CN116904364A - 产2-乙酰基吡咯的乳酸片球菌及应用 - Google Patents
产2-乙酰基吡咯的乳酸片球菌及应用 Download PDFInfo
- Publication number
- CN116904364A CN116904364A CN202310919331.4A CN202310919331A CN116904364A CN 116904364 A CN116904364 A CN 116904364A CN 202310919331 A CN202310919331 A CN 202310919331A CN 116904364 A CN116904364 A CN 116904364A
- Authority
- CN
- China
- Prior art keywords
- pediococcus acidilactici
- acetylpyrrole
- producing
- fermentation
- temperature
- 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
- IGJQUJNPMOYEJY-UHFFFAOYSA-N 2-acetylpyrrole Chemical compound CC(=O)C1=CC=CN1 IGJQUJNPMOYEJY-UHFFFAOYSA-N 0.000 title claims abstract description 68
- 241000191998 Pediococcus acidilactici Species 0.000 title claims abstract description 41
- 238000000855 fermentation Methods 0.000 claims abstract description 17
- 230000004151 fermentation Effects 0.000 claims abstract description 17
- 235000021107 fermented food Nutrition 0.000 claims abstract description 8
- 238000004321 preservation Methods 0.000 claims abstract description 7
- 235000013555 soy sauce Nutrition 0.000 claims abstract description 7
- 238000002360 preparation method Methods 0.000 claims abstract description 5
- 235000019990 fruit wine Nutrition 0.000 claims abstract description 4
- 235000015278 beef Nutrition 0.000 claims description 10
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 8
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims description 7
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 7
- 239000000843 powder Substances 0.000 claims description 5
- 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 4
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 claims description 4
- 238000012258 culturing Methods 0.000 claims description 4
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims description 4
- 239000008103 glucose Substances 0.000 claims description 4
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 4
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 4
- 229940099596 manganese sulfate Drugs 0.000 claims description 4
- 235000007079 manganese sulphate Nutrition 0.000 claims description 4
- 239000011702 manganese sulphate Substances 0.000 claims description 4
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 claims description 4
- 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 4
- 229940040526 anhydrous sodium acetate Drugs 0.000 claims description 3
- 210000004556 brain Anatomy 0.000 claims description 3
- 235000008429 bread Nutrition 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
- 235000013355 food flavoring agent Nutrition 0.000 claims description 3
- 238000004519 manufacturing process Methods 0.000 claims description 3
- 239000011780 sodium chloride Substances 0.000 claims description 3
- 239000012137 tryptone Substances 0.000 claims description 3
- 239000004480 active ingredient Substances 0.000 claims description 2
- 239000003795 chemical substances by application Substances 0.000 claims description 2
- 238000011081 inoculation Methods 0.000 claims description 2
- 230000000813 microbial effect Effects 0.000 claims description 2
- 229920000136 polysorbate Polymers 0.000 claims description 2
- 239000001963 growth medium Substances 0.000 abstract description 7
- 240000007154 Coffea arabica Species 0.000 abstract description 6
- 235000016213 coffee Nutrition 0.000 abstract description 6
- 235000013353 coffee beverage Nutrition 0.000 abstract description 6
- 235000013305 food Nutrition 0.000 abstract description 6
- 235000014101 wine Nutrition 0.000 abstract description 6
- 235000013599 spices Nutrition 0.000 abstract description 4
- 235000007466 Corylus avellana Nutrition 0.000 abstract description 3
- 239000000654 additive Substances 0.000 abstract description 3
- 230000000996 additive effect Effects 0.000 abstract description 3
- 235000014571 nuts Nutrition 0.000 abstract description 3
- 235000013616 tea Nutrition 0.000 abstract description 3
- 241000723382 Corylus Species 0.000 abstract description 2
- 238000013124 brewing process Methods 0.000 abstract description 2
- 235000015067 sauces Nutrition 0.000 abstract description 2
- 241001122767 Theaceae Species 0.000 abstract 1
- 235000020097 white wine Nutrition 0.000 abstract 1
- 239000002609 medium Substances 0.000 description 11
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 9
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 description 6
- 230000001580 bacterial effect Effects 0.000 description 6
- 150000001875 compounds Chemical class 0.000 description 6
- 238000001514 detection method Methods 0.000 description 6
- 238000002290 gas chromatography-mass spectrometry Methods 0.000 description 6
- 238000004458 analytical method Methods 0.000 description 5
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 5
- 239000007788 liquid Substances 0.000 description 5
- 241000894006 Bacteria Species 0.000 description 4
- 238000009630 liquid culture Methods 0.000 description 4
- 238000000034 method Methods 0.000 description 4
- 239000000047 product Substances 0.000 description 4
- 108020004465 16S ribosomal RNA Proteins 0.000 description 3
- 241000186660 Lactobacillus Species 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 238000000605 extraction Methods 0.000 description 3
- 239000000796 flavoring agent Substances 0.000 description 3
- 235000019634 flavors Nutrition 0.000 description 3
- 239000012634 fragment Substances 0.000 description 3
- 229940039696 lactobacillus Drugs 0.000 description 3
- 238000009629 microbiological culture Methods 0.000 description 3
- 239000007787 solid Substances 0.000 description 3
- 239000006228 supernatant Substances 0.000 description 3
- 244000269722 Thea sinensis Species 0.000 description 2
- 230000004913 activation Effects 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 239000002054 inoculum Substances 0.000 description 2
- 239000004310 lactic acid Substances 0.000 description 2
- 235000014655 lactic acid Nutrition 0.000 description 2
- 238000001819 mass spectrum Methods 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- SJWFXCIHNDVPSH-UHFFFAOYSA-N octan-2-ol Chemical compound CCCCCCC(C)O SJWFXCIHNDVPSH-UHFFFAOYSA-N 0.000 description 2
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 description 2
- 229920000053 polysorbate 80 Polymers 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 238000012163 sequencing technique Methods 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 241000894007 species Species 0.000 description 2
- 230000001954 sterilising effect Effects 0.000 description 2
- 238000004659 sterilization and disinfection Methods 0.000 description 2
- 238000003860 storage Methods 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- MOMFXATYAINJML-UHFFFAOYSA-N 2-Acetylthiazole Chemical group CC(=O)C1=NC=CS1 MOMFXATYAINJML-UHFFFAOYSA-N 0.000 description 1
- 229920001817 Agar Polymers 0.000 description 1
- 240000009226 Corylus americana Species 0.000 description 1
- 235000001543 Corylus americana Nutrition 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- 239000001888 Peptone Substances 0.000 description 1
- 108010080698 Peptones Proteins 0.000 description 1
- 235000011229 Prunus domestica subsp. syriaca Nutrition 0.000 description 1
- 244000169641 Spondias dulcis Species 0.000 description 1
- 235000005138 Spondias dulcis Nutrition 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 239000011543 agarose gel Substances 0.000 description 1
- 230000003698 anagen phase Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000000975 bioactive effect Effects 0.000 description 1
- 230000005540 biological transmission Effects 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
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- 238000001212 derivatisation Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 238000010586 diagram Methods 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
- 239000003814 drug Substances 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 239000005454 flavour additive Substances 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 238000004817 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
- 150000002500 ions Chemical class 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 238000004949 mass spectrometry Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 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
- 239000003960 organic solvent Substances 0.000 description 1
- 235000019319 peptone Nutrition 0.000 description 1
- 238000004987 plasma desorption mass spectroscopy Methods 0.000 description 1
- 229920000435 poly(dimethylsiloxane) Polymers 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 150000003233 pyrroles Chemical class 0.000 description 1
- 125000000168 pyrrolyl group Chemical group 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 239000001632 sodium acetate Substances 0.000 description 1
- 235000017281 sodium acetate Nutrition 0.000 description 1
- 238000003836 solid-state method Methods 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 230000003068 static effect Effects 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 239000001393 triammonium citrate Substances 0.000 description 1
- 235000011046 triammonium citrate Nutrition 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- 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
- 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
- A21D8/045—Methods for preparing dough; Treating dough prior to baking treating dough with microorganisms or enzymes with a leaven or a composition containing acidifying bacteria
-
- 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
- A23L27/20—Synthetic spices, flavouring agents or condiments
- A23L27/24—Synthetic spices, flavouring agents or condiments prepared by fermentation
-
- 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
- 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
- C12G3/024—Preparation of other alcoholic beverages by fermentation of fruits other than botanical genus Vitis
-
- 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
- C12P17/10—Nitrogen as only ring hetero atom
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- 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)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Microbiology (AREA)
- Food Science & Technology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Biomedical Technology (AREA)
- Virology (AREA)
- Nutrition Science (AREA)
- Medicinal Chemistry (AREA)
- Polymers & Plastics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
本发明属于生物技术领域，具体涉及产2‑乙酰基吡咯的乳酸片球菌(Pediococcus acidilactici)及应用。所述乳酸片球菌保藏编号为：CGMCC No.26365，该菌株在发酵培养基中所产的2‑乙酰基吡咯含量达0.203mg/L，能应用于白酒、果酒、酱油等多种发酵食品制备中，且能用作食用香精添加剂，用于咖啡、茶叶、榛子、坚果类，以及用于食品用香料的生产。同时所提供的乳酸片球菌具有耐高温性能，能适应于中高温大曲制备及酱酒酿造过程中的高温发酵环境并能在其中发挥功能。
Description
技术领域
本发明属于生物技术领域，具体涉及一株从酒曲中分离得到的耐高温且产2-乙酰基吡咯的乳酸片球菌及应用。
背景技术
2-乙酰基吡咯，英文名为2-Acetyl pyrrole，常温常压下为浅灰至淡黄色细小结晶，在水中有一定的溶解度，易溶于醇类有机溶剂。2-乙酰基吡咯属于吡咯类衍生物，具有一定的碱性，常用作有机合成和医药化学中间体，多用于有机功能材料分子和含有吡咯结构的生物活性分子的结构修饰和衍生化。此外，该化合物呈面包香气，可用作食用香精添加剂，用于咖啡、茶叶、榛子、坚果类，以及用于食品用香料的生产。
除了作为食品香精添加剂，现有研究中已发现2-乙酰基吡咯对发酵食品香气体系的贡献作用，以及微生物的添加对于食品中2-乙酰基吡咯含量的影响。
如戴鑫(戴鑫.基于气相色谱—质谱的黄酒香气分析和酒龄，产地鉴别[D].上海应用技术学院，2014.)对34个不同黄酒样品进行香气成分检测分析，有19种香气成分为重要致香成分，其中包括了2-乙酰基吡咯。黄梅(黄梅.酿酒酵母对半干法加工咖啡风味品质的影响研究[D].宁夏大学,2022.)研究了5种不同酿酒酵母对咖啡风味的调节作用，CMCC1557菌的添加提高了对咖啡香气体系的贡献作用较大的化合物含量，其中包含了2-乙酰基吡咯。杨峥等(杨峥,邓旭.酱油中香气成分的剖析[J].食品与发酵工业,1990(2):7.)对低盐固态法制的酱油香气成分进行分析，共鉴定出50种化合物，2-乙酰基吡咯作为此类酱油的主要风味化合物之一。
然而，现有研究中关于产2-乙酰基吡咯的菌种较少，开发更多产2-乙酰基吡咯并温度适应范围较广的菌种对于2-乙酰基吡咯在发酵食品、香精香料、医药领域的应用均有重要的意义。
发明内容
针对现有研究中关于产2-乙酰基吡咯的菌种较少的技术问题，本发明从泸州老窖中温大曲中分离筛选得到一株耐高温且可产2-乙酰基吡咯的乳酸片球菌(Pediococcusacidilactici)。
所述乳酸片球菌筛选于泸州老窖中温大曲中，保藏编号为：CGMCC No.26365，已于2022年12月30日保藏在中国微生物菌种保藏管理委员会普通微生物中心(CGMCC)，名称为：乳酸片球菌LZLJ2-5(Pediococcus acidilactici)。
本发明产2-乙酰基吡咯的乳酸片球菌LZLJ2-5的16S rDNA序列如SEQ ID NO:1所示：
SEQ ID NO:1
CCACCGGCTTTGGGTGTTACAAACTCTCATGGTGTGACGGGCGGTGTGTACAAGGCCCGGGAACGTATTCACCGCGGCATGCTGATCCGCGATTACTAGCGATTCCGACTTCGTGTAGGCGAGTTGCAGCCTACAGTCCGAACTGAGAATGGTTTTAAGAGATTAGCTAAACCTCGCGGTTTCGCGACTCGTTGTACCATCCATTGTAGCACGTGTGTAGCCCAGGTCATAAGGGGCATGATGATTTGACGTCGTCCCCACCTTCCTCCGGTTTGTCACCGGCAGTCTCACTAGAGTGCCCAACTGAATGCTGGCAACTAGTAATAAGGGTTGCGCTCGTTGCGGGACTTAACCCAACATCTCACGACACGAGCTGACGACAACCATGCACCACCTGTCATTCTGTCCCCGAAGGGAACGCCTAATCTCTTAGGTTGGCAGAAGATGTCAAGACCTGGTAAGGTTCTTCGCGTAGCTTCGAATTAAACCACATGCTCCACCGCTTGTGCGGGCCCCCGTCAATTCTTTTGAGTTTCAACCTTGCGGTCGTACTCCCCAGGCGGATTACTTAATGCGTTAGCTGCAGCACTGAAGGGCGGAAACCCTCCAACACTTAGTAATCATCGTTTACGGCATGGACTACCAGGGTATCTAATCCTGTTCGCTACCCATGCTTTCGAGCCTCAGCGTCAGTTACAGACCAGACAGCCGCCTTCGCCACTGGTGTTCTTCCATATATCTACGCATTTCACCGCTACACATGGAGTTCCACTGTCCTCTTCTGCACTCAAGTCTCCCAGTTTCCAATGCACTTCTTCGGTTGAGCCGAAGGCTTTCACATTAGACTTAAAAGACCGCCTGCGCTCGCTTTACGCCCAATAAATCCGGATAACGCTTGCCACCTACGTATTACCGCGGCTGCTGGCACGTAGTTAGCCGTGGCTTTCTGGTTAAATACCGTCACTGGGTGAACAGTTACTCTCACCCACGTTCTTCTTTAACAACAGAGCTTTACGAGCCGAAACCCTTCTTCACTCACGCGGCGTTGCTCCATCAGACTTGCGTCCATTGTGGAAGATTCCCTACTGCTGCCTCCCGTAGGAGTCTGGGCCGTGTCTCAGTCCCAATGTGGCCGATTACCCTCTCAGGTCGGCTACGCATCATCGCCTTGGTGAGCCGTTACCTCACCAACTAGCTAATGCGCCGCGGGTCCATCCAGAAGTGATAGCAGAGCCATCTTTTAAAAGAAAACCAGGCGGTTTTCTCTGTTATACGGTATTAGCATCTGTTTCCAGGTGTTATCCCCTGCTTCTGGGCAGGTTACCCACGTGTTACTCACCCGTCCGCCACTCACTTCGTGTTAAAATCTCATTCAGTGCAAGCACCTCATAATCAATTAACGGAAGTT
本发明还提供所述乳酸片球菌在产2-乙酰基吡咯方面的应用。
本发明还提供一种微生物菌剂，其以所述乳酸片球菌为活性成分。
本发明还提供所述乳酸片球菌在制备含2-乙酰基吡咯的发酵食品、中高温大曲或香精香料添加剂上的应用。
其中，所述发酵食品包括果酒、白酒、酱油或面包中至少一种。
具体地，所述应用是乳酸片球菌LZLJ2-5在发酵培养基中40-60℃静置培养24-48h，pH值为7.0。
所述乳酸片球菌LZLJ2-5的接种量为1-10v/v％。
更具体地，所述发酵培养基的组成为每升培养基中含有16g葡萄糖，10g胰蛋白胨，2.5g酵母粉，5g牛肉膏，3.85g牛脑，4.9g牛心，0.5g吐温80，1g柠檬酸铵，2.5g氯化钠，2.5g无水乙酸钠，0.05g硫酸镁，0.025g硫酸锰，1.25g磷酸氢二钠，1g磷酸氢二钾。
采用上述发酵工艺，该菌株所产的2-乙酰基吡咯含量为0.203mg/L。
有益效果：本发明从泸州老窖中温大曲中筛选分离出一株能产2-乙酰基吡咯的乳酸片球菌，保藏编号为CGMCC No.26365，其性能稳定，在发酵培养基中所产的2-乙酰基吡咯含量达0.203mg/L，能应用于白酒、果酒、酱油等多种发酵食品制备中，也能用作食用香精添加剂，用于咖啡、茶叶、榛子、坚果类，以及用于食品用香料的生产。同时所提供的乳酸片球菌具有耐高温性能，能适应于中高温大曲制备及酱酒酿造过程中的高温发酵环境并能在其中发挥功能。
附图说明
图1为实施例2的乳酸片球菌产2-乙酰基吡咯GC-MS图；
图2为实施例2的乳酸片球菌所产的2-乙酰基吡咯GC-MS分子碎片质谱图；
图3为2-乙酰基吡咯标准品GC-MS分子碎片质谱图。
本发明产2-乙酰基吡咯的乳酸片球菌，已于2022年12月30日保藏在中国微生物菌种保藏管理委员会普通微生物中心(CGMCC)，保藏编号为CGMCC No.26365，保藏地址为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：乳酸片球菌(Pediococcus acidilactici)。
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，pH7.0，115℃下高压蒸汽灭菌20分钟。
实施例1乳酸菌分离和筛选
取20g泸州老窖中温大曲样品，放入装有180mL无菌蒸馏水的锥形瓶中，振荡速率150r/min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50℃培养36-48h，并及时观察；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；将纯化完成后的每种菌株的单菌落挑入5mL于MRS液体培养基中，置于50℃静置培养20-24h，吸取1mL菌液至保菌管中，加入0.5mL60％的无菌甘油溶液，重悬，置于-80℃保存以备进一步分析。
实施例2乳酸菌挥发性代谢产物检测实验
(1)检测样品制备：将甘油保存管中筛选得到的菌株溶解后接种于5mL MRS液体培养基中，并于50℃条件下静置培养12-20h，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)挥发性物质测定
采用顶空固相微萃取法：
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50/30μmDVB/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1、2、3所示，从分离的菌株中筛选出一株能产2-乙酰基吡咯的乳酸菌，选择该菌株做进一步研究。
实施例3目标乳酸菌分子鉴定
纯化筛选得到的乳酸菌，细胞呈圆球状，革兰氏阳性，在MRS固态培养基上为圆形乳白色菌落，表面光滑，凸起，边缘整齐。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：
27F，SEQ ID NO:2：(5′-AGAGTTTGATCCTGGCTCAG-3′)
1492R，SEQ ID NO:3：(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，确定菌株种属信息，鉴定其为乳酸片球菌(Pediococcusacidilactici)，并命名为乳酸片球菌LZLJ2-5，于2022年12月30日保藏在中国微生物菌种保藏管理委员会普通微生物中心(CGMCC)，保藏编号为CGMCC No.26365。
实施例4乳酸片球菌LZLJ2-5发酵培养
将乳酸片球菌LZLJ2-5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50℃进行培养24h，pH值为7.0。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-乙酰基吡咯的含量。在静置培养乳酸片球菌24h后，所产2-乙酰基吡咯产量为0.203mg/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 (8)
1.产2-乙酰基吡咯的乳酸片球菌(Pediococcus acidilactici)，其特征在于：保藏编号为：CGMCC No.26365。
2.一种微生物菌剂，其特征在于：以权利要求1所述乳酸片球菌为活性成分。
3.权利要求1所述乳酸片球菌在产2-乙酰基吡咯方面的应用。
4.权利要求1所述乳酸片球菌在制备含2-乙酰基吡咯的发酵食品、中高温大曲或香精香料添加剂上的应用。
5.根据权利要求4所述应用，其特征在于：所述发酵食品包括果酒、白酒、酱油或面包中至少一种。
6.权利要求2-5任一项所述应用，其特征在于：所述乳酸片球菌发酵产2-乙酰基吡咯的方法包括以下步骤：乳酸片球菌在发酵培养基中40-60℃静置培养24-48h，pH值为7.0，即可。
7.根据权利要求6所述应用，其特征在于：所述乳酸片球菌的接种量为1-10v/v％。
8.根据权利要求7所述应用，其特征在于：所述发酵培养基的组成为每升培养基中含有16g葡萄糖，10g胰蛋白胨，2.5g酵母粉，5g牛肉膏，3.85g牛脑，4.9g牛心，0.5g吐温80，1g柠檬酸铵，2.5g氯化钠，2.5g无水乙酸钠，0.05g硫酸镁，0.025g硫酸锰，1.25g磷酸氢二钠，1g磷酸氢二钾。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310919331.4A CN116904364A (zh) | 2023-07-25 | 2023-07-25 | 产2-乙酰基吡咯的乳酸片球菌及应用 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310919331.4A CN116904364A (zh) | 2023-07-25 | 2023-07-25 | 产2-乙酰基吡咯的乳酸片球菌及应用 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN116904364A true CN116904364A (zh) | 2023-10-20 |
Family
ID=88364523
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202310919331.4A Pending CN116904364A (zh) | 2023-07-25 | 2023-07-25 | 产2-乙酰基吡咯的乳酸片球菌及应用 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN116904364A (zh) |
-
2023
- 2023-07-25 CN CN202310919331.4A patent/CN116904364A/zh active Pending
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN112342168B (zh) | 一株地衣芽孢杆菌及其促进酱油发酵增香的应用 | |
| CN111172079B (zh) | 任清芽孢杆菌新菌种及其应用 | |
| CN110846260B (zh) | 一株降低生物胺的肉葡萄球菌及其应用 | |
| CN114456979B (zh) | 一种在发酵食品中促进风味物质产生的类肠膜魏斯氏菌及其应用 | |
| CN114507610B (zh) | 一株产酱香酿酒酵母菌及其应用 | |
| CN111019860A (zh) | 一株降低生物胺的乳酸片球菌及其应用 | |
| CN110669683B (zh) | 一种酿酒酵母菌及其在制备烟用香料中的应用 | |
| CN117343871B (zh) | 一种鲍氏不动杆菌、酯化液及在黄水处理中的应用 | |
| CN116904364A (zh) | 产2-乙酰基吡咯的乳酸片球菌及应用 | |
| CN116574655A (zh) | 一株具有良好产风味能力的索诺拉沙漠芽孢杆菌 | |
| CN116555077A (zh) | 一株戊糖乳杆菌sj-2及其发酵应用 | |
| CN113817650B (zh) | 一株产吡嗪高温放线菌、分离筛选及应用 | |
| WO2017197993A1 (zh) | 一种液化沙雷氏菌及其转化合成洋茉莉醛的方法 | |
| CN116410874A (zh) | 一株发酵毕赤酵母及其在制备发酵酸肉中的应用 | |
| CN109735473B (zh) | 发酵法制水溶性姜黄素 | |
| CN116656561A (zh) | 融合魏斯氏菌及其应用 | |
| CN115948303B (zh) | 同时产苯乙醛和d-苯基乳酸的副布氏慢生乳杆菌及其用途 | |
| 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 | |
| CN116676231A (zh) | 一株产反式-2-癸烯酸的食窦魏斯氏菌及其应用 | |
| CN116970522A (zh) | 一株产棕榈酸乙酯的食丙二醇慢生乳杆菌及其应用 | |
| CN117887623A (zh) | 产2,6-二叔丁基-4-甲基苯酚的芬氏纤维微细菌及应用 | |
| CN115717114A (zh) | 一种可产多种醛类香味物质的泰国肠球菌及其应用 | |
| CN112662597B (zh) | 一株高产三甲基吡嗪和四甲基吡嗪的短小芽孢杆菌 | |
| CN116836869A (zh) | 一株产正癸醇的食窦魏斯氏菌及其应用 | |
| CN116891815A (zh) | 一株产棕榈酸甲酯的类肠膜魏斯氏菌及其应用 |
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 |