CN117778210A - Candida glabrata strain 196 and application, product and method thereof - Google Patents
Candida glabrata strain 196 and application, product and method thereof Download PDFInfo
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- CN117778210A CN117778210A CN202311829435.2A CN202311829435A CN117778210A CN 117778210 A CN117778210 A CN 117778210A CN 202311829435 A CN202311829435 A CN 202311829435A CN 117778210 A CN117778210 A CN 117778210A
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- Prior art keywords
- ethyl
- fermentation
- strain
- acid
- methyl
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- 241000222126 [Candida] glabrata Species 0.000 title claims abstract description 57
- 208000032343 candida glabrata infection Diseases 0.000 title claims abstract description 57
- 238000000034 method Methods 0.000 title claims abstract description 14
- 238000000855 fermentation Methods 0.000 claims abstract description 130
- 230000004151 fermentation Effects 0.000 claims abstract description 129
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims abstract description 31
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 claims abstract description 31
- 238000004321 preservation Methods 0.000 claims abstract description 28
- 239000000126 substance Substances 0.000 claims abstract description 20
- XFRVVPUIAFSTFO-UHFFFAOYSA-N 1-Tridecanol Chemical compound CCCCCCCCCCCCCO XFRVVPUIAFSTFO-UHFFFAOYSA-N 0.000 claims abstract description 18
- 241000222120 Candida <Saccharomycetales> Species 0.000 claims abstract description 16
- OHEFFKYYKJVVOX-UHFFFAOYSA-N sulcatol Chemical compound CC(O)CCC=C(C)C OHEFFKYYKJVVOX-UHFFFAOYSA-N 0.000 claims abstract description 16
- NMRPBPVERJPACX-UHFFFAOYSA-N octan-3-ol Chemical compound CCCCCC(O)CC NMRPBPVERJPACX-UHFFFAOYSA-N 0.000 claims abstract description 12
- RUOPINZRYMFPBF-UHFFFAOYSA-N pentane-1,3-diol Chemical compound CCC(O)CCO RUOPINZRYMFPBF-UHFFFAOYSA-N 0.000 claims abstract description 10
- 239000007858 starting material Substances 0.000 claims abstract description 10
- JRTBBCBDKSRRCY-UHFFFAOYSA-N 3,7-dimethyloct-6-en-3-ol Chemical compound CCC(C)(O)CCC=C(C)C JRTBBCBDKSRRCY-UHFFFAOYSA-N 0.000 claims abstract description 8
- NMRPBPVERJPACX-QMMMGPOBSA-N 3-Octanol Natural products CCCCC[C@@H](O)CC NMRPBPVERJPACX-QMMMGPOBSA-N 0.000 claims abstract description 6
- MMXKVMNBHPAILY-UHFFFAOYSA-N ethyl laurate Chemical compound CCCCCCCCCCCC(=O)OCC MMXKVMNBHPAILY-UHFFFAOYSA-N 0.000 claims description 32
- MDHYEMXUFSJLGV-UHFFFAOYSA-N phenethyl acetate Chemical compound CC(=O)OCCC1=CC=CC=C1 MDHYEMXUFSJLGV-UHFFFAOYSA-N 0.000 claims description 30
- KBPLFHHGFOOTCA-UHFFFAOYSA-N 1-Octanol Chemical compound CCCCCCCCO KBPLFHHGFOOTCA-UHFFFAOYSA-N 0.000 claims description 28
- HBEDSQVIWPRPAY-UHFFFAOYSA-N 2,3-dihydrobenzofuran Chemical compound C1=CC=C2OCCC2=C1 HBEDSQVIWPRPAY-UHFFFAOYSA-N 0.000 claims description 28
- 239000000758 substrate Substances 0.000 claims description 28
- WWZKQHOCKIZLMA-UHFFFAOYSA-N octanoic acid Chemical compound CCCCCCCC(O)=O WWZKQHOCKIZLMA-UHFFFAOYSA-N 0.000 claims description 26
- REIUXOLGHVXAEO-UHFFFAOYSA-N pentadecan-1-ol Chemical compound CCCCCCCCCCCCCCCO REIUXOLGHVXAEO-UHFFFAOYSA-N 0.000 claims description 24
- 239000001963 growth medium Substances 0.000 claims description 22
- DNIAPMSPPWPWGF-UHFFFAOYSA-N Propylene glycol Chemical compound CC(O)CO DNIAPMSPPWPWGF-UHFFFAOYSA-N 0.000 claims description 21
- MTHSVFCYNBDYFN-UHFFFAOYSA-N diethylene glycol Chemical compound OCCOCCO MTHSVFCYNBDYFN-UHFFFAOYSA-N 0.000 claims description 21
- 241000894006 Bacteria Species 0.000 claims description 18
- -1 geranyl acetonate Chemical compound 0.000 claims description 18
- 239000000463 material Substances 0.000 claims description 18
- 235000019674 grape juice Nutrition 0.000 claims description 17
- ALVGHPMGQNBJRC-UHFFFAOYSA-N pentadecan-2-ol Chemical compound CCCCCCCCCCCCCC(C)O ALVGHPMGQNBJRC-UHFFFAOYSA-N 0.000 claims description 16
- YOMSJEATGXXYPX-UHFFFAOYSA-N 2-methoxy-4-vinylphenol Chemical compound COC1=CC(C=C)=CC=C1O YOMSJEATGXXYPX-UHFFFAOYSA-N 0.000 claims description 15
- IGIDLTISMCAULB-YFKPBYRVSA-N (3s)-3-methylpentanoic acid Chemical compound CC[C@H](C)CC(O)=O IGIDLTISMCAULB-YFKPBYRVSA-N 0.000 claims description 14
- JSNRRGGBADWTMC-UHFFFAOYSA-N (6E)-7,11-dimethyl-3-methylene-1,6,10-dodecatriene Chemical compound CC(C)=CCCC(C)=CCCC(=C)C=C JSNRRGGBADWTMC-UHFFFAOYSA-N 0.000 claims description 14
- BBMCTIGTTCKYKF-UHFFFAOYSA-N 1-heptanol Chemical compound CCCCCCCO BBMCTIGTTCKYKF-UHFFFAOYSA-N 0.000 claims description 14
- XKWSWANXMRXDES-UHFFFAOYSA-N 3-methylbutyl octanoate Chemical compound CCCCCCCC(=O)OCCC(C)C XKWSWANXMRXDES-UHFFFAOYSA-N 0.000 claims description 14
- JELGPLUONQGOHF-UHFFFAOYSA-N Hexadec-9-ensaeure-ethylester Natural products CCCCCCC=CCCCCCCCC(=O)OCC JELGPLUONQGOHF-UHFFFAOYSA-N 0.000 claims description 14
- IGIDLTISMCAULB-UHFFFAOYSA-N anteisohexanoic acid Natural products CCC(C)CC(O)=O IGIDLTISMCAULB-UHFFFAOYSA-N 0.000 claims description 14
- OWBTYPJTUOEWEK-UHFFFAOYSA-N butane-2,3-diol Chemical compound CC(O)C(C)O OWBTYPJTUOEWEK-UHFFFAOYSA-N 0.000 claims description 14
- BQOFWKZOCNGFEC-UHFFFAOYSA-N carene Chemical compound C1C(C)=CCC2C(C)(C)C12 BQOFWKZOCNGFEC-UHFFFAOYSA-N 0.000 claims description 14
- KHAVLLBUVKBTBG-UHFFFAOYSA-N dec-9-enoic acid Chemical compound OC(=O)CCCCCCCC=C KHAVLLBUVKBTBG-UHFFFAOYSA-N 0.000 claims description 14
- MGWAVDBGNNKXQV-UHFFFAOYSA-N diisobutyl phthalate Chemical compound CC(C)COC(=O)C1=CC=CC=C1C(=O)OCC(C)C MGWAVDBGNNKXQV-UHFFFAOYSA-N 0.000 claims description 14
- GFJVXXWOPWLRNU-UHFFFAOYSA-N ethenyl formate Chemical compound C=COC=O GFJVXXWOPWLRNU-UHFFFAOYSA-N 0.000 claims description 14
- BKOJTZORTHALGP-UHFFFAOYSA-N ethyl 9-decenoate Chemical compound CCOC(=O)CCCCCCCC=C BKOJTZORTHALGP-UHFFFAOYSA-N 0.000 claims description 14
- JELGPLUONQGOHF-MDZDMXLPSA-N ethyl 9-hexadecenoate Chemical compound CCCCCC\C=C\CCCCCCCC(=O)OCC JELGPLUONQGOHF-MDZDMXLPSA-N 0.000 claims description 14
- RGXWDWUGBIJHDO-UHFFFAOYSA-N ethyl decanoate Chemical compound CCCCCCCCCC(=O)OCC RGXWDWUGBIJHDO-UHFFFAOYSA-N 0.000 claims description 14
- TVQGDYNRXLTQAP-UHFFFAOYSA-N ethyl heptanoate Chemical compound CCCCCCC(=O)OCC TVQGDYNRXLTQAP-UHFFFAOYSA-N 0.000 claims description 14
- PPXUHEORWJQRHJ-UHFFFAOYSA-N ethyl isovalerate Chemical compound CCOC(=O)CC(C)C PPXUHEORWJQRHJ-UHFFFAOYSA-N 0.000 claims description 14
- YYZUSRORWSJGET-UHFFFAOYSA-N ethyl octanoate Chemical compound CCCCCCCC(=O)OCC YYZUSRORWSJGET-UHFFFAOYSA-N 0.000 claims description 14
- MLFHJEHSLIIPHL-UHFFFAOYSA-N isoamyl acetate Chemical compound CC(C)CCOC(C)=O MLFHJEHSLIIPHL-UHFFFAOYSA-N 0.000 claims description 14
- KQNPFQTWMSNSAP-UHFFFAOYSA-N isobutyric acid Chemical compound CC(C)C(O)=O KQNPFQTWMSNSAP-UHFFFAOYSA-N 0.000 claims description 14
- 238000004519 manufacturing process Methods 0.000 claims description 14
- GYHFUZHODSMOHU-UHFFFAOYSA-N nonanal Chemical compound CCCCCCCCC=O GYHFUZHODSMOHU-UHFFFAOYSA-N 0.000 claims description 14
- IDHBLVYDNJDWNO-UHFFFAOYSA-N propyl octanoate Chemical compound CCCCCCCC(=O)OCCC IDHBLVYDNJDWNO-UHFFFAOYSA-N 0.000 claims description 14
- XMUJIPOFTAHSOK-UHFFFAOYSA-N undecan-2-ol Chemical compound CCCCCCCCCC(C)O XMUJIPOFTAHSOK-UHFFFAOYSA-N 0.000 claims description 14
- OBETXYAYXDNJHR-UHFFFAOYSA-N alpha-ethylcaproic acid Natural products CCCCC(CC)C(O)=O OBETXYAYXDNJHR-UHFFFAOYSA-N 0.000 claims description 13
- KSMVZQYAVGTKIV-UHFFFAOYSA-N decanal Chemical compound CCCCCCCCCC=O KSMVZQYAVGTKIV-UHFFFAOYSA-N 0.000 claims description 12
- DOIRQSBPFJWKBE-UHFFFAOYSA-N dibutyl phthalate Chemical compound CCCCOC(=O)C1=CC=CC=C1C(=O)OCCCC DOIRQSBPFJWKBE-UHFFFAOYSA-N 0.000 claims description 12
- 235000014787 Vitis vinifera Nutrition 0.000 claims description 11
- XYFCBTPGUUZFHI-UHFFFAOYSA-N Phosphine Chemical compound P XYFCBTPGUUZFHI-UHFFFAOYSA-N 0.000 claims description 10
- HNZUNIKWNYHEJJ-FMIVXFBMSA-N geranyl acetone Chemical compound CC(C)=CCC\C(C)=C\CCC(C)=O HNZUNIKWNYHEJJ-FMIVXFBMSA-N 0.000 claims description 10
- 239000003795 chemical substances by application Substances 0.000 claims description 9
- XCLIHDJZGPCUBT-UHFFFAOYSA-N dimethylsilanediol Chemical compound C[Si](C)(O)O XCLIHDJZGPCUBT-UHFFFAOYSA-N 0.000 claims description 9
- CZCBTSFUTPZVKJ-NXEZZACHSA-N (2S,4R)-rose oxide Chemical compound C[C@@H]1CCO[C@H](C=C(C)C)C1 CZCBTSFUTPZVKJ-NXEZZACHSA-N 0.000 claims description 8
- AFHJYKBGDDJSRR-UHFFFAOYSA-N 1-propan-2-yloxypropan-2-ol Chemical compound CC(C)OCC(C)O AFHJYKBGDDJSRR-UHFFFAOYSA-N 0.000 claims description 8
- AWNIQMQADACLCJ-CMDGGOBGSA-N Ethyl 4-decenoate Chemical compound CCCCC\C=C\CCC(=O)OCC AWNIQMQADACLCJ-CMDGGOBGSA-N 0.000 claims description 8
- PTEYJUIKYIKULL-UHFFFAOYSA-N Ethyl pentadecanoate Chemical compound CCCCCCCCCCCCCCC(=O)OCC PTEYJUIKYIKULL-UHFFFAOYSA-N 0.000 claims description 8
- OKDGZLITBCRLLJ-UHFFFAOYSA-N dodecan-3-ol Chemical compound CCCCCCCCCC(O)CC OKDGZLITBCRLLJ-UHFFFAOYSA-N 0.000 claims description 8
- 239000002068 microbial inoculum Substances 0.000 claims description 8
- CXENHBSYCFFKJS-UHFFFAOYSA-N (3E,6E)-3,7,11-Trimethyl-1,3,6,10-dodecatetraene Natural products CC(C)=CCCC(C)=CCC=C(C)C=C CXENHBSYCFFKJS-UHFFFAOYSA-N 0.000 claims description 7
- ADLXTJMPCFOTOO-BQYQJAHWSA-N (E)-non-2-enoic acid Chemical compound CCCCCC\C=C\C(O)=O ADLXTJMPCFOTOO-BQYQJAHWSA-N 0.000 claims description 7
- CSZZMFWKAQEMPB-UHFFFAOYSA-N 1-methoxybutan-2-ol Chemical compound CCC(O)COC CSZZMFWKAQEMPB-UHFFFAOYSA-N 0.000 claims description 7
- UXZIDIYMFIBDKT-UHFFFAOYSA-N 1-methyl-3-prop-1-en-2-ylcyclohexene Chemical compound CC(=C)C1CCCC(C)=C1 UXZIDIYMFIBDKT-UHFFFAOYSA-N 0.000 claims description 7
- NZRNIYUJHYKFBO-UHFFFAOYSA-N 3-chloro-2-methylpentan-2-ol Chemical compound CCC(Cl)C(C)(C)O NZRNIYUJHYKFBO-UHFFFAOYSA-N 0.000 claims description 7
- WBLZUCOIBUDNBV-UHFFFAOYSA-N 3-nitropropanoic acid Chemical compound OC(=O)CC[N+]([O-])=O WBLZUCOIBUDNBV-UHFFFAOYSA-N 0.000 claims description 7
- 241000219095 Vitis Species 0.000 claims description 7
- 235000009754 Vitis X bourquina Nutrition 0.000 claims description 7
- 235000012333 Vitis X labruscana Nutrition 0.000 claims description 7
- ZCZSIDMEHXZRLG-UHFFFAOYSA-N acetic acid heptyl ester Natural products CCCCCCCOC(C)=O ZCZSIDMEHXZRLG-UHFFFAOYSA-N 0.000 claims description 7
- XAPCMTMQBXLDBB-UHFFFAOYSA-N butanoic acid hexyl ester Natural products CCCCCCOC(=O)CCC XAPCMTMQBXLDBB-UHFFFAOYSA-N 0.000 claims description 7
- 229930006737 car-3-ene Natural products 0.000 claims description 7
- SHZIWNPUGXLXDT-UHFFFAOYSA-N ethyl hexanoate Chemical compound CCCCCC(=O)OCC SHZIWNPUGXLXDT-UHFFFAOYSA-N 0.000 claims description 7
- DWMKJZMYLQUIDA-UHFFFAOYSA-N ethyl oct-7-enoate Chemical compound CCOC(=O)CCCCCC=C DWMKJZMYLQUIDA-UHFFFAOYSA-N 0.000 claims description 7
- 229930009668 farnesene Natural products 0.000 claims description 7
- HNZUNIKWNYHEJJ-UHFFFAOYSA-N geranyl acetone Natural products CC(C)=CCCC(C)=CCCC(C)=O HNZUNIKWNYHEJJ-UHFFFAOYSA-N 0.000 claims description 7
- JPXGPRBLTIYFQG-UHFFFAOYSA-N heptan-4-yl acetate Chemical compound CCCC(CCC)OC(C)=O JPXGPRBLTIYFQG-UHFFFAOYSA-N 0.000 claims description 7
- GOKKOFHHJFGZHW-UHFFFAOYSA-N hexyl propanoate Chemical compound CCCCCCOC(=O)CC GOKKOFHHJFGZHW-UHFFFAOYSA-N 0.000 claims description 7
- VNWKTOKETHGBQD-UHFFFAOYSA-N methane Natural products C VNWKTOKETHGBQD-UHFFFAOYSA-N 0.000 claims description 7
- DILOFCBIBDMHAY-UHFFFAOYSA-N methyl 2-(3,4-dimethoxyphenyl)acetate Chemical compound COC(=O)CC1=CC=C(OC)C(OC)=C1 DILOFCBIBDMHAY-UHFFFAOYSA-N 0.000 claims description 7
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 claims description 7
- 230000000813 microbial effect Effects 0.000 claims description 7
- LQAVWYMTUMSFBE-UHFFFAOYSA-N pent-4-en-1-ol Chemical compound OCCCC=C LQAVWYMTUMSFBE-UHFFFAOYSA-N 0.000 claims description 7
- BSCCSDNZEIHXOK-UHFFFAOYSA-N phenyl carbamate Chemical compound NC(=O)OC1=CC=CC=C1 BSCCSDNZEIHXOK-UHFFFAOYSA-N 0.000 claims description 7
- WLAMNBDJUVNPJU-UHFFFAOYSA-N 2-methylbutyric acid Chemical compound CCC(C)C(O)=O WLAMNBDJUVNPJU-UHFFFAOYSA-N 0.000 claims description 6
- JCEAFZUEUSBESW-UHFFFAOYSA-N 4-(4-formylphenyl)benzoic acid Chemical compound C1=CC(C(=O)O)=CC=C1C1=CC=C(C=O)C=C1 JCEAFZUEUSBESW-UHFFFAOYSA-N 0.000 claims description 6
- UXDDRFCJKNROTO-UHFFFAOYSA-N Glycerol 1,2-diacetate Chemical compound CC(=O)OCC(CO)OC(C)=O UXDDRFCJKNROTO-UHFFFAOYSA-N 0.000 claims description 6
- YYBMOGCOPQVSLQ-SAPNQHFASA-N [(2e)-3,7-dimethylocta-2,6-dienyl] octanoate Chemical compound CCCCCCCC(=O)OC\C=C(/C)CCC=C(C)C YYBMOGCOPQVSLQ-SAPNQHFASA-N 0.000 claims description 6
- YYBMOGCOPQVSLQ-UHFFFAOYSA-N geranyl octanoate Natural products CCCCCCCC(=O)OCC=C(C)CCC=C(C)C YYBMOGCOPQVSLQ-UHFFFAOYSA-N 0.000 claims description 6
- CDOSHBSSFJOMGT-UHFFFAOYSA-N linalool Chemical compound CC(C)=CCCC(C)(O)C=C CDOSHBSSFJOMGT-UHFFFAOYSA-N 0.000 claims description 6
- 230000004580 weight loss Effects 0.000 claims description 6
- GIEMHYCMBGELGY-UHFFFAOYSA-N 10-undecen-1-ol Chemical compound OCCCCCCCCCC=C GIEMHYCMBGELGY-UHFFFAOYSA-N 0.000 claims description 5
- JCTXKRPTIMZBJT-UHFFFAOYSA-N 2,2,4-trimethylpentane-1,3-diol Chemical compound CC(C)C(O)C(C)(C)CO JCTXKRPTIMZBJT-UHFFFAOYSA-N 0.000 claims description 5
- GISVICWQYMUPJF-UHFFFAOYSA-N 2,4-Dimethylbenzaldehyde Chemical compound CC1=CC=C(C=O)C(C)=C1 GISVICWQYMUPJF-UHFFFAOYSA-N 0.000 claims description 5
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- FOTKYAAJKYLFFN-UHFFFAOYSA-N decane-1,10-diol Chemical compound OCCCCCCCCCCO FOTKYAAJKYLFFN-UHFFFAOYSA-N 0.000 claims description 5
- 125000001495 ethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 claims description 5
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- 229910000073 phosphorus hydride Inorganic materials 0.000 claims description 5
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- 125000000026 trimethylsilyl group Chemical group [H]C([H])([H])[Si]([*])(C([H])([H])[H])C([H])([H])[H] 0.000 claims description 5
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Abstract
The invention discloses a Candida glabrata (Candida glabra) strain 196 and application, a product and a method thereof, belonging to the technical field of microorganisms. The invention provides a Candida glabrata (Candida glabrata) strain 196, the preservation number of which is CCTCC NO: M2023928, and the invention also provides the application of the strain 196 in the aspects of producing, regulating aroma substances and/or fermenting and/or brewing, and a fermentation process, a brewing starter and a wine brewing method based on the strain 196. Compared with the commercial saccharomyces cerevisiae strain F15, the strain 196 of the invention can produce aroma substances which cannot be produced by the F15 single-strain fermentation, including 1-tridecanol, 1, 3-pentanediol, 6-methyl-5-hepten-2-ol, dihydro linalool, 3-octanol and the like.
Description
Technical Field
The invention belongs to the technical field of microorganisms, and particularly relates to a Candida glabrata (Candida glabrata) strain 196, and application, a product and a method thereof.
Background
Candida glabrata is a microorganism of Candida genus (Candida), also called Candida glabrata, and is a pathogenic fungus. Candida glabrata has been reported to be used for brewing vinegar.
At present, no report of aroma substances such as TRANS-4-decenoic acid ethyl ester, (2S-cis) -tetrahydro-4-methyl-2- (2-methyl-1-propenyl) -2H-pyran, 2-pentadecanol, 3-dodecanol, 3-chloro-2-methyl-2-amyl alcohol, dimethyl silanediol, 1-tridecanol, 1, 3-pentanediol, 6-methyl-5-hepten-2-ol, ALL TRANS-3,7, 11-15-tetramethyl-2, 6,10, 14-hexadecen-1-ol, dihydro linalool and the like is seen in the fermentation and brewing of wine by candida glabrata.
Disclosure of Invention
The invention provides a Candida glabrata (Candida glabrata) strain 196, application, product and method thereof, and aims to solve the technical problems of filling up the blank of brewing wine by Candida glabrata, producing and regulating aroma substances and developing more new applications of Candida glabrata.
The technical scheme of the invention is as follows:
a Candida glabrata (Candida glabrata) strain 196 has a preservation number of CCTCC NO: M2023928.
Use of Candida glabrata strain 196 with a accession number of cctccc No. M2023928 for production, regulation of aroma substances and/or fermentation and/or brewing.
The fragrance material is selected from the group consisting of ethyl heptanoate, ethyl octanoate, ethyl laurate, ethyl isovalerate, ethyl decanoate, ethyl n-hexanoate, ethyl 2-enolate, ethyl 7-octenoate, ethyl 9-hexadecenoate, ethyl 9-decenoate, geranyl acetonate, methyl triacontate C30, phenethyl acetate, heptyl acetate, isopentyl acetate, diethylene glycol, n-heptanol, octanol, 2-undecanol, (2R, 3R) - (-) -2, 3-butanediol, 1-isopropoxy-2-propanol, 1-methoxy-2-butanol, propylene glycol, 4-penten-1-ol, 4-vinyl-2-methoxyphenol, 2, 3-dihydrobenzofuran, (1-acetoxy-3-hydroxypropion-2-yl) acetate, isopentyl octanoate, hexyl propionate, hexyl butyrate, geranyl octanoate, phenyl carbamate, propyl octanoate, vinyl formate, diisobutyl phthalate, pentadecanol, decanal, nonanal, 3-methylpentanoic acid, ethyl- (Z) -4-decanoate, 62-decanoate, methyl-2-propanol, n-decanoate, 4-hydroxy-10-butanete, n-1-methyl-4-decanoate, n-acetyl-2-methoxymethyl-butanoate, n-4-decanoate, n-methyl-10-decanoate, n-butyl acetate, n-decanoate, n-methyl-10-decanoate, n-butyl acetate, n-methyl-decanoate, n-methyl-10-decanoate, TRANS-2-nonenoic acid, 3-nitropropionic acid, 9-decenoic acid, 3-carene, farnesene, 3-isopropenyl-1-methylcyclohexene, 6, 10-dimethyl-5, 9-undecadiene-2-one, ethyl laurate, ethyl 9-hexadecenoate, ethyl 9-decenoate, phenethyl acetate, octanol, 4-vinyl-2-methoxyphenol, 2, 3-dihydrobenzofuran, isobutyl 2, 4-trimethylpentanediol, vinyl formate, 3-methylpentanoic acid, octanoic acid, 2-methyl-propionic acid, large Ma Shitong, TRANS-4-decenoic acid ethyl ester, (2S-cis) -tetrahydro4-methyl-2- (2-methyl-1-propenyl) -2H-pyran, 2-pentadecanol, 3-dodecanol, 3-chloro-2-methyl-2-pentanol, dimethylsilanediol, 1-tridecanol, 1, 3-pentanediol, 6-methyl-5-hepten-2-ol, ALL-3, 11, 15-tetramethyl-2-butanediol, 4-dimethylbutyrate, 2-dimethylbutyrate, 4-dimethylbenzoic acid, 2-dimethylbutyrate, 4-dimethylbenzene, 2-dimethylbenzene-n-4-dimethylbutyrate;
preferably, the production refers to the single bacterial fermentation to produce TRANS-4-decenoic acid ethyl ester, (2S-cis) -tetrahydro-4-methyl-2- (2-methyl-1-propenyl) -2H-pyran, 2-pentadecanol, 3-dodecanol, 3-chloro-2-methyl-2-pentanol, dimethylsilanediol, 1-tridecanol, 1, 3-pentanediol, 6-methyl-5-hepten-2-ol, ALL TRANS-3,7, 11-15-tetramethyl-2, 6,10, 14-hexadecanetetraen-1-ol, dihydrolinalool, 3-octanol, n-nonyl chloroformate, 2, 4-trimethyl-1, 3-pentanediol monoisobutyrate, dibutyl phthalate, 2, 4-dimethylbenzaldehyde, 2-methylbutanoic acid, terpinene; or, mixed bacteria fermentation to produce ethyl 3-phenylpropionate, ethyl 4-acetoxybutyrate, ethyl pentadecanoate, (trimethylsilyl) oxy ] phosphine ] acetate trimethylsilyl ester, 10-undecen-1-ol, 1, 10-decanediol, 1-pentadecanol, rose ether, isobutyl decanoate, n-propyl decanoate, trans-2-nonenoic acid, 3-nitropropionic acid, 9-decenoic acid, 3-carene, farnesene, 3-isopropenyl-1-methylcyclohexene, 6, 10-dimethyl-5, 9-undecadien-2-one;
preferably, the adjustment means: during mixed fermentation, the ethyl laurate, the ethyl 9-hexadecenoate, the ethyl 9-decenoate, the phenethyl acetate, the octanol, the 4-vinyl-2-methoxyphenol, the 2, 3-dihydrobenzofuran, the 2, 4-trimethylpentanediol isobutyl ester, the vinyl formate, the 3-methylpentanoic acid, the octanoic acid, the 2-methyl-propionic acid and the large Ma Shitong are improved, or reducing ethyl heptanoate, ethyl octanoate, ethyl laurate, ethyl isovalerate, ethyl decanoate, ethyl n-hexanoate, ethyl hex-2-enoate, ethyl 7-octenoate, ethyl 9-hexadecenoate, ethyl 9-decenoate, geranyl acetone, methyl triacontate C30, phenethyl acetate, heptyl acetate, isopentyl acetate, diethylene glycol, n-heptanol, octanol, 2-undecanol, (2R, 3R) - (-) -2, 3-butanediol, 1-isopropoxy-2-propanol, 1-methoxy-2-butanol, propylene glycol, 4-penten-1-ol, 4-vinyl-2-methoxyphenol, 2, 3-dihydrobenzofuran, (1-acetoxy-3-hydroxyprop-2-yl) acetate, isopentyl octanoate, hexyl propionate, hexyl butyrate, geranyl octanoate, phenyl carbamate, propyl octanoate, vinyl formate, diisobutyl phthalate, pentadecanol, decanal, nonanal, 3-methylpentanoic acid, ethyl- (Z) -4-pentenoic acid, ma Shitong, linalool, antioxidant, 4-hydroxy-2-butanone;
preferably, the single-fungus fermentation means: fermentation with Candida glabra strain 196 alone;
preferably, the mixed bacteria fermentation means: the strain 196 of Candida glabra and Saccharomyces cerevisiae F15 are mixed and fermented.
The fermentation is selected from: single-bacteria fermentation or mixed-bacteria fermentation;
preferably, the mixed bacteria fermentation means: mixed fermentation of Candida glabrata (Candida glabra) strain 196 and saccharomyces cerevisiae F15;
preferably, the brewing means brewing wine.
A brewing starter, its fermentation active substance comprises Candida glabrata strain 196 with a preservation number of CCTCC NO: M2023928.
The fermentation active further comprises Saccharomyces cerevisiae;
preferably, the brewing starter is a microbial inoculum;
preferably, the brewing starter further comprises auxiliary materials acceptable by a microbial inoculum;
preferably, the auxiliary materials acceptable by the microbial inoculum comprise culture medium materials of bacteria; the culture medium materials of the bacteria include, but are not limited to: starch, sucrose, peptone and water.
A fermentation process adopts Candida glabrata (Candida glabrata) strain 196 with a preservation number of CCTCC NO: M2023928 for fermentation.
The fermentation is selected from single-bacteria fermentation or mixed-bacteria fermentation;
preferably, the single-fungus fermentation refers to fermentation by inoculating a strain 196 into a substrate;
preferably, the mixed fermentation refers to inoculating a strain 196 and saccharomyces cerevisiae into a substrate for fermentation;
preferably, the saccharomyces cerevisiae is selected from: f15;
preferably, the concentration of the access strain is 10 6 -10 7 CFU/ml, preferably 10 6 CFU/ml；
Preferably, the temperature of the fermentation is 18 ℃ ± 2 ℃;
preferably, the fermentation is terminated until the substrate weight loss is no longer changed for three consecutive days;
preferably, the substrate is grape juice or culture medium;
preferably, the medium is YPD medium.
A method for brewing wine comprises fermenting Candida glabrata strain 196 with preservation number of CCTCC NO: M2023928 with grape juice as substrate.
The fermentation is selected from single-bacteria fermentation or mixed-bacteria fermentation;
preferably, the single-fungus fermentation refers to fermentation by inoculating a strain 196 into a substrate;
preferably, the mixed fermentation refers to inoculating a strain 196 and saccharomyces cerevisiae into a substrate for fermentation;
preferably, the saccharomyces cerevisiae is selected from: f15;
preferably, the concentration of the access strain is 10 6 -10 7 CFU/ml, preferably 10 6 CFU/ml；
Preferably, the temperature of the fermentation is 18 ℃ ± 2 ℃;
preferably, the fermentation is terminated until the substrate weight loss is no longer changed for three consecutive days;
preferably, the substrate is grape juice.
The beneficial effects of the invention are as follows:
the invention screens out a strain, and the strain can generate a large amount of rich aroma substances through fermentation experiments, and compared with the conventional commercial saccharomyces cerevisiae strain, the strain can reduce the content of certain aroma substances to a large extent through aroma substance component identification experiments, for example: ethyl heptanoate, ethyl octanoate, ethyl laurate, ethyl isovalerate, ethyl decanoate, ethyl n-hexanoate, ethyl hex-2-enoate, ethyl 7-octenoate, ethyl 9-hexadecenoate, ethyl 9-decenoate, geranyl acetone, methyl triacontate C30, phenethyl acetate, heptyl acetate, isoamyl acetate, diethylene glycol, n-heptanol, octanol, 2-undecanol, (2 r,3 r) - (-) -2, 3-butanediol, 1-isopropoxy-2-propanol, 1-methoxy-2-butanol, propylene glycol, 4-penten-1-ol, 4-vinyl-2-methoxyphenol, 2, 3-dihydrobenzofuran, (1-acetoxy-3-hydroxyprop-2-yl) acetate, isopentyl octanoate, hexyl propionate, hexyl butyrate, geraniol octanoate, phenyl carbamate, propyl octanoate, vinyl formate, diisobutyl phthalate, pentadecanol, decanol, nonanal, 3-methylpentanoic acid, octanoic acid, ethyl- (Z) -4-decenoic acid, ma Shitong, aromatic alcohol, 4-hydroxy butanone.
Wherein, the content of the phenethyl acetate, the n-hexanol, the 4-vinyl-2-methoxyphenol and the octanoic acid is below a threshold value when the strain of the invention is fermented singly, and people cannot smell the strain, so that the flavor of the wine is changed. The content of octanoic acid is below a threshold value when the strain and the F15 strain are fermented singly respectively, people cannot sniff, the content of octanoic acid generated when the strain and the F15 strain are mixed and fermented is far above the threshold value, people can sniff, and the change of the content can cause the quality of the fragrance of the wine to be changed.
However, fragrance materials such as ethyl 3-phenylpropionate, ethyl 4-acetoxybutyrate, ethyl pentadecanoate, (trimethylsilyl) oxy ] phosphine ] trimethylsilyl acetate, 10-undecen-1-ol, 1, 10-decanediol, 1-pentadecanol, rose ether, isobutyl decanoate, n-propyl decanoate, trans-2-nonenoic acid, 3-nitropropionic acid, 9-decenoic acid, 3-carene, farnesene, 3-isopropenyl-1-methylcyclohexene, 6, 10-dimethyl-5, 9-undecadiene-2-one are not produced in the single-cell fermentation of the strain of the present invention and the F15 strain, respectively, but such fragrance materials can be produced in the mixed-cell fermentation of the two.
In addition, the strains of the invention are capable of being multiplied by the amount of certain aroma substances produced by commercial s.cerevisiae strain F15, for example, ethyl laurate, ethyl 9-hexadecenoate, ethyl 9-decenoate, phenethyl acetate, octanol, 4-vinyl-2-methoxyphenol, 2, 3-dihydrobenzofuran, 2, 4-trimethylpentanediol isobutyl ester, vinyl formate, 3-methylpentanoic acid, octanoic acid, 2-methyl-propionic acid, large Ma Shitong, after fermentation in combination therewith; wherein, the content of octanoic acid is not more than a threshold value when being fermented by F15 strain single bacteria, but the content of octanoic acid after mixed bacteria fermentation is multiplied and exceeds the threshold value, and the quality of the grape wine flavor is changed. Wherein, the content of the ethyl laurate can not be detected during the single-strain fermentation of the strain, and the ethyl laurate can not be smelled by people, so that the flavor of the wine is changed.
Meanwhile, compared with the commercial saccharomyces cerevisiae strain F15, the strain single-strain fermentation can also generate aroma substances which cannot be generated by the F15 single-strain fermentation, for example: TRANS-4-decenoic acid ethyl ester, (2S-cis) -tetrahydro4-methyl-2- (2-methyl-1-propenyl) -2H-pyran, 2-pentadecanol, 3-dodecanol, 3-chloro-2-methyl-2-pentanol, dimethylsilanediol, 1-tridecanol, 1, 3-pentanediol, 6-methyl-5-hepten-2-ol, ALL TRANS-3,7, 11-15-tetramethyl-2, 6,10, 14-hexadecen-1-ol, dihydrolinalool, 3-octanol, n-nonyl chloroformate, 2, 4-trimethyl-1, 3-pentanediol monoisobutyrate, dibutyl phthalate, 2, 4-dimethylbenzaldehyde, 2-methylbutanoic acid, terpinene; wherein the yield of the dimethyl silanediol is up to 22.80 mug/L, the yield of the 1, 3-pentanediol is up to 114.28 mug/L, the yield of the dihydro linalool is up to 35.90 mug/L, the yield of the dibutyl phthalate is up to 39.52 mug/L, and the yield of the 2-methylbutyric acid is up to 33.28 mug/L. The strain is identified as Candida glabrata (Candida glabrata). The applicant designated 196 and sent the strain for preservation.
Preservation number: CCTCC NO: M2023928
Classification naming: candida glabrata 196
Latin name: candida glabra 196
Preservation date: 2023, 6 and 5 days
Preservation unit: china center for type culture Collection
Preservation address: chinese, wuhan, university of Wuhan.
Detailed Description
The present invention will be described in further detail with reference to specific examples and experimental examples, but the embodiments of the present invention are not limited thereto, and the scope of the present invention is not limited thereto. Unless otherwise specified, various reagent consumables used in the examples and experimental examples of the invention are commercially available, and related experimental steps are common operations in the field and have technical meanings which can be conventionally understood by those skilled in the art.
Sources of biological materials
1. Grape variety used in experimental example 1 of the present invention: the common grape varieties of chang jersey, dan-tiu, melo, marbeck, cabernet Sauvignon, meile, bai Shi nan, jiamei and Sang Jiaowei Sai are all commercially available.
2. Saccharomyces cerevisiae F15 used in Experimental example 2 of the present invention was purchased from Laffort, inc.
3. Ma Selan wine grape used in Experimental example 2 of the present invention is commercially available.
Group 1 example, strain 196 of the invention
The present example provides a Candida glabrata (Candida glabrata) strain 196 with a preservation number of cctccc No. M2023928.
Any of culturing, propagating, fermenting, enriching, producing, preparing, using, inoculating, amplifying, transforming, modifying, transforming, selling, offering to sell the act of the Candida glabrata strain 196 with the preservation number of cctccc No. M2023928 and/or the act of combining the Candida glabrata strain 196 with other saccharomyces cerevisiae with the preservation number of cctccc No. M2023928 falls within the scope of the present invention.
According to the actual production requirement, the person skilled in the art can combine the conventional technical means or the basic common sense of the production process in the field of microbial agents (for example, encyclopedia of preparation technology, research and application of microbial agents and the like) to perform conventional selection or adjustment on preparation auxiliary materials, so that Candida glabrata strain 196 with the preservation number of CCTCC No. M2023928 is prepared into microbial agent products with different dosage forms, different storage conditions and different shelf lives, which is free from technical barriers for the person skilled in the art and can be easily achieved.
Group 2 example, use of strain 196 of the invention
The present set of examples provides the use of Candida glabrata strain 196 with a accession number cctccc No. M2023928 for the production, regulation of aroma substances and/or fermentation and/or brewing.
In the specific embodiment of the present invention, the fragrance material is selected from the group consisting of ethyl heptanoate, ethyl octanoate, ethyl laurate, ethyl isovalerate, ethyl decanoate, ethyl n-hexanoate, ethyl 2-enoate, ethyl 7-octenoate, ethyl 9-hexadecenoate, ethyl 9-decenoate, geranyl acetonate, methyl triacontate C30, phenethyl acetate, heptyl acetate, isopentyl acetate, diethylene glycol, n-heptanol, octanol, 2-undecanol, (2R, 3R) - (-) -2, 3-butanediol, 1-isopropoxy-2-propanol, 1-methoxy-2-butanol, propylene glycol, 4-penten-1-ol, 4-vinyl-2-methoxyphenol, 2, 3-dihydrobenzofuran, (1-acetoxy-3-hydroxyprop-2-yl) acetate, isopentyl octanoate, hexyl propionate, hexyl butyrate, geranyl octanoate, phenyl carbamate, propyl octanoate, vinyl formate, diisobutyl phthalate, pentadecanol, decanal, nonanal, 3-methylpentanoic acid, ethyl- (Z) -4-decenyl, 62, 1-isopropoxy-2-propanol, 1-methoxy-butanol, 2-propanediol, 4-pentenoic acid, 4-hydroxy-2-methyl-butanone, methyl-10-n-10-ethyl decanoate, n-10-methyl-10-decanoate, n-methyl-1-hydroxy-ethyl decanoate, n-10-yl, methyl-n-ethyl decanoate, n-1-10-hydroxy-yl acetate, methyl-n-ethyl decanoate, n-methyl-10-yl acetate, methyl decanoate, methyl-n, N-propyl decanoate, TRANS-2-nonenoic acid, 3-nitropropionic acid, 9-decenoic acid, 3-carene, farnesene, 3-isopropenyl-1-methylcyclohexene, 6, 10-dimethyl-5, 9-undecadiene-2-one, ethyl laurate, ethyl 9-hexadecenoate, ethyl 9-decenoate, phenethyl acetate, octanol, 4-vinyl-2-methoxyphenol, 2, 3-dihydrobenzofuran, 2, 4-trimethylpentanediol isobutyl formate, vinyl formate, 3-methylpentanoic acid, octanoic acid, 2-methyl-propionic acid, mega Ma Shitong, TRANS-4-decenoic acid ethyl ester, (2S-cis) -tetrahydro4-methyl-2- (2-methyl-1-propenyl) -2H-pyran, 2-pentadecanol, 3-dodecanol, 3-chloro-2-methyl-2-pentanol, dimethylsilanediol, 1-tridecanol, 1, 3-pentanediol, 6-methyl-5-hept-ol, ALL-3, 7, 4-trimethylpentanediol isobutyl formate, methyl-2, 4-hexadecanoate, 4-methyl-2-n-dimethylbutyrate, 4-methyl-2, 4-dimethylbutyrate;
preferably, the production refers to the single bacterial fermentation to produce TRANS-4-decenoic acid ethyl ester, (2S-cis) -tetrahydro-4-methyl-2- (2-methyl-1-propenyl) -2H-pyran, 2-pentadecanol, 3-dodecanol, 3-chloro-2-methyl-2-pentanol, dimethylsilanediol, 1-tridecanol, 1, 3-pentanediol, 6-methyl-5-hepten-2-ol, ALL TRANS-3,7, 11-15-tetramethyl-2, 6,10, 14-hexadecanetetraen-1-ol, dihydrolinalool, 3-octanol, n-nonyl chloroformate, 2, 4-trimethyl-1, 3-pentanediol monoisobutyrate, dibutyl phthalate, 2, 4-dimethylbenzaldehyde, 2-methylbutanoic acid, terpinene; or, mixed bacteria fermentation to produce ethyl 3-phenylpropionate, ethyl 4-acetoxybutyrate, ethyl pentadecanoate, (trimethylsilyl) oxy ] phosphine ] acetate trimethylsilyl ester, 10-undecen-1-ol, 1, 10-decanediol, 1-pentadecanol, rose ether, isobutyl decanoate, n-propyl decanoate, trans-2-nonenoic acid, 3-nitropropionic acid, 9-decenoic acid, 3-carene, farnesene, 3-isopropenyl-1-methylcyclohexene, 6, 10-dimethyl-5, 9-undecadien-2-one;
preferably, the adjustment means: during mixed fermentation, the ethyl laurate, the ethyl 9-hexadecenoate, the ethyl 9-decenoate, the phenethyl acetate, the octanol, the 4-vinyl-2-methoxyphenol, the 2, 3-dihydrobenzofuran, the 2, 4-trimethylpentanediol isobutyl ester, the vinyl formate, the 3-methylpentanoic acid, the octanoic acid, the 2-methyl-propionic acid and the large Ma Shitong are improved, or reducing ethyl heptanoate, ethyl octanoate, ethyl laurate, ethyl isovalerate, ethyl decanoate, ethyl n-hexanoate, ethyl hex-2-enoate, ethyl 7-octenoate, ethyl 9-hexadecenoate, ethyl 9-decenoate, geranyl acetone, methyl triacontate C30, phenethyl acetate, heptyl acetate, isopentyl acetate, diethylene glycol, n-heptanol, octanol, 2-undecanol, (2R, 3R) - (-) -2, 3-butanediol, 1-isopropoxy-2-propanol, 1-methoxy-2-butanol, propylene glycol, 4-penten-1-ol, 4-vinyl-2-methoxyphenol, 2, 3-dihydrobenzofuran, (1-acetoxy-3-hydroxyprop-2-yl) acetate, isopentyl octanoate, hexyl propionate, hexyl butyrate, geranyl octanoate, phenyl carbamate, propyl octanoate, vinyl formate, diisobutyl phthalate, pentadecanol, decanal, nonanal, 3-methylpentanoic acid, ethyl- (Z) -4-pentenoic acid, ma Shitong, linalool, antioxidant, 4-hydroxy-2-butanone;
preferably, the single-fungus fermentation means: fermentation with Candida glabra strain 196 alone;
preferably, the mixed bacteria fermentation means: the strain 196 of Candida glabra and Saccharomyces cerevisiae F15 are mixed and fermented.
In some embodiments, the fermentation is selected from: single-bacteria fermentation or mixed-bacteria fermentation;
preferably, the mixed bacteria fermentation means: mixed fermentation of Candida glabrata (Candida glabra) strain 196 and saccharomyces cerevisiae F15;
preferably, the brewing means brewing wine.
Group 3 example, brewing leaven of the present invention
The present set of examples provides a brewing starter. All embodiments of this group share the following common features: the fermentation active substances of the brewing starter comprise Candida glabrata (Candida glabrata) strain 196 with the preservation number of CCTCC NO: M2023928.
In a further embodiment, the fermentation active further comprises saccharomyces cerevisiae;
preferably, the brewing starter is a microbial inoculum;
preferably, the brewing starter further comprises auxiliary materials acceptable by a microbial inoculum;
preferably, the auxiliary materials acceptable by the microbial inoculum comprise culture medium materials of bacteria; the culture medium materials of the bacteria include, but are not limited to: starch, sucrose, peptone and water.
In a more specific embodiment, the microbial agent acceptable adjuvant is selected from the group consisting of: food additives, solvents, propellants, solubilizing agents, co-solvents, emulsifiers, colorants, binders, disintegrants, fillers, lubricants, wetting agents, osmotic pressure regulators, stabilizers, glidants, flavoring agents, preservatives, suspending agents, coating materials, fragrances, anti-binding agents, integration agents, permeation enhancers, pH modifiers, buffers, plasticizers, surfactants, foaming agents, defoamers, thickeners, inclusion agents, humectants, absorbents, diluents, flocculants, deflocculants, filter aids, release retarders, and the like.
According to the invention, the edible auxiliary materials can be selected and formulated by combining with conventional technical means (for example, encyclopedia of preparation technology, technical research and application of microbial agents and the like) in the field of microbial agent preparation according to different requirements in practical production application, and the Candida glabra (Candida glabra) strain 196 with the preservation number of CCTCC NO: M2023928 can be prepared into different dosage forms, such as powder, granules, liquid and the like.
Group 4 example, fermentation Process of the invention
The present set of embodiments provides a fermentation process. All embodiments of this group share the following common features: fermentation was performed using Candida glabrata strain 196 with a accession number of cctccc No. M2023928.
In some embodiments, the fermentation is selected from single or mixed fermentation;
preferably, the single-fungus fermentation refers to fermentation by inoculating a strain 196 into a substrate;
preferably, the mixed fermentation refers to inoculating a strain 196 and saccharomyces cerevisiae into a substrate for fermentation;
preferably, the saccharomyces cerevisiae is selected from: f15;
preferably, the concentration of the access strain is 10 6 -10 7 CFU/ml, preferably 10 6 CFU/ml；
Preferably, the temperature of the fermentation is 18 ℃ ± 2 ℃;
preferably, the fermentation is terminated until the substrate weight loss is no longer changed for three consecutive days;
preferably, the substrate is grape juice or culture medium;
preferably, the medium is YPD medium.
Group 5 example, method of brewing wine of the present invention
The embodiment of the group provides a wine brewing method. All embodiments of this group share the following common features: a Candida glabrata strain 196 with a preservation number of CCTCC NO: M2023928 is adopted to ferment and brew with grape juice as a substrate.
In specific embodiments, the fermentation is selected from single-or mixed-fermentation;
preferably, the single-fungus fermentation refers to fermentation by inoculating a strain 196 into a substrate;
preferably, the mixed fermentation refers to inoculating a strain 196 and saccharomyces cerevisiae into a substrate for fermentation;
preferably, the saccharomyces cerevisiae is selected from: f15;
preferably, the concentration of the access strain is 10 6 -10 7 CFU/ml, preferably 10 6 CFU/ml；
Preferably, the temperature of the fermentation is 18 ℃ ± 2 ℃;
preferably, the fermentation is terminated until the substrate weight loss is no longer changed for three consecutive days;
preferably, the substrate is grape juice.
Experimental example 1 screening procedure of Candida glabrata strain 196 according to the invention
Collecting Ningxia Hovenia shandong foot grapes of different production places and different varieties, removing rotten, moldy and damaged fruits and sundries, weighing 10.0g of uniform and complete grape fruits, adding into 90mL of sterile YPD liquid culture medium, oscillating for 10min to prepare bacterial suspension, coating the bacterial suspension with proper dilution on the YPD solid culture medium added with 100mg/L chloramphenicol by adopting a gradient dilution method, culturing for 2-3 d at 28 ℃, and carrying out repeated streak purification on the YPD solid culture medium according to colony morphology to obtain purified bacterial strains. After the purified strain is activated for 24 hours in YPD culture medium, 1mL of bacterial liquid is taken to extract yeast genome according to a specification of a Biospin fungus genome DNA extraction kit, primers NL-1 and NL-4 described in Chinese patent No. 202110107456.8 are used for PCR amplification, the gene sequence of a 26S rDNA fragment is obtained through sequencing, known standard strain information with higher homology is obtained through BLAST comparison of NCBI, and strain with the similarity of more than 99% is subjected to species identification, so that strain species information is finally obtained.
The strain is screened for the variety of wine grapes and the information of the production area are shown in the following table 1:
TABLE 1
| Grape variety | Production area | Source | Sample number (copy) |
| Radix seu herba Gei Albae | Foot producing area of Helan mountain | Acquisition of | 25 |
| Dan (pill) | Foot producing area of Helan mountain | Acquisition of | 25 |
| Mello | Foot producing area of Helan mountain | Acquisition of | 25 |
| Ma Erbei g | Foot producing area of Helan mountain | Acquisition of | 25 |
| Cabernet Sauvignon | Foot producing area of Helan mountain | Acquisition of | 25 |
| Meile medicine | Foot producing area of Helan mountain | Acquisition of | 25 |
| Bai Shi south | Foot producing area of Helan mountain | Acquisition of | 25 |
| Jiamei (good beauty) | Foot producing area of Helan mountain | Acquisition of | 25 |
| Sang Jiaowei plug | Foot producing area of Helan mountain | Acquisition of | 25 |
A total of 36 Candida glabrata (Candida glabrata) strains were obtained in this experimental example. Respectively mixing 18 strains of Candida glabrata (Candida glabrata) at an initial concentration of 10 6 The CFU/mL is inoculated into sterilized Wedelia grape juice (100 ℃ for 10 min), the fermented product is fermented for 18 days at 18+/-2 ℃ and the aroma content is measured, so that a Candida glabrata strain 196 with the best aroma producing effect is finally obtained, and the strain is sent to be preserved, wherein the preservation information is as follows:
preservation number: CCTCC NO: M2023928
Classification naming: candida glabrata 196
Latin name: candida glabra 196
Preservation date: 2023, 6 and 5 days
Preservation unit: china center for type culture Collection
Preservation address: chinese, wuhan, university of Wuhan.
Experimental example 2 fermentation experiment of aroma substances produced by the inventive Strain 196
Grape variety: ma Selan wine grape
The grape juice production process comprises the following steps: crushing the harvested Ma Selan wine grapes, fermenting with peel, and adding sulfur dioxide (50 mg/L K) 2 S 2 O 5 ) And 20mg/L pectase (more than or equal to 500U/mg), inhibiting bacterial diseases and improving juice yield.
Grape juice fermentation conditions: the macerated must was divided into 3 groups (196 groups, F15 groups, f15+196 groups).
Group 196 must at an initial concentration of 10 6 CFU/mL was inoculated with Candida glabrata strain 196;
grape juice of group F15At an initial concentration of 10 6 CFU/mL is accessed into Saccharomyces cerevisiae F15;
the grape juice of F15+196 group has an initial concentration of 10 6 CFU/mL was simultaneously inoculated with Saccharomyces cerevisiae F15 and Candida glabrata strain 196.
After each group of the strains are combined, fermentation is carried out, the fermentation temperature is 18+/-2 ℃, and the fermentation is stopped when the weight of the grape juice is lost for three consecutive days. All wine samples were centrifuged at 7500rpm for 8 minutes and the supernatant was stored at 4 ℃.
The detection method of each aroma substance comprises the following steps: a headspace-solid phase microextraction method-gas phase mass spectrometry (HS-SPME-GC/MS) is adopted. An accurate measurement of 8mL of wine sample was added to a headspace bottle containing 1.5g NaCl, while 394.08. Mu.g/L4-methyl-1-pentanol (internal standard) was capped and sealed. The CAR/DVB/PDMS extraction fiber is inserted, the extraction fiber is desorbed for 3min at 250 ℃ at the GC inlet after being adsorbed for 30min at 45 ℃ for GC-MS analysis. Chromatographic column: an InertCap WAX polar column (60 m 0.25mm,0.25 μm); the temperature-raising program is as follows: keeping the temperature at 40 ℃ for 5min, raising the temperature to 120 ℃ at 3 ℃/min, raising the temperature to 230 ℃ at 8 ℃/min, and keeping the temperature for 10min; the carrier gas (He) flow rate was 0.8mL/min, without split flow. An electron bombardment ion source; electron energy 70eV; the temperature of the transmission line is 275 ℃; the ion source temperature is 230 ℃; the activation voltage is 1.5V; filament flow 0.25mA; mass scanning range m/z is 33-450. Compound quantitative analysis was performed using an external standard quantitative method.
The same batch of grape juice produced by grapes of the same variety is taken as a substrate, under the same fermentation condition, 3 groups of grape juice with the same quantity are respectively subjected to fermentation brewing treatment, and the content of each aroma substance (unit: mug/L, meaning: the content of the aroma substance in each liter of grape wine) is detected by a headspace-solid phase microextraction method-gas phase mass spectrometry method to obtain the following table 2:
TABLE 2
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In table 1 above, "-" indicates that the relevant aroma is not detected.
Experimental example 3 YPD fermentation experiment of aroma substances produced by the Strain 196 of the present invention
1. Group and detection sample preparation:
(one) 196 group: the 196 glycerol storage tube of the strain is dissolved and inoculated into 5mL YPD culture medium, and is cultured for 12-20h at 30 ℃, and after three generations of activation, the strain is inoculated into 400mL fermentation culture medium according to an inoculum size of 1% (volume ratio), the culture temperature is 30 ℃, and the rotation speed of a shaking table is 120rpm, and the strain is cultured for 72h. After fermentation, obtaining fermentation broth, centrifuging at 10000rpm for 5min, and collecting supernatant.
(II) group F15: the Saccharomyces cerevisiae F15 glycerol storage tube is dissolved and inoculated into 5mL of YPD culture medium, and is cultured for 12-20h at 30 ℃, and after three generations of activation, the culture medium is inoculated into 400mL of fermentation culture medium according to an inoculum size of 1% (volume ratio), wherein the culture temperature is 30 ℃, and the rotation speed of a shaking table is 120rpm, and the culture is carried out for 72h. After fermentation, obtaining fermentation broth, centrifuging at 10000rpm for 5min, and collecting supernatant.
(III) 196+F15 group: the 196 glycerol storage tube and the Saccharomyces cerevisiae F15 glycerol storage tube of the strain are respectively dissolved and then inoculated into 5mL of YPD culture medium, and are cultured for 12-20h at 30 ℃, and after three generations of activation, the strain is inoculated into 400mL of fermentation culture medium according to 1% (volume ratio) of inoculum size, and the culture temperature is 30 ℃, the rotation speed of a shaking table is 120rpm, and the strain is cultured for 72h. After fermentation, obtaining fermentation broth, centrifuging at 10000rpm for 5min, and collecting supernatant.
2. Volatile metabolite detection
The volatile metabolites in the samples were detected by the HS-SPME/GC-MS method described in Experimental example 2, the 4-methyl-1-pentanol was used as an internal standard, the results of the compound search were matched with the NIST standard spectrum library, the similarity was 80% or more, and the content of each volatile aroma was calculated. The test results are shown in table 3 below:
TABLE 3 Table 3
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Claims (10)
1. A Candida glabrata (Candida glabrata) strain 196 is characterized by having a preservation number of CCTCC NO: M2023928.
2. Use of Candida glabrata strain 196 with a accession number of cctccc No. M2023928 for production, regulation of aroma substances and/or fermentation and/or brewing.
3. The preservation number of claim 2 is CCTCC NO: use of a Candida glabrata strain 196 of M2023928 for the production, modulation of aroma substances and/or fermentation and/or brewing of wine, characterized in that, the aroma is selected from ethyl heptanoate, ethyl octanoate, ethyl laurate, ethyl isovalerate, ethyl decanoate, ethyl n-hexanoate, ethyl 2-enoate, ethyl 7-octenoate, ethyl 9-hexadecenoate, ethyl 9-decenoate, geranyl acetonate, methyl triacontate C30, phenethyl acetate, heptyl acetate, isopentyl acetate, diethylene glycol, n-heptanol, octanol, 2-undecanol, (2R, 3R) - (-) -2, 3-butanediol, 1-isopropoxy-2-propanol, 1-methoxy-2-butanol, propylene glycol, 4-penten-1-ol, 4-vinyl-2-methoxyphenol, 2, 3-dihydrobenzofuran, (1-acetoxy-3-hydroxyprop-2-yl) acetate, isopentyl octanoate, hexyl propionate, hexyl butyrate, geranyl octanoate, phenyl carbamate, propyl octanoate, vinyl formate, diisobutyl phthalate, pentadecanol, decanal, nonanal, 3-methylpentanoic acid, ethyl- (Z) -4-pentenoic acid, ethyl-37-35, ethyl 2-methoxybutanone, ethyl 2-butanone, ethyl 2-propanoate, ethyl 2-butanone, n-2-butanone, (trimethylsilyl) oxy ] phosphine ] acetic acid trimethylsilyl ester, 10-undecen-1-ol, 1, 10-decanediol, 1-pentadecanol, rose ether, isobutyl decanoate, n-propyl decanoate, TRANS-2-nonenoic acid, 3-nitropropionic acid, 9-decenoic acid, 3-carene, farnesene, 3-isopropenyl-1-methylcyclohexene, 6, 10-dimethyl-5, 9-undecen-2-one, ethyl laurate, ethyl 9-hexadecenoate, ethyl 9-decenoate, phenethyl acetate, octanol, 4-vinyl-2-methoxyphenol, 2, 3-dihydrobenzofuran 2, 4-trimethylpentanediol isobutyl formate, vinyl formate, 3-methylpentanoic acid, octanoic acid, 2-methyl-propionic acid, large Ma Shitong, TRANS-4-decenoic acid ethyl ester, (2S-cis) -tetrahydro4-methyl-2- (2-methyl-1-propenyl) -2H-pyran, 2-pentadecanol, 3-dodecanol, 3-chloro-2-methyl-2-pentanol, dimethylsilanediol, 1-tridecanol, 1, 3-pentanediol, 6-methyl-5-hepten-2-ol, ALL TRANS-3,7, 11-15-tetramethyl-2, 6,10, 14-hexadecen-1-ol, dihydro linalool, 3-octanol, n-nonyl chloroformate, 2, 4-trimethyl-1, 3-pentanediol monoisobutyrate, dibutyl phthalate, 2, 4-dimethylbenzaldehyde, 2-methylbutyric acid, terpinene;
and/or, said producing means that the single bacteria fermentation produces TRANS-4-decenoic acid ethyl ester, (2S-cis) -tetrahydro-4-methyl-2- (2-methyl-1-propenyl) -2H-pyran, 2-pentadecanol, 3-dodecanol, 3-chloro-2-methyl-2-pentanol, dimethylsilanediol, 1-tridecanol, 1, 3-pentanediol, 6-methyl-5-hepten-2-ol, ALL TRANS-3,7, 11-15-tetramethyl-2, 6,10, 14-hexadecanetetraen-1-ol, dihydrolinalool, 3-octanol, n-nonyl chloroformate, 2, 4-trimethyl-1, 3-pentanediol monoisobutyrate, dibutyl phthalate, 2, 4-dimethylbenzaldehyde, 2-methylbutanoic acid, terpinene; or, mixed bacteria fermentation to produce ethyl 3-phenylpropionate, ethyl 4-acetoxybutyrate, ethyl pentadecanoate, (trimethylsilyl) oxy ] phosphine ] acetate trimethylsilyl ester, 10-undecen-1-ol, 1, 10-decanediol, 1-pentadecanol, rose ether, isobutyl decanoate, n-propyl decanoate, trans-2-nonenoic acid, 3-nitropropionic acid, 9-decenoic acid, 3-carene, farnesene, 3-isopropenyl-1-methylcyclohexene, 6, 10-dimethyl-5, 9-undecadien-2-one;
and/or, the adjusting means: during mixed fermentation, the ethyl laurate, the ethyl 9-hexadecenoate, the ethyl 9-decenoate, the phenethyl acetate, the octanol, the 4-vinyl-2-methoxyphenol, the 2, 3-dihydrobenzofuran, the 2, 4-trimethylpentanediol isobutyl ester, the vinyl formate, the 3-methylpentanoic acid, the octanoic acid, the 2-methyl-propionic acid and the large Ma Shitong are improved, or reducing ethyl heptanoate, ethyl octanoate, ethyl laurate, ethyl isovalerate, ethyl decanoate, ethyl n-hexanoate, ethyl hex-2-enoate, ethyl 7-octenoate, ethyl 9-hexadecenoate, ethyl 9-decenoate, geranyl acetone, methyl triacontate C30, phenethyl acetate, heptyl acetate, isopentyl acetate, diethylene glycol, n-heptanol, octanol, 2-undecanol, (2R, 3R) - (-) -2, 3-butanediol, 1-isopropoxy-2-propanol, 1-methoxy-2-butanol, propylene glycol, 4-penten-1-ol, 4-vinyl-2-methoxyphenol, 2, 3-dihydrobenzofuran, (1-acetoxy-3-hydroxyprop-2-yl) acetate, isopentyl octanoate, hexyl propionate, hexyl butyrate, geranyl octanoate, phenyl carbamate, propyl octanoate, vinyl formate, diisobutyl phthalate, pentadecanol, decanal, nonanal, 3-methylpentanoic acid, ethyl- (Z) -4-pentenoic acid, ma Shitong, linalool, antioxidant, 4-hydroxy-2-butanone;
and/or, the single-fungus fermentation means: fermentation with Candida glabra strain 196 alone;
and/or, the mixed bacteria fermentation means: the strain 196 of Candida glabra and Saccharomyces cerevisiae F15 are mixed and fermented.
4. Use of Candida glabrata strain 196 with a preservation number of cctccc NO: M2023928 according to claim 2 or 3 for the production, regulation of aroma substances and/or fermentation and/or brewing, wherein the fermentation is selected from the group consisting of: single-bacteria fermentation or mixed-bacteria fermentation;
and/or, the mixed bacteria fermentation means: mixed fermentation of Candida glabrata (Candida glabra) strain 196 and saccharomyces cerevisiae F15;
and/or, brewing refers to brewing wine.
5. A brewing starter is characterized in that the fermentation active substance comprises Candida glabrata (Candida glabrata) strain 196 with a preservation number of CCTCCNO: M2023928.
6. A brewery starter according to claim 5, wherein said fermentation active further comprises saccharomyces cerevisiae;
and/or, the brewing ferment is a microbial agent;
and/or, the brewing ferment further comprises auxiliary materials acceptable by microbial inoculum;
and/or, the auxiliary materials acceptable by the microbial inoculum comprise culture medium substances of bacteria; the culture medium materials of the bacteria include, but are not limited to: starch, sucrose, peptone and water.
7. A fermentation process is characterized in that Candida glabrata (Candida glabrata) strain 196 with a preservation number of CCTCC NO: M2023928 is adopted for fermentation.
8. A fermentation process according to claim 6, wherein the fermentation is selected from single-or mixed-bacteria fermentation;
and/or, the single-fungus fermentation refers to inoculating a strain 196 into a substrate for fermentation;
and/or, the mixed fermentation refers to inoculating a strain 196 and saccharomyces cerevisiae into a substrate for fermentation;
and/or, the saccharomyces cerevisiae is selected from: f15;
and/or the concentration of the access strain is 10 6 -10 7 CFU/ml, preferably 10 6 CFU/ml；
And/or the fermentation temperature is 18 ℃ +/-2 ℃;
and/or stopping fermentation when the substrate weight loss is no longer changed for three consecutive days;
and/or the substrate is grape juice or culture medium;
and/or, the culture medium is YPD culture medium.
9. A method for brewing grape wine is characterized in that a wine with a preservation number of CCTCC NO: M is adopted
A Candida glabrata strain 196 of 2023928 is fermented and brewed by taking grape juice as a substrate.
10. A method of brewing wine according to claim 9, wherein said fermentation is selected from single or mixed fermentation;
and/or, the single-fungus fermentation refers to inoculating a strain 196 into a substrate for fermentation;
and/or, the mixed fermentation refers to inoculating a strain 196 and saccharomyces cerevisiae into a substrate for fermentation;
and/or, the saccharomyces cerevisiae is selected from: f15;
and/or the concentration of the access strain is 10 6 -10 7 CFU/ml, preferably 10 6 CFU/ml；
And/or the fermentation temperature is 18 ℃ +/-2 ℃;
and/or stopping fermentation when the substrate weight loss is no longer changed for three consecutive days;
and/or, the substrate is grape juice.
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