CN117925426A - Rhodotorula mucilaginosa (Rhodotorula mucilaginosa) strain 147 and application, product and method thereof - Google Patents
Rhodotorula mucilaginosa (Rhodotorula mucilaginosa) strain 147 and application, product and method thereof Download PDFInfo
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- CN117925426A CN117925426A CN202311835325.7A CN202311835325A CN117925426A CN 117925426 A CN117925426 A CN 117925426A CN 202311835325 A CN202311835325 A CN 202311835325A CN 117925426 A CN117925426 A CN 117925426A
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- 241000223254 Rhodotorula mucilaginosa Species 0.000 title claims abstract description 84
- 238000000034 method Methods 0.000 title claims abstract description 14
- 238000000855 fermentation Methods 0.000 claims abstract description 123
- 230000004151 fermentation Effects 0.000 claims abstract description 122
- 238000004321 preservation Methods 0.000 claims abstract description 32
- 239000000126 substance Substances 0.000 claims abstract description 27
- 238000004519 manufacturing process Methods 0.000 claims abstract description 19
- 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 16
- KHAVLLBUVKBTBG-UHFFFAOYSA-N dec-9-enoic acid Chemical compound OC(=O)CCCCCCCC=C KHAVLLBUVKBTBG-UHFFFAOYSA-N 0.000 claims abstract description 16
- JAGZUIGGHGTFHO-UHFFFAOYSA-N Ethyl 3-phenylpropanoate Chemical compound CCOC(=O)CCC1=CC=CC=C1 JAGZUIGGHGTFHO-UHFFFAOYSA-N 0.000 claims abstract description 15
- 239000007858 starting material Substances 0.000 claims abstract description 11
- HIGQPQRQIQDZMP-UHFFFAOYSA-N geranil acetate Natural products CC(C)=CCCC(C)=CCOC(C)=O HIGQPQRQIQDZMP-UHFFFAOYSA-N 0.000 claims abstract description 10
- FXHIOQYSZLPDAR-UHFFFAOYSA-N 1-(1,3-dioxolan-2-yl)propan-2-one Chemical compound CC(=O)CC1OCCO1 FXHIOQYSZLPDAR-UHFFFAOYSA-N 0.000 claims abstract description 9
- PTEYJUIKYIKULL-UHFFFAOYSA-N Ethyl pentadecanoate Chemical compound CCCCCCCCCCCCCCC(=O)OCC PTEYJUIKYIKULL-UHFFFAOYSA-N 0.000 claims abstract description 8
- CTXGTHVAWRBISV-UHFFFAOYSA-N 2-hydroxyethyl dodecanoate Chemical compound CCCCCCCCCCCC(=O)OCCO CTXGTHVAWRBISV-UHFFFAOYSA-N 0.000 claims abstract description 7
- HIGQPQRQIQDZMP-FLIBITNWSA-N neryl acetate Chemical compound CC(C)=CCC\C(C)=C/COC(C)=O HIGQPQRQIQDZMP-FLIBITNWSA-N 0.000 claims abstract description 7
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- 239000000758 substrate Substances 0.000 claims description 28
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- WQEPLUUGTLDZJY-UHFFFAOYSA-N pentadecanoic acid Chemical compound CCCCCCCCCCCCCCC(O)=O WQEPLUUGTLDZJY-UHFFFAOYSA-N 0.000 claims description 24
- 239000001963 growth medium Substances 0.000 claims description 22
- JGHZJRVDZXSNKQ-UHFFFAOYSA-N methyl octanoate Chemical compound CCCCCCCC(=O)OC JGHZJRVDZXSNKQ-UHFFFAOYSA-N 0.000 claims description 22
- MTHSVFCYNBDYFN-UHFFFAOYSA-N diethylene glycol Chemical compound OCCOCCO MTHSVFCYNBDYFN-UHFFFAOYSA-N 0.000 claims description 21
- LSKONYYRONEBKA-UHFFFAOYSA-N 2-Dodecanone Chemical compound CCCCCCCCCCC(C)=O LSKONYYRONEBKA-UHFFFAOYSA-N 0.000 claims description 20
- 235000019674 grape juice Nutrition 0.000 claims description 17
- KBPLFHHGFOOTCA-UHFFFAOYSA-N 1-Octanol Chemical compound CCCCCCCCO KBPLFHHGFOOTCA-UHFFFAOYSA-N 0.000 claims description 16
- HSJKGGMUJITCBW-UHFFFAOYSA-N 3-hydroxybutanal Chemical compound CC(O)CC=O HSJKGGMUJITCBW-UHFFFAOYSA-N 0.000 claims description 16
- XKWSWANXMRXDES-UHFFFAOYSA-N 3-methylbutyl octanoate Chemical compound CCCCCCCC(=O)OCCC(C)C XKWSWANXMRXDES-UHFFFAOYSA-N 0.000 claims description 16
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- 239000005641 Methyl octanoate Substances 0.000 claims description 14
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- MMXKVMNBHPAILY-UHFFFAOYSA-N ethyl laurate Chemical compound CCCCCCCCCCCC(=O)OCC MMXKVMNBHPAILY-UHFFFAOYSA-N 0.000 claims description 14
- 239000000463 material Substances 0.000 claims description 14
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- IGIDLTISMCAULB-UHFFFAOYSA-N 3-methylvaleric acid Chemical compound CCC(C)CC(O)=O IGIDLTISMCAULB-UHFFFAOYSA-N 0.000 claims description 11
- KQNPFQTWMSNSAP-UHFFFAOYSA-N isobutyric acid Chemical compound CC(C)C(O)=O KQNPFQTWMSNSAP-UHFFFAOYSA-N 0.000 claims description 11
- DSVGICPKBRQDDX-UHFFFAOYSA-N 1,3-diacetoxypropane Chemical compound CC(=O)OCCCOC(C)=O DSVGICPKBRQDDX-UHFFFAOYSA-N 0.000 claims description 10
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- 235000014787 Vitis vinifera Nutrition 0.000 claims description 10
- SZXQTJUDPRGNJN-UHFFFAOYSA-N dipropylene glycol Chemical compound OCCCOCCCO SZXQTJUDPRGNJN-UHFFFAOYSA-N 0.000 claims description 10
- KMPQIYXXLIFPKA-UHFFFAOYSA-N ethyl 4-acetyloxybutanoate Chemical compound CCOC(=O)CCCOC(C)=O KMPQIYXXLIFPKA-UHFFFAOYSA-N 0.000 claims description 10
- SHZIWNPUGXLXDT-UHFFFAOYSA-N ethyl hexanoate Chemical compound CCCCCC(=O)OCC SHZIWNPUGXLXDT-UHFFFAOYSA-N 0.000 claims description 10
- ZAJNGDIORYACQU-UHFFFAOYSA-N methyl n-octyl ketone Natural products CCCCCCCCC(C)=O ZAJNGDIORYACQU-UHFFFAOYSA-N 0.000 claims description 10
- 239000002068 microbial inoculum Substances 0.000 claims description 9
- GLZPCOQZEFWAFX-UHFFFAOYSA-N Geraniol Chemical compound CC(C)=CCCC(C)=CCO GLZPCOQZEFWAFX-UHFFFAOYSA-N 0.000 claims description 8
- FHLGUOHLUFIAAA-UHFFFAOYSA-N Linalyl butyrate Chemical compound CCCC(=O)OC(C)(C=C)CCC=C(C)C FHLGUOHLUFIAAA-UHFFFAOYSA-N 0.000 claims description 8
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- JELGPLUONQGOHF-UHFFFAOYSA-N ethyl hexadec-9-enoate Chemical compound CCCCCCC=CCCCCCCCC(=O)OCC JELGPLUONQGOHF-UHFFFAOYSA-N 0.000 claims description 7
- YYZUSRORWSJGET-UHFFFAOYSA-N ethyl octanoate Chemical compound CCCCCCCC(=O)OCC YYZUSRORWSJGET-UHFFFAOYSA-N 0.000 claims description 7
- WWZKQHOCKIZLMA-UHFFFAOYSA-M octanoate Chemical compound CCCCCCCC([O-])=O WWZKQHOCKIZLMA-UHFFFAOYSA-M 0.000 claims description 7
- 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 6
- HBEDSQVIWPRPAY-UHFFFAOYSA-N 2,3-dihydrobenzofuran Chemical compound C1=CC=C2OCCC2=C1 HBEDSQVIWPRPAY-UHFFFAOYSA-N 0.000 claims description 6
- 241000219095 Vitis Species 0.000 claims description 6
- 235000009754 Vitis X bourquina Nutrition 0.000 claims description 6
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- OBETXYAYXDNJHR-UHFFFAOYSA-N alpha-ethylcaproic acid Natural products CCCCC(CC)C(O)=O OBETXYAYXDNJHR-UHFFFAOYSA-N 0.000 claims description 6
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 claims description 6
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- IDHBLVYDNJDWNO-UHFFFAOYSA-N propyl octanoate Chemical compound CCCCCCCC(=O)OCCC IDHBLVYDNJDWNO-UHFFFAOYSA-N 0.000 claims description 5
- 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 4
- AFHJYKBGDDJSRR-UHFFFAOYSA-N 1-propan-2-yloxypropan-2-ol Chemical compound CC(C)OCC(C)O AFHJYKBGDDJSRR-UHFFFAOYSA-N 0.000 claims description 4
- GLZPCOQZEFWAFX-YFHOEESVSA-N Geraniol Natural products CC(C)=CCC\C(C)=C/CO GLZPCOQZEFWAFX-YFHOEESVSA-N 0.000 claims description 4
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- UXDDRFCJKNROTO-UHFFFAOYSA-N Glycerol 1,2-diacetate Chemical compound CC(=O)OCC(CO)OC(C)=O UXDDRFCJKNROTO-UHFFFAOYSA-N 0.000 claims description 4
- GFJVXXWOPWLRNU-UHFFFAOYSA-N ethenyl formate Chemical compound C=COC=O GFJVXXWOPWLRNU-UHFFFAOYSA-N 0.000 claims description 4
- JELGPLUONQGOHF-MDZDMXLPSA-N ethyl 9-hexadecenoate Chemical compound CCCCCC\C=C\CCCCCCCC(=O)OCC JELGPLUONQGOHF-MDZDMXLPSA-N 0.000 claims description 4
- DWMKJZMYLQUIDA-UHFFFAOYSA-N ethyl oct-7-enoate Chemical compound CCOC(=O)CCCCCC=C DWMKJZMYLQUIDA-UHFFFAOYSA-N 0.000 claims description 4
- 229940113087 geraniol Drugs 0.000 claims description 4
- HNZUNIKWNYHEJJ-FMIVXFBMSA-N geranyl acetone Chemical compound CC(C)=CCC\C(C)=C\CCC(C)=O HNZUNIKWNYHEJJ-FMIVXFBMSA-N 0.000 claims description 4
- HNZUNIKWNYHEJJ-UHFFFAOYSA-N geranyl acetone Natural products CC(C)=CCCC(C)=CCCC(C)=O HNZUNIKWNYHEJJ-UHFFFAOYSA-N 0.000 claims description 4
- BSCCSDNZEIHXOK-UHFFFAOYSA-N phenyl carbamate Chemical compound NC(=O)OC1=CC=CC=C1 BSCCSDNZEIHXOK-UHFFFAOYSA-N 0.000 claims description 4
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Abstract
The invention discloses Rhodotorula mucilaginosa strain 147 and application, a product and a method thereof, belonging to the technical field of microorganisms. The invention provides a rhodotorula glutinis (Rhodotorula mucilaginosa) strain 147 with a preservation number of CCTCC NO: M2023934, and also provides application of the strain 147 in production, aroma substance regulation and/or fermentation and/or brewing, and a fermentation process, a brewing starter and a wine brewing method based on the strain 147. In contrast to commercial s.cerevisiae strain F15, strain 147 of the present invention may produce aroma substances that F15 cannot produce, including ethyl 3-phenylpropionate, ethyl pentadecanoate, nerol acetate, dihydrolinalool, ethylene glycol laurate, 9-decenoic acid, 1- (1, 3-dioxolan-2-yl) acetone, and the like.
Description
Technical Field
The invention belongs to the technical field of microorganisms, and particularly relates to a rhodotorula mucilaginosa (Rhodotorula mucilaginosa) strain 147, and application, a product and a method thereof.
Background
Rhodotorula mucilaginosa is a microorganism of Rhodotorula genus, and has main characteristics of China origin: heterotrophy of chemical energy; mesophilic; acid-chemotaxis; facultative anaerobism. The 5Be' wort agar was incubated at 30℃for 3 days, and the colonies were round, orange-yellow, slightly glossy, sticky and full-edged. Sausage-shaped, elongated, individual spherical, a few more 1.2-3.5X4.2-15 μm. The asexual propagation mode is polygonal bud. Is obtained through SICC 2.506 screening. The main application of the yeast is research and production, and the specific production application is feed yeast.
At present, reports of fermenting rhodotorula mucilaginosa, brewing wine, producing and regulating aroma substances are not yet seen.
Disclosure of Invention
The invention provides a rhodotorula mucilaginosa (Rhodotorula mucilaginosa) strain 147, application, product and method thereof, aiming at solving the technical problems of filling up the blank of brewing wine by rhodotorula mucilaginosa and producing and adjusting aroma substances and developing more new applications of rhodotorula mucilaginosa.
The technical scheme of the invention is as follows:
a rhodotorula mucilaginosa (Rhodotorula mucilaginosa) strain 147 has a preservation number of CCTCC NO: M2023934.
The application of rhodotorula mucilaginosa (Rhodotorula mucilaginosa) strain 147 with preservation number of CCTCC NO: M2023934 in the aspects of production, aroma substance regulation and/or fermentation and/or brewing.
The aroma substances are selected from lauric acid ethyl ester, ethyl caproate, 7-octenoate ethyl ester, 9-hexadecenoate ethyl ester, isoamyl octanoate, 3-methyl valeric acid, octanoic acid, (E) -7, 11-dimethyl-3-methylene-1, 6, 10-dodecatriene, 3-phenylpropionic acid ethyl ester, 4-acetoxybutyric acid ethyl ester, tridecanoic acid ethyl ester, (2S-cis) -tetrahydrogen-4-methyl-2- (2-methyl-1-propenyl) -2H-pyran, dipropylene glycol, 10-undecen-1-ol, dihydro linalool, 1-pentadecanol, 1, 3-propylene glycol diacetate, butyric acid-1-vinyl-1, 5-dimethyl-4-hexenyl ester 3-hydroxybutyraldehyde, 2-dodecanone, ethyl isovalerate, geranyl acetone, methyl triacontanoate C30, diethylene glycol, octanol 1-isopropoxy-2-propanol, (1-acetoxy-3-hydroxyprop-2-yl) acetate, methyl octanoate, geraniol octanoate, phenyl carbamate 1-isopropoxy-2-propanol, (1-acetoxy-3-hydroxyprop-2-yl) acetate methyl octanoate, geraniol octanoate, phenyl carbamate, 2, 4-trimethyl-1, 3-pentanediol monoisobutyrate, nα, nω -dibenzyloxycarbonyl-L-arginine, 9-decenoic acid, 1- (1, 3-dioxolan-2-yl) acetone;
Preferably, the production refers to the production of ethyl 3-phenylpropionate, ethyl 4-acetoxybutyrate, ethyl tridecanoate, (2S-cis) -tetrahydro4-methyl-2- (2-methyl-1-propenyl) -2H-pyran, dipropylene glycol, 10-undecen-1-ol, dihydrolinalool, 1-pentadecanol, 1, 3-propanediol diacetate, 1-vinyl-1, 5-dimethyl-4-hexenyl butyrate, 3-hydroxybutyraldehyde, 2-dodecanone, ethyl 3-phenylpropionate, pentadecanoate, nerol acetate, dihydrolinalool, ethylene glycol laurate, 2, 4-trimethyl-1, 3-pentanediol monoisobutyrate, nα, nω -dibenzyloxycarbonyl-L-arginine, 9-decenoic acid, 1- (1, 3-dioxolan-2-yl) acetone;
Preferably, the adjustment means: increasing ethyl 3-phenylpropionate, ethyl 4-acetoxybutyrate, ethyl tridecanoate, (2S-cis) -tetrahydro-4-methyl-2- (2-methyl-1-propenyl) -2H-pyran, dipropylene glycol, 10-undecen-1-ol, dihydrolinalool, 1-pentadecanol, 1, 3-propanediol diacetate, butyric acid-1-vinyl-1, 5-dimethyl-4-hexenyl ester, 3-hydroxybutyraldehyde, 2-dodecanone during mixed fermentation, or increasing ethyl laurate, ethyl n-hexanoate, 7-octenoate, ethyl 9-hexadecenoate, isopentyl octanoate, 3-methylpentanoate, octanoate, (E) -7, 11-dimethyl-3-methylene-1, 6, 10-dodecene during single fermentation, decreasing ethyl isovalerate, geranyl acetonate, methyl triacontate C30, diethylene glycol, octanol, 1-isopropoxy-2-hydroxy-2-yl butyrate, (1-acetoxy-3-hydroxypropionate), geranyl acetate, methyl octanoate, 2-methylpropanoate, 2-methyl octanoate, 2-propanoate, methyl octanoate, 2-ethyl octanoate, 2- (2-hexadecenoate) octanoate during single fermentation;
Preferably, the single-fungus fermentation means: fermenting with Rhodotorula mucilaginosa strain 147 alone;
Preferably, the mixed bacteria fermentation means: fermenting with Rhodotorula mucilaginosa strain 147 and Saccharomyces cerevisiae F15.
The fermentation is selected from: single-bacteria fermentation or mixed-bacteria fermentation;
Preferably, the mixed bacteria fermentation means: mixing and fermenting rhodotorula mucilaginosa (Rhodotorula mucilaginosa) strain 147 and Saccharomyces cerevisiae F15;
Preferably, the brewing means brewing wine.
A brewing starter, its fermentation active substance comprises Rhodotorula mucilaginosa (Rhodotorula mucilaginosa) strain 147 with a preservation number of CCTCC NO: M2023934.
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 rhodotorula mucilaginosa (Rhodotorula mucilaginosa) strain 147 with a preservation number of CCTCC NO: M2023934 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 147 into a substrate;
preferably, the mixed fermentation means that a strain 147 and saccharomyces cerevisiae are inoculated into a substrate for fermentation;
preferably, the saccharomyces cerevisiae is selected from: f15 or F33;
Preferably, the concentration of the access strain is 10 6-107 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 Rhodotorula mucilaginosa (Rhodotorula mucilaginosa) strain 147 with preservation number of CCTCC NO: M2023934 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 147 into a substrate;
preferably, the mixed fermentation means that a strain 147 and saccharomyces cerevisiae are inoculated into a substrate for fermentation;
preferably, the saccharomyces cerevisiae is selected from: f15 or F33;
Preferably, the concentration of the access strain is 10 6-107 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 F15 strain, the strain can greatly improve the content of certain aroma substances through aroma substance component identification experiments, for example: ethyl laurate, ethyl n-hexanoate, ethyl 7-octenoate, ethyl 9-hexadecenoate, isoamyl octanoate, 3-methylpentanoic acid, octanoic acid, (E) -7, 11-dimethyl-3-methylene-1, 6, 10-dodecatriene; the isoamyl octanoate and octanoic acid can be raised above a threshold value, so that people can sniff.
And aroma substances such as ethyl 3-phenylpropionate, ethyl 4-acetoxybutyrate, ethyl tridecanoate, (2S-cis) -tetrahydrochysene-4-methyl-2- (2-methyl-1-propenyl) -2H-pyran, dipropylene glycol, 10-undecen-1-ol, dihydrolinalool, 1-pentadecanol, 1, 3-propanediol diacetate, butyric acid-1-vinyl-1, 5-dimethyl-4-hexenyl ester, 3-hydroxybutyraldehyde and 2-dodecanone are not produced or produced in single bacterial fermentation of 147 strain and F15 strain respectively, but can produce high content of the aroma substances in mixed bacterial fermentation of the 147 strain and the F15 strain.
For certain aroma substances produced by the single-fungus fermentation of the Saccharomyces cerevisiae strain F15, the single-fungus fermentation of the strain of the invention does not produce at all or falls below a threshold value, so that people cannot sniff, for example: ethyl isovalerate, geranyl acetone, methyl triacontanoate C30, diethylene glycol, octanol, 1-isopropoxy-2-propanol, (1-acetoxy-3-hydroxyprop-2-yl) acetate, methyl octanoate, geraniol octanoate, phenyl carbamate, propyl octanoate, vinyl formate, pentadecanol, 2-methyl-propionic acid, ethyl- (Z) -4-decenoic acid, 4-hydroxy-2-butanone.
In addition, the strains of the invention are capable of being multiplied after fermentation with their mixture, for example octanol, 4-vinyl-2-methoxyphenol, 2, 3-dihydrobenzofuran, vinyl formate, 3-methylpentanoic acid, octanoic acid, 2-methyl-propionic acid, compared to the content of certain aroma substances produced by commercial Saccharomyces cerevisiae strain F15.
Meanwhile, compared with the commercial saccharomyces cerevisiae strain F15, the strain of the invention can also generate aroma substances which cannot be generated by the F15, such as: ethyl 3-phenylpropionate, ethyl pentadecanoate, nerol acetate, dihydrolinalool, ethylene glycol laurate, 2, 4-trimethyl-1, 3-pentanediol monoisobutyrate, nα, nω -dibenzyloxycarbonyl-L-arginine, 9-decenoic acid, 1- (1, 3-dioxolan-2-yl) acetone, wherein the yield of 1- (1, 3-dioxolan-2-yl) acetone is up to 557.1879648 μg/L, the yield of ethyl 3-phenylpropionate is up to 266.65 μg/L, and the yield of dihydrolinalool is up to 66.17 μg/L. The strain is identified as rhodotorula mucilaginosa (Rhodotorula mucilaginosa). The applicant designated 147 and sent the strain for preservation.
Preservation number: CCTCC NO: M2023934
Classification naming: rhodotorula mucilaginosa 147
Latin name: rhodotorula mucilaginosa 147A 147
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 long jersey, the Saimei, the Meiluo, the Pinctada, the Chardonnay, the Bai Shi south, the Jiamei and the Xian Ping are all common grape varieties and 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 147 of the present invention
The embodiment provides a rhodotorula mucilaginosa (Rhodotorula mucilaginosa) strain 147 with a preservation number of CCTCC NO: M2023934.
Any of the actions of culturing, propagating, fermenting, enriching, producing, preparing, using, inoculating, amplifying, transforming, modifying, selling, offering to sell rhodotorula mucilaginosa (Rhodotorula mucilaginosa) strain 147 with a preservation number of CCTCC NO: M2023934 and/or the actions of combining rhodotorula mucilaginosa (Rhodotorula mucilaginosa) strain 147 with a preservation number of CCTCC NO: M2023934 with other Saccharomyces cerevisiae 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 common general knowledge 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 rhodotorula mucilaginosa (Rhodotorula mucilaginosa) strain 147 with the preservation number of CCTCC NO: M2023934 is prepared into microbial agent products with different dosage forms, different storage conditions and different shelf lives, which is free from technical barriers and can be easily achieved by the person skilled in the art.
Group 2 example, use of the strain 147 of the invention
The present set of examples provides the use of rhodotorula mucilaginosa (Rhodotorula mucilaginosa) strain 147 with a preservation number of CCTCC NO: M2023934 in the production, regulation of aroma substances and/or fermentation and/or brewing.
In some embodiments of the present invention, in some embodiments, the aroma substances are selected from lauric acid ethyl ester, ethyl caproate, 7-octenoate ethyl ester, 9-hexadecenoate ethyl ester, isoamyl octanoate, 3-methyl valeric acid, octanoic acid, (E) -7, 11-dimethyl-3-methylene-1, 6, 10-dodecatriene, 3-phenylpropionic acid ethyl ester, 4-acetoxybutyric acid ethyl ester, tridecanoic acid ethyl ester, (2S-cis) -tetrahydrogen-4-methyl-2- (2-methyl-1-propenyl) -2H-pyran, dipropylene glycol, 10-undecen-1-ol, dihydro linalool, 1-pentadecanol, 1, 3-propylene glycol diacetate, butyric acid-1-vinyl-1, 5-dimethyl-4-hexenyl ester 3-hydroxybutyraldehyde, 2-dodecanone, ethyl isovalerate, geranyl acetone, methyl triacontanoate C30, diethylene glycol, octanol, 1-isopropoxy-2-propanol, (1-acetoxy-3-hydroxyprop-2-yl) acetate, methyl octanoate, geranyl octanoate, phenyl carbamate, ethyl acetate, ethyl octanoate, methyl octanoate, ethyl octanoate, methyl propyl octanoate, vinyl formate, pentadecanol, 2-methyl-propionic acid, ethyl- (Z) -4-decenoic acid, 4-hydroxy-2-butanone, 4-vinyl-2-methoxyphenol, 2, 3-dihydrobenzofuran, ethyl pentadecanoate, nerol acetate, ethylene glycol laurate, ethyl acetate, and methyl, ethyl acrylate, and methyl, and ethyl acrylate, methyl acrylate, and methyl acrylate, and methyl acrylate and, 2, 4-trimethyl-1, 3-pentanediol monoisobutyrate, nα, nω -dibenzyloxycarbonyl-L-arginine, 9-decenoic acid, 1- (1, 3-dioxolan-2-yl) acetone;
Preferably, the production refers to the production of ethyl 3-phenylpropionate, ethyl 4-acetoxybutyrate, ethyl tridecanoate, (2S-cis) -tetrahydro4-methyl-2- (2-methyl-1-propenyl) -2H-pyran, dipropylene glycol, 10-undecen-1-ol, dihydrolinalool, 1-pentadecanol, 1, 3-propanediol diacetate, 1-vinyl-1, 5-dimethyl-4-hexenyl butyrate, 3-hydroxybutyraldehyde, 2-dodecanone, ethyl 3-phenylpropionate, pentadecanoate, nerol acetate, dihydrolinalool, ethylene glycol laurate, 2, 4-trimethyl-1, 3-pentanediol monoisobutyrate, nα, nω -dibenzyloxycarbonyl-L-arginine, 9-decenoic acid, 1- (1, 3-dioxolan-2-yl) acetone;
Preferably, the adjustment means: increasing ethyl 3-phenylpropionate, ethyl 4-acetoxybutyrate, ethyl tridecanoate, (2S-cis) -tetrahydro-4-methyl-2- (2-methyl-1-propenyl) -2H-pyran, dipropylene glycol, 10-undecen-1-ol, dihydrolinalool, 1-pentadecanol, 1, 3-propanediol diacetate, butyric acid-1-vinyl-1, 5-dimethyl-4-hexenyl ester, 3-hydroxybutyraldehyde, 2-dodecanone during mixed fermentation, or increasing ethyl laurate, ethyl n-hexanoate, 7-octenoate, ethyl 9-hexadecenoate, isopentyl octanoate, 3-methylpentanoate, octanoate, (E) -7, 11-dimethyl-3-methylene-1, 6, 10-dodecene during single fermentation, decreasing ethyl isovalerate, geranyl acetonate, methyl triacontate C30, diethylene glycol, octanol, 1-isopropoxy-2-hydroxy-2-yl butyrate, (1-acetoxy-3-hydroxypropionate), geranyl acetate, methyl octanoate, 2-methylpropanoate, 2-methyl octanoate, 2-propanoate, methyl octanoate, 2-ethyl octanoate, 2- (2-hexadecenoate) octanoate during single fermentation;
Preferably, the single-fungus fermentation means: fermenting with Rhodotorula mucilaginosa strain 147 alone;
Preferably, the mixed bacteria fermentation means: fermenting with Rhodotorula mucilaginosa strain 147 and Saccharomyces cerevisiae F15.
In other embodiments, the fermentation is selected from: single-bacteria fermentation or mixed-bacteria fermentation;
Preferably, the mixed bacteria fermentation means: mixing and fermenting rhodotorula mucilaginosa (Rhodotorula mucilaginosa) strain 147 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 rhodotorula mucilaginosa (Rhodotorula mucilaginosa) strain 147 with the preservation number of CCTCC NO: M2023934.
In some embodiments, 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 man skilled in the art can select and blend the edible auxiliary materials and prepare rhodotorula mucilaginosa (Rhodotorula mucilaginosa) strain 147 with the preservation number of CCTCC NO: M2023934 into different dosage forms, such as powder, granules, liquid and the like, according to different requirements in practical production and application and in combination with conventional technical means in the field of microbial preparation (for example, encyclopedia of preparation technology, research and application of microbial preparation 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: the rhodotorula mucilaginosa (Rhodotorula mucilaginosa) strain 147 with the preservation number of CCTCC NO: M2023934 is adopted for fermentation.
In some embodiments, the fermentation is selected from single or mixed fermentation;
Preferably, the single-fungus fermentation refers to fermentation by inoculating a strain 147 into a substrate;
preferably, the mixed fermentation means that a strain 147 and saccharomyces cerevisiae are inoculated into a substrate for fermentation;
preferably, the saccharomyces cerevisiae is selected from: f15 or F33;
Preferably, the concentration of the access strain is 10 6-107 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: rhodotorula mucilaginosa (Rhodotorula mucilaginosa) strain 147 with preservation number of CCTCC NO: M2023934 is adopted to ferment and brew with grape juice as 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 147 into a substrate;
preferably, the mixed fermentation means that a strain 147 and saccharomyces cerevisiae are inoculated into a substrate for fermentation;
preferably, the saccharomyces cerevisiae is selected from: f15 or F33;
Preferably, the concentration of the access strain is 10 6-107 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 Rhodotorula mucilaginosa (Rhodotorula mucilaginosa) Strain 147 according to the present 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, PCR amplification is carried out by using primers NL-1 and NL-4 described in Chinese patent No. 202110107456.8, a gene sequence of a 26S rDNA fragment is obtained by sequencing, known standard strain information with higher homology is obtained by 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(s) | Sample number (copy) |
| Radix seu herba Gei Albae | Foot producing area of Helan mountain | Acquisition of | 25 |
| Saimei (Saimei Rong) | Foot producing area of Helan mountain | Acquisition of | 25 |
| Mello | Foot producing area of Helan mountain | Acquisition of | 25 |
| Pinli bead | Foot producing area of Helan mountain | Acquisition of | 25 |
| Chardonnay | 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 |
| Xian Pink | Foot producing area of Helan mountain | Acquisition of | 25 |
In this experimental example, a total of 26 Rhodotorula mucilaginosa strains were obtained. Respectively inoculating 12 Rhodotorula mucilaginosa strains into sterilized Wedelir grape juice (100 ℃ for 10 min) according to the initial concentration of 10 6 CFU/mL, fermenting at 18+/-2 ℃ for 18 days, measuring the fragrance content of the strain, finally obtaining a rhodotorula mucilaginosa (Rhodotorula mucilaginosa) strain 147 with the best fragrance producing effect, and delivering the strain to be preserved, wherein the preservation information is as follows:
Preservation number: CCTCC NO: M2023934
Classification naming: rhodotorula mucilaginosa 147
Latin name: rhodotorula mucilaginosa 147A 147
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 strain 147 of the present invention
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 2S2O5) and 20mg/L pectase (more than or equal to 500U/mg) to inhibit bacterial diseases and improve juice yield.
Grape juice fermentation conditions: the macerated must was divided into 3 groups (147 groups, F15 groups, f15+147 groups).
The grape juice of 147 group is inoculated with rhodotorula mucilaginosa (Rhodotorula mucilaginosa) strain 147 according to the initial concentration of 10 6 CFU/mL;
the grape juice of the F15 group is inoculated into Saccharomyces cerevisiae F15 according to the initial concentration of 10 6 CFU/mL;
the F15+147 group must was inoculated with both Saccharomyces cerevisiae F15 and Rhodotorula mucilaginosa (Rhodotorula mucilaginosa) strain 147 at an initial concentration of 10 6 CFU/mL.
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/L of 4-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: inertCap WAX polarity chromatography 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
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 147 of the present invention
1. Group and detection sample preparation:
Group (one) 147: the strain 147 glycerol storage tube of the invention 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 147 glycerol storage tube 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) 147+F15 group: the strain 147 glycerol save tube and the saccharomyces cerevisiae F15 glycerol save tube are respectively dissolved and then inoculated into 5mL YPD culture medium, and are cultured for 12-20h at 30 ℃, and after three generations of activation, the strain 147 glycerol save tube and the saccharomyces cerevisiae F15 glycerol save tube are inoculated into 400mL fermentation culture medium according to 1% (volume ratio), 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 rhodotorula mucilaginosa (Rhodotorula mucilaginosa) strain 147 is characterized in that the preservation number is CCTCC NO: M2023934.
2. The application of rhodotorula mucilaginosa (Rhodotorula mucilaginosa) strain 147 with preservation number of CCTCC NO: M2023934 in the aspects of production, aroma substance regulation and/or fermentation and/or brewing.
3. The preservation number of claim 2 is CCTCC NO: the use of rhodotorula mucilaginosa (Rhodotorula mucilaginosa) strain 147 of M2023934 for the production, regulation of aroma substances and/or fermentation and/or brewing, characterized in that, the aroma substances are selected from lauric acid ethyl ester, ethyl caproate, 7-octenoate ethyl ester, 9-hexadecenoate ethyl ester, isoamyl octanoate, 3-methyl valeric acid, octanoic acid, (E) -7, 11-dimethyl-3-methylene-1, 6, 10-dodecatriene, 3-ethyl phenylpropionate, 4-acetoxybutyric acid ethyl ester, tridecanoate ethyl ester, (2S-cis) -tetrahydrogen-4-methyl-2- (2-methyl-1-propenyl) -2H-pyran, dipropylene glycol, 10-undecen-1-ol, dihydro linalool, 1-pentadecanol, and 1, 3-propanediol diacetate, 1-vinyl-1, 5-dimethyl-4-hexenyl butyrate, 3-hydroxybutyraldehyde, 2-dodecanone, ethyl isovalerate, geranyl acetone, methyl triacontate C30, diethylene glycol, octanol, 1-isopropoxy-2-propanol, (1-acetoxy-3-hydroxyprop-2-yl) acetate, methyl octanoate, geraniol octanoate, phenyl carbamate, propyl octanoate, vinyl formate, pentadecanol, 2-methyl-propionic acid, ethyl- (Z) -4-decenoic acid, 4-hydroxy-2-butanone, 4-vinyl-2-methoxyphenol, 2, 3-dihydrobenzofuran, ethyl pentadecanoate, nerol acetate, ethylene glycol laurate, 2, 4-trimethyl-1, 3-pentanediol monoisobutyrate, nα, nω -dibenzyloxycarbonyl-L-arginine, 9-decenoic acid, 1- (1, 3-dioxolan-2-yl) acetone;
And/or, said producing refers to single or mixed fermentation to produce ethyl 3-phenylpropionate, ethyl 4-acetoxybutyrate, ethyl tridecanoate, (2S-cis) -tetrahydro4-methyl-2- (2-methyl-1-propenyl) -2H-pyran, dipropylene glycol, 10-undecen-1-ol, dihydrolinalool, 1-pentadecanol, 1, 3-propanediol diacetate, butyric acid-1-vinyl-1, 5-dimethyl-4-hexenyl ester, 3-hydroxybutyraldehyde, 2-dodecanone, ethyl 3-phenylpropionate, pentadecanoate, nerol acetate, dihydrolinalool, ethylene glycol laurate, 2, 4-trimethyl-1, 3-pentanediol monoisobutyrate, nα, nω -dibenzyloxycarbonyl-L-arginine, 9-decenoic acid, 1- (1, 3-dioxolan-2-yl) acetone;
And/or, the adjusting means: increasing ethyl 3-phenylpropionate, ethyl 4-acetoxybutyrate, ethyl tridecanoate, (2S-cis) -tetrahydro-4-methyl-2- (2-methyl-1-propenyl) -2H-pyran, dipropylene glycol, 10-undecen-1-ol, dihydrolinalool, 1-pentadecanol, 1, 3-propanediol diacetate, butyric acid-1-vinyl-1, 5-dimethyl-4-hexenyl ester, 3-hydroxybutyraldehyde, 2-dodecanone during mixed fermentation, or increasing ethyl laurate, ethyl n-hexanoate, 7-octenoate, ethyl 9-hexadecenoate, isopentyl octanoate, 3-methylpentanoate, octanoate, (E) -7, 11-dimethyl-3-methylene-1, 6, 10-dodecene during single fermentation, decreasing ethyl isovalerate, geranyl acetonate, methyl triacontate C30, diethylene glycol, octanol, 1-isopropoxy-2-hydroxy-2-yl butyrate, (1-acetoxy-3-hydroxypropionate), geranyl acetate, methyl octanoate, 2-methylpropanoate, 2-methyl octanoate, 2-propanoate, methyl octanoate, 2-ethyl octanoate, 2- (2-hexadecenoate) octanoate during single fermentation;
and/or, the single-fungus fermentation means: fermenting with Rhodotorula mucilaginosa strain 147 alone;
And/or, the mixed bacteria fermentation means: fermenting with Rhodotorula mucilaginosa strain 147 and Saccharomyces cerevisiae F15.
4. Use of rhodotorula mucilaginosa (Rhodotorula mucilaginosa) strain 147 with a preservation number of CCTCC NO: M2023934 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: mixing and fermenting rhodotorula mucilaginosa (Rhodotorula mucilaginosa) strain 147 and Saccharomyces cerevisiae F15;
and/or, brewing refers to brewing wine.
5. A brewing starter is characterized in that the fermentation active substance comprises rhodotorula mucilaginosa (Rhodotorula mucilaginosa) strain 147 with a preservation number of CCTCCNO: M2023934.
6. A brewery starter according to claim 5, wherein said fermentation active further comprises saccharomyces cerevisiae;
Preferably, the brewing starter is a microbial inoculum;
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 rhodotorula mucilaginosa (Rhodotorula mucilaginosa) strain 147 with the preservation number of CCTCC NO: M2023934 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 fermentation by inoculating a strain 147 into a substrate;
and/or, the mixed fermentation refers to inoculating a strain 147 and saccharomyces cerevisiae into a substrate for fermentation;
and/or, the saccharomyces cerevisiae is selected from: f15 or F33;
and/or the concentration of the access strain is 10 6-107 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 rhodotorula mucilaginosa (Rhodotorula mucilaginosa) strain 147 with the preservation number of CCTCC NO: M2023934 is adopted to perform fermentation brewing 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 fermentation by inoculating a strain 147 into a substrate;
and/or, the mixed fermentation refers to inoculating a strain 147 and saccharomyces cerevisiae into a substrate for fermentation;
and/or, the saccharomyces cerevisiae is selected from: f15 or F33;
and/or the concentration of the access strain is 10 6-107 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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