CN114606165A - Microbial agent for accelerating secretion and rapidly converting alcohol-rich oil of cigar and preparation method thereof - Google Patents
Microbial agent for accelerating secretion and rapidly converting alcohol-rich oil of cigar and preparation method thereof Download PDFInfo
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- CN114606165A CN114606165A CN202210373808.9A CN202210373808A CN114606165A CN 114606165 A CN114606165 A CN 114606165A CN 202210373808 A CN202210373808 A CN 202210373808A CN 114606165 A CN114606165 A CN 114606165A
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- 235000019506 cigar Nutrition 0.000 title claims abstract description 69
- 230000000813 microbial effect Effects 0.000 title claims abstract description 48
- 230000028327 secretion Effects 0.000 title claims abstract description 38
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 title claims abstract description 26
- 238000002360 preparation method Methods 0.000 title claims abstract description 15
- 244000005700 microbiome Species 0.000 claims abstract description 32
- 239000003795 chemical substances by application Substances 0.000 claims abstract description 29
- 238000000034 method Methods 0.000 claims abstract description 28
- 230000009466 transformation Effects 0.000 claims abstract description 28
- 230000001737 promoting effect Effects 0.000 claims abstract description 19
- 239000004519 grease Substances 0.000 claims abstract description 18
- 239000007788 liquid Substances 0.000 claims abstract description 18
- 239000002068 microbial inoculum Substances 0.000 claims abstract description 18
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims abstract description 16
- 235000015278 beef Nutrition 0.000 claims abstract description 13
- 238000000855 fermentation Methods 0.000 claims abstract description 13
- 230000004151 fermentation Effects 0.000 claims abstract description 13
- 239000000843 powder Substances 0.000 claims abstract description 13
- 238000012258 culturing Methods 0.000 claims abstract description 11
- 239000001963 growth medium Substances 0.000 claims abstract description 11
- 230000001580 bacterial effect Effects 0.000 claims abstract description 10
- 238000003756 stirring Methods 0.000 claims abstract description 9
- 238000009777 vacuum freeze-drying Methods 0.000 claims abstract description 9
- 239000006229 carbon black Substances 0.000 claims abstract description 8
- 238000000227 grinding Methods 0.000 claims abstract description 8
- 150000001875 compounds Chemical class 0.000 claims abstract description 7
- 238000001035 drying Methods 0.000 claims abstract description 7
- 238000007873 sieving Methods 0.000 claims abstract description 7
- 238000001179 sorption measurement Methods 0.000 claims abstract description 7
- NLYAJNPCOHFWQQ-UHFFFAOYSA-N kaolin Chemical class O.O.O=[Al]O[Si](=O)O[Si](=O)O[Al]=O NLYAJNPCOHFWQQ-UHFFFAOYSA-N 0.000 claims abstract description 6
- 239000000203 mixture Substances 0.000 claims abstract description 6
- HDTRYLNUVZCQOY-UHFFFAOYSA-N α-D-glucopyranosyl-α-D-glucopyranoside Natural products OC1C(O)C(O)C(CO)OC1OC1C(O)C(O)C(O)C(CO)O1 HDTRYLNUVZCQOY-UHFFFAOYSA-N 0.000 claims abstract description 5
- HDTRYLNUVZCQOY-WSWWMNSNSA-N Trehalose Natural products O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-WSWWMNSNSA-N 0.000 claims abstract description 5
- HDTRYLNUVZCQOY-LIZSDCNHSA-N alpha,alpha-trehalose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-LIZSDCNHSA-N 0.000 claims abstract description 5
- 239000003223 protective agent Substances 0.000 claims abstract description 5
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 4
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 4
- 230000008569 process Effects 0.000 claims description 20
- 241000894006 Bacteria Species 0.000 claims description 9
- 239000001888 Peptone Substances 0.000 claims description 9
- 108010080698 Peptones Proteins 0.000 claims description 9
- 235000019319 peptone Nutrition 0.000 claims description 9
- 244000063299 Bacillus subtilis Species 0.000 claims description 6
- 235000014469 Bacillus subtilis Nutrition 0.000 claims description 6
- 241000233866 Fungi Species 0.000 claims description 6
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 6
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 claims description 6
- 239000008367 deionised water Substances 0.000 claims description 6
- 229910021641 deionized water Inorganic materials 0.000 claims description 6
- 239000002609 medium Substances 0.000 claims description 6
- 239000008055 phosphate buffer solution Substances 0.000 claims description 6
- VWDWKYIASSYTQR-UHFFFAOYSA-N sodium nitrate Chemical compound [Na+].[O-][N+]([O-])=O VWDWKYIASSYTQR-UHFFFAOYSA-N 0.000 claims description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 6
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims description 5
- 150000002632 lipids Chemical class 0.000 claims description 5
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 4
- 239000002131 composite material Substances 0.000 claims description 4
- 241000235575 Mortierella Species 0.000 claims description 3
- 229930006000 Sucrose Natural products 0.000 claims description 3
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 3
- 239000011790 ferrous sulphate Substances 0.000 claims description 3
- 235000003891 ferrous sulphate Nutrition 0.000 claims description 3
- BAUYGSIQEAFULO-UHFFFAOYSA-L iron(2+) sulfate (anhydrous) Chemical compound [Fe+2].[O-]S([O-])(=O)=O BAUYGSIQEAFULO-UHFFFAOYSA-L 0.000 claims description 3
- 229910000359 iron(II) sulfate Inorganic materials 0.000 claims description 3
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 3
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 3
- 238000002156 mixing Methods 0.000 claims description 3
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 3
- 235000019796 monopotassium phosphate Nutrition 0.000 claims description 3
- 239000001103 potassium chloride Substances 0.000 claims description 3
- 235000011164 potassium chloride Nutrition 0.000 claims description 3
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 claims description 3
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 claims description 3
- 238000000926 separation method Methods 0.000 claims description 3
- 239000004317 sodium nitrate Substances 0.000 claims description 3
- 235000010344 sodium nitrate Nutrition 0.000 claims description 3
- 239000005720 sucrose Substances 0.000 claims description 3
- 238000000967 suction filtration Methods 0.000 claims description 3
- 239000011780 sodium chloride Substances 0.000 claims description 2
- 230000001131 transforming effect Effects 0.000 claims description 2
- 239000007787 solid Substances 0.000 abstract description 5
- 238000001914 filtration Methods 0.000 abstract 1
- 239000003921 oil Substances 0.000 description 35
- 241000208125 Nicotiana Species 0.000 description 10
- 235000002637 Nicotiana tabacum Nutrition 0.000 description 10
- 230000000052 comparative effect Effects 0.000 description 7
- 238000004519 manufacturing process Methods 0.000 description 7
- YZXBAPSDXZZRGB-DOFZRALJSA-N arachidonic acid Chemical compound CCCCC\C=C/C\C=C/C\C=C/C\C=C/CCCC(O)=O YZXBAPSDXZZRGB-DOFZRALJSA-N 0.000 description 6
- 235000014113 dietary fatty acids Nutrition 0.000 description 5
- 239000000194 fatty acid Substances 0.000 description 5
- 229930195729 fatty acid Natural products 0.000 description 5
- 150000004665 fatty acids Chemical class 0.000 description 5
- 238000006243 chemical reaction Methods 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 235000019197 fats Nutrition 0.000 description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- 235000021342 arachidonic acid Nutrition 0.000 description 3
- 229940114079 arachidonic acid Drugs 0.000 description 3
- 235000020777 polyunsaturated fatty acids Nutrition 0.000 description 3
- 239000002994 raw material Substances 0.000 description 3
- 230000000391 smoking effect Effects 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 241000193830 Bacillus <bacterium> Species 0.000 description 2
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 2
- 150000001720 carbohydrates Chemical class 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- XBDQKXXYIPTUBI-UHFFFAOYSA-N dimethylselenoniopropionate Natural products CCC(O)=O XBDQKXXYIPTUBI-UHFFFAOYSA-N 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 230000012010 growth Effects 0.000 description 2
- 150000007524 organic acids Chemical class 0.000 description 2
- 238000012163 sequencing technique Methods 0.000 description 2
- 238000011282 treatment Methods 0.000 description 2
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 241000195585 Chlamydomonas Species 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 241000192125 Firmicutes Species 0.000 description 1
- 102000004882 Lipase Human genes 0.000 description 1
- 108090001060 Lipase Proteins 0.000 description 1
- 239000004367 Lipase Substances 0.000 description 1
- 241000306282 Umbelopsis isabellina Species 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- JAZBEHYOTPTENJ-JLNKQSITSA-N all-cis-5,8,11,14,17-icosapentaenoic acid Chemical compound CC\C=C/C\C=C/C\C=C/C\C=C/C\C=C/CCCC(O)=O JAZBEHYOTPTENJ-JLNKQSITSA-N 0.000 description 1
- DTOSIQBPPRVQHS-PDBXOOCHSA-N alpha-linolenic acid Chemical compound CC\C=C/C\C=C/C\C=C/CCCCCCCC(O)=O DTOSIQBPPRVQHS-PDBXOOCHSA-N 0.000 description 1
- 235000020661 alpha-linolenic acid Nutrition 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 235000014633 carbohydrates Nutrition 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 239000001569 carbon dioxide Substances 0.000 description 1
- 229910002092 carbon dioxide Inorganic materials 0.000 description 1
- 230000010261 cell growth Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 235000019504 cigarettes Nutrition 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 229960005135 eicosapentaenoic acid Drugs 0.000 description 1
- JAZBEHYOTPTENJ-UHFFFAOYSA-N eicosapentaenoic acid Natural products CCC=CCC=CCC=CCC=CCC=CCCCC(O)=O JAZBEHYOTPTENJ-UHFFFAOYSA-N 0.000 description 1
- 235000020673 eicosapentaenoic acid Nutrition 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 239000002360 explosive Substances 0.000 description 1
- VZCCETWTMQHEPK-UHFFFAOYSA-N gamma-Linolensaeure Natural products CCCCCC=CCC=CCC=CCCCCC(O)=O VZCCETWTMQHEPK-UHFFFAOYSA-N 0.000 description 1
- VZCCETWTMQHEPK-QNEBEIHSSA-N gamma-linolenic acid Chemical compound CCCCC\C=C/C\C=C/C\C=C/CCCCC(O)=O VZCCETWTMQHEPK-QNEBEIHSSA-N 0.000 description 1
- 235000020664 gamma-linolenic acid Nutrition 0.000 description 1
- 229960002733 gamolenic acid Drugs 0.000 description 1
- 229930195733 hydrocarbon Natural products 0.000 description 1
- 150000002430 hydrocarbons Chemical class 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
- 229960004488 linolenic acid Drugs 0.000 description 1
- KQQKGWQCNNTQJW-UHFFFAOYSA-N linolenic acid Natural products CC=CCCC=CCC=CCCCCCCCC(O)=O KQQKGWQCNNTQJW-UHFFFAOYSA-N 0.000 description 1
- 235000019421 lipase Nutrition 0.000 description 1
- 230000037353 metabolic pathway Effects 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 230000000877 morphologic effect Effects 0.000 description 1
- 235000005985 organic acids Nutrition 0.000 description 1
- 238000003825 pressing Methods 0.000 description 1
- 235000019260 propionic acid Nutrition 0.000 description 1
- IUVKMZGDUIUOCP-BTNSXGMBSA-N quinbolone Chemical compound O([C@H]1CC[C@H]2[C@H]3[C@@H]([C@]4(C=CC(=O)C=C4CC3)C)CC[C@@]21C)C1=CCCC1 IUVKMZGDUIUOCP-BTNSXGMBSA-N 0.000 description 1
- 239000010499 rapseed oil Substances 0.000 description 1
- 230000005070 ripening Effects 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 230000035807 sensation Effects 0.000 description 1
- 235000019615 sensations Nutrition 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 239000000779 smoke Substances 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 239000013589 supplement Substances 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 230000009044 synergistic interaction Effects 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
-
- A—HUMAN NECESSITIES
- A24—TOBACCO; CIGARS; CIGARETTES; SIMULATED SMOKING DEVICES; SMOKERS' REQUISITES
- A24B—MANUFACTURE OR PREPARATION OF TOBACCO FOR SMOKING OR CHEWING; TOBACCO; SNUFF
- A24B15/00—Chemical features or treatment of tobacco; Tobacco substitutes, e.g. in liquid form
- A24B15/18—Treatment of tobacco products or tobacco substitutes
- A24B15/20—Biochemical treatment
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/02—Separating microorganisms from their culture media
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
- C12N1/16—Yeasts; Culture media therefor
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- 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
- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
- C12N11/10—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a carbohydrate
-
- 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
- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/14—Enzymes or microbial cells immobilised on or in an inorganic carrier
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- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Genetics & Genomics (AREA)
- Biotechnology (AREA)
- Biochemistry (AREA)
- Biomedical Technology (AREA)
- General Health & Medical Sciences (AREA)
- Microbiology (AREA)
- General Engineering & Computer Science (AREA)
- Medicinal Chemistry (AREA)
- Virology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Mycology (AREA)
- Botany (AREA)
- Inorganic Chemistry (AREA)
- Molecular Biology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention provides a microbial agent for accelerating secretion and rapid transformation of alcohol-rich oil of cigar and a preparation method thereof, wherein the method comprises the following steps: microorganism enrichment, namely obtaining cigar alcohol-cultured microorganism enrichment, and continuously culturing the microorganism enrichment by adopting a beef extract protein culture medium and a Chao's culture medium to obtain a grease secretion and transformation promoting microorganism fermentation liquid; drying the modified kaolin, the white carbon black and the diatomite, and grinding to obtain microbial inoculum carrier powder; adding trehalose protective agent into the grease secretion promoting and transformation microbial fermentation liquor, stirring to obtain a compound bacterial liquid, adding bacterial agent carrier powder, stirring uniformly, and standing for adsorption; and filtering the adsorbed mixture to separate solid from liquid to obtain filter residue, and performing vacuum freeze drying, grinding and sieving on the filter residue to obtain the microbial agent for promoting oil secretion transformation.
Description
Technical Field
The invention relates to the field of microbial bacteria, and particularly relates to a microbial agent for accelerating secretion and rapid transformation of alcohol-fed oil of cigar and a preparation method thereof.
Background
The domestic cigar market shows explosive growth, cigars become a new growth point of Chinese tobacco economy, but the raw material supply and the production process of the domestic cigars seriously limit the development of cigar production, and the important process for improving the appearance characteristics, the physical characteristics, the internal components and the taste varieties of cigar leaves has an important improvement effect on the quality of the cigar leaves. In the cigar production process, the change of the type and the content of the tobacco leaf oil has a vital effect on the style and the taste of the cigars. The tobacco oil is a key factor for the formation of the surface gloss of the cigar coat, and the glossy light sensation is a mark of the cigar ripening and the first impression of the cigar quality judged by consumers.
Tobacco oil, also known as leaf or fat, is generally a soft semi-liquid or liquid substance contained in the cells of tobacco leaves, and this substance reflects on the appearance of the tobacco leaves and has a feeling of greasiness and fullness, and its content is directly related to the quality of the tobacco leaves. The main oil and fat substances of different cigar tobacco raw materials at home and abroad comprise a plurality of polyunsaturated fatty acids, arachidonic acid, organic acid and the like. Under specific conditions, some microorganisms in nature can synthesize and store a large amount of oil in vivo by using carbohydrates, hydrocarbons, carbon dioxide and the like as carbon sources, and all microorganisms which can accumulate oil in cells and exceed 20% of dry cell weight (w/w, wherein w/w represents mass ratio) are called as oleaginous microorganisms. The oil-producing microorganism can produce single cell oil by using saccharides. In addition, the single-cell oil component produced by the microbial fermentation of the oil is consistent with the rape oil, and even the content of high-value gamma-linolenic acid, arachidonic acid, eicosapentaenoic acid and the like can be increased, so that the oil content in the processes of the fermentation and the alcohol culture of the cigar is increased, and the quality and the taste of the cigar are improved.
The method for producing the grease by using the microorganisms has the advantages that the grease fatty acid exists in the thallus, the yield is influenced by the thallus yield and the fatty acid content, the method is suitable for the condition that a large amount of cell growth conditions and fatty acid accumulation conditions are often inconsistent, and most of polyunsaturated fatty acids except linolenic acid cannot be industrially produced. Therefore, the breeding and preparation of the microbial bacteria for promoting the secretion and transformation of the alcohol-rich oil of the cigar are used as necessary supplements for improving the quality of the cigar, so that the microbial strain has wide market prospect and is an important key technology for realizing the high-efficiency oil increasing of the cigar microorganisms.
Disclosure of Invention
The invention provides a microbial agent for accelerating secretion and rapid transformation of alcohol-rich oil of cigar and a preparation method thereof, and aims to solve the problems in the prior art.
In order to achieve the above purpose, the embodiment of the invention provides a microbial agent for accelerating secretion and rapid transformation of alcohol-rich oil of cigar and a preparation method thereof.
Designing a cigar alcohol-culturing experiment, and determining a microorganism succession rule, promoting oil secretion and converting key microorganisms in the cigar alcohol-culturing process by adopting 16sRNA + ITS sequencing, metabolome, metagenome and transcriptome; in order to improve the oil increasing performance in the alcohol-culturing process of the cigar, thereby improving the quality and the smoking taste of the cigar, the invention provides a preparation method of a microbial agent for accelerating secretion and rapid conversion of alcohol-culturing oil of the cigar, which comprises the following steps:
s1, selecting phosphate buffer solution to carry out microbial enrichment to obtain cigar alcohol-cultured microbial enrichment, and continuously culturing the obtained microbial enrichment by adopting a beef extract protein culture medium and a Chao' S culture medium to obtain a grease secretion-promoting transformation microbial fermentation liquid;
s2, drying the modified kaolin, white carbon black and diatomite, and grinding and sieving by a 100-mesh sieve to obtain microbial inoculum carrier powder;
s3, adding the trehalose protective agent into the grease secretion-promoting transformed microorganism fermentation liquor, stirring to obtain a compound bacterial liquid, adding the bacterial agent carrier powder, stirring uniformly, and standing for adsorption for 12 hours;
s4, carrying out suction filtration and adsorption on the mixture for 12 hours to carry out solid-liquid separation to obtain filter residue, and carrying out vacuum freeze drying, grinding and sieving on the filter residue to obtain the microbial agent for promoting grease secretion and transformation.
Further, the beef extract peptone medium is as follows: 3.0g of beef extract, 10.0g of peptone and 5.0g of NaCl5, and the volume is adjusted to 1000mL by using deionized water, and the pH value is 7.5.
Further, the Chaudhur culture medium is: 3g of sodium nitrate, 1g of monopotassium phosphate, 0.5g of magnesium sulfate, 0.5g of potassium chloride, 0.01g of ferrous sulfate and 30g of sucrose, wherein the volume is determined to 1000mL by using deionized water, and the pH value is 7.0-7.2.
Further, the temperature of the microorganism enrichment culture process is 20-25 ℃; the temperature of the continuous culture process is 20-25 ℃, and the temperature is 170-180rpm for 120-160 h.
Further, the microbial inoculum carrier powder comprises the following components: white carbon black: the ratio of diatomite is 2:1: 1; and mixing the composite bacterial liquid and the microbial inoculum carrier powder according to a solid-liquid ratio of 1: 2-2.5.
Further, the vacuum freeze drying temperature is-20 ℃, and the drying time is 48-56 h.
Further, the lipid secretion promoting and transformation microbial fermentation liquor is prepared by enriching microbes in the alcohol culture process of cigars by adopting a phosphate buffer solution, and continuously culturing at 20-25 ℃ by using the beef extract peptone and the Chao's medium to respectively obtain the fungi and bacteria for promoting the lipid secretion and transformation, and performing the steps of: 1-1.5 in proportion.
Based on a general inventive concept, the invention also provides a microbial agent for accelerating secretion and rapid transformation of the alcohol-rich oil of the cigar, which is obtained by the preparation method, wherein the microbial agent comprises the following microorganisms in percentage by weight: 30-40% of bacillus subtilis, 30-40% of oleaginous yeast, 10-15% of mortierella isabellina and 2-10% of chlamydomonas.
Furthermore, the effective viable count of the microbial inoculum is 7.5-9 multiplied by 109CFU/g。
The invention selects different culture mediums from cigar alcohol-cultured sample tobacco for enrichment and screening, obtains fermentation liquor through enlarged culture, adsorbs microorganisms in the fermentation liquor through a specific microbial inoculum carrier, and selects a specific drying mode (vacuum freeze drying) to obtain the solid microbial inoculum with high strain survival rate, long storage time and high cigar oil increasing efficiency.
The scheme of the invention has the following beneficial effects:
(1) according to the invention, based on the microbial enrichment in the alcohol culture process of the cigar, the fungus and bacterium composite microbial agent is obtained by continuously culturing a beef extract protein culture medium and a Chao's culture medium, and the composite microbial agent contains more types of microorganisms which are all microorganisms for promoting secretion and transformation of grease, so that the secretion and transformation of grease can be promoted in the alcohol culture process of the cigar, and the quality and the taste of the cigar are improved. The bacillus subtilis is also called bacillus subtilis, is aerobic and endogenous anti-spore rod-shaped gram-positive bacteria, and is one of the strains widely applied in the bacillus. Bacillus can secrete highly active lipase, promote the secretion and transformation of fat, and produce various organic acids such as: acetic acid, propionic acid, various polyunsaturated fatty acids, arachidonic acid, and the like. The oil-producing yeast can promote the secretion and transformation of oil by using organic matters in cigar. The enzymes and metabolites produced by different microorganisms are different, and the multiple microorganisms simultaneously participate in the synergistic interaction effect, so that the secretion and conversion of the grease are promoted, and the quality and the taste of the cigar are improved.
(2) The solid microbial compound inoculant is obtained by mixing modified kaolin, white carbon black and diatomite according to a certain proportion to serve as a microbial carrier, protecting microorganisms with a trehalose protective agent, uniformly stirring the modified kaolin, the white carbon black and the diatomite, placing the mixture for a period of time to enable the microbial carrier to fully adsorb the microorganisms, and adopting a vacuum freeze drying method.
(3) The invention defines key microorganisms for converting organic matters into fatty acid in the alcohol culture process of cigar and metabolic pathways for converting the fatty acid, adopts a biological fermentation technology to perform propagation, selects a proper microbial inoculum carrier, and combines a vacuum freeze drying technology and a plate-and-frame filter pressing technology to prepare the solid powder microbial inoculum. The prepared solid microbial inoculum is applied to the process of the alcohol cultivation of the cigar, so that the conversion of the oil and fat of the cigar is improved, and the quality and the smoking taste of the cigar are improved.
Detailed Description
In order to make the technical problems, technical solutions and advantages to be solved by the present invention clearer, the following detailed description is given with reference to specific embodiments.
Unless otherwise defined, all terms of art used hereinafter have the same meaning as commonly understood by one of ordinary skill in the art. The terminology used herein is for the purpose of describing particular embodiments only and is not intended to limit the scope of the present invention. Unless otherwise specifically stated, various raw materials, reagents, instruments, equipment and the like used in the present invention may be purchased from commercial sites or may be prepared by existing methods.
The invention provides a microbial agent for accelerating secretion and rapid transformation of alcohol-rich oil of cigar and a preparation method thereof, aiming at the existing problems.
In the embodiment of the invention, the cigar is selected from cigarette production areas such as Hainan, Yunnan and the like, a cigar alcohol culture experiment is designed, 16sRNA + ITS sequencing, a metabolome, a metagenome and a transcriptome are adopted, the microbial succession rule, the grease secretion promotion and key microbe conversion in the cigar alcohol culture process are determined, and the microbial enrichment in the cigar alcohol culture process is enriched by phosphate buffer solution, wherein in the embodiment of the invention, the beef extract peptone culture medium is as follows: 3.0g of beef extract, 10.0g of peptone and 5.0g of NaCl, and using deionized water to fix the volume to 1000mL, wherein the pH value is 7.5; the Chao's medium is: 3g of sodium nitrate, 1g of monopotassium phosphate, 0.5g of magnesium sulfate, 0.5g of potassium chloride, 0.01g of ferrous sulfate and 30g of sucrose, wherein the volume is determined to 1000mL by using deionized water, and the pH value is 7.0-7.2.
Examples
The microbial agent for accelerating secretion and rapidly converting the cigar alcohol-nourishing grease is prepared by the following method:
selecting cigars in smoke producing areas such as Hainan, Yunnan and the like as the experimental sample, adopting phosphate buffer solution to enrich microorganisms in the alcohol culture process of the sample, and continuously culturing the cigars by using the beef extract peptone and the Chao's medium at 20-25 ℃ to respectively obtain fungi and bacteria for promoting oil secretion and transformation, wherein the fungi and the bacteria are cultured according to the ratio of 2: 1-1.5 to obtain the compound bacterial liquid for promoting the secretion and transformation of the grease.
Drying the modified kaolin, white carbon black and diatomite, and grinding and sieving by a 100-mesh sieve to obtain microbial inoculum carrier powder; adding a trehalose 2% (w/v) protective agent into the compound bacterial liquid for promoting oil secretion and transforming microorganisms, stirring, adding the microbial agent carrier powder, stirring uniformly, and standing for adsorption for 12 hours. And (3) carrying out suction filtration and adsorption on the mixture for 12 hours to carry out solid-liquid separation to obtain filter residue, and carrying out vacuum freeze drying and grinding on the filter residue and sieving the filter residue with a 100-mesh sieve to obtain the compound microbial agent for promoting oil secretion and transformation.
The microbial inoculum comprises the following microorganisms in percentage by weight: 30-40% of bacillus subtilis; 30-40% of oleaginous yeast; 10-15% of Mortierella pusilla; 2-10% of the genus corteobacteria. The effective viable count of the microbial inoculum is 7.5-9 multiplied by 109CFU/g。
Comparative example
The cigar tobacco leaves are used as the microorganism enrichment, and other steps are the same as the embodiment, so that the treating microbial inoculum is obtained. The effective bacteria number of the microbial inoculum is not 5.0-6.4 multiplied by 108CFU/g, microbial composition: 20-30% of bacillus subtilis; oleaginous yeast 20-40%; 15-30% of Mortierella pusilla;
application example
Selecting cigars produced in the same production area (Hainan production area) and putting the cigars into a cigar alcohol culture box for alcohol culture, wherein the alcohol culture conditions are as follows: the temperature is 20-25 ℃, and the humidity is 60-70%. The control and example treatments were set up in two, six replicates each, three of which were tested and the remainder were observed and reviewed. According to related data, a microbial agent for promoting oil secretion and transformation is not added and is used as a comparative example CK, a microbial agent for promoting oil secretion and transformation is used as an example Sample, the proportion of the microbial agent added in the example is 1%, samples are respectively taken at 0, 8, 15, 30 and 60 days, and the content and the change of the cigar oil in the comparative example and the cigar oil in the example are detected through a transcriptome and a metabolomete. And comparing the difference of the two treatments by observing the surface appearance characteristics of the cigar in the mellow culture process, and judging the mouthfeel of the cigar on the 60 th day.
Basic situation of cigar alcohol curing process
From the above table, it can be seen that the comparative example is obviously different from the examples, the examples have the advantages that the types and the contents of the grease are higher than the comparative example in the whole alcohol-feeding process of the cigar, the morphological characteristics of the cigar are obviously different, the surface of the comparative example is rough than the surface of the example, and the smoking evaluation feeling of the examples is better than the comparative example. The result shows that the embodiment added with the microbial agent for promoting the oil secretion transformation has obvious effect on the oil secretion transformation in the alcohol culture of the cigar, thereby improving the quality and the mouth sensitivity of the cigar.
While the foregoing is directed to the preferred embodiment of the present invention, it will be understood by those skilled in the art that various changes and modifications may be made without departing from the spirit and scope of the invention as defined in the appended claims.
Claims (9)
1. A preparation method of a microbial agent for accelerating secretion and rapid transformation of alcohol-rich oil of cigars is characterized by comprising the following steps:
s1, selecting phosphate buffer solution to carry out microbial enrichment to obtain cigar alcohol-cultured microbial enrichment, and continuously culturing the obtained microbial enrichment by adopting a beef extract protein culture medium and a Chao' S culture medium to obtain a grease secretion-promoting transformation microbial fermentation liquid;
s2, drying the modified kaolin, white carbon black and diatomite, and grinding and sieving by a 100-mesh sieve to obtain microbial inoculum carrier powder;
s3, adding the trehalose protective agent into the grease secretion-promoting transformed microorganism fermentation liquor, stirring to obtain a compound bacterial liquid, adding the bacterial agent carrier powder, stirring uniformly, and standing for adsorption for 12 hours;
s4, carrying out suction filtration and adsorption on the mixture for 12 hours to carry out solid-liquid separation to obtain filter residue, and carrying out vacuum freeze drying, grinding and sieving on the filter residue to obtain the microbial agent for promoting grease secretion and transformation.
2. The method of claim 1, wherein the beef extract peptone medium is: 3.0g of beef extract, 10.0g of peptone and 5.0g of NaCl, and the volume is adjusted to 1000mL by using deionized water, wherein the pH is 7.5.
3. The method according to claim 1, wherein the culture medium is: 3g of sodium nitrate, 1g of monopotassium phosphate, 0.5g of magnesium sulfate, 0.5g of potassium chloride, 0.01g of ferrous sulfate and 30g of sucrose, wherein the volume is determined to 1000mL by using deionized water, and the pH value is 7.0-7.2.
4. The preparation method according to claim 1, wherein the temperature of the microorganism enrichment culture process is 20-25 ℃; the temperature of the continuous culture process is 20-25 ℃, and the temperature is 170-180rpm for 120-160 h.
5. The preparation method according to claim 1, wherein the microbial inoculum carrier powder comprises the following components: white carbon black: the ratio of diatomite is 2:1: 1; and mixing the composite bacterial liquid and the microbial inoculum carrier powder according to a solid-liquid ratio of 1: 2-2.5.
6. The method according to claim 1, wherein the vacuum freeze-drying temperature is-20 ℃ and the drying time is 48 to 56 hours.
7. The preparation method according to claim 1, wherein the lipid secretion promoting transformed microorganism fermentation broth is enriched by microorganisms in a cigar alcohol-culturing process by using a phosphate buffer solution, and is continuously cultured by using the beef extract peptone and a Chao's medium at 20-25 ℃ to obtain the lipid secretion promoting transformed fungi and bacteria respectively, and the fungi and the bacteria are cultured according to a ratio of 2: 1-1.5 in proportion.
8. The microbial agent for accelerating secretion and rapidly transforming the cigar alcohol-nourishing grease obtained by the preparation method of any one of claims 1 to 7, wherein the microbial agent comprises the following microorganisms in percentage by weight: 30-40% of bacillus subtilis; 30-40% of oleaginous yeast; 10-15% of Mortierella pusilla; 2-10% of the genus corteobacteria.
9. The microbial agent for accelerating secretion and quickly converting alcohol-rich lipid in cigar according to claim 8, wherein the effective viable count of the microbial agent is 7.5-9 x 109CFU/g。
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