and deposit number is CCTCC M
2017370, preservation address Wuhan, China, Wuhan University.
Brief description of the drawings
Fig. 1 is tyrosine concentration standard curve.
Embodiment
In embodiment, yellow rice wine industry is disclosed with saccharomyces cerevisiae (Saccharomyces cerevisiae) N85
It is entitled in 2016《The research of metabolic engineering brewing yellow rice wine yeast low yield urethanes》Paper in.Glucose vinegar
Bacillus (Gluconacetobacter) Q1, it is preserved in China typical culture collection center, preservation on July 23rd, 2014
Numbering is CCTCCM2014353；Lactobacillus delbruockii subspecies bulgaricus (Lactobacillus delbrueckii
Subsp.bulgaricus strain) B1 screened from cheese, has been preserved in Chinese Typical Representative culture guarantor on June 26th, 2017
Tibetan center, deposit number CCTCCM2017370；Lactobacillus delbruockii subspecies bulgaricus (Lactobacillus
Delbrueckii subsp.bulgaricus strain) B2 is in the religion of Agricultural University of the Inner Mongol dairy products biotechnology
Preservation in Yu Bu key labs lactic acid bacteria culturers resources bank (LABCC), deposit number are LABCC IMAU20094 DBTE
ML12-2-2。
The thalline condition of culture that the embodiment of table 1 is related to
Organic acidity test：Ferment enzyme liquid to be measured is taken, after diluting ten times with ultra-pure water, using syringe by liquid assimilating, is led to
After crossing organic acid filter membrane, inject in liquid phase bottle.And calibrated using nitration mixture standard specimen.Use the high performance liquid chromatography of Agilent 1200
Instrument, carry out the measurement of kinds of organic acids and quantity.
DPPH radical scavenging activities determine：Ferment sample is diluted 5 times, takes 2mL samples to be added to 2mL0.1mg/mL's
In DPPH- ethanol solutions, fully mix, 25 DEG C of lucifuges react 30min, using deionized water as reference solution, under the conditions of 517nm
Absorbance is determined, blank group replaces sample with 2mL absolute ethyl alcohols.
DPPH clearance rates (/ %)=[1- (A1-A2)/A0]×100
In formula：A0For the absorbance of blank group, A1For the absorbance of sample solution, A2For the absorbance of sample copy bottom tube.
The measure of reducing power：It is measured using potassium ferricyanide reducing process.Ferment sample to be measured is diluted 5 times, takes 1mL dilute
Release sample to be added in the test tube equipped with 1mL phosphate buffers (0.2mol/L, pH6.6), it is 1% to add 2mL mass fractions
The potassium ferricyanide, 50 DEG C of water-bath accurate response 30min are placed in, add the trichloroacetic acid that 2mL mass fractions are 10%, 3000r/
Min centrifuges 10min, takes 2ml supernatants immediately, adds ferric trichloride and 1mL deionized waters that 1mL mass fractions are 0.5%, quiet
After putting 5min, absorbance is determined under the conditions of 700nm, absorbance is bigger, shows that reducing power is stronger, and same measure repeats two
It is secondary.
The measure of prolease activity：The measure of tyrosine standard curve：Take 6 test tubes to draw 0 respectively, 20,40,60,80,
100ug/mL tyrosine solution 1mL, each sodium carbonate liquor 5ml for adding 0.4mol/L, add and diluted 3 times of forint examination
Agent 1mL.Shake up, be placed in 40 DEG C of water-bath insulation color development 20min, absorbance is determined under the conditions of wavelength 660nm.Measure three
It is secondary, average.Using absorbance as ordinate, tyrosine concentration is abscissa, draws standard curve.1mL is added in test tube
Ferment enzyme liquid, it is placed in 40 DEG C of water-baths and preheats 5 minutes, then adds 1mL2% casein solution, accurate clock reaction
10min, 2mL 0.4mol/L solution of trichloroacetic acid is added immediately, is filtered after 15min with filter paper, is taken 1mL filtrates, add
5mL0.4mol/L sodium carbonate liquor and 1mL Folin reagents, 40 DEG C of insulation color development 20min, it is close to determine light under wavelength 660nm
Angle value.Tyrosine concentration can be converted into according to standard curve.Prolease activity unit definition：At 40 DEG C, under the conditions of pH3.6, often
The enzyme amount that minute caseinhydrolysate discharges 1 microgram tyrosine is defined as 1 protease unit.
In formula：
X is the tyrosine concentration in reaction solution；
V is reaction solution cumulative volume (mL)；
T is the reaction time (min)；
N is enzyme liquid extension rate；
W is enzyme liquid volume (mL).
The measure of bacteriostasis：It is measured using Odontothrips loti.Sterilized agar medium is heated to melt completely
Change, be poured in culture dish, get over 20mL per ware, 4~6 Oxford cups are put in every ware media surface after solidification.Meanwhile it will go out
Semisolid culturemedium (bacterium culture medium) heating of bacterium is melted completely, is dispensed into sterilized 50mL centrifuge tubes, often pipe is about
20mL.Treat that semisolid culturemedium is cooled to 45 DEG C or so, be respectively connected to extension rate as 103、104、105、106Bacterium solution
100uL, after being well mixed, carefully pour into and put well in the culture dish of Oxford cup, to avoid pouring into culture medium in Oxford cup.Wait to train
After supporting base solidification, Oxford cup is carefully taken out, idle loop circular one by one is formed on culture dish, is added in idle loop
100uL measuring samples or control sample, in 37 DEG C of 16~18h of quiescent culture.
Volatile materials determines：It is measured using GC-MS.
Embodiment 1
1. prepared by ferment raw material：After dragon fruit, apple, tomato and soya bean are cleaned with hot water, the work of cleaning is placed in
Typhoon is done, and be cut into small pieces shape；Each 100g of every kind of fruits and vegetables is weighed, uniformly mixing, is put into round, adds vegetables and fruits material quality
The glucose of percentage 15%, the water that vegetables and fruits material quality percentage is 100% is added, is stirred.
2. inoculating strain：
Activated strains：Saccharomyces cerevisiae N85, gluconacetobacter Q1 are taken out from -80 DEG C of refrigerators, streak inoculation is in corresponding respectively
Culture medium.
Obtain thalline：Single bacterium colony on picking solid medium is inoculated into fluid nutrient medium, and culture to logarithm respectively is given birth to
For a long time, 10000rpm centrifuges 10min and abandons supernatant, with sterile saline with physiological saline：Bacterium solution 1:1 ratio is resuspended, and makes bacterium
It is dense up to 1~5 × 107CFU/mL。
Inoculation：By volume, 1mL bacterium solutions/100g fruits and vegetables quality saccharomyces cerevisiae N85 and 2mL bacterium solution/100g fruits and vegetables matter is added
The gluconacetobacter Q1 of amount, 37 DEG C of constant-temperature enclosed fermentations are placed in, stirring in every 24 hours is once.
3. obtain ferment enzyme liquid：Ferment and terminated fermentation by the 15th day, centrifuged through 10000r/min, supernatant is enzyme liquid.
Embodiment 2
1. prepared by ferment raw material：After dragon fruit, apple, tomato and soya bean are cleaned with hot water, the work of cleaning is placed in
Typhoon is done, and be cut into small pieces shape；Each 100g of every kind of fruits and vegetables is weighed, uniformly mixing, is put into round, adds vegetables and fruits material quality
Percentage is 15% glucose, adds the water that vegetables and fruits material quality percentage is 100%, stirs.
2. inoculating strain：
Activated strains：By saccharomyces cerevisiae N85 and lactobacillus delbruockii subspecies bulgaricus B1, taken out from -80 DEG C of refrigerators, line
It is inoculated in corresponding culture medium.
Obtain thalline：Single bacterium colony on picking solid medium is inoculated into fluid nutrient medium, and culture to logarithm respectively is given birth to
For a long time, 10000rpm centrifuges 10min and abandons supernatant, with sterile saline with physiological saline：Bacterium solution 1:1 ratio is resuspended, and makes bacterium
It is dense up to 1~5 × 107CFU/mL。
Inoculation：Add the saccharomyces cerevisiae N85 of 1mL bacterium solutions/100g fruits and vegetables quality, be placed in 37 DEG C it is incubated, keep unidirectional
Exhaust, and stirring in 24 hours is once.Ferment by the 4th day, the Lactobacillus delbrueckii Bao Jiali of addition 5mL bacterium solutions/100g fruits and vegetables quality
Subspecies B1,30 DEG C of constant-temperature enclosed fermentations.
3. obtain ferment enzyme liquid：Ferment and terminated fermentation by the 15th day, centrifuged through 10000r/min, supernatant is enzyme liquid.
Embodiment 3.
1. prepared by ferment raw material：After dragon fruit, apple, tomato and soya bean are cleaned with hot water, the work of cleaning is placed in
Typhoon is done, and be cut into small pieces shape；Each 100g of every kind of fruits and vegetables is weighed, uniformly mixing, is put into round, adds vegetables and fruits material quality
Percentage is 15% glucose, adds the water that vegetables and fruits material quality percentage is 100%, stirs.
2. inoculating strain：
Activated strains：By saccharomyces cerevisiae N85 and lactobacillus delbruockii subspecies bulgaricus B2, taken out from -80 DEG C of refrigerators, line
It is inoculated in corresponding culture medium.
Obtain thalline：Single bacterium colony on picking solid medium is inoculated into fluid nutrient medium, and culture to logarithm respectively is given birth to
For a long time, 10000rpm centrifuges 10min and abandons supernatant, with sterile saline with physiological saline：Bacterium solution 1:1 ratio is resuspended, and makes bacterium
It is dense up to 1~5 × 107CFU/mL。
Inoculation：Add the saccharomyces cerevisiae N85 of 1mL bacterium solutions/100g fruits and vegetables quality, be placed in 37 DEG C it is incubated, keep unidirectional
Exhaust, and stirring in 24 hours is once.Ferment by the 4th day, the Lactobacillus delbrueckii Bao Jiali of addition 5mL bacterium solutions/100g fruits and vegetables quality
Subspecies B2,30 DEG C of constant-temperature enclosed fermentations.
3. obtain ferment enzyme liquid：Ferment and terminated fermentation by the 15th day, centrifuged through 10000r/min, supernatant is enzyme liquid.
Embodiment 4.
1. prepared by ferment raw material：After dragon fruit, apple, tomato and soya bean are cleaned with hot water, the work of cleaning is placed in
Typhoon is done, and be cut into small pieces shape；Each 100g of every kind of fruits and vegetables is weighed, uniformly mixing, is put into round, adds vegetables and fruits material quality
Percentage is 15% glucose, adds the water that vegetables and fruits material quality percentage is 100%, stirs.
2. inoculating strain：
Activated strains：By saccharomyces cerevisiae N85, gluconacetobacter Q1 and lactobacillus delbruockii subspecies bulgaricus B1, from -80 DEG C
Refrigerator takes out, and streak inoculation is in corresponding culture medium.
Obtain thalline：Single bacterium colony on picking solid medium is inoculated into fluid nutrient medium, and culture to logarithm respectively is given birth to
For a long time, 10000rpm centrifuges 10min and abandons supernatant, with sterile saline with physiological saline：Bacterium solution 1:1 ratio is resuspended, and makes bacterium
It is dense up to 1~5 × 107CFU/mL。
Inoculation：Add the Portugal of saccharomyces cerevisiae N85 and 2mL bacterium solution/100g fruits and vegetables quality of 1mL bacterium solutions/100g fruits and vegetables quality
Sweet and sour bacillus Q1,37 DEG C of incubated, holding one-way exhausts are placed in, and stirring in 24 hours is once.Ferment by the 4th day, add 5mL
The lactobacillus delbruockii subspecies bulgaricus B1 of bacterium solution/100g fruits and vegetables quality, 30 DEG C of constant-temperature enclosed fermentations.
3. obtain ferment enzyme liquid：Ferment and terminated fermentation by the 15th day, centrifuged through 10000r/min, supernatant is enzyme liquid.
Embodiment 5.
1. prepared by ferment raw material：After dragon fruit, apple, tomato and soya bean are cleaned with hot water, the work of cleaning is placed in
Typhoon is done, and be cut into small pieces shape；Each 100g of every kind of fruits and vegetables is weighed, uniformly mixing, is put into round, adds vegetables and fruits material quality
Percentage is 15% glucose, adds the water that vegetables and fruits material quality percentage is 100%, stirs.
2. inoculating strain：
Activated strains：By saccharomyces cerevisiae N85, gluconacetobacter Q1 and lactobacillus delbruockii subspecies bulgaricus B2, from -80 DEG C
Refrigerator takes out, and streak inoculation is in corresponding culture medium.
Obtain thalline：Single bacterium colony on picking solid medium is inoculated into fluid nutrient medium, and culture to logarithm respectively is given birth to
For a long time, 10000rpm centrifuges 10min and abandons supernatant, with sterile saline with physiological saline：Bacterium solution 1:1 ratio is resuspended, and makes bacterium
It is dense up to 1~5 × 107CFU/mL。
Inoculation：Add the Portugal of saccharomyces cerevisiae N85 and 2mL bacterium solution/100g fruits and vegetables quality of 1mL bacterium solutions/100g fruits and vegetables quality
Sweet and sour bacillus Q1,37 DEG C of incubated, holding one-way exhausts are placed in, and stirring in 24 hours is once.Ferment by the 4th day, add 5mL
The lactobacillus delbruockii subspecies bulgaricus B2 of bacterium solution/100g fruits and vegetables quality, 30 DEG C of constant-temperature enclosed fermentations.
3. obtain ferment enzyme liquid：Ferment and terminated fermentation by the 15th day, centrifuged through 10000r/min, supernatant is enzyme liquid.
Reference examples
1. prepared by ferment raw material：After dragon fruit, apple, tomato and soya bean are cleaned with hot water, the work of cleaning is placed in
Typhoon is done, and be cut into small pieces shape；Each 100g of every kind of fruits and vegetables is weighed, uniformly mixing, is put into round, adds vegetables and fruits material quality
Percentage is 15% glucose, adds the water that vegetables and fruits material quality percentage is 100%, stirs.
2. inoculating strain：
Activated strains：Saccharomyces cerevisiae N85 is taken out from -80 DEG C of refrigerators, streak inoculation is in corresponding culture medium.
Obtain thalline：Single bacterium colony on picking solid medium is inoculated into fluid nutrient medium, and culture to logarithm respectively is given birth to
For a long time, 10000rpm centrifuges 10min and abandons supernatant, with sterile saline with physiological saline：Bacterium solution 1:1 ratio is resuspended, and makes bacterium
It is dense up to 1~5 × 107CFU/mL。
Inoculation：The saccharomyces cerevisiae N85 of 1mL bacterium solutions/100g fruits and vegetables quality is added, is placed in 37 DEG C of constant-temperature enclosed fermentations, is kept
One-way exhaust, and stirring in 24 hours is once.
3. obtain ferment enzyme liquid：Ferment and terminated fermentation by the 15th day, centrifuged through 10000r/min, supernatant is enzyme liquid.
The ferment prepared to embodiment 1~5 and reference examples carries out inoxidizability, organic acid content, protease activity, antibacterial energy
Power and volatile materials measure.
Inoxidizability ability measurement result is as shown in table 2, the ferment after lactobacillus delbruockii subspecies bulgaricus B1 reinforcings,
Oxidation resistance improves, and wherein reducing power improves 43.3%, DPPH clearance rates and brought up to 93.09% by 88.25%, and adds
Common lactobacillus delbruockii subspecies bulgaricus B2 ferment reducing power raising amount and DPPH clearance rates is only 21.2% He
91.07%.
The inoxidizability ability of ferment under the different embodiments of table 2
Organic acid content testing result is as shown in table 3, the ferment after lactobacillus delbruockii subspecies bulgaricus B1 reinforcings, has
Machine acid content increases before being relatively not added with, and wherein lactic acid content improves 36.1%, and acetic acid content improves 35.2%；Add
Add the acetic acid of lactobacillus delbruockii subspecies bulgaricus B2 ferment and lactic acid content to show as reducing, be unfavorable for product storage and
Guarantee the quality.
The organic acid content (g/L) of ferment under the different embodiments of table 3
Protease activity measurement result is as shown in table 4, tyrosine standard curve such as Fig. 1, sub- through Lactobacillus delbrueckii
Ferment after kind B1 reinforcings, protease activity improve 11.9%.
The protease activity of ferment under the different embodiments of table 4
Bacteriostasis measurement result is as shown in table 5, the ferment after lactobacillus delbruockii subspecies bulgaricus B1 reinforcings, performance
Going out the inhibitory action to bacillus subtilis, Escherichia coli and Human fetal cardiomyocytes, antibacterial circle diameter is respectively 15.3mm,
16.6mm 14.3mm；And the ferment for adding lactobacillus delbruockii subspecies bulgaricus B2 does not have bacteriostasis.
The bacteriostasis (d/mm) of ferment under the different embodiments of table 5
Volatile materials measurement result is as shown in table 6, the ferment after lactobacillus delbruockii subspecies bulgaricus B1 reinforcings, waves
In volatile material, ethanol content reduces 20.1%, and suitable more crowds eat, and ethyl acetate content improves 85.1%, second
Sour pentyl ester content improves 89.1%, adds fruit aroma；Add the ferment second of common lactobacillus delbruockii subspecies bulgaricus
Alcohol content adds 6.0% compared to lactobacillus delbruockii subspecies bulgaricus B1, and ethyl acetate content reduces 70.9%, flavor
It is bad.
The volatile matter content (g/L) of ferment under the different embodiments of table 6.
Although the present invention is disclosed as above with preferred embodiment, it is not limited to the present invention, any to be familiar with this skill
The people of art, without departing from the spirit and scope of the present invention, it can all do various change and modification, therefore the protection model of the present invention
Enclose being defined of being defined by claims.
Claims (10)
- It is 1. a kind of while improve pectase feature and the method for flavor, it is characterised in that methods described be to using fruits and vegetables as Inoculation yeast bacterium, gluconacetobacter and lactobacillus are fermented in the ferment of raw material.
- 2. according to the method for claim 1, it is characterised in that the saccharomycete is saccharomyces cerevisiae；The gluconacetobacter For xylose gluconacetobacter；The lactobacillus is lactobacillus delbruockii subspecies bulgaricus.
- 3. according to the method for claim 1, it is characterised in that the saccharomycete is saccharomyces cerevisiae N85, the glucose vinegar bar Bacterium is xylose gluconacetobacter Q1；The lactobacillus is lactobacillus delbruockii subspecies bulgaricus (Lactobacillus Delbrueckii subsp.bulgaricus strain) B1, it is preserved in Chinese Typical Representative culture on June 26th, 2017 Collection, deposit number are CCTCC M 2017370, preservation address Wuhan, China, Wuhan University.
- 4. a kind of ferment, it is characterised in that formed using vegetables and/or fruit as raw material through microbial fermentation, contain thalline quantity ≥1×108Saccharomycete, gluconacetobacter and lactobacillus.
- 5. ferment according to claim 4, it is characterised in that the raw material includes dragon fruit, apple, tomato or soya bean At least one of.
- 6. the ferment according to claim 4 or 5, it is characterised in that be prepared according to the following steps：(1) added into raw material former Expect the glucose, the water of material quality 90~110% that mass percent is 15~20%, stir；(2) by volume, connect The saccharomycete and 1~2% gluconacetobacter of kind of step (1) mixed liquor 0.5~1%, are placed in 35~37 DEG C of cultures, and every 20~24 Hour stirring once, is fermented to the 3rd~4 day, the lactobacillus of inoculation 3~5%, 28~30 DEG C of anaerobic cultures, ferment to the 12nd~ Terminate fermentation within 15 days；The saccharomycete, gluconacetobacter and or lactobacillus concentration be 1.0 × 107~9.9 × 109CFU/mL。
- 7. ferment according to claim 6, it is characterised in that saccharomycete is saccharomyces cerevisiae N85, and gluconacetobacter is xylose Gluconacetobacter Q1, lactobacillus are lactobacillus delbruockii subspecies bulgaricus B1 or lactobacillus delbruockii subspecies bulgaricus B2.
- 8. a kind of preparation method of ferment, it is characterised in that methods described comprises the following steps：(1) it is former to fruit and/or vegetables The glucose, the water with raw material phase homogenous quantities that material quality percentage is 15~20% are added in material, is stirred；(2) body is pressed Product meter, the saccharomycete and 1~2% gluconacetobacter of inoculation step (1) mixed liquor 0.5~1%, is placed in 35~37 DEG C of cultures, Stirring in every 20~24 hours once, is fermented to the 3rd~4 day, the lactobacillus delbruockii subspecies bulgaricus B1 of inoculation 3~5%, 28~ 30 DEG C of anaerobic cultures, ferment and terminated fermentation to the 12nd~15 day；The saccharomycete, gluconacetobacter and or Lactobacillus delbrueckii protect plus Leah subspecies B1 concentration is 1.0 × 107~9.9 × 109CFU/mL；The lactobacillus is lactobacillus delbruockii subspecies bulgaricus B1, China typical culture collection center is preserved on June 26th, 2017, deposit number is CCTCC M 2017370, is protected Hide address Wuhan, China, Wuhan University.
- 9. the food containing claim 4-5, the extract of 7 any ferment or concentrate.
- 10. one plant of lactobacillus delbruockii subspecies bulgaricus (Lactobacillus delbrueckii subsp.bulgaricus Strain) B1, it is characterised in that be preserved in China typical culture collection center on June 26th, 2017, deposit number is CCTCC M 2017370, preservation address Wuhan, China, Wuhan University.
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