CN116656561A - 融合魏斯氏菌及其应用 - Google Patents
融合魏斯氏菌及其应用 Download PDFInfo
- Publication number
- CN116656561A CN116656561A CN202310697229.4A CN202310697229A CN116656561A CN 116656561 A CN116656561 A CN 116656561A CN 202310697229 A CN202310697229 A CN 202310697229A CN 116656561 A CN116656561 A CN 116656561A
- Authority
- CN
- China
- Prior art keywords
- weissella
- fusion
- fermentation
- acetylfuran
- culture
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 230000004927 fusion Effects 0.000 title claims abstract description 35
- 241000202221 Weissella Species 0.000 claims abstract description 35
- IEMMBWWQXVXBEU-UHFFFAOYSA-N 2-acetylfuran Chemical compound CC(=O)C1=CC=CO1 IEMMBWWQXVXBEU-UHFFFAOYSA-N 0.000 claims abstract description 28
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims abstract description 16
- 238000004321 preservation Methods 0.000 claims abstract description 13
- 235000021107 fermented food Nutrition 0.000 claims abstract description 7
- 238000000855 fermentation Methods 0.000 claims description 19
- 230000004151 fermentation Effects 0.000 claims description 19
- 239000001963 growth medium Substances 0.000 claims description 14
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 12
- 235000015278 beef Nutrition 0.000 claims description 12
- 239000002609 medium Substances 0.000 claims description 10
- 238000009630 liquid culture Methods 0.000 claims description 9
- 239000000843 powder Substances 0.000 claims description 9
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 7
- 238000000034 method Methods 0.000 claims description 7
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 6
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 claims description 6
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims description 6
- 239000000284 extract Substances 0.000 claims description 6
- 239000008103 glucose Substances 0.000 claims description 6
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 6
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 6
- 229940099596 manganese sulfate Drugs 0.000 claims description 6
- 235000007079 manganese sulphate Nutrition 0.000 claims description 6
- 239000011702 manganese sulphate Substances 0.000 claims description 6
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 claims description 6
- YWYZEGXAUVWDED-UHFFFAOYSA-N triammonium citrate Chemical compound [NH4+].[NH4+].[NH4+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O YWYZEGXAUVWDED-UHFFFAOYSA-N 0.000 claims description 6
- 241000186675 Weissella confusa Species 0.000 claims description 5
- 238000004519 manufacturing process Methods 0.000 claims description 5
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 claims description 5
- 229920000053 polysorbate 80 Polymers 0.000 claims description 5
- 230000004913 activation Effects 0.000 claims description 4
- 239000001888 Peptone Substances 0.000 claims description 3
- 108010080698 Peptones Proteins 0.000 claims description 3
- 229940040526 anhydrous sodium acetate Drugs 0.000 claims description 3
- 210000004556 brain Anatomy 0.000 claims description 3
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 claims description 3
- 235000019319 peptone Nutrition 0.000 claims description 3
- 238000002360 preparation method Methods 0.000 claims description 3
- 239000001632 sodium acetate Substances 0.000 claims description 3
- 235000017281 sodium acetate Nutrition 0.000 claims description 3
- 239000011780 sodium chloride Substances 0.000 claims description 3
- 230000003068 static effect Effects 0.000 claims description 3
- 239000001393 triammonium citrate Substances 0.000 claims description 3
- 235000011046 triammonium citrate Nutrition 0.000 claims description 3
- 239000012137 tryptone Substances 0.000 claims description 3
- 239000003795 chemical substances by application Substances 0.000 claims description 2
- 238000012258 culturing Methods 0.000 claims description 2
- 239000003205 fragrance Substances 0.000 claims description 2
- 238000011081 inoculation Methods 0.000 claims description 2
- 230000000813 microbial effect Effects 0.000 claims description 2
- 239000012450 pharmaceutical intermediate Substances 0.000 claims description 2
- 239000002994 raw material Substances 0.000 claims description 2
- 229920000136 polysorbate Polymers 0.000 claims 1
- 229960005486 vaccine Drugs 0.000 claims 1
- 244000005700 microbiome Species 0.000 abstract description 5
- 235000013599 spices Nutrition 0.000 abstract description 3
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 15
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 description 13
- 239000007788 liquid Substances 0.000 description 8
- 230000001580 bacterial effect Effects 0.000 description 7
- 238000001514 detection method Methods 0.000 description 5
- 238000002290 gas chromatography-mass spectrometry Methods 0.000 description 5
- 108020004465 16S ribosomal RNA Proteins 0.000 description 4
- LZCLXQDLBQLTDK-UHFFFAOYSA-N ethyl 2-hydroxypropanoate Chemical compound CCOC(=O)C(C)O LZCLXQDLBQLTDK-UHFFFAOYSA-N 0.000 description 4
- 239000000047 product Substances 0.000 description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- 241000894006 Bacteria Species 0.000 description 3
- 238000000605 extraction Methods 0.000 description 3
- 235000013305 food Nutrition 0.000 description 3
- 239000012634 fragment Substances 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 238000012216 screening Methods 0.000 description 3
- 239000006228 supernatant Substances 0.000 description 3
- 235000014101 wine Nutrition 0.000 description 3
- NGDNVOAEIVQRFH-UHFFFAOYSA-N 2-nonanol Chemical compound CCCCCCCC(C)O NGDNVOAEIVQRFH-UHFFFAOYSA-N 0.000 description 2
- KWOLFJPFCHCOCG-UHFFFAOYSA-N Acetophenone Chemical compound CC(=O)C1=CC=CC=C1 KWOLFJPFCHCOCG-UHFFFAOYSA-N 0.000 description 2
- 229920001817 Agar Polymers 0.000 description 2
- FINHMKGKINIASC-UHFFFAOYSA-N Tetramethylpyrazine Chemical compound CC1=NC(C)=C(C)N=C1C FINHMKGKINIASC-UHFFFAOYSA-N 0.000 description 2
- 239000008272 agar Substances 0.000 description 2
- IOJUPLGTWVMSFF-UHFFFAOYSA-N benzothiazole Chemical compound C1=CC=C2SC=NC2=C1 IOJUPLGTWVMSFF-UHFFFAOYSA-N 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- 229940116333 ethyl lactate Drugs 0.000 description 2
- ZSIAUFGUXNUGDI-UHFFFAOYSA-N hexan-1-ol Chemical compound CCCCCCO ZSIAUFGUXNUGDI-UHFFFAOYSA-N 0.000 description 2
- 239000002054 inoculum Substances 0.000 description 2
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 2
- 238000001819 mass spectrum Methods 0.000 description 2
- 238000009629 microbiological culture Methods 0.000 description 2
- 235000013336 milk Nutrition 0.000 description 2
- 239000008267 milk Substances 0.000 description 2
- 210000004080 milk Anatomy 0.000 description 2
- ZWRUINPWMLAQRD-UHFFFAOYSA-N nonan-1-ol Chemical compound CCCCCCCCCO ZWRUINPWMLAQRD-UHFFFAOYSA-N 0.000 description 2
- SJWFXCIHNDVPSH-UHFFFAOYSA-N octan-2-ol Chemical compound CCCCCCC(C)O SJWFXCIHNDVPSH-UHFFFAOYSA-N 0.000 description 2
- BDERNNFJNOPAEC-UHFFFAOYSA-N propan-1-ol Chemical compound CCCO BDERNNFJNOPAEC-UHFFFAOYSA-N 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- 238000012163 sequencing technique Methods 0.000 description 2
- 239000002689 soil Substances 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 230000001954 sterilising effect Effects 0.000 description 2
- 238000004659 sterilization and disinfection Methods 0.000 description 2
- MIDXCONKKJTLDX-UHFFFAOYSA-N 3,5-dimethylcyclopentane-1,2-dione Chemical compound CC1CC(C)C(=O)C1=O MIDXCONKKJTLDX-UHFFFAOYSA-N 0.000 description 1
- 244000144725 Amygdalus communis Species 0.000 description 1
- 235000011437 Amygdalus communis Nutrition 0.000 description 1
- 241000228212 Aspergillus Species 0.000 description 1
- 241000228197 Aspergillus flavus Species 0.000 description 1
- 241000228245 Aspergillus niger Species 0.000 description 1
- 241000193744 Bacillus amyloliquefaciens Species 0.000 description 1
- 241000194108 Bacillus licheniformis Species 0.000 description 1
- 240000000560 Citrus x paradisi Species 0.000 description 1
- 241000235058 Komagataella pastoris Species 0.000 description 1
- 241000186660 Lactobacillus Species 0.000 description 1
- 235000007688 Lycopersicon esculentum Nutrition 0.000 description 1
- 241000235395 Mucor Species 0.000 description 1
- 240000003768 Solanum lycopersicum Species 0.000 description 1
- 244000269722 Thea sinensis Species 0.000 description 1
- 240000006365 Vitis vinifera Species 0.000 description 1
- 235000014787 Vitis vinifera Nutrition 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 239000011543 agarose gel Substances 0.000 description 1
- 235000020224 almond Nutrition 0.000 description 1
- 230000003698 anagen phase Effects 0.000 description 1
- 235000013405 beer Nutrition 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 235000008429 bread Nutrition 0.000 description 1
- 235000013736 caramel Nutrition 0.000 description 1
- 239000012159 carrier gas Substances 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 235000016213 coffee Nutrition 0.000 description 1
- 235000013353 coffee beverage Nutrition 0.000 description 1
- 235000009508 confectionery Nutrition 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- 238000003113 dilution method Methods 0.000 description 1
- 239000004205 dimethyl polysiloxane Substances 0.000 description 1
- 235000013870 dimethyl polysiloxane Nutrition 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 239000000796 flavoring agent Substances 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 238000004817 gas chromatography Methods 0.000 description 1
- 235000009569 green tea Nutrition 0.000 description 1
- 238000003988 headspace gas chromatography Methods 0.000 description 1
- 238000001319 headspace solid-phase micro-extraction Methods 0.000 description 1
- 239000001307 helium Substances 0.000 description 1
- 229910052734 helium Inorganic materials 0.000 description 1
- SWQJXJOGLNCZEY-UHFFFAOYSA-N helium atom Chemical compound [He] SWQJXJOGLNCZEY-UHFFFAOYSA-N 0.000 description 1
- 239000000543 intermediate Substances 0.000 description 1
- 230000006799 invasive growth in response to glucose limitation Effects 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 239000004310 lactic acid Substances 0.000 description 1
- 235000014655 lactic acid Nutrition 0.000 description 1
- 229940039696 lactobacillus Drugs 0.000 description 1
- 238000004949 mass spectrometry Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- 239000013028 medium composition Substances 0.000 description 1
- 230000000877 morphologic effect Effects 0.000 description 1
- 229930014626 natural product Natural products 0.000 description 1
- 235000014571 nuts Nutrition 0.000 description 1
- CXQXSVUQTKDNFP-UHFFFAOYSA-N octamethyltrisiloxane Chemical compound C[Si](C)(C)O[Si](C)(C)O[Si](C)(C)C CXQXSVUQTKDNFP-UHFFFAOYSA-N 0.000 description 1
- 238000004987 plasma desorption mass spectroscopy Methods 0.000 description 1
- 229920000435 poly(dimethylsiloxane) Polymers 0.000 description 1
- 235000013606 potato chips Nutrition 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 238000002470 solid-phase micro-extraction Methods 0.000 description 1
- 235000013555 soy sauce Nutrition 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 229920002994 synthetic fiber Polymers 0.000 description 1
- 235000013311 vegetables Nutrition 0.000 description 1
- 235000021419 vinegar Nutrition 0.000 description 1
- 239000000052 vinegar Substances 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
- 239000009029 wenxiao Substances 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
-
- A—HUMAN NECESSITIES
- A21—BAKING; EDIBLE DOUGHS
- A21D—TREATMENT, e.g. PRESERVATION, OF FLOUR OR DOUGH, e.g. BY ADDITION OF MATERIALS; BAKING; BAKERY PRODUCTS; PRESERVATION THEREOF
- A21D13/00—Finished or partly finished bakery products
-
- A—HUMAN NECESSITIES
- A21—BAKING; EDIBLE DOUGHS
- A21D—TREATMENT, e.g. PRESERVATION, OF FLOUR OR DOUGH, e.g. BY ADDITION OF MATERIALS; BAKING; BAKERY PRODUCTS; PRESERVATION THEREOF
- A21D8/00—Methods for preparing or baking dough
- A21D8/02—Methods for preparing dough; Treating dough prior to baking
- A21D8/04—Methods for preparing dough; Treating dough prior to baking treating dough with microorganisms or enzymes
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L27/00—Spices; Flavouring agents or condiments; Artificial sweetening agents; Table salts; Dietetic salt substitutes; Preparation or treatment thereof
- A23L27/50—Soya sauce
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12G—WINE; PREPARATION THEREOF; ALCOHOLIC BEVERAGES; PREPARATION OF ALCOHOLIC BEVERAGES NOT PROVIDED FOR IN SUBCLASSES C12C OR C12H
- C12G3/00—Preparation of other alcoholic beverages
- C12G3/02—Preparation of other alcoholic beverages by fermentation
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12J—VINEGAR; PREPARATION OR PURIFICATION THEREOF
- C12J1/00—Vinegar; Preparation or purification thereof
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P17/00—Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms
- C12P17/02—Oxygen as only ring hetero atoms
- C12P17/04—Oxygen as only ring hetero atoms containing a five-membered hetero ring, e.g. griseofulvin, vitamin C
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- General Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Food Science & Technology (AREA)
- Microbiology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Biomedical Technology (AREA)
- Virology (AREA)
- Medicinal Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nutrition Science (AREA)
- Polymers & Plastics (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
本发明属于微生物技术领域，具体涉及产2‑乙酰基呋喃的融合魏斯氏菌及其应用。针对现有研究中未发现能产2‑乙酰基呋喃的融合魏斯氏菌，本发明从酒曲中分离筛选得到一株能产2‑乙酰基呋喃的融合魏斯氏菌新菌株，其保藏编号为：CGMCC No.26364。该菌株不仅可应用于发酵食品行业，还可应用于生物来源的香精香料、医疗行业，促进行业发展。
Description
技术领域
本发明属于微生物技术领域，具体涉及产2-乙酰基呋喃的融合魏斯氏菌及其应用。
背景技术
2-乙酰基呋喃黄棕色液体，呈杏仁、坚果、酵香、牛奶和甜的焦糖似香气。天然品存在于咖啡、西红柿、啤酒、绿茶、葡萄干和马铃薯片的挥发性香成分中。可用作有机合成原料、医药中间体、食品用香料。
现有研究中，发现可通过微生物的调节，提高发酵食品挥发性物质中2-乙酰基呋喃的含量。如汤晓宏等(汤晓宏,胡文效,魏彦锋,等.葡萄酒野生酿酒酵母的筛选及其生物特性的研究[J].山东大学学报：理学版,2014,49(3):7.)从葡萄果实表皮和葡萄园土壤中分离纯化得到一株酿酒酵母PJ16，其所酿葡萄酒的香气成分正己醇、1-壬醇、2-壬醇、乳酸乙酯、乙酰苯、苯并噻唑、2-乙酰基呋喃浓度远高于对照组商品酵母所酿葡萄酒。曾德勇(曾德勇.酱香型白酒生产中功能微生物的优化组合及其应用研究[D].贵州大学.)通过优化功能细菌(解淀粉芽孢杆菌、地衣芽孢杆菌)与酵母菌(酿酒酵母、毕赤酵母)以及曲霉(黑曲霉、毛霉、黄曲霉)之间的比例，制备强化大曲，其酒样中正丙醇、乙酸、乳酸乙酯、2,3,5,6-四甲基吡嗪、己酸酯类羟乙基、2-乙酰呋喃等挥发性物质有了明显的提高。
融合魏斯氏菌是一种广泛存在于发酵食品、肉类、牛奶、蔬菜和土壤的乳酸菌，常用于食品行业中以改善食品的性质。但现有研究中未发现有能产2-乙酰基呋喃的融合魏斯氏菌。
发明内容
针对现有研究中未发现能产2-乙酰基呋喃的融合魏斯氏菌，本发明从酒曲中分离筛选得到一株能产2-乙酰基呋喃的融合魏斯氏菌新菌株，其不仅可应用于发酵食品行业，还可应用于生物来源的香精香料、医疗行业，促进行业发展。
本发明首先提供能产2-乙酰基呋喃的融合魏斯氏菌(Weissella confusa)，其保藏编号为：CGMCC No.26364。
保藏时间为2022年12月30日，保藏单位为中国微生物菌种保藏管理委员会普通微生物中心(CGMCC)，地址：北京市朝阳区北辰西路1号院3号中国科学院微生物研究所，邮编：100101。名称为：融合魏斯氏菌LZLJ2-4(Weissella confusa)。
其中，上述融合魏斯氏菌LZLJ2-4的16S rDNA序列如SEQ ID NO:1所示：
GGGATAATGCGGACCTGCTATACATGCAGTCGAACGCTTTGTGGTTCAACTGATTTGAAGAGCTTAGCTCAGATATGACGATGGACATTGCAAAGAGTGGCGAACGGGTGAGTAACACGTGGGAAACCTACCTCTTAGCAGGGGATAACATTTGGAAACAGATGCTAATACCGTATAACAATGACAACCGCATGGTTGTTATTTAAAAGATGGTTCTGCTATCACTAAGAGATGGTCCCGCGGTGCATTAGCTAGTTGGTAAGGTAATGGCTTACCAAGGCGATGATGCATAGCCGAGTTGAGAGACTGATCGGCCACAATGGGACTGAGACACGGCCCATACTCCTACGGGAGGCAGCAGTAGGGAATCTTCCACAATGGGCGAAAGCCTGATGGAGCAACGCCGCGTGTGTGATGAAGGGTTTCGGCTCGTAAAACACTGTTGTAAGAGAAGAATGACATTGAGAGTAACTGTTCAATGTGTGACGGTATCTTACCAGAAAGGAACGGCTAAATACGTGCCAGCAGCCGCGGTAATACGTATGTTCCAAGCGTTATCCGGATTTATTGGGCGTAAAGCGAGCGCAGACGGTTATTTAAGTCTGAAGTGAAAGCCCTCAGCTCAACTGAGGAATTGCTTTGGAAACTGGATGACTTGAGTGCAGTAGAGGAAAGTGGAACTCCATGTGTAGCGGTGAAATGCGTAGATATATGGAAGAACACCAGTGGCGAAGGCGGCTTTCTGGACTGTAACTGACGTTGAGGCTCGAAAGTGTGGGTAGCAAACAGGATTAGATACCCTGGTAGTCCACACCGTAAACGATGAGTGCTAGGTGTTTGAGGGTTTCCGCCCTTAAGTGCCGCAGCTAACGCATTAAGCACTCCGCCTGGGGAGTACGACCGCAAGGTTGAAACTCAAAGGAATTGACGGGGACCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAACGCGAAGAACCTTACCAGGTCTTGACATCCCTTGACAACTCCAGAGATGGAGCGTTCCCTTCGGGGACAAGGTGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTATTACTAGTTGCCAGCATTCAGTTGGGCACTCTAGTGAGACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAATCATCATGCCCCTTATGACCTGGGCTACACACGTGCTACAATGGCGTATACAACGAGTTGCCAACCCGCGAGGGTGAGCTAATCTCTTAAAGTACGTCTCAGTTCGGATTGTAGGCTGCAACTCGCCTACATGAAGTCGGAATCGCTAGTAATCGCGGATCAGCACGCCGCGGTGAATACGTTCCCGGGTCTTGTACACACCGCCCGTCACACCATGAGAGTTTGTAACACCCAAAGCCGGTGGGGTAACCTTCGGGAGCCAGCCGTCTAATGGAGGTCACAAATCGG。
本发明还提供了一种微生物菌剂，其以所述的融合魏斯氏菌为活性成分。
本发明还提供上述融合魏斯氏菌在产2-乙酰基呋喃方面的应用。
本发明还提供上述融合魏斯氏菌在白酒酿造领域的应用。
本发明还提供上述融合魏斯氏菌在利用2-乙酰基呋喃制备有机合成原料、医药中间体、香精香料、发酵食品上的应用。
具体地，所述应用中，融合魏斯氏菌发酵产2-乙酰基呋喃的方法包括以下步骤：融合魏斯氏菌接种于MRS液体培养基培养活化后，接种至发酵培养基发酵中，发酵培养即可。
其中，所述MRS液体培养基成分为：葡萄糖20g/L，蛋白胨10g/L，酵母粉4g/L，牛肉浸粉5g/L，吐温80 1g/L，柠檬酸三铵2g/L，醋酸钠5g/L，硫酸镁0.2g/L，硫酸锰0.05g/L，磷酸氢二钾1g/L。
其中，所述液体培养基培养条件为30℃-37℃静置培养20h-24h。
其中，所述融合魏斯氏菌的接种至发酵培养基的接种量为2v/v％。
其中，所述发酵培养条件为：30℃-37℃静置培养24h-48h。
其中，所述发酵培养基的组成为葡萄糖16g/L，胰蛋白胨10g/L，酵母粉2.5g/L，牛肉膏5g/L，牛脑3.85g/L，牛心4.9g/L，吐温80 0.5g/L，柠檬酸铵1g/L，氯化钠2.5g/L，无水乙酸钠2.5g/L，硫酸镁0.05g/L，硫酸锰0.025g/L，磷酸氢二钠1.25g/L，磷酸氢二钾1g/L。
采用上述发酵工艺，融合魏斯氏菌所产2-乙酰基呋喃含量为0.096mg/L。
有益效果：本发明从市售的酒曲中分离筛选得到一株能产2-乙酰基呋喃的融合魏斯氏菌，其保藏编号为：CGMCC No.26364。命名为融合魏斯氏菌LZLJ2-4。该融合魏斯氏菌，性能稳定，发酵工艺简单，不破坏原有发酵过程中微生物的平衡，且产2-乙酰基呋喃含量为0.096mg/L，能广泛应用于酒类、酱油、食醋、面包等多种发酵食品制备中。
附图说明
图1为实施例1的融合魏斯氏菌菌落形态图；
图2为实施例2中融合魏斯氏菌所产的2-乙酰基呋喃GC-MS分子碎片质谱图；
图3为2-乙酰基呋喃标准品GC-MS分子碎片质谱图。
本发明的融合魏斯氏菌于2022年12月30日保藏，其保藏编号为：CGMCC No.26364。保藏单位为中国微生物菌种保藏管理委员会普通微生物中心(CGMCC)，地址：北京市朝阳区北辰西路1号院3号中国科学院微生物研究所，邮编：100101。分类命名为：融合魏斯氏菌(Weissella confusa)。
具体实施方式
下面将结合实施例对本发明的方案进行解释。本领域技术人员将会理解，下面的实施例仅用于说明本发明，而不应视为限定本发明的范围。实施例中未注明具体技术或条件的，按照本领域内的文献所描述的技术或条件或者按照产品说明书进行。所用试剂或仪器未注明生产厂商者，均为可以通过市购获得的常规产品。
实施例中涉及的培养基配方：
MRS液体培养基：葡萄糖20g/L，蛋白胨10g/L，酵母粉4g/L，牛肉浸粉5g/L，吐温801g/L，柠檬酸三铵2g/L，醋酸钠5g/L，硫酸镁0.2g/L，硫酸锰0.05g/L，磷酸氢二钾1g/L，pH7.0，115℃下高压蒸汽灭菌20分钟。
MRS固体培养基：在MRS液体培养基基础上添加15g/L的琼脂，115℃下高压蒸汽灭菌20分钟。
发酵培养基：葡萄糖16g/L，胰蛋白胨10g/L，酵母粉2.5g/L，牛肉膏5g/L，牛脑3.85g/L，牛心4.9g/L，吐温80 0.5g/L，柠檬酸铵1g/L，氯化钠2.5g/L，无水乙酸钠2.5g/L，硫酸镁0.05g/L，硫酸锰0.025g/L，磷酸氢二钠1.25g/L，磷酸氢二钾1g/L，pH 6.0，115℃下高压蒸汽灭菌20分钟。
实施例1：菌株分离纯化
1)分离方法：取20g酒曲样品，放入装有180mL无菌蒸馏水的锥形瓶中，恒温摇床振荡10min将样品充分打散、混匀。取1mL样品悬液，采用倍比稀释法，稀释至10-2～10-7，从每个梯度的稀释液中吸取100μL，均匀涂布于MRS固体培养基平板上，平行制备两块平板，倒置，放于37℃厌氧下培养36-48h，并及时观察。
2)划线纯化：将长出菌落的平板取出，挑取不同菌落形态的单菌落，进行二次划线，直至纯化出所有单菌落。MRS琼脂培养基上菌落较小，湿润，白色，中间凸起，表面光滑，如图1所示。
3)菌种保藏：将纯化完成后的每种菌株的单菌落挑入5mL MRS液体培养基中，置于37℃厌氧下静置培养20-24h，吸取1mL菌液至保菌管中，加入0.5mL 60％的无菌甘油溶液，重悬，置于-80℃保存。
实施例2：产2-乙酰基呋喃能力检测
1)准备待测菌液：
筛选得到的菌株甘油保存管溶解后，分别将其接种于MRS液体培养基中，并于37℃下静置培养24h，活化三代后按2％(体积比)接种量接入50mL MRS液体培养基中进行培养24h，得到待测菌液。
(2)HS-SPME/GC-MS检测2-乙酰基呋喃方法：
运用顶空固相微萃取/气相色谱-质谱联用仪技术：取上清液，加入顶空瓶中，加入饱和NaCl溶液，将制备好的样品于60℃保温平衡5min后，用50/30μm DVB/CAR/PDMS萃取头在60℃萃取50min，萃取结束后在GC进样口250℃解吸5min。化合物检索结果与NIST标准谱库进行匹配，相似度达到80％以上确认为目的化合物。
GC-MS检测色谱条件：
气相色谱条件：HP-INNOWAX色谱柱(60m×0.25mm×0.25μm)；升温程序：起始温度40℃，保持5min，以4℃/min升到100℃，再以6℃/min升至230℃，
保持10min，载气为高纯氦气(1.0mL/min)；进样口温度250℃，不分流。
质谱条件：电子电离源，电子能量70eV；电子倍增器电压350V；离子源温度230℃；传输线温度250℃；质量范围40～450m/z。
通过上述检测方法，如图2、3所示，从分离的菌株中筛选出一株可产2-乙酰基呋喃的菌株，选择该菌株做进一步研究。
实施例3：菌株鉴定方法
纯化筛选得到的菌株，取对数生长期新鲜菌液，离心收集菌体，采用细菌基因组抽提试剂盒提取基因组DNA。采用乳酸菌通用引物27F/1492R扩增其16S rDNA全长序列，具体如下：
SEQ ID NO:2：27F(5′-AGAGTTTGATCCTGGCTCAG-3′)
SEQ ID NO:3：1492R(5′-GGTTACCTTGTTACGACTT-3′)
反应体系(50μL)
反应程序
PCR产物用1.0％的琼脂糖凝胶电泳分离检验，电压约11V/cm，电泳时间20min。
PCR产物的测序由上海生工生物技术公司完成。测序获得16S rDNA片段的基因序列通过NCBI的BLAST比对，其16S rDNA序列如SEQ ID NO:1所示，并根据菌株形态学特征确定菌株种属信息，鉴定其为融合魏斯氏菌(Weissella confusa)，并命名为融合魏斯氏菌LZLJ2-4。
实施例4：融合魏斯氏菌发酵培养
将融合魏斯氏菌LZLJ2-4甘油保存管溶解后接种于MRS液体培养基中，并于37℃静置培养24h，活化三代后按2％(体积比)接种量接入300mL发酵培养基中进行培养24h。发酵结束后得到菌液，10000rpm离心5min后取上清。检测方法同实施例2中的GC-MS检测方法，以0.822mg/mL的2-辛醇为内标物，以原始培养基为空白对照，计算2-乙酰基呋喃的含量。在静置培养融合魏斯氏菌24h后，所产2-乙酰基呋喃产量为0.096mg/L。
需要说明的是，本说明书中描述的具体特征、结构、材料或者特点可以在任一个或多个实施例中以合适的方式结合。此外，在不相互矛盾的情况下，本领域的技术人员可以将本说明书中描述的不同实施例以及不同实施例的特征进行结合和组合。
Claims (10)
1.融合魏斯氏菌(Weissella confusa)，其特征在于：保藏编号为：CGMCC No.26364。
2.一种微生物菌剂，其特征在于：以权利要求1所述的融合魏斯氏菌为活性成分。
3.权利要求1所述融合魏斯氏菌在产2-乙酰基呋喃方面的应用。
4.权利要求1所述融合魏斯氏菌在白酒酿造领域的应用。
5.权利要求1所述融合魏斯氏菌在利用2-乙酰基呋喃制备有机合成原料、医药中间体、香精香料或发酵食品上的应用。
6.根据权利要求3-5任一项所述应用，其特征在于：所述融合魏斯氏菌发酵产2-乙酰基呋喃的方法包括以下步骤：融合魏斯氏菌接种于MRS液体培养基培养活化后，接种至发酵培养基发酵中，发酵培养即可。
7.根据权利要求6所述的应用，其特征在于：所述MRS液体培养基成分为：葡萄糖20g/L，蛋白胨10g/L，酵母粉4g/L，牛肉浸粉5g/L，吐温80 1g/L，柠檬酸三铵2g/L，醋酸钠5g/L，硫酸镁0.2g/L，硫酸锰0.05g/L，磷酸氢二钾1g/L。
8.根据权利要求6所述的应用，其特征在于：所述液体培养基培养条件为30℃-37℃静置培养20h-24h。
9.根据权利要求6所述的应用，其特征在于：所述融合魏斯氏菌的接种至发酵培养基的接种量为2v/v％；所述发酵培养条件为：30℃-37℃静置培养24h-48h。
10.根据权利要求6所述的应用，其特征在于：所述发酵培养基的组成为葡萄糖16g/L，胰蛋白胨10g/L，酵母粉2.5g/L，牛肉膏5g/L，牛脑3.85g/L，牛心4.9g/L，吐温80 0.5g/L，柠檬酸铵1g/L，氯化钠2.5g/L，无水乙酸钠2.5g/L，硫酸镁0.05g/L，硫酸锰0.025g/L，磷酸氢二钠1.25g/L，磷酸氢二钾1g/L。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310697229.4A CN116656561A (zh) | 2023-06-13 | 2023-06-13 | 融合魏斯氏菌及其应用 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310697229.4A CN116656561A (zh) | 2023-06-13 | 2023-06-13 | 融合魏斯氏菌及其应用 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN116656561A true CN116656561A (zh) | 2023-08-29 |
Family
ID=87715037
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202310697229.4A Pending CN116656561A (zh) | 2023-06-13 | 2023-06-13 | 融合魏斯氏菌及其应用 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN116656561A (zh) |
-
2023
- 2023-06-13 CN CN202310697229.4A patent/CN116656561A/zh active Pending
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN110951629B (zh) | 低产酒精的产香酵母及其筛选方法及应用 | |
| CN112342168B (zh) | 一株地衣芽孢杆菌及其促进酱油发酵增香的应用 | |
| CN116138429B (zh) | 矮小哈萨克斯坦酵母xj-65及其在辣椒发酵中的应用 | |
| CN113388533A (zh) | 一株发酵性能良好且具有产香功能的汉逊德巴利酵母菌及其筛选方法 | |
| CN110669683B (zh) | 一种酿酒酵母菌及其在制备烟用香料中的应用 | |
| CN114606152B (zh) | 一株贝莱斯芽孢杆菌、微生物菌剂及其应用 | |
| CN117343871B (zh) | 一种鲍氏不动杆菌、酯化液及在黄水处理中的应用 | |
| CN117229929A (zh) | 一株高产酯异常威克汉姆酵母菌及其应用 | |
| CN116656561A (zh) | 融合魏斯氏菌及其应用 | |
| CN113122470A (zh) | 一种高产苯甲酸甲酯和邻氨基苯甲酸甲酯的菌株筛选和应用 | |
| CN105296370B (zh) | 一株用于提高蜂蜜饮品风味物质含量的酵母菌及其应用 | |
| CN115747085B (zh) | 一株产多种吡嗪的酵母菌哈萨克斯坦酵母及其应用 | |
| CN112662597B (zh) | 一株高产三甲基吡嗪和四甲基吡嗪的短小芽孢杆菌 | |
| CN117660213B (zh) | 一株伯顿丝孢毕赤酵母y09-3及其应用 | |
| CN115820432B (zh) | 一株产香云芝栓孔菌及其应用 | |
| CN116904364A (zh) | 产2-乙酰基吡咯的乳酸片球菌及应用 | |
| CN116286413B (zh) | 一株酱油上色和产香的近玫色锁掷孢酵母及其应用 | |
| CN110305912B (zh) | 从茉莉小皮伞菌株中提取香料的方法及提取的香料在烟草中的用途 | |
| CN116970522A (zh) | 一株产棕榈酸乙酯的食丙二醇慢生乳杆菌及其应用 | |
| CN116891815A (zh) | 一株产棕榈酸甲酯的类肠膜魏斯氏菌及其应用 | |
| CN116083288A (zh) | 产2,3-二氢-2,2,6-三甲基苯甲醛的肠膜明串珠菌及其应用 | |
| CN116555124A (zh) | 大曲岩石芽孢杆菌及其应用 | |
| CN116218743A (zh) | 一株可同时产2-壬醇及油酸乙酯的食丙二醇慢生乳杆菌及其应用 | |
| CN117229939A (zh) | 一株植物乳杆菌及其在辣椒低盐发酵中的应用 | |
| CN115820469A (zh) | 布氏慢生乳杆菌菌株及其用途 |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |