CN106119166B - One plant of Switzerland lactic acid bacteria and its application - Google Patents
One plant of Switzerland lactic acid bacteria and its application Download PDFInfo
- Publication number
- CN106119166B CN106119166B CN201610542256.4A CN201610542256A CN106119166B CN 106119166 B CN106119166 B CN 106119166B CN 201610542256 A CN201610542256 A CN 201610542256A CN 106119166 B CN106119166 B CN 106119166B
- Authority
- CN
- China
- Prior art keywords
- vinegar
- lactic acid
- fermentation
- lactobacillus helveticus
- preservation
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 title claims abstract description 76
- 239000004310 lactic acid Substances 0.000 title claims abstract description 38
- 235000014655 lactic acid Nutrition 0.000 title claims abstract description 38
- 241000894006 Bacteria Species 0.000 title claims abstract description 23
- 239000000052 vinegar Substances 0.000 claims abstract description 41
- 235000021419 vinegar Nutrition 0.000 claims abstract description 41
- 240000002605 Lactobacillus helveticus Species 0.000 claims abstract description 20
- 238000004321 preservation Methods 0.000 claims abstract description 13
- 244000005700 microbiome Species 0.000 claims abstract description 6
- 238000000855 fermentation Methods 0.000 claims description 22
- 230000004151 fermentation Effects 0.000 claims description 22
- 235000013967 Lactobacillus helveticus Nutrition 0.000 claims description 15
- 230000001580 bacterial effect Effects 0.000 claims description 15
- 229940054346 lactobacillus helveticus Drugs 0.000 claims description 15
- 240000007594 Oryza sativa Species 0.000 claims description 8
- 235000007164 Oryza sativa Nutrition 0.000 claims description 8
- 238000000034 method Methods 0.000 claims description 8
- 235000009566 rice Nutrition 0.000 claims description 8
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 6
- 235000020265 peanut milk Nutrition 0.000 claims description 6
- 235000015096 spirit Nutrition 0.000 claims description 6
- 239000007788 liquid Substances 0.000 claims description 5
- 238000002360 preparation method Methods 0.000 claims description 5
- 235000013339 cereals Nutrition 0.000 claims description 4
- 238000011081 inoculation Methods 0.000 claims description 4
- 239000002054 inoculum Substances 0.000 claims description 4
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims description 3
- 239000004382 Amylase Substances 0.000 claims description 2
- 108010065511 Amylases Proteins 0.000 claims description 2
- 102000013142 Amylases Human genes 0.000 claims description 2
- 235000019418 amylase Nutrition 0.000 claims description 2
- 108010089934 carbohydrase Proteins 0.000 claims description 2
- 235000013312 flour Nutrition 0.000 claims description 2
- 238000002156 mixing Methods 0.000 claims description 2
- 235000020183 skimmed milk Nutrition 0.000 claims description 2
- 230000001954 sterilising effect Effects 0.000 claims description 2
- 235000015099 wheat brans Nutrition 0.000 claims description 2
- 238000013124 brewing process Methods 0.000 claims 1
- 239000000796 flavoring agent Substances 0.000 abstract description 9
- 235000019634 flavors Nutrition 0.000 abstract description 9
- 239000000047 product Substances 0.000 description 13
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 10
- 241000196324 Embryophyta Species 0.000 description 8
- 239000002253 acid Substances 0.000 description 7
- 238000004519 manufacturing process Methods 0.000 description 7
- 108020004465 16S ribosomal RNA Proteins 0.000 description 4
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 4
- 229960000583 acetic acid Drugs 0.000 description 4
- 239000001963 growth medium Substances 0.000 description 4
- 239000002609 medium Substances 0.000 description 4
- 235000013305 food Nutrition 0.000 description 3
- 238000005259 measurement Methods 0.000 description 3
- 238000012216 screening Methods 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- 102000016938 Catalase Human genes 0.000 description 2
- 108010053835 Catalase Proteins 0.000 description 2
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 229910000019 calcium carbonate Inorganic materials 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 235000019441 ethanol Nutrition 0.000 description 2
- 230000002906 microbiologic effect Effects 0.000 description 2
- 239000000843 powder Substances 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 238000012163 sequencing technique Methods 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 235000012976 tarts Nutrition 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 229920001817 Agar Polymers 0.000 description 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- 238000007400 DNA extraction Methods 0.000 description 1
- 206010013911 Dysgeusia Diseases 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 229920002527 Glycogen Polymers 0.000 description 1
- 241000186660 Lactobacillus Species 0.000 description 1
- 238000012408 PCR amplification Methods 0.000 description 1
- 208000004880 Polyuria Diseases 0.000 description 1
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 238000002479 acid--base titration Methods 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 238000000246 agarose gel electrophoresis Methods 0.000 description 1
- 230000001476 alcoholic effect Effects 0.000 description 1
- 235000015278 beef Nutrition 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 210000001124 body fluid Anatomy 0.000 description 1
- 239000010839 body fluid Substances 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 239000007795 chemical reaction product Substances 0.000 description 1
- 239000006071 cream Substances 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 230000007850 degeneration Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 230000035619 diuresis Effects 0.000 description 1
- 125000000816 ethylene group Chemical group [H]C([H])([*:1])C([H])([H])[*:2] 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 235000012055 fruits and vegetables Nutrition 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 229940096919 glycogen Drugs 0.000 description 1
- 229910052736 halogen Inorganic materials 0.000 description 1
- 150000002367 halogens Chemical class 0.000 description 1
- 230000002440 hepatic effect Effects 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 230000007794 irritation Effects 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 229940039696 lactobacillus Drugs 0.000 description 1
- 230000002475 laxative effect Effects 0.000 description 1
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 1
- 235000019341 magnesium sulphate Nutrition 0.000 description 1
- 229940099596 manganese sulfate Drugs 0.000 description 1
- 239000011702 manganese sulphate Substances 0.000 description 1
- 235000007079 manganese sulphate Nutrition 0.000 description 1
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 description 1
- 235000013622 meat product Nutrition 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 238000009629 microbiological culture Methods 0.000 description 1
- 239000005445 natural material Substances 0.000 description 1
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 description 1
- 229920000053 polysorbate 80 Polymers 0.000 description 1
- 108010009004 proteose-peptone Proteins 0.000 description 1
- 230000003014 reinforcing effect Effects 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 230000001932 seasonal effect Effects 0.000 description 1
- 239000001632 sodium acetate Substances 0.000 description 1
- 235000017281 sodium acetate Nutrition 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 235000014214 soft drink Nutrition 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 230000000638 stimulation Effects 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 238000012549 training Methods 0.000 description 1
- YWYZEGXAUVWDED-UHFFFAOYSA-N triammonium citrate Chemical compound [NH4+].[NH4+].[NH4+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O YWYZEGXAUVWDED-UHFFFAOYSA-N 0.000 description 1
- 239000001393 triammonium citrate Substances 0.000 description 1
- 235000011046 triammonium citrate Nutrition 0.000 description 1
Classifications
-
- 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
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12J—VINEGAR; PREPARATION OR PURIFICATION THEREOF
- C12J1/00—Vinegar; Preparation or purification thereof
- C12J1/04—Vinegar; Preparation or purification thereof from alcohol
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Wood Science & Technology (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Tropical Medicine & Parasitology (AREA)
- Virology (AREA)
- Biomedical Technology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Distillation Of Fermentation Liquor, Processing Of Alcohols, Vinegar And Beer (AREA)
Abstract
The invention discloses one plant of Lactobacillus helveticus Lactobacillus helveticus HS1-8, it is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, preservation place is Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3 Institute of Microorganism, Academia Sinica, the number that preservation is registered on the books is CGMCC NO.12225, and the deposit date is March 18 in 2016.The application of the lactic acid bacteria can significantly improve the content of lactic acid in vinegar, improve product special flavour, improve product quality.
Description
Technical field
The present invention relates to one plant of Switzerland lactic acid bacteria and its application, specially one plant is suitable for vinegar brewing and food can be improved
The Lactobacillus helveticus (Lactobacillus helveticus HS1-8) of lactic acid content, belongs to microbe application and vinegar in vinegar
Brewing field.
Background technique
Lactic acid bacteria distributes widely in nature, closely related with human lives, has different physiological roles.With the modern times
The good characteristic of microbiological development, food grade lactic acid bacterium causes the concern of microbial world, and lactic acid bacteria is widely used to cream
The food such as product, meat products, fruit and vegetable product, soft drink.Application of the lactic acid bacteria in brewery industry is also more and more closed
Note.
Lactic acid is soft with tart flavour, irritation is small, can mitigate the tart flavour of acetic acid stimulation, while having and dropping in alimentary canal
Solution metabolism, generates energy, can play sharp diuresis and laxative action, the synthesis to hepatic glycogen, adjusting cellular redox state with
Body fluid plays good action.Zhenjiang vinegar is one of traditional four big vinegar in China, has mellowness dense, the spy of long times of aftertaste
Point.Fixed acid is flavor substance important in zhenjiang vinegar, and " GB/T 18623-2011 geography symbol product Zhenjiang is fragrant
Vinegar " in require fixed acid (in terms of lactic acid) content >=1.60g/100ml of superfine zhenjiang vinegar.It is non-volatile in zhenjiang vinegar
Acid based on lactic acid, lactic acid content can the total fixed acid of Zhan 80% or more.
In actual Vinegar Fermentation production process, other than dominating the acetic acid bacteria of acetic fermentation, lactic acid bacteria is also to make
Common, important microorganism during making, and play an important role to the flavor and quality of product.During traditional zymotic
The main source of lactic acid bacteria be natural material bring into and with kind when bring into, between different seasons, different batches have it is larger
Difference, especially the content of lactic acid is obviously relatively low in winter.In addition, the part lactic acid bacteria brought into is during the fermentation because inadaptable
The special brewing environment such as high ethano, high acetic acid and can not rise in value.Therefore, the lactic acid of the special brewing environment of suitable vinegar is filtered out
Bacterium, the brewing applied to vinegar have great importance.
The present invention is directed to the above subject, and the present invention provides one plant of Lactobacillus helveticus for being suitably applied vinegar brewing, the bacterium
Strain may be directly applied to the brewing of vinegar, can significantly improve the content of lactic acid in vinegar, improve product special flavour, improve product product
Matter.
Summary of the invention
Present invention separation screening from the distiller's wort of zhenjiang vinegar production process obtains one plant and is more suited to vinegar brewing
The Lactobacillus helveticus used.The application of the lactic acid bacteria can significantly improve the content of lactic acid in vinegar, improve product special flavour, improve and produce
Product quality.
Specifically, the Lactobacillus helveticus Lactobacillus helveticus HS1-8, is preserved in China Microbiological bacterium
Kind preservation administration committee common micro-organisms center, preservation place are Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3 Chinese science
Institute of microbiology, institute, the number that preservation is registered on the books are CGMCC NO.12225, and the deposit date is March 18 in 2016.
It is separated, purifying and identification method are as follows:
(1) it separates
Distiller's wort sample of the 10ml after being inoculated with the special yeast of zhenjiang vinegar by alcoholic fermentation is taken, 1ml sample is taken to be added
Into 9ml sterile saline, after being mixed evenly on eddy mixer, then the solution that takes 1ml to dilute is added to the life of 9ml
It manages in salt water, and so on.It chooses 3 suitable concentration to be successively applied on the MRS solid plate added with calcium carbonate, in 37
48h is cultivated in DEG C anaerobic culture box.
Isolation medium (MRS culture medium): casein peptone 10.0g, glucose 20.0g, yeast powder 4.0g, powdered beef
8.0g, sodium acetate 5.0g, Triammonium citrate 2.0g, dipotassium hydrogen phosphate 2.0g, magnesium sulfate 0.2g, manganese sulfate 0.05g, Tween 80
1.0g, distilled water 1000mL adjust pH6.2 ± 0.2.Solid medium adds 15~20g/L agar powder.Screening and culturing medium addition
15~20g/L calcium carbonate.
(2) bacterial strain purifies
According to the size of microbe colony, color, gloss, transparency etc., biggish doubtful lactic acid bacteria is enclosed in the dissolution of picking calcium
Bacterium colony, and purified 2 times using the method for scribing line in the flat lining out of MRS.The scribed single colonie of picking carries out Gram's staining
And catalase, choose Gram-positive, after the strain of negative catalase is crossed on inclined-plane, the training of 37 DEG C of anaerobism
Feeding 48h is placed on 4 DEG C of refrigerators and saves.
(3) bacterial strain primary dcreening operation
The bacterial strain of preservation is inoculated into respectively in the MRS culture medium containing 9%vol ethyl alcohol, measurement is sent out afterwards for 24 hours for 37 DEG C of cultures
The pH value of zymotic fluid chooses the bacterial strain that pH value is lower than 4.0.
Simultaneously by the MRS culture medium of the strain inoculated to pH value 4.0,37 DEG C of cultures for 24 hours, measure the pH value of fermentation liquid.
One plant of resistance to ethyl alcohol, acidproof, the preferable bacterial strain HS1-8 of characteristic of lactic acid production are obtained, colonial morphology is as shown in Figure 1.
(4) physio-biochemical characteristics
1 physio-biochemical characteristics of table
Note: "+" represents the positive, and "-" represents feminine gender, and " W " growth is weak.
(5) bacterial strain molecular biology identification
Detect the sequence of the 16S rRNA of bacterial strain HS1-8.
Using the bacterial genomes DNA extraction kit of Sangon Biotech (Shanghai) Co., Ltd., this hair is extracted
The genomic DNA of the bright bacterial strain HS1-8 screened, PCR amplification primer are mentioned by Sangon Biotech (Shanghai) Co., Ltd.
For sequencing primer is as follows:
27F:AGAGTTTGATCCTGGCTCAG
1492R:GGTTACCTTGTTACGACTT
PCR reaction cycle condition:
The detection of PCR product: taking 5ul reaction product, is detected using 1% agarose gel electrophoresis.If PCR success,
There is band appearance at about 1500bp.
Sequencing analysis: PCR product being entrusted into Sangon Biotech (Shanghai) Co., Ltd.) limited liability company is sequenced, and sequence is as follows
Literary 16S rRNA sequence.The sequence blast program measured compares analysis in ncbi database.As a result bacterial strain HS1- of the present invention
The 16S rRNA homology of 8 and Lactobacillus helveticus (Lactobacillus helveticus) DSM20075 is 99%.
The 16S rRNA sequence of bacterial strain HS1-8
CGTGCCTAATACATGCAAGTCGAGCGAGCAGAACCAGCAGATTTACTTCGGTAATGACGCTGGGGACGC
GAGCGGCGGATGGGTGAGTAACACGTGGGGAACCTGCCCCATAGTCTGGGATACCACTTGGAAACAGGTGCTAATAC
CGGATAAGAAAGCAGATCGCATGATCAGCTTATAAAAGGCGGCGTAAGCTGTCGCTATGGGATGGCCCCGCGGTGCA
TTAGCTAGTTGGTAAGGCAATGGCTTACCAAGGCAATGATGCATAGCCGAGTTGAGAGACTGATCGGCCACATTGGG
ACTGAGACACGGCCCAAACTCCTACGGGAGGCAGCAGTAGGGAATCTTCCACAATGGACGCAAGTCTGATGGAGCAA
CGCCGCGTGAGTGAAGAAGGTTTTCGGATCGTAAAGCTCTGTTGTTGGTGAAGAAGGATAGAGGTAGTAACTGGCCT
TTATTTGACGGTAATCAACCAGAAAGTCACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGT
TGTCCGGATTTATTGGGCGTAAAGCGAGCGCAGGCGGAAGAATAAGTCTGATGTGAAAGCCCTCGGCTTAACCGAGG
AACTGCATCGGAAACTGTTTTTCTTGAGTGCAGAAGAGGAGAGTGGAATTCCATGTGTAGCGGTGGAATGCGTAGAT
ATATGGAAGAACACCAGTGGCGAAGGCGACTCTCTGGTCTGCAACTGACGCTGAGGCTCGAAAGCATGGGTAGCGAA
CAGGATTAGATACCCTGGTAGTCCATGCCGTAAACGATGAGTGCTAAGTGTTGGGAGGTTTCCGCCTCTCAGTGCTG
CAGCTAACGCATTAAGCACTCCGCCTGGGGAGTACGACCGCAAGGTTGAAACTCAAAGGAATTGACGGGGGCCCGCA
CAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAACGCGAAGAACCTTACCAGGTCTTGACATCTAGTGCCATCCTAA
GAGATTAGGAGTTCCCTTCGGGGACGCTAAGACAGGTGGTGCATGGCTGTCGTCAGCTCGTGTCGTGAGATGTTGGG
TTAAGTCCCGCAACGAGCGCAACCCTTGTTATTAGTTGCCAGCATTAAGTTGGGCACTCTAATGAGACTGCCGGTGA
TAAACCGGAGGAAGGTGGGGATGACGTCAAGTCATCATGCCCCTTATGACCTGGGCTACACACGTGCTACAATGGAC
AGTACAACGAGAAGCGAGCCTGCGAAGGCAAGCGAATCTCTGAAAGCTGTTCTCAGTTCGGACTGCAGTCTGCAACT
CGACTGCACGAAGCTGGAATCGCTAGTAATCGCGGATCAGAACGCCGCGGTGAATACGTTCCCGGGCCTTGTACACA
CCGCCCGTCACACCATGGAAGTCTGCAATGCCCAAAGCCGGTGGCCTAACCTTCGGAAGGAGCCGTCTAAGGCAGGG
CAGATGACTGGGGTGAAGTCGTAACAAGGTAGCCGTAGGAGAACCTGCGGTTGGA
It based on the above results, is Lactobacillus helveticus HS1-8 by Strain Designation of the present invention, in 2016
It is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on March 18, in, preservation place is Beijing
No. 3 Institute of Microorganism, Academia Sinica, institute of city, North Star West Road, Chaoyang District 1, the number that preservation is registered on the books is CGMCC
NO.12225。
Present invention further proposes application of the above-mentioned Lactobacillus helveticus in vinegar production.
By experiments have shown that, Lactobacillus helveticus of the invention can effectively improve containing for lactic acid in zhenjiang vinegar vinegar fermented grain
Amount, effectively improves the flavor and quality of zhenjiang vinegar.
The utility model has the advantages that the present invention provides the Lactobacillus helveticus of one plant of suitable vinegar brewing, the wine of vinegar can be directly made an addition to
During making, by the use of the lactic acid bacteria, lactic acid caused by can improving because of the problems such as seasonal temperature variation, old spawn degeneration
The problem of content reduces.By the use of the lactic acid bacteria, the relative abundance and zhenjiang vinegar of lactic acid bacteria in vinegar fermented grain can be significantly improved
The content of middle lactic acid improves the flavor and quality of zhenjiang vinegar.
Detailed description of the invention
Fig. 1 is the colonial morphology figure of Lactobacillus helveticus Lactobacillus helveticus HS1-8；
Fig. 2 is the changes of contents schematic diagram of total acid after inoculation, when show-through and envelope unstrained spirits in fermentation process；
Fig. 3 is the changes of contents schematic diagram of lactic acid after inoculation, when show-through and envelope unstrained spirits in fermentation process.
Specific embodiment
Present invention separation screening from the distiller's wort of zhenjiang vinegar production process obtains one plant and is more suited to vinegar brewing
The Lactobacillus helveticus used, the Lactobacillus helveticus Lactobacillus helveticus HS1-8, is preserved in China Microbiological
Culture presevation administration committee common micro-organisms center, preservation place are section, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3 China
Institute of microbiology, institute, the number that preservation is registered on the books are CGMCC NO.12225, and the deposit date is on March 18th, 2016.
The Lactobacillus helveticus Lactobacillus helveticus HS1- is described in detail below by specific embodiment
8 application in zhenjiang vinegar production.
1. the expansion culture of bacterial strain
The preparation of primary seed solution: according to 5% inoculum concentration, the Lactobacillus helveticus HS1-8 of purifying is inoculated into liquid MRS
In culture medium, 37 DEG C of culture 20h.
The preparation of secondary seed solution: selecting 50L liquid fermentation fermentor, and the processed Rice & peanut milk of 35L, rice flour content in Rice & peanut milk is added
15%~25%, the treatment process of the Rice & peanut milk is as follows: 100 DEG C of high temperature are gelatinized 30min, and the amylase of 6~8u/ml of addition is kept
85~90 DEG C of liquefaction 30min, the carbohydrase for then adding 100~300u/ml keep 55~60 DEG C of saccharification 30min.Add 2 ‰
After skimmed milk power, 115 DEG C of sterilizing 20min.Primary seed solution is accessed according to 5% inoculum concentration.37 DEG C of temperature of control, pressure in tank
For 0.05MPa, mixing speed 60r/min, it is 6.0 that it is constant, which to control medium pH value, with NaOH solution, and fermentation is for 24 hours.Fermentation knot
The viable count of Shu Hou, lactic acid bacteria reach 109cfu/ml。
2. vinegar fermented grain inoculation fermentation
6 400kg large cylinders are chosen, distiller's wort 250kg, wheat bran 90kg, big chaff 50kg is added in each cylinder.Select 3 cylinders for examination
Group is tested, 11L second order fermentation seed liquor is accessed in each cylinder, remaining 3 cylinders are control group.It is carried out according to zhenjiang vinegar brewage process
Band kind turns over unstrained spirits fermentation.
3. the detection of total acid and lactic acid content
In fermentation process, halogen after being inoculated with, when show-through and envelope unstrained spirits is taken to be detected.
Total acid (with Acetometer) uses determination of acid-basetitration, and measurement result is as shown in Figure 2.
Lactic acid uses high performance liquid chromatography, referring to " GB/T 18623-2011 geography symbol product zhenjiang vinegar " Appendix B
Middle method, measurement result are as shown in Figure 3.
During acetic fermentation, the total acid content of test group and control group does not have significant difference.During the fermentation,
The lactic acid content of test group is apparently higher than control group, and test group lactic acid content improves 11.4% than control group when sealing unstrained spirits.
Bacterial strain HS1-8 of the invention can preferably be applied to as lactobacillus forced strain in the production of zhenjiang vinegar, lead to
The reinforcing for crossing the bacterial strain can significantly improve lactic acid content in zhenjiang vinegar, improve product quality.
Claims (1)
1. a kind of application of Lactobacillus helveticus in zhenjiang vinegar brewing, which is characterized in that its classification naming isLactobacillus helveticusHS1-8 is preserved in Chinese microorganism strain preservation management committee on March 18th, 2016
Member's meeting common micro-organisms center, preservation place are Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3 Chinese Academy of Sciences microbe research
Institute, the number that preservation is registered on the books are CGMCC NO.12225, and the Lactobacillus helveticus is in the special yeast preparation of zhenjiang vinegar
It is isolated in distiller's wort, it can directly make an addition in the brewing process of vinegar, specific steps are as follows:
(1) the expansion culture of bacterial strain:
The preparation of primary seed solution: according to 5% inoculum concentration, the Lactobacillus helveticus HS1-8 of purifying is inoculated into liquid MRS culture
In base, 37 DEG C of culture 20h；
The preparation of secondary seed solution: selecting 50L liquid fermentation fermentor, and the processed Rice & peanut milk of 35L is added, and rice flour content 15% in Rice & peanut milk ~
25%, the treatment process of the Rice & peanut milk is as follows: 100 DEG C of high temperature are gelatinized 30min, and the amylase of 6 ~ 8u/ml of addition is kept for 85 ~ 90 DEG C
Liquefy 30min, and the carbohydrase for then adding 100 ~ 300 u/ml keeps 55 ~ 60 DEG C of saccharification 30min, the skimmed milk power of addition 2 ‰
Afterwards, 115 DEG C of sterilizing 20min；
Primary seed solution is accessed according to 5% inoculum concentration, controls 37 DEG C of temperature, pressure is 0.05 MPa, mixing speed 60 in tank
R/min, it is 6.0 that it is constant, which to control medium pH value, with NaOH solution, and for 24 hours, after fermentation, the viable count of lactic acid bacteria reaches for fermentation
109cfu/ml；
(2) vinegar fermented grain inoculation fermentation:
Large cylinder is chosen, is added distiller's wort 250kg in each cylinder, wheat bran 90kg, big chaff 50kg access 11L second order fermentation in each cylinder
Seed liquor carries out band kind according to zhenjiang vinegar brewage process, turns over unstrained spirits fermentation.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610542256.4A CN106119166B (en) | 2016-07-11 | 2016-07-11 | One plant of Switzerland lactic acid bacteria and its application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610542256.4A CN106119166B (en) | 2016-07-11 | 2016-07-11 | One plant of Switzerland lactic acid bacteria and its application |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN106119166A CN106119166A (en) | 2016-11-16 |
| CN106119166B true CN106119166B (en) | 2019-10-29 |
Family
ID=57282446
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610542256.4A Active CN106119166B (en) | 2016-07-11 | 2016-07-11 | One plant of Switzerland lactic acid bacteria and its application |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106119166B (en) |
Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108315277B (en) * | 2018-03-19 | 2021-01-05 | 金华银河生物科技有限公司 | Lactobacillus plantarum and method for freeze-drying lactic acid bacteria |
| CN109112083B (en) * | 2018-08-01 | 2020-08-04 | 江南大学 | A kind of Lactobacillus helveticus with high caprylic acid and application thereof |
| CN110819576B (en) * | 2019-12-16 | 2020-08-11 | 江苏恒顺醋业股份有限公司 | Binary composite leaven and application thereof |
| CN115725444A (en) * | 2022-08-05 | 2023-03-03 | 江苏恒顺醋业股份有限公司 | Lactobacillus helveticus bacterium for high yield of phenyllactic acid and application thereof |
| CN116083322A (en) * | 2023-03-07 | 2023-05-09 | 江苏恒顺醋业股份有限公司 | A strain of Lactobacillus and its application in fruit wine |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102329762A (en) * | 2011-11-01 | 2012-01-25 | 内蒙古农业大学 | Lactobacillus helveticus H9 and application thereof |
| CN103865749B (en) * | 2014-04-09 | 2015-11-04 | 福建师范大学 | A kind of brewing method of Fujian red yeast rice vinegar |
-
2016
- 2016-07-11 CN CN201610542256.4A patent/CN106119166B/en active Active
Also Published As
| Publication number | Publication date |
|---|---|
| CN106119166A (en) | 2016-11-16 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| WO2021120619A1 (en) | Bacillus safensis, microbial agent, and applications thereof and vinegar preparation method | |
| CN106190893B (en) | The preparation method and application of one plant of lactobacillus fermenti for being suitable for vinegar brewing and its bacterium powder | |
| CN106754508B (en) | A strain of Bacillus subtilis and its application in soy sauce fermentation and aroma enhancement | |
| CN106119166B (en) | One plant of Switzerland lactic acid bacteria and its application | |
| CN110257285A (en) | One plant with effectively degrading nitrite and the strong lactobacillus plantarum for producing acid function and its application | |
| KR102804252B1 (en) | Saccharopolyspora and its applications in biogenic amine reduction | |
| CN108913628A (en) | One plant of Sonora desert bacillus and its application | |
| CN111961615A (en) | Saccharopolyspora capable of reducing biogenic amine and application thereof | |
| CN102643767A (en) | Lactobacillus plantarum and application thereof in fermenting and ensiling sweet potato stem and leaf | |
| CN110004090B (en) | A kind of Leuconostoc membranaceus and its application in fermented sauerkraut | |
| CN102936573A (en) | Garlic endophyte for accelerating black garlic fermentation process and application thereof in black garlic fermentation | |
| CN116555060B (en) | Saccharomyces cerevisiae CMRC 14S and its applications | |
| CN109136129B (en) | A strain of Lactobacillus acidophilus NCU426 | |
| CN110408571A (en) | One bacillus coagulans and its application | |
| CN113308418A (en) | Lactobacillus chaff for fermentation and fermentation preparation process thereof | |
| CN110093285A (en) | A strain of acid-resistant fermented Lactobacillus and its application | |
| CN116179401B (en) | Lactobacillus plantarum ZF605 and application thereof | |
| CN106754507B (en) | A kind of compound flavor bacterial agent and preparation method thereof and direct-throwing application in soy sauce flavor enhancement | |
| CN111979148A (en) | Saccharopolyspora composition and application thereof in food | |
| CN113249268B (en) | Saccharopolyspora rosea for reducing biogenic amine and application thereof | |
| CN110885767B (en) | Lactococcus lactis subsp. hollisae with good antioxidant activity and application thereof | |
| CN113308419B (en) | Lactobacillus chaff for fermentation and application thereof | |
| CN109401998B (en) | A strain of Lactobacillus Mindena degrading biogenic amines and its application | |
| CN113801800A (en) | Saccharomyces cerevisiae and application thereof | |
| CN113699069A (en) | Strain HSCY2073, separation and screening thereof and application of strain to improvement of flavor quality of vinegar |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |