CN112725107A - Mixed fermentation process based on Klatomyces and saccharomyces cerevisiae - Google Patents
Mixed fermentation process based on Klatomyces and saccharomyces cerevisiae Download PDFInfo
- Publication number
- CN112725107A CN112725107A CN202110191367.6A CN202110191367A CN112725107A CN 112725107 A CN112725107 A CN 112725107A CN 202110191367 A CN202110191367 A CN 202110191367A CN 112725107 A CN112725107 A CN 112725107A
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- strain
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- fermentation
- saccharomyces cerevisiae
- saccharomyces
- Prior art date
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- Granted
Links
- 240000004808 Saccharomyces cerevisiae Species 0.000 title claims abstract description 123
- 238000000855 fermentation Methods 0.000 title claims abstract description 114
- 230000004151 fermentation Effects 0.000 title claims abstract description 114
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 title claims abstract description 108
- 241000235649 Kluyveromyces Species 0.000 claims abstract description 12
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- YHVUVJYEERGYNU-UHFFFAOYSA-N 4',8-Di-Me ether-5,7,8-Trihydroxy-3-(4-hydroxybenzyl)-4-chromanone Natural products COC1(C)CC(O)OC(C)C1O YHVUVJYEERGYNU-UHFFFAOYSA-N 0.000 claims abstract description 5
- 239000000758 substrate Substances 0.000 claims description 33
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims description 21
- 241000235070 Saccharomyces Species 0.000 claims description 21
- 239000007222 ypd medium Substances 0.000 claims description 19
- 235000019674 grape juice Nutrition 0.000 claims description 17
- 240000000560 Citrus x paradisi Species 0.000 claims description 16
- 239000001963 growth medium Substances 0.000 claims description 16
- 238000004519 manufacturing process Methods 0.000 claims description 16
- RAHZWNYVWXNFOC-UHFFFAOYSA-N Sulphur dioxide Chemical compound O=S=O RAHZWNYVWXNFOC-UHFFFAOYSA-N 0.000 claims description 14
- 238000011081 inoculation Methods 0.000 claims description 14
- 239000011550 stock solution Substances 0.000 claims description 10
- 239000002245 particle Substances 0.000 claims description 9
- 241000222290 Cladosporium Species 0.000 claims description 8
- 230000004913 activation Effects 0.000 claims description 8
- 230000004580 weight loss Effects 0.000 claims description 8
- 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 7
- 239000001888 Peptone Substances 0.000 claims description 7
- 108010080698 Peptones Proteins 0.000 claims description 7
- 108010059820 Polygalacturonase Proteins 0.000 claims description 7
- 229940041514 candida albicans extract Drugs 0.000 claims description 7
- 108010093305 exopolygalacturonase Proteins 0.000 claims description 7
- 239000008103 glucose Substances 0.000 claims description 7
- 239000002609 medium Substances 0.000 claims description 7
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- 239000000843 powder Substances 0.000 claims description 7
- 239000012138 yeast extract Substances 0.000 claims description 7
- 239000002054 inoculum Substances 0.000 claims description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 6
- 241001362614 Crassa Species 0.000 claims description 5
- 230000008859 change Effects 0.000 claims description 5
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- 238000004321 preservation Methods 0.000 claims description 4
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- 238000002360 preparation method Methods 0.000 claims description 3
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- 239000000126 substance Substances 0.000 description 15
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- 238000000034 method Methods 0.000 description 9
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- 239000002994 raw material Substances 0.000 description 5
- KBPLFHHGFOOTCA-UHFFFAOYSA-N 1-Octanol Chemical compound CCCCCCCCO KBPLFHHGFOOTCA-UHFFFAOYSA-N 0.000 description 4
- BBMCTIGTTCKYKF-UHFFFAOYSA-N 1-heptanol Chemical compound CCCCCCCO BBMCTIGTTCKYKF-UHFFFAOYSA-N 0.000 description 4
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- ICKWICRCANNIBI-UHFFFAOYSA-N 2,4-di-tert-butylphenol Chemical compound CC(C)(C)C1=CC=C(O)C(C(C)(C)C)=C1 ICKWICRCANNIBI-UHFFFAOYSA-N 0.000 description 4
- YIWUKEYIRIRTPP-UHFFFAOYSA-N 2-ethylhexan-1-ol Chemical compound CCCCC(CC)CO YIWUKEYIRIRTPP-UHFFFAOYSA-N 0.000 description 4
- QIMMUPPBPVKWKM-UHFFFAOYSA-N 2-methylnaphthalene Chemical compound C1=CC=CC2=CC(C)=CC=C21 QIMMUPPBPVKWKM-UHFFFAOYSA-N 0.000 description 4
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- XKWSWANXMRXDES-UHFFFAOYSA-N 3-methylbutyl octanoate Chemical compound CCCCCCCC(=O)OCCC(C)C XKWSWANXMRXDES-UHFFFAOYSA-N 0.000 description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 4
- ROWKJAVDOGWPAT-UHFFFAOYSA-N Acetoin Chemical compound CC(O)C(C)=O ROWKJAVDOGWPAT-UHFFFAOYSA-N 0.000 description 4
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- QSJXEFYPDANLFS-UHFFFAOYSA-N Diacetyl Chemical compound CC(=O)C(C)=O QSJXEFYPDANLFS-UHFFFAOYSA-N 0.000 description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 4
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 description 4
- AMQJEAYHLZJPGS-UHFFFAOYSA-N N-Pentanol Chemical compound CCCCCO AMQJEAYHLZJPGS-UHFFFAOYSA-N 0.000 description 4
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- MWKFXSUHUHTGQN-UHFFFAOYSA-N decan-1-ol Chemical compound CCCCCCCCCCO MWKFXSUHUHTGQN-UHFFFAOYSA-N 0.000 description 4
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- LZCLXQDLBQLTDK-UHFFFAOYSA-N ethyl 2-hydroxypropanoate Chemical compound CCOC(=O)C(C)O LZCLXQDLBQLTDK-UHFFFAOYSA-N 0.000 description 4
- OMSUIQOIVADKIM-UHFFFAOYSA-N ethyl 3-hydroxybutyrate Chemical compound CCOC(=O)CC(C)O OMSUIQOIVADKIM-UHFFFAOYSA-N 0.000 description 4
- XIRNKXNNONJFQO-UHFFFAOYSA-N ethyl hexadecanoate Chemical compound CCCCCCCCCCCCCCCC(=O)OCC XIRNKXNNONJFQO-UHFFFAOYSA-N 0.000 description 4
- SHZIWNPUGXLXDT-UHFFFAOYSA-N ethyl hexanoate Chemical compound CCCCCC(=O)OCC SHZIWNPUGXLXDT-UHFFFAOYSA-N 0.000 description 4
- MMKRHZKQPFCLLS-UHFFFAOYSA-N ethyl myristate Chemical compound CCCCCCCCCCCCCC(=O)OCC MMKRHZKQPFCLLS-UHFFFAOYSA-N 0.000 description 4
- YYZUSRORWSJGET-UHFFFAOYSA-N ethyl octanoate Chemical compound CCCCCCCC(=O)OCC YYZUSRORWSJGET-UHFFFAOYSA-N 0.000 description 4
- ZSIAUFGUXNUGDI-UHFFFAOYSA-N hexan-1-ol Chemical compound CCCCCCO ZSIAUFGUXNUGDI-UHFFFAOYSA-N 0.000 description 4
- MLFHJEHSLIIPHL-UHFFFAOYSA-N isoamyl acetate Chemical compound CC(C)CCOC(C)=O MLFHJEHSLIIPHL-UHFFFAOYSA-N 0.000 description 4
- PHTQWCKDNZKARW-UHFFFAOYSA-N isoamylol Chemical compound CC(C)CCO PHTQWCKDNZKARW-UHFFFAOYSA-N 0.000 description 4
- ZXEKIIBDNHEJCQ-UHFFFAOYSA-N isobutanol Chemical compound CC(C)CO ZXEKIIBDNHEJCQ-UHFFFAOYSA-N 0.000 description 4
- CPJRRXSHAYUTGL-UHFFFAOYSA-N isopentenyl alcohol Chemical compound CC(=C)CCO CPJRRXSHAYUTGL-UHFFFAOYSA-N 0.000 description 4
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- WVDDGKGOMKODPV-UHFFFAOYSA-N Benzyl alcohol Chemical compound OCC1=CC=CC=C1 WVDDGKGOMKODPV-UHFFFAOYSA-N 0.000 description 3
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- DHKHKXVYLBGOIT-UHFFFAOYSA-N 1,1-Diethoxyethane Chemical compound CCOC(C)OCC DHKHKXVYLBGOIT-UHFFFAOYSA-N 0.000 description 2
- 239000005968 1-Decanol Substances 0.000 description 2
- YOMSJEATGXXYPX-UHFFFAOYSA-N 2-methoxy-4-vinylphenol Chemical compound COC1=CC(C=C)=CC=C1O YOMSJEATGXXYPX-UHFFFAOYSA-N 0.000 description 2
- WLAMNBDJUVNPJU-UHFFFAOYSA-N 2-methylbutyric acid Chemical compound CCC(C)C(O)=O WLAMNBDJUVNPJU-UHFFFAOYSA-N 0.000 description 2
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- XVSZRAWFCDHCBP-UHFFFAOYSA-N 3-methylbutyl hexanoate Chemical compound CCCCCC(=O)OCCC(C)C XVSZRAWFCDHCBP-UHFFFAOYSA-N 0.000 description 2
- GHBSPIPJMLAMEP-UHFFFAOYSA-N 6-pentyloxan-2-one Chemical compound CCCCCC1CCCC(=O)O1 GHBSPIPJMLAMEP-UHFFFAOYSA-N 0.000 description 2
- IKHGUXGNUITLKF-UHFFFAOYSA-N Acetaldehyde Chemical compound CC=O IKHGUXGNUITLKF-UHFFFAOYSA-N 0.000 description 2
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 2
- 235000014493 Crataegus Nutrition 0.000 description 2
- 241001092040 Crataegus Species 0.000 description 2
- DKMROQRQHGEIOW-UHFFFAOYSA-N Diethyl succinate Chemical compound CCOC(=O)CCC(=O)OCC DKMROQRQHGEIOW-UHFFFAOYSA-N 0.000 description 2
- JAGZUIGGHGTFHO-UHFFFAOYSA-N Ethyl 3-phenylpropanoate Chemical compound CCOC(=O)CCC1=CC=CC=C1 JAGZUIGGHGTFHO-UHFFFAOYSA-N 0.000 description 2
- NHXSTXWKZVAVOQ-UHFFFAOYSA-N Ethyl furoate Chemical compound CCOC(=O)C1=CC=CO1 NHXSTXWKZVAVOQ-UHFFFAOYSA-N 0.000 description 2
- ICMAFTSLXCXHRK-UHFFFAOYSA-N Ethyl pentanoate Chemical compound CCCCC(=O)OCC ICMAFTSLXCXHRK-UHFFFAOYSA-N 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 229960000583 acetic acid Drugs 0.000 description 2
- OBETXYAYXDNJHR-UHFFFAOYSA-N alpha-ethylcaproic acid Natural products CCCCC(CC)C(O)=O OBETXYAYXDNJHR-UHFFFAOYSA-N 0.000 description 2
- GONOPSZTUGRENK-UHFFFAOYSA-N benzyl(trichloro)silane Chemical compound Cl[Si](Cl)(Cl)CC1=CC=CC=C1 GONOPSZTUGRENK-UHFFFAOYSA-N 0.000 description 2
- POIARNZEYGURDG-FNORWQNLSA-N beta-damascenone Chemical compound C\C=C\C(=O)C1=C(C)C=CCC1(C)C POIARNZEYGURDG-FNORWQNLSA-N 0.000 description 2
- OWBTYPJTUOEWEK-UHFFFAOYSA-N butane-2,3-diol Chemical compound CC(O)C(C)O OWBTYPJTUOEWEK-UHFFFAOYSA-N 0.000 description 2
- QMVPMAAFGQKVCJ-UHFFFAOYSA-N citronellol Chemical compound OCCC(C)CCC=C(C)C QMVPMAAFGQKVCJ-UHFFFAOYSA-N 0.000 description 2
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- 229940067592 ethyl palmitate Drugs 0.000 description 2
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- 239000007788 liquid Substances 0.000 description 2
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- UAXOELSVPTZZQG-UHFFFAOYSA-N tiglic acid Natural products CC(C)=C(C)C(O)=O UAXOELSVPTZZQG-UHFFFAOYSA-N 0.000 description 2
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- 108090000790 Enzymes Proteins 0.000 description 1
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- 229920002633 Kraton (polymer) Polymers 0.000 description 1
- FEWJPZIEWOKRBE-UHFFFAOYSA-N Tartaric acid Natural products [H+].[H+].[O-]C(=O)C(O)C(O)C([O-])=O FEWJPZIEWOKRBE-UHFFFAOYSA-N 0.000 description 1
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- 239000001273 butane Substances 0.000 description 1
- 239000001569 carbon dioxide Substances 0.000 description 1
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- 239000012159 carrier gas Substances 0.000 description 1
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- 238000004587 chromatography analysis Methods 0.000 description 1
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- 229960002626 clarithromycin Drugs 0.000 description 1
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- 238000012258 culturing Methods 0.000 description 1
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- 235000013870 dimethyl polysiloxane Nutrition 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 238000000556 factor analysis Methods 0.000 description 1
- 239000000835 fiber Substances 0.000 description 1
- 235000011389 fruit/vegetable juice Nutrition 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- 238000000265 homogenisation Methods 0.000 description 1
- 238000009776 industrial production Methods 0.000 description 1
- 238000004949 mass spectrometry Methods 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- IJDNQMDRQITEOD-UHFFFAOYSA-N n-butane Chemical compound CCCC IJDNQMDRQITEOD-UHFFFAOYSA-N 0.000 description 1
- OFBQJSOFQDEBGM-UHFFFAOYSA-N n-pentane Natural products CCCCC OFBQJSOFQDEBGM-UHFFFAOYSA-N 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- CXQXSVUQTKDNFP-UHFFFAOYSA-N octamethyltrisiloxane Chemical compound C[Si](C)(C)O[Si](C)(C)O[Si](C)(C)C CXQXSVUQTKDNFP-UHFFFAOYSA-N 0.000 description 1
- 230000008520 organization Effects 0.000 description 1
- WVDDGKGOMKODPV-ZQBYOMGUSA-N phenyl(114C)methanol Chemical compound O[14CH2]C1=CC=CC=C1 WVDDGKGOMKODPV-ZQBYOMGUSA-N 0.000 description 1
- 238000004987 plasma desorption mass spectroscopy Methods 0.000 description 1
- 229920000435 poly(dimethylsiloxane) Polymers 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 238000004445 quantitative analysis Methods 0.000 description 1
- 230000000630 rising effect Effects 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 239000007790 solid phase Substances 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 150000008163 sugars Chemical class 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 235000002906 tartaric acid Nutrition 0.000 description 1
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Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12G—WINE; PREPARATION THEREOF; ALCOHOLIC BEVERAGES; PREPARATION OF ALCOHOLIC BEVERAGES NOT PROVIDED FOR IN SUBCLASSES C12C OR C12H
- C12G1/00—Preparation of wine or sparkling wine
- C12G1/02—Preparation of must from grapes; Must treatment and fermentation
- C12G1/0203—Preparation of must from grapes; Must treatment and fermentation by microbiological or enzymatic treatment
-
- 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
- C12N1/14—Fungi; Culture media therefor
- C12N1/16—Yeasts; 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
- 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
- C12N1/14—Fungi; Culture media therefor
- C12N1/16—Yeasts; Culture media therefor
- C12N1/18—Baker's yeast; Brewer's yeast
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12G—WINE; PREPARATION THEREOF; ALCOHOLIC BEVERAGES; PREPARATION OF ALCOHOLIC BEVERAGES NOT PROVIDED FOR IN SUBCLASSES C12C OR C12H
- C12G2200/00—Special features
- C12G2200/05—Use of particular microorganisms in the preparation of wine
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Genetics & Genomics (AREA)
- Biotechnology (AREA)
- Mycology (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Health & Medical Sciences (AREA)
- General Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- Microbiology (AREA)
- Botany (AREA)
- Medicinal Chemistry (AREA)
- Tropical Medicine & Parasitology (AREA)
- Virology (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses a mixed fermentation process based on cladinose and saccharomyces cerevisiae, belonging to the technical field of microorganisms. The mixed fermentation process based on the Kluyveromyces and the Saccharomyces cerevisiae is characterized in that the Kluyveromyces and the Saccharomyces cerevisiae are subjected to mixed fermentation. Compared with a single-bacterium fermented product, the mixed-bacterium fermented product has more unique fragrance, can produce a wine with unique flavor, fragrance and taste, fills the consumer category and expands the consumption selection.
Description
Technical Field
The invention belongs to the technical field of beverage fermentation, and particularly relates to a mixed fermentation process based on Kluyveromyces and saccharomyces cerevisiae.
Background
Wine fermentation is a complex biochemical process, and klaton yeast plays a very critical role in the fermentation process, such as the conversion of sugars to ethanol, carbon dioxide and other secondary metabolites in the thousands. A large number of scientific studies show that the quality of the wine is highly dependent on the metabolic activity and fermentation behavior of different cladonone yeasts, and the different cladonone yeasts have important contributions to the chemical composition, the organoleptic properties, the flavor characteristics and the like of the wine. Saccharomyces cerevisiae is the strain which is widely applied in the industrial production of wine so far, has the advantages of ensuring the risk of deterioration in the fermentation process of wine, and having good fermentation power, but has the problems of single flavor characteristic, serious homogenization phenomenon and the like of wine. Therefore, in order to pursue wine style specialization and make the aroma characteristics more representative, diverse and complex, brewers often adopt a method of mixing and fermenting saccharomyces cerevisiae and non-saccharomyces cerevisiae, especially some native klaton yeasts with strong adaptability and representativeness, so as to improve and enhance the flavor quality of wine.
Non-saccharomyces cerevisiae has become an option for improving the quality of wine. Numerous studies have shown that non-saccharomyces cerevisiae is able to produce enzymes as well as some of our desired secondary metabolites, thus improving wine aroma and flavor characteristics, and is able to control the growth of some undesirable species in wine, but has the disadvantage of insufficient fermentation power. The mixed fermentation of the non-saccharomyces cerevisiae and the saccharomyces cerevisiae can improve the aroma diversity and complexity of the wine, can make up for the problem of insufficient fermentation power of the non-saccharomyces cerevisiae, and is an effective method for improving the aroma quality of the wine.
The related reports of fermentation by the yeast Saccharomyces crataegensis are extremely rare at present in the field, and the related reports on the mixed fermentation process of the yeast Saccharomyces crataegensis and Saccharomyces cerevisiae are not found at present in the field.
Disclosure of Invention
Based on the above requirements and blanks in the field, the invention provides a mixed fermentation process based on the Kluyveromyces crataegensis and the Saccharomyces cerevisiae, and compared with single-strain fermentation of the Saccharomyces cerevisiae, the mixed fermentation process has the advantages that various aroma substances which can affect the flavor of wine are remarkably improved.
The technical scheme of the invention is as follows:
a mixed fermentation process based on Klatomyces and Saccharomyces cerevisiae is characterized in that the Klatomyces and the Saccharomyces cerevisiae are subjected to mixed fermentation.
Preferably, the yeast is a yeast species of the species Saccharomyces crataegensis strain YC 30; the collection number of the cladonospora crassa strain YC30 is CCTCC M2021089.
Preferably, the Saccharomyces cerevisiae refers to Saccharomyces cerevisiae strain F33.
The mixed fermentation process based on the Kluyveromyces and the Saccharomyces cerevisiae comprises the following steps: the activated strain and the activated strain are inoculated into a substrate for secondary fermentation for fermentation.
Preferably, said strain refers to: a strain of Saccharomyces crassoensis, strain YC30, or a strain of Saccharomyces cerevisiae, strain F33;
preferably, the activated strain refers to a strain inoculated in a culture medium for culture;
preferably, the culture temperature is 24-30 ℃, preferably 28 ℃; the culture time is 20-30h, preferably 24h, the rotation speed is 120-250rpm, preferably 150 rpm;
preferably, the inoculation refers to inoculating the initial strain stock solution into the culture medium according to the inoculation amount of 2-4%, preferably 3% in volume ratio;
preferably, the medium is a YPD medium; preferably, the YPD medium comprises the following components in mass-to-volume ratio: 0.5-3.5%, preferably 1%, of a yeast extract powder of klaton, 1.0-3.0%, preferably 2%, of peptone, 1.0-5.0%, preferably 2%, of glucose, and the balance water;
preferably, the initial strain stock solution refers to the strain stock in 15-35% by volume, preferably 25% glycerol/YPD medium;
more preferably, the activation of the strain is performed 1-3 times, preferably 2 times.
The activated strain refers to: the activated yeast strain of the cladosporium clavatum crataegensis YC30, or the activated saccharomyces cerevisiae strain F33;
preferably, the fermentation of the activated substrate into the secondary fermentation refers to: firstly inoculating an activated cladinose yeast Saccharomyces crataegensis strain YC30 to a substrate for fermentation for 0-96h, preferably 0h, 24h, 48h and 96h, and then inoculating an activated saccharomyces cerevisiae strain F33 for continuous fermentation;
preferably, the fermentation temperature is 18 ℃ +/-2 ℃;
preferably, the total inoculum size of the activated yeast strain Saccharomyces crataegensis YC30, and the activated strain of Saccharomyces cerevisiae F33 is 106-107CFU, preferably 6X 106CFU；
Preferably, the fermentation is terminated when the substrate weight loss does not change for 3 consecutive days;
preferably, the substrate is grape juice.
A production method of wine is characterized in that grape juice is used as a substrate, and mixed fermentation is carried out on Klatin yeast and saccharomyces cerevisiae;
the yeast is Saccharomyces crabayanus strain YC 30; the collection number of the cladonospora crassa strain YC30 is CCTCC M2021089.
The saccharomyces cerevisiae refers to saccharomyces cerevisiae strain F33.
The production method of the wine comprises the following steps: the activated strain and the activated strain are inoculated into a substrate for secondary fermentation for fermentation.
Preferably, said strain refers to: a strain of Saccharomyces crassoensis, strain YC30, or a strain of Saccharomyces cerevisiae, strain F33;
preferably, the activated strain refers to a strain inoculated in a culture medium for culture;
preferably, the culture temperature is 24-30 ℃, preferably 28 ℃; the culture time is 20-30h, preferably 24h, the rotation speed is 120-250rpm, preferably 150 rpm;
preferably, the inoculation refers to inoculating the initial strain preservation solution into the culture medium according to the inoculation amount of 3% by volume;
preferably, the medium is a YPD medium; preferably, the YPD medium comprises the following components in mass-to-volume ratio: 0.5-3.5%, preferably 1%, of a yeast extract powder of klaton, 1.0-3.0%, preferably 2%, of peptone, 1.0-5.0%, preferably 2%, of glucose, and the balance water;
preferably, the initial strain stock solution refers to the strain stock in 15-35% by volume, preferably 25% glycerol/YPD medium;
more preferably, the activation of the strain is performed 1-3 times, preferably 2 times.
The activated strain refers to: the activated yeast strain of the cladosporium clavatum crataegensis YC30, or the activated saccharomyces cerevisiae strain F33;
preferably, the fermentation of the activated substrate into the secondary fermentation refers to: firstly inoculating an activated cladinose yeast Saccharomyces crataegensis strain YC30 to a substrate for fermentation for 0-96h, preferably 0h, 24h, 48h or 96h, and then inoculating an activated saccharomyces cerevisiae strain F33 for continuous fermentation;
preferably, the fermentation temperature is 18 ℃ +/-2 ℃;
preferably, the total inoculum size of the activated yeast strain Saccharomyces crataegensis YC30 and the activated strain Saccharomyces cerevisiae F33 is 106-107CFU, preferably 6X 106CFU；
Preferably, the fermentation is terminated when the substrate weight loss is not changed for 3 consecutive days.
The wine production method further comprises the following steps: preparing grape juice;
the preparation of the grape juice comprises the following steps: squeezing grape fruit grains at low temperature;
preferably, the harvested grape fruit particles are pressed with ice by an air bag press;
preferably, pressing is performed while adding sulfur dioxide or K2S2O5And, pectinase;
preferably, sulfur dioxide or K2S2O5The addition amount of (B) is 30-100mg/L, preferably 50 mg/L; the addition amount of pectinase is 10-30mg/L, preferably 20 mg/L;
preferably, the grape fruit pieces are uniformly sized fruit pieces;
preferably, the grape fruit particles are grape fruit particles collected when the temperature of the mature grapes continuously drops below 24 hours to 8 ℃.
The wine is characterized by being produced by the wine production method.
The invention adopts the Klattorney yeast Saccharomyces crataegensis and Saccharomyces cerevisiae to carry out mixed fermentation, and the yield of most aroma substances in dozens of aroma substances generated is improved, for example, 1-methylnaphthalene is improved by 64.7 percent, 2-ethylhexanol is improved by 1.33 times, 1-pentanol is improved by 14 times, n-butanol is improved by 7.25 times, ethyl lactate is improved by 31.67 times, acetaldehyde is improved by 1.35 times, diethyl succinate is improved by 26.33 times, ethyl 3-hydroxybutyrate is improved by 1.1 times, 2-methylbutyric acid is improved by 15.7 times, isoamyl hexanoate is improved by 45.6 percent, ethyl acetate is improved by 51.75 times, n-hexanol is improved by 1.17 times, 2, 4-di-tert-butylphenol is improved by 9.52 times, hexanoic acid is improved by 20.36 times, 9-decenoic acid is improved by 19.23 times, linalool is improved by 14.25 times, 2, 3-butanediol is improved by 52.67 percent, benzyl alcohol is improved by 88.14 percent, and 1, 3-propylene glycol monoethyl ether is improved by 43.77 times, ethyl myristate by 97.8%, damascenone by 53.5%, ethyl acetate by 11 times, ethyl palmitate by 12.3 times, 4-vinyl-2-methoxyphenol by 47.4%, octanol by 1.1 times, citronella alcohol by 84.1%, ethyl hexanoate by 23.8 times, isoamyl acetate by 10.2 times, octanoic acid by 8.58 times, isobutanol by 55.7%, 1, 1-diethoxyethane by 1.83 times, ethyl octanoate by 51.1%, glacial acetic acid by 72.5%, and isoamyl alcohol by 39.8%; meanwhile, aroma substances which cannot be produced by single-strain fermentation of saccharomyces cerevisiae are also produced, such as: delta-decalactone, 2-methylnaphthalene, ethyl 2-furoate, nonanal, 3-methyl-3-buten-1-ol, isobutyl acetate, isoamyl octanoate, n-heptanol, ethyl valerate, butyric acid, 2, 3-butanedione, 3-hydroxy-2-butanone, 1-decanol, isovaleraldehyde, dodecanol, ethyl 3-phenylpropionate. The production of the aroma substances or the increase of the yield of the aroma substances can exert certain influence on the flavor and aroma of the beverage obtained by fermentation, so that the mixed fermentation product presents more unique aroma compared with a single-bacterium fermentation product, a wine beverage with unique flavor, aroma and taste can be produced, the consumer class is enriched, and the consumption selection is expanded.
Detailed Description
The following detailed description of the present invention will be made with reference to specific examples, but the scope of the present invention is not limited thereto.
Sources and documentations of biological materials
The yeast Saccharomyces crabayanus strain YC30 used in the experimental examples was a new strain selected in the applicant's laboratory with the following deposit information:
naming: YC30
And (4) classification name: clarithromycin yeast
The name of Latin is: saccharomyces crataegensis
The preservation number is as follows: CCTCC M2021089
The preservation organization: china center for type culture Collection
The preservation date is as follows: 1 month 15 days 2021;
saccharomyces cerevisiae F33 was a commercial strain purchased from Lafford (Laffort) France.
The grape variety used was a wital ice grape, purchased from Ningxia Bagges drunk intersomatic wine village, Inc.
Group 1 example, Mixed fermentation Process of the invention
The embodiment of the group provides a mixed fermentation process based on Klatomyces and Saccharomyces cerevisiae. All embodiments of this group share the following common features: and (3) carrying out mixed fermentation on the Kluyveromyces and the saccharomyces cerevisiae.
Those skilled in the art can use any combination of yeast from the group of Saccharomyces cerevisiae and Saccharomyces cerevisiae to perform fermentation and production in accordance with the teachings of the present invention. Target products of fermentation include, but are not limited to: wine drink, fermented milk, bread, etc.
In a preferred embodiment, the yeast is a yeast species Saccharomyces crataegensis strain YC 30; the collection number of the cladonospora crassa strain YC30 is CCTCC M2021089.
In a specific embodiment, the Saccharomyces cerevisiae is Saccharomyces cerevisiae strain F33.
According to the teaching of the present invention, other commercial strains can be selected by those skilled in the art for mixed fermentation, and besides F33, there are many commercial strains on the market, such as Saccharomyces cerevisiae V1116, Saccharomyces cerevisiae VL1, Saccharomyces cerevisiae X16, etc., which can be used for mixed fermentation with the Kluyveromyces crataegensis strain YC30 of the present invention, to achieve similar technical effects as the present invention.
In some embodiments, the mixed fermentation process based on the Kluyveromyces and the Saccharomyces cerevisiae comprises the following steps: the activated strain and the activated strain are inoculated into a substrate for secondary fermentation for fermentation.
Preferably, said strain refers to: a strain of Saccharomyces crassoensis, strain YC30, or a strain of Saccharomyces cerevisiae, strain F33;
preferably, the activated strain refers to a strain inoculated in a culture medium for culture;
preferably, the culture temperature is 24-30 ℃, preferably 28 ℃; the culture time is 20-30h, preferably 24h, the rotation speed is 120-250rpm, preferably 150 rpm;
preferably, the inoculation refers to inoculating the initial strain stock solution into the culture medium according to the inoculation amount of 2-4%, preferably 3% in volume ratio;
preferably, the medium is a YPD medium; preferably, the YPD medium comprises the following components in mass-to-volume ratio: 0.5-3.5%, preferably 1%, of a yeast extract powder of klaton, 1.0-3.0%, preferably 2%, of peptone, 1.0-5.0%, preferably 2%, of glucose, and the balance water;
preferably, the initial strain stock solution refers to the strain stock in 15-35% by volume, preferably 25% glycerol/YPD medium;
more preferably, the activation of the strain is performed 1-3 times, preferably 2 times.
In other embodiments, the activated strain refers to: the activated yeast strain of the cladosporium clavatum crataegensis YC30, or the activated saccharomyces cerevisiae strain F33;
preferably, the fermentation of the activated substrate into the secondary fermentation refers to: firstly inoculating an activated cladinose yeast Saccharomyces crataegensis strain YC30 to a substrate for fermentation for 0-96h, preferably 0h, 24h, 48h or 96h, and then inoculating an activated saccharomyces cerevisiae strain F33 for continuous fermentation;
preferably, the fermentation temperature is 18 ℃ +/-2 ℃;
preferably, the total inoculum size of the activated yeast strain Saccharomyces crataegensis YC30 and the activated strain Saccharomyces cerevisiae F33 is 106-107CFU, preferably 6X 106CFU；
Preferably, the fermentation is terminated when the substrate weight loss does not change for 3 consecutive days;
preferably, the substrate is grape juice.
Group 2 example, Process for the production of wine according to the invention
The present group of embodiments provides a method of producing wine. The present group of embodiments all have the following common features: taking grape juice as a substrate, and carrying out mixed fermentation on the Kluyveromyces and the saccharomyces cerevisiae.
In some preferred embodiments, the yeast Saccharomyces crataegus crataegensis refers to the yeast Saccharomyces crataegus crataegensis strain YC 30; the collection number of the cladonospora crassa strain YC30 is CCTCC M2021089.
In a specific embodiment, the Saccharomyces cerevisiae is Saccharomyces cerevisiae strain F33.
In other specific embodiments, the wine production method comprises: the activated strain and the activated strain are inoculated into a substrate for secondary fermentation for fermentation.
Preferably, said strain refers to: a strain of Saccharomyces crassoensis, strain YC30, or a strain of Saccharomyces cerevisiae, strain F33;
preferably, the activated strain refers to a strain inoculated in a culture medium for culture;
preferably, the culture temperature is 24-30 ℃, preferably 28 ℃; the culture time is 20-30h, preferably 24h, the rotation speed is 120-250rpm, preferably 150 rpm;
preferably, the inoculation refers to inoculating the initial strain stock solution into the culture medium according to the inoculation amount of 2-4%, preferably 3% in volume ratio;
preferably, the medium is a YPD medium; preferably, the YPD medium comprises the following components in mass-to-volume ratio: 0.5-3.5%, preferably 1%, of a yeast extract powder of klaton, 1.0-3.0%, preferably 2%, of peptone, 1.0-5.0%, preferably 2%, of glucose, and the balance water;
preferably, the initial strain stock solution refers to the strain stock in 15-35% by volume, preferably 25% glycerol/YPD medium;
more preferably, the activation of the strain is performed 1-3 times, preferably 2 times.
In specific embodiments, the activated strain refers to: the activated yeast strain of the cladosporium clavatum crataegensis YC30, or the activated saccharomyces cerevisiae strain F33;
preferably, the fermentation of the activated substrate into the secondary fermentation refers to: firstly inoculating an activated cladinose yeast Saccharomyces crataegensis strain YC30 to a substrate for fermentation for 0-96h, preferably 0h, 24h, 48h or 96h, and then inoculating an activated saccharomyces cerevisiae strain F33 for continuous fermentation;
when the time for inoculating the activated cladosporium Saccharomyces crataegensis strain YC30 to the substrate for fermentation is 0h, the activated cladosporium crataegensis strain YC30 and the activated saccharomyces cerevisiae strain F33 can be simultaneously inoculated to the substrate for fermentation until the substrate weight loss is not changed any more for 3 days, and the fermentation is stopped.
Preferably, the fermentation temperature is 18 ℃ +/-2 ℃;
preferably, the total inoculum size of the activated yeast strain Saccharomyces crataegensis YC30 and the activated strain Saccharomyces cerevisiae F33 is 106-107CFU, preferably 6X 106CFU；
Preferably, the fermentation is terminated when the substrate weight loss is not changed for 3 consecutive days.
In a further embodiment, the method of wine production further comprises: preparing grape juice;
the preparation of the grape juice comprises the following steps: squeezing grape fruit grains at low temperature;
preferably, the harvested grape fruit particles are pressed with ice by an air bag press;
preferably, pressing is performed while adding sulfur dioxide or K2S2O5And, pectinase;
preferably, sulfur dioxide or K2S2O5The addition amount of (B) is 30-100mg/L, preferably 50 mg/L; the addition amount of pectinase is 10-30mg/L, preferably 20 mg/L;
preferably, the grape fruit pieces are uniformly sized fruit pieces;
preferably, the grape fruit particles are grape fruit particles collected when the temperature of the mature grapes continuously drops below 24 hours to 8 ℃.
Group 3 examples, wine of the invention
The present group of embodiments provides a wine. All embodiments of this group share the following common features: produced by the wine production method of any one of the group 2 examples.
The wine of the invention produces the following aroma substances which cannot be produced by the wine fermented by single bacteria: the wine is prepared from the following raw materials, wherein the raw materials comprise butane decalactone, 2-methylnaphthalene, 2-furoic acid ethyl ester, nonanal, 3-methyl-3-butene-1-ol, isobutyl acetate, isoamyl octanoate, n-heptanol, ethyl valerate, butyric acid, 2, 3-butanedione, 3-hydroxy-2-butanone, 1-decanol, isovaleraldehyde, dodecanol and 3-phenylpropionic acid ethyl ester, and the contents of the following aroma substances are much higher than those of a single-bacterium fermented wine: 1-methylnaphthalene, 2-ethylhexanol, 1-pentanol, n-butanol, ethyl lactate, acetaldehyde, diethyl succinate, ethyl 3-hydroxybutyrate, 2-methylbutyric acid, isoamyl hexanoate, ethyl acetate, n-hexanol, 2, 4-di-tert-butylphenol, hexanoic acid, 9-decenoic acid, linalool, 2, 3-butanediol, benzyl alcohol, 1, 3-propanediol monoethyl ether, ethyl myristate, damascenone, phenethyl acetate, ethyl palmitate, 4-vinyl-2-methoxyphenol, octanol, citronellol, ethyl hexanoate, isoamyl acetate, octanoic acid, isobutanol, 1-diethoxyethane, ethyl octanoate, glacial acetic acid, isoamyl alcohol.
Experimental example, mixed bacteria fermentation process and fermentation data of the invention
1. Bacterial strains
The strains used in this experiment were: commercial s.cerevisiae F33 and the isolated and selected strain of the yeast Saccharomyces crataegensis YC30 according to the invention.
2. Grape juice
The Weidai ice grape raw material is planted in Yuquan Yingning county, Yinchuan city, Ningxia Bagges Zuius boundary wine village GmbH (E106.02 degree, N38.24 degree). The grape vines are planted in 2013, small-canopy-frame cultivation is adopted, the plant row spacing is 1.0m multiplied by 2.0m, the grape vines are harvested in 2017, the grape vines are not harvested after being mature, the grape vines are harvested when the temperature is reduced to be below 24 hours-8 ℃, small fruit grains are removed, and ice grape fruits with the same size are randomly selected and squeezed at low temperature. Pressing harvested ice grape with ice by air bag press while adding sulfur dioxide (50mg/L K)2S2O5) And 20mg/L of pectinase (more than or equal to 500U/mg), inhibits bacteria and improves the juice yield. The squeezed grape juice has sugar content of 432g/dm3Acidity of 4.65g/dm3(tartaric acid) pH 4.21.
3. Fermentation operation
2 strains: saccharomyces cerevisiae F33 strain, Saccharomyces clavulans crataegensis strain YC30 were all stored in 25% by volume glycerol/YPD medium prior to use. YPD cultureThe nutrient medium is 1% of kraton yeast extract powder, 2% of peptone and 2% of glucose. Respectively inoculating the strain with 3% of volume ratio into a 50mL triangular flask containing 40mLYPD culture medium and a 250mL triangular flask containing 150mLYPD culture medium, culturing at 28 deg.C and 150rpm for 24h to obtain the 1 st activated bacterial liquid, respectively inoculating the strain with 3% of volume ratio into a 50mL triangular flask containing 40mLYPD culture medium and a 250mL triangular flask containing 150mLYPD culture medium, and repeating the above culture to complete the 2 nd passage activation, thus obtaining the activated bacterial liquid. Firstly inoculating activated Saccharomyces crataensis crataegensis strain YC30 into collected grape juice, inoculating into S.cerevisiae F33 strain at inoculation ratio of YC30/F33 of 2:1 after 0 hr (namely, inoculating YC30 and F33 at the same time), or 24 hr, or 48 hr, or 96 hr, controlling total inoculation amount of strain at 6 × 106CFU, using S.cerevisiae F33 pure fermented grape juice as blank control, fermenting at 18 + -2 deg.C, and stopping fermentation when grape juice weight loss is not changed for three consecutive days. All wine samples were centrifuged at 7500rpm for 8 minutes and the supernatant was stored at 4 ℃.
4. Method for quantifying aroma substance
Headspace-solid phase microextraction-gas mass spectrometry (HS-SPME-GC/MS) was used. An 8mL sample of wine was accurately weighed into a headspace bottle containing 1.5g NaCl, while 394.08. mu.g/L4-methyl-1-pentanol (internal standard) was capped and sealed. Inserting CAR/DVB/PDMS extraction fiber, adsorbing at 45 deg.C for 30min, desorbing at 250 deg.C for 3min, and performing GC-MS analysis. A chromatographic column: InertCap WAX polar chromatography column (60m × 0.25mm, 0.25 μm); the temperature rising procedure is as follows: maintaining at 40 deg.C for 5min, heating to 120 deg.C at 3 deg.C/min, heating to 230 deg.C at 8 deg.C/min, and maintaining for 10 min; the flow rate of the carrier gas (He) was 0.8mL/min, and was not split. Electron bombardment ion source; electron energy 70 eV; the transmission line temperature was 275 ℃; the ion source temperature is 230 ℃; the activation voltage is 1.5V; the filament flow is 0.25 mA; the mass scanning range m/z is 33-450. Compound quantitative analysis was performed using external standard quantitation method.
5. Data analysis method
All samples were averaged in 3 replicates respectively. Single-factor analysis of variance (ANOVA) and Duncan's multi-range test (P <0.05) were performed using SPSS 22.0 for Windows (SPSS inc., Chicago, IL, US); unscamblebler 9.7(CAMOASA, Norway) was analyzed by partial least squares.
The final statistical interpretation gives the following Table 1, where the data are in μ g/L, meaning: the content of the aroma substances in each liter of wine.
TABLE 1
The aroma threshold value in the above table 1 refers to the lower limit value of the lowest concentration at which a human can smell the substance, and the aroma description and the threshold value are subject to the relevant literature reports that the aroma description and the threshold value of the aroma substance are recorded, and the aroma substance without the aroma description in the table is the literature that the relevant literature about the aroma description of the substance is not searched. "F33" in the header refers to data obtained by single fermentation using the commercial strain Saccharomyces cerevisiae F33, and "F33 _ YC 30" refers to data obtained by mixed fermentation using the commercial strains Saccharomyces cerevisiae F33 and Saccharomyces clavulans crataegenesis strain YC 30.
F33 refers to the data obtained by inoculating only Saccharomyces cerevisiae F33 for fermentation;
sc +48H F33 refers to data obtained by inoculating a cladinose strain YC30 for fermentation for 48h and then inoculating saccharomyces cerevisiae F33 for continuous fermentation;
sc +24H F33 refers to data obtained by inoculating a cladinose strain YC30 for fermentation for 24h and then inoculating saccharomyces cerevisiae F33 for continuous fermentation;
sc +96H F33 refers to data obtained by inoculating a cladinose strain YC30 for fermentation for 96h and then inoculating saccharomyces cerevisiae F33 for continuous fermentation;
sc + F33 refers to data obtained from fermentation by simultaneously inoculating a strain of Kluyveromyces Clarithrombus YC30 and a strain of Saccharomyces cerevisiae F33.
As is well known in the field of fermentation, factors influencing fermentation are numerous and complex, and the components of a fermented product can change due to the change of factors such as raw material batches, raw material components, fermentation conditions, temperature, time and the like, so that the condition that single-strain fermentation is higher than mixed-strain fermentation on certain aroma components can occur, which is a normal phenomenon in the field.
Claims (10)
1. A mixed fermentation process based on Klatomyces and Saccharomyces cerevisiae is characterized in that the Klatomyces and the Saccharomyces cerevisiae are subjected to mixed fermentation.
2. The fermentation process of claim 1, wherein the yeast is a yeast strain YC 30; the preservation number of the cladinose Saccharomyces crataegensis strain YC30 is CCTCC M2021089; preferably, the Saccharomyces cerevisiae refers to Saccharomyces cerevisiae strain F33.
3. The mixed fermentation process based on the yeast Saccharomyces crataegensis and Saccharomyces cerevisiae as claimed in claim 1, which comprises the following steps: the activated strain and the activated strain are inoculated into a substrate for secondary fermentation for fermentation.
Preferably, said strain refers to: a strain of Saccharomyces crassoensis, strain YC30, or a strain of Saccharomyces cerevisiae, strain F33;
preferably, the activated strain refers to a strain inoculated in a culture medium for culture;
preferably, the culture temperature is 24-30 ℃, preferably 28 ℃; the culture time is 20-30h, preferably 24h, the rotation speed is 120-250rpm, preferably 150 rpm;
preferably, the inoculation refers to inoculating the initial strain stock solution into the culture medium according to the inoculation amount of 2-4%, preferably 3% in volume ratio;
preferably, the medium is a YPD medium; preferably, the YPD medium comprises the following components in mass-to-volume ratio: 0.5-3.5%, preferably 1%, of a yeast extract powder of klaton, 1.0-3.0%, preferably 2%, of peptone, 1.0-5.0%, preferably 2%, of glucose, and the balance water;
preferably, the initial strain stock solution refers to the strain stock in 15-35% by volume, preferably 25% glycerol/YPD medium;
more preferably, the activation of the strain is performed 1-3 times, preferably 2 times.
4. The mixed fermentation process based on the Kluyveromyces and the Saccharomyces cerevisiae as claimed in claim 3, wherein the activated strains refer to: the activated yeast strain of the cladosporium clavatum crataegensis YC30, or the activated saccharomyces cerevisiae strain F33;
preferably, the fermentation of the activated substrate into the secondary fermentation refers to: firstly inoculating an activated cladinose yeast Saccharomyces crataegensis strain YC30 to a substrate for fermentation for 0-96h, preferably 0h, 24h, 48h or 96h, and then inoculating an activated saccharomyces cerevisiae strain F33 for continuous fermentation;
preferably, the fermentation temperature is 18 ℃ +/-2 ℃;
preferably, the total inoculum size of the activated yeast strain Saccharomyces crataegensis YC30, and the activated strain of Saccharomyces cerevisiae F33 is 106-107CFU, preferably 6X 106CFU；
Preferably, the fermentation is terminated when the substrate weight loss does not change for 3 consecutive days;
preferably, the substrate is grape juice.
5. A production method of wine is characterized in that grape juice is used as a substrate, and mixed fermentation is carried out on Klatin yeast and saccharomyces cerevisiae;
the yeast is Saccharomyces crabayanus strain YC 30; the collection number of the cladonospora crassa strain YC30 is CCTCC M2021089.
6. A wine production method according to claim 5, characterised in that the Saccharomyces cerevisiae is Saccharomyces cerevisiae strain F33.
7. A wine production process according to claim 5 or 6, comprising: the activated strain and the activated strain are inoculated into a substrate for secondary fermentation for fermentation.
Preferably, said strain refers to: a strain of Saccharomyces crassoensis, strain YC30, or a strain of Saccharomyces cerevisiae, strain F33;
preferably, the activated strain refers to a strain inoculated in a culture medium for culture;
preferably, the culture temperature is 24-30 ℃, preferably 28 ℃; the culture time is 20-30h, preferably 24h, the rotation speed is 120-250rpm, preferably 150 rpm;
preferably, the inoculation refers to inoculating the initial strain preservation solution into the culture medium according to the inoculation amount of 3% by volume;
preferably, the medium is a YPD medium; preferably, the YPD medium comprises the following components in mass-to-volume ratio: 0.5-3.5%, preferably 1%, of a yeast extract powder of klaton, 1.0-3.0%, preferably 2%, of peptone, 1.0-5.0%, preferably 2%, of glucose, and the balance water;
preferably, the initial strain stock solution refers to the strain stock in 15-35% by volume, preferably 25% glycerol/YPD medium;
more preferably, the activation of the strain is performed 1-3 times, preferably 2 times.
8. A wine production process according to claim 7, wherein the activated strain is: the activated yeast strain of the cladosporium clavatum crataegensis YC30, or the activated saccharomyces cerevisiae strain F33;
preferably, the fermentation of the activated substrate into the secondary fermentation refers to: firstly inoculating an activated Kluyveromyces crataegensis strain YC30 to a substrate for fermentation for 0-96h, preferably 48h, and then inoculating an activated Saccharomyces cerevisiae strain F33 for continuous fermentation;
preferably, the fermentation temperature is 18 ℃ +/-2 ℃;
preferably, the total inoculum size of the activated yeast strain Saccharomyces crataegensis YC30 and the activated strain Saccharomyces cerevisiae F33 is 106-107CFU, preferably 6X 106CFU；
Preferably, the fermentation is terminated when the substrate weight loss is not changed for 3 consecutive days.
9. A wine production process according to any one of claims 5 to 8, further comprising: preparing grape juice;
the preparation of the grape juice comprises the following steps: squeezing grape fruit grains at low temperature;
preferably, the harvested grape fruit particles are pressed with ice by an air bag press;
preferably, pressing is performed while adding sulfur dioxide or K2S2O5And, pectinase;
preferably, sulfur dioxide or K2S2O5The addition amount of (B) is 30-100mg/L, preferably 50 mg/L; the addition amount of pectinase is 10-30mg/L, preferably 20 mg/L;
preferably, the grape fruit pieces are uniformly sized fruit pieces;
preferably, the grape fruit particles are grape fruit particles collected when the temperature of the mature grapes continuously drops below 24 hours to 8 ℃.
10. Wine produced by the wine production method of any one of claims 5 to 9.
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| JP2004254554A (en) * | 2003-02-25 | 2004-09-16 | Mitsubishi Chemicals Corp | Method of producing optically active 2-methoxy-1-(4-trifluoromethyl-phenyl)ethanol |
| CN106987531A (en) * | 2017-05-05 | 2017-07-28 | 湖南农业大学 | One plant of grape wine laminating adhesive spore yeast and its application in monoterpene production |
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| JP2004254554A (en) * | 2003-02-25 | 2004-09-16 | Mitsubishi Chemicals Corp | Method of producing optically active 2-methoxy-1-(4-trifluoromethyl-phenyl)ethanol |
| CN106987531A (en) * | 2017-05-05 | 2017-07-28 | 湖南农业大学 | One plant of grape wine laminating adhesive spore yeast and its application in monoterpene production |
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