CN106520377A - Preparation method of fermented tobacco leaf extract capable of reducing irritation and application of fermented tobacco leaf extract to recombinant tobacco leaves - Google Patents
Preparation method of fermented tobacco leaf extract capable of reducing irritation and application of fermented tobacco leaf extract to recombinant tobacco leaves Download PDFInfo
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- CN106520377A CN106520377A CN201611086848.6A CN201611086848A CN106520377A CN 106520377 A CN106520377 A CN 106520377A CN 201611086848 A CN201611086848 A CN 201611086848A CN 106520377 A CN106520377 A CN 106520377A
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- Prior art keywords
- tobacco leaf
- extract
- extracting solution
- leaf extract
- seed culture
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- 235000002637 Nicotiana tabacum Nutrition 0.000 title claims abstract description 80
- 241000208125 Nicotiana Species 0.000 title claims abstract description 45
- 229940055329 tobacco leaf extract Drugs 0.000 title claims abstract description 38
- 238000002360 preparation method Methods 0.000 title claims abstract description 20
- 230000007794 irritation Effects 0.000 title abstract 3
- 238000011218 seed culture Methods 0.000 claims abstract description 52
- 241000894006 Bacteria Species 0.000 claims abstract description 50
- 239000000284 extract Substances 0.000 claims abstract description 46
- 235000019504 cigarettes Nutrition 0.000 claims abstract description 17
- 241000588746 Raoultella planticola Species 0.000 claims abstract description 13
- 240000001080 Grifola frondosa Species 0.000 claims abstract description 12
- 235000007710 Grifola frondosa Nutrition 0.000 claims abstract description 12
- 241000563903 Bacillus velezensis Species 0.000 claims abstract description 11
- 238000002156 mixing Methods 0.000 claims abstract description 10
- 244000183278 Nephelium litchi Species 0.000 claims abstract description 7
- 238000001914 filtration Methods 0.000 claims abstract description 7
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 135
- 239000012531 culture fluid Substances 0.000 claims description 38
- 244000061176 Nicotiana tabacum Species 0.000 claims description 35
- 235000013305 food Nutrition 0.000 claims description 35
- 241000193755 Bacillus cereus Species 0.000 claims description 34
- 239000001963 growth medium Substances 0.000 claims description 31
- 239000007788 liquid Substances 0.000 claims description 31
- 230000006798 recombination Effects 0.000 claims description 30
- 238000005215 recombination Methods 0.000 claims description 30
- 241000196324 Embryophyta Species 0.000 claims description 27
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 22
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 20
- 238000000855 fermentation Methods 0.000 claims description 19
- 230000004151 fermentation Effects 0.000 claims description 19
- 239000002609 medium Substances 0.000 claims description 17
- 230000004936 stimulating effect Effects 0.000 claims description 14
- OWEGMIWEEQEYGQ-UHFFFAOYSA-N 100676-05-9 Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC2C(OC(O)C(O)C2O)CO)O1 OWEGMIWEEQEYGQ-UHFFFAOYSA-N 0.000 claims description 13
- 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 13
- GUBGYTABKSRVRQ-PICCSMPSSA-N Maltose Natural products O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@@H](CO)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-PICCSMPSSA-N 0.000 claims description 13
- 229920001817 Agar Polymers 0.000 claims description 12
- 239000001888 Peptone Substances 0.000 claims description 12
- 108010080698 Peptones Proteins 0.000 claims description 12
- 239000008272 agar Substances 0.000 claims description 12
- 239000000706 filtrate Substances 0.000 claims description 12
- 239000012467 final product Substances 0.000 claims description 12
- 239000008103 glucose Substances 0.000 claims description 12
- 238000009630 liquid culture Methods 0.000 claims description 12
- 239000000203 mixture Substances 0.000 claims description 12
- 235000019319 peptone Nutrition 0.000 claims description 12
- 239000011780 sodium chloride Substances 0.000 claims description 11
- 230000000638 stimulation Effects 0.000 claims description 9
- 238000010992 reflux Methods 0.000 claims description 8
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 8
- 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 7
- 229930006000 Sucrose Natural products 0.000 claims description 7
- 239000005720 sucrose Substances 0.000 claims description 7
- 244000061456 Solanum tuberosum Species 0.000 claims description 6
- 235000002595 Solanum tuberosum Nutrition 0.000 claims description 6
- 239000012530 fluid Substances 0.000 claims description 6
- 238000007654 immersion Methods 0.000 claims description 6
- 239000001965 potato dextrose agar Substances 0.000 claims description 6
- 239000002244 precipitate Substances 0.000 claims description 6
- 241000193830 Bacillus <bacterium> Species 0.000 claims description 5
- 238000000605 extraction Methods 0.000 claims description 3
- 238000000034 method Methods 0.000 abstract description 15
- 102000004190 Enzymes Human genes 0.000 abstract description 10
- 108090000790 Enzymes Proteins 0.000 abstract description 10
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- 230000008569 process Effects 0.000 abstract description 5
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 abstract description 4
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 abstract description 4
- 229910052799 carbon Inorganic materials 0.000 abstract description 4
- 239000000758 substrate Substances 0.000 abstract description 4
- 229910052757 nitrogen Inorganic materials 0.000 abstract description 2
- 235000015742 Nephelium litchi Nutrition 0.000 abstract 1
- 239000000796 flavoring agent Substances 0.000 abstract 1
- 235000019634 flavors Nutrition 0.000 abstract 1
- 229960001031 glucose Drugs 0.000 description 11
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 10
- 230000000694 effects Effects 0.000 description 7
- UGFAIRIUMAVXCW-UHFFFAOYSA-N Carbon monoxide Chemical compound [O+]#[C-] UGFAIRIUMAVXCW-UHFFFAOYSA-N 0.000 description 6
- 239000003546 flue gas Substances 0.000 description 6
- 240000005373 Panax quinquefolius Species 0.000 description 4
- 239000002253 acid Substances 0.000 description 4
- 239000000654 additive Substances 0.000 description 4
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- 238000004519 manufacturing process Methods 0.000 description 3
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- 239000005017 polysaccharide Substances 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 235000013599 spices Nutrition 0.000 description 3
- MGADZUXDNSDTHW-UHFFFAOYSA-N 2H-pyran Chemical compound C1OC=CC=C1 MGADZUXDNSDTHW-UHFFFAOYSA-N 0.000 description 2
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 2
- SIKJAQJRHWYJAI-UHFFFAOYSA-N Indole Chemical compound C1=CC=C2NC=CC2=C1 SIKJAQJRHWYJAI-UHFFFAOYSA-N 0.000 description 2
- 102000004317 Lyases Human genes 0.000 description 2
- 108090000856 Lyases Proteins 0.000 description 2
- 108091005804 Peptidases Proteins 0.000 description 2
- 239000004365 Protease Substances 0.000 description 2
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 description 2
- 150000007513 acids Chemical class 0.000 description 2
- 230000009471 action Effects 0.000 description 2
- 150000001298 alcohols Chemical class 0.000 description 2
- 150000001299 aldehydes Chemical class 0.000 description 2
- PYMYPHUHKUWMLA-LMVFSUKVSA-N aldehydo-D-ribose Chemical compound OC[C@@H](O)[C@@H](O)[C@@H](O)C=O PYMYPHUHKUWMLA-LMVFSUKVSA-N 0.000 description 2
- SRBFZHDQGSBBOR-UHFFFAOYSA-N beta-D-Pyranose-Lyxose Natural products OC1COC(O)C(O)C1O SRBFZHDQGSBBOR-UHFFFAOYSA-N 0.000 description 2
- 238000009395 breeding Methods 0.000 description 2
- 230000001488 breeding effect Effects 0.000 description 2
- 238000006555 catalytic reaction Methods 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 238000005660 chlorination reaction Methods 0.000 description 2
- 235000009508 confectionery Nutrition 0.000 description 2
- 230000002255 enzymatic effect Effects 0.000 description 2
- 230000032050 esterification Effects 0.000 description 2
- 238000005886 esterification reaction Methods 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 230000007062 hydrolysis Effects 0.000 description 2
- 238000006460 hydrolysis reaction Methods 0.000 description 2
- 230000006872 improvement Effects 0.000 description 2
- 150000002576 ketones Chemical class 0.000 description 2
- 150000002632 lipids Chemical class 0.000 description 2
- 230000004060 metabolic process Effects 0.000 description 2
- 244000005700 microbiome Species 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
- 150000002989 phenols Chemical class 0.000 description 2
- 150000003222 pyridines Chemical class 0.000 description 2
- 229910052708 sodium Inorganic materials 0.000 description 2
- 239000011734 sodium Substances 0.000 description 2
- 150000003505 terpenes Chemical class 0.000 description 2
- 235000007586 terpenes Nutrition 0.000 description 2
- 108010082340 Arginine deiminase Proteins 0.000 description 1
- FBPFZTCFMRRESA-FSIIMWSLSA-N D-Glucitol Natural products OC[C@H](O)[C@H](O)[C@@H](O)[C@H](O)CO FBPFZTCFMRRESA-FSIIMWSLSA-N 0.000 description 1
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 1
- QNAYBMKLOCPYGJ-UHFFFAOYSA-N D-alpha-Ala Natural products CC([NH3+])C([O-])=O QNAYBMKLOCPYGJ-UHFFFAOYSA-N 0.000 description 1
- FBPFZTCFMRRESA-JGWLITMVSA-N D-glucitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-JGWLITMVSA-N 0.000 description 1
- SRBFZHDQGSBBOR-SOOFDHNKSA-N D-ribopyranose Chemical compound O[C@@H]1COC(O)[C@H](O)[C@@H]1O SRBFZHDQGSBBOR-SOOFDHNKSA-N 0.000 description 1
- 241000192125 Firmicutes Species 0.000 description 1
- 108010010803 Gelatin Proteins 0.000 description 1
- 229920002527 Glycogen Polymers 0.000 description 1
- QNAYBMKLOCPYGJ-UWTATZPHSA-N L-Alanine Natural products C[C@@H](N)C(O)=O QNAYBMKLOCPYGJ-UWTATZPHSA-N 0.000 description 1
- ONIBWKKTOPOVIA-BYPYZUCNSA-N L-Proline Chemical compound OC(=O)[C@@H]1CCCN1 ONIBWKKTOPOVIA-BYPYZUCNSA-N 0.000 description 1
- QNAYBMKLOCPYGJ-REOHCLBHSA-N L-alanine Chemical compound C[C@H](N)C(O)=O QNAYBMKLOCPYGJ-REOHCLBHSA-N 0.000 description 1
- 229930195725 Mannitol Natural products 0.000 description 1
- OVRNDRQMDRJTHS-UHFFFAOYSA-N N-acelyl-D-glucosamine Natural products CC(=O)NC1C(O)OC(CO)C(O)C1O OVRNDRQMDRJTHS-UHFFFAOYSA-N 0.000 description 1
- OVRNDRQMDRJTHS-FMDGEEDCSA-N N-acetyl-beta-D-glucosamine Chemical compound CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O OVRNDRQMDRJTHS-FMDGEEDCSA-N 0.000 description 1
- 102100021079 Ornithine decarboxylase Human genes 0.000 description 1
- 102000052812 Ornithine decarboxylases Human genes 0.000 description 1
- 108700005126 Ornithine decarboxylases Proteins 0.000 description 1
- NGFMICBWJRZIBI-JZRPKSSGSA-N Salicin Natural products O([C@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@H](CO)O1)c1c(CO)cccc1 NGFMICBWJRZIBI-JZRPKSSGSA-N 0.000 description 1
- 238000004026 adhesive bonding Methods 0.000 description 1
- 229960003767 alanine Drugs 0.000 description 1
- NGFMICBWJRZIBI-UHFFFAOYSA-N alpha-salicin Natural products OC1C(O)C(O)C(CO)OC1OC1=CC=CC=C1CO NGFMICBWJRZIBI-UHFFFAOYSA-N 0.000 description 1
- 229940024606 amino acid Drugs 0.000 description 1
- 235000001014 amino acid Nutrition 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- PYMYPHUHKUWMLA-UHFFFAOYSA-N arabinose Natural products OCC(O)C(O)C(O)C=O PYMYPHUHKUWMLA-UHFFFAOYSA-N 0.000 description 1
- GUBGYTABKSRVRQ-QUYVBRFLSA-N beta-maltose Chemical compound OC[C@H]1O[C@H](O[C@H]2[C@H](O)[C@@H](O)[C@H](O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@@H]1O GUBGYTABKSRVRQ-QUYVBRFLSA-N 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
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- 230000008859 change Effects 0.000 description 1
- 238000013329 compounding Methods 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 229940096919 glycogen Drugs 0.000 description 1
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 description 1
- PZOUSPYUWWUPPK-UHFFFAOYSA-N indole Natural products CC1=CC=CC2=C1C=CN2 PZOUSPYUWWUPPK-UHFFFAOYSA-N 0.000 description 1
- RKJUIXBNRJVNHR-UHFFFAOYSA-N indolenine Natural products C1=CC=C2CC=NC2=C1 RKJUIXBNRJVNHR-UHFFFAOYSA-N 0.000 description 1
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical class CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 1
- 239000000594 mannitol Substances 0.000 description 1
- 235000010355 mannitol Nutrition 0.000 description 1
- 229960001855 mannitol Drugs 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 238000000199 molecular distillation Methods 0.000 description 1
- 239000000178 monomer Substances 0.000 description 1
- 150000002772 monosaccharides Chemical class 0.000 description 1
- 229950006780 n-acetylglucosamine Drugs 0.000 description 1
- NGFMICBWJRZIBI-UJPOAAIJSA-N salicin Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1=CC=CC=C1CO NGFMICBWJRZIBI-UJPOAAIJSA-N 0.000 description 1
- 229940120668 salicin Drugs 0.000 description 1
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- 238000002864 sequence alignment Methods 0.000 description 1
- 230000000391 smoking effect Effects 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 235000010356 sorbitol Nutrition 0.000 description 1
- 229960002920 sorbitol Drugs 0.000 description 1
- 239000007921 spray Substances 0.000 description 1
- 238000005728 strengthening Methods 0.000 description 1
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Classifications
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
- C11B9/00—Essential oils; Perfumes
- C11B9/02—Recovery or refining of essential oils from raw materials
- C11B9/025—Recovery by solvent extraction
-
- 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
- A24B15/24—Treatment of tobacco products or tobacco substitutes by extraction; Tobacco extracts
- A24B15/241—Extraction of specific substances
-
- A—HUMAN NECESSITIES
- A24—TOBACCO; CIGARS; CIGARETTES; SIMULATED SMOKING DEVICES; SMOKERS' REQUISITES
- A24B—MANUFACTURE OR PREPARATION OF TOBACCO FOR SMOKING OR CHEWING; TOBACCO; SNUFF
- A24B3/00—Preparing tobacco in the factory
- A24B3/14—Forming reconstituted tobacco products, e.g. wrapper materials, sheets, imitation leaves, rods, cakes; Forms of such products
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
- C11B9/00—Essential oils; Perfumes
- C11B9/02—Recovery or refining of essential oils from raw materials
- C11B9/022—Refining
-
- 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
- C12P1/00—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes
- C12P1/04—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes by using bacteria
Abstract
The invention discloses a preparation method of fermented tobacco leaf extract capable of reducing irritation and application of the fermented tobacco leaf extract to recombinant tobacco leaves. The method comprises the following steps: mixing a raoultella planticola seed culture solution and a bacillus methylotrophicus seed culture solution at the weight ratio of 1 to (1-3) to obtain a mixed bacterium solution; inoculating the mixed bacterium solution into a mixed extracting solution according to the volume ratio of 0.5%-2% and fermenting to obtain a fermented solution; and after filtering, decompressing and concentrating to obtain extract with the relative density of 1.0522-1.0833, so as to obtain the fermented tobacco leaf extract capable of reducing the irritation. According to the preparation method disclosed by the invention, the mixed extracting solution formed by a tobacco leaf extracting solution, a grifola frondosa extracting solution and a lychee seed extracting solution is used as a carbon source, a nitrogen source and a substrate and is a source substance of cigarettes; a microbial enzyme system can easily utilize the mixed extracting solution to ferment to obtain a cigarette flavor; and the method disclosed by the invention is simple in technological process, and the raw materials are original tobacco leaves of cigarette products, so that the method has an industrialized application prospect.
Description
Technical field
The present invention relates to recombination tobacco leaf additive technology field, specifically refers to a kind of fermenting tobacco leaf extract for reducing and stimulating
Preparation method and its application in recombination tobacco leaf.
Background technology
Recombination tobacco leaf is mainly added by offal, fragment, cabo or discarded tobacco leaf gluing also known as reconstituted tobacco or homogenizing Nicotiana tabacum L.
The composition such as agent and other additives.Water-soluble substanceses in Nicotiana tabacum L. and cabo are used by domestic existing paper process thin slice production technology
Water is extracted, and Jing after solid-liquid separation, makes paper substrate again, and return after liquid portion is then concentrated and be coated onto paper substrate after solid part lease making slurrying
On.This production technology is the physics regrouping process of a raw material to a great extent.For its chemical composition, using should
Recombination tobacco leaf obtained by method is little with the Nicotiana tabacum L. as raw material, cabo difference.So cause recombination tobacco leaf exist miscellaneous QI weight,
Zest is big and the suction taste defect such as mouthfeel discomfort, have impact on using effect and addition of the recombination tobacco leaf in cigarette product.State
Interior common practice is to adopt the method for feeding to improve part weak point, wherein being also adopted by tobacco extract as addition
Agent carries out flavoring and casing to recombination tobacco leaf, but this traditional chemical method has to the improvement degree of quality of tobacco of recombinating very much
Limit, it is difficult to which some for solving the problems, such as recombination tobacco leaf quality are profound.For this phenomenon, improve the quality of recombination tobacco leaf and make
One of the key subjects in this field are had become with value.
The more tobacco extract of cigarette is typically obtained using traditional method, i.e., using the leaf group of monomer Nicotiana tabacum L. or compounding
Obtained by water, alcohol extraction or molecular distillation for raw material, the tobacco extract odor characteristic for so extracting is weaker, right after addition
The improvement of Medicated cigarette has certain limitation.Recombination tobacco leaf natural biological spice is produced using microbial strains fermented tobacco extractive,
Have not been reported.
Microorganism can produce huge high activity enzyme system, such as polysaccharide hydrolysis enzyme, protease, esterification in breeding
Enzyme, redox enzymes and lyases etc., the collaboration of complicated metabolism in enzymatic catalysis, chemical action and microbial body are made
With under, the effect such as decomposed as raw material with Nicotiana tabacum L., degraded, being aoxidized, being reduced, being polymerized, being coupled, being converted forms complicated low point
Sub- compound, including various perfume compounds, such as alcohols, aldehydes, ketone, acids, lipid, phenols, pyran, pyridines
With terpenes etc., these aroma substances are the aroma components of Nicotiana tabacum L. or the additive for Nicotiana tabacum L., and the cigarette perfume of recombination tobacco leaf is entered
Row characteristic strengthening, improve mouthfeel.Tobacco aromatics using is produced using microbial fermentation technology, natural green is environmentally friendly, technical process
It is simple to operation, do not produce toxic by-products, pollution-free etc..In prior art, using Nicotiana tabacum L. of the raw Raoul bacterium of plant to stimulation
Extract carries out the method for bioconversion production tobacco aromatics using and has no report.
The content of the invention
The suction taste defect such as the present invention has miscellaneous QI weight for current recombination tobacco leaf, zest is big and mouthfeel is uncomfortable, there is provided
A kind of preparation method of fermenting tobacco leaf extract for reducing stimulating and its application in recombination tobacco leaf.By planting made by the present invention
Raw Raoul bacterium and food methanol bacillus cereuss collective effect bioconversion generate the fermenting tobacco leaf extract for reducing that flue gas stimulates, and change
Being apt to recombination tobacco leaf stimulates the uncomfortable suction taste defect of big, mouthfeel, can use with other essence spice for cigarette mixing preparations.
The present invention provides a kind of preparation method of the fermenting tobacco leaf extract for reducing and stimulating, and it comprises the following steps：
1) Nicotiana tabacum L. is crushed to into 60～80 mesh, then by mistake after offal and water 1: 30～50 1～3h of reflux, extract, in mass ratio
Filter, repeats 1～3 reflux, extract, filtrate, and merging filtrate obtains final product Nicotiana tabacum L. extracting solution, and sterilizes；
2) weigh by ratio of weight and the number of copies 40～80 parts Nicotiana tabacum L. extracting solution, 20～40 parts of Grifola frondosa extracting solution and 5～20
The litchi nucleus extract fluid of part, mix homogeneously obtain mixed extract；
3) plant raw Raoul bacterium actication of culture：Raw Raoul bacterium Raoultella planticola VP4-4 will be planted
CCTCC NO:M 2012005 is inoculated on the raw Raoul bacterium slant medium of plant, at 25～38 DEG C, after 24～72h of quiescent culture
Activated spawn is obtained, is placed in refrigerator；
4) prepare and plant raw Raoul bacterium seed culture fluid：By step 3) in gained activated spawn, be inoculated into plant Sheng Lawu
That bacterium seed culture medium, under the conditions of 25～38 DEG C, 100～150r/min shaken cultivation 24h～72h is continuously transferred 1～3 time,
It is obtained and plants raw Raoul bacterium seed culture fluid；
5) eat methanol bacillus cereuss slant culture：Will food methanol bacillus methylotrophicus
VJ4-1 CCTCC NO:M 2012004 is inoculated in potato dextrose agar slant culture medium, 25～35 DEG C, pH5～9
Under the conditions of, 24～72h of quiescent culture；
6) prepare food methanol bacillus cereuss seed culture fluid：With step 5) obtained by slant culture, be inoculated into food methanol
Bacillus cereuss seed culture medium, under the conditions of 25～35 DEG C, 100～150rpm shaken cultivation 24h～72h, continuously switching 1～4
It is secondary, food methanol bacillus cereuss seed culture fluid is obtained；
7) raw Raoul bacterium seed culture fluid will be planted and food methanol bacillus cereuss seed culture fluid is mixed by weight 1: 1～3
Close, obtain mixed bacteria liquid；
8) ferment：Mixed bacteria liquid 0.5～2% is inoculated in above-mentioned mixed extract by volume, postvaccinal mixing
The tinning amount of extracting solution be 25～45%, pH5～8,30～40 DEG C of temperature, 100～200r/min of rotating speed, not lucifuge condition issue
Ferment 3～7 days, obtains fermentation liquid；
9) it is 80～90% ethanol by volume 1: 2～6 by fermentation liquid and volume parts, precipitates overnight, 4000～
10000r/min is centrifuged 10～30min, is evaporated to the extractum that relative density is 1.0522～1.0833, obtains final product drop after filtration
The fermenting tobacco leaf extract of low stimulation.
Preferably, the mixed extract is carried by 50～70 parts of Nicotiana tabacum L. extracting solution, 25～35 parts of Grifola frondosa
Take the litchi nucleus extract fluid composition of liquid and 8～18 parts.
Preferably, the mixed extract is by 60 parts of Nicotiana tabacum L. extracting solution, 30 parts of Grifola frondosa extracting solution and 10
The litchi nucleus extract fluid composition of part.
Preferably, the step 3) in, the life Raoul bacterium slant medium of planting is：Beerwort：120mL,
Sucrose：20g/L, agar：15g/L.
Preferably, the step 4) in, the sub- liquid culture medium of raw Raoul strain of planting is consisted of：Maltose：
50g/L, peptone：10g/L, Sodium Chloride：5g/L.
Preferably, the step 5) in, potato glucose slant medium：Rhizoma Solani tuber osi immersion 200mL/L, Portugal
Grape sugar 20g/L, agar 13g/L；
Preferably, the step 6) in, food methanol bacillus cereuss seed liquid culture medium is consisted of：Maltose 50g/
L, peptone 10g/L, Sodium Chloride 5g/L.
Present invention also offers the application of the fermenting tobacco leaf extract extractum of stimulation obtained by a kind of said method, is reduced, it is described
Reduce, in the fermenting tobacco leaf extract extractum addition recombination tobacco leaf for stimulating, adding cigarette fermenting tobacco leaf extract in recombination tobacco leaf
Mass ratio is 0.08～0.12%.
Raoul bacterium, the entitled Raoultella planticola VP4-4 of the bacterial strain, in January 11 in 2012 are given birth in above-mentioned plant
It is preserved in Wuhan University's China typical culture collection center day, preserving number is CCTCC NO:M 2012005, plants raw Raoul
Bacterium Raoultella planticola VP4-4CCTCC NO:The biological property of M 2012005 is：Gram negative bacteria, energy
Produce acid, make milk liquefy and generate indole generation, E.C. 4.1.1.18 experiment to be positive；ODC Ornithine decarboxylase experiment, H2S are produced
Raw, gelatin liquefaction is negative；Can be grown for sole carbon source with monosaccharide, polysaccharide, esters and aminoacid,
The Molecular Identification for planting raw Raoul bacterium Raoultella planticola VP4-4 is characterized as：Its 16SrRNA sequence
Total length 1408bp, 16S rRNA sequences reach 99.563% with the similarity of Raoultella planticola.
Above-mentioned food methanol bacillus cereuss, the entitled Bacillus methylotrophicus VJ4-1 of the bacterial strain, in 2012
January 11 was preserved in Wuhan University's China typical culture collection center, deposit number CCTCC NO:M 2012004, which is biological
Be characterized in that：It is gram positive bacteria, can grows in 10%NaCl, have arginine dihydrolase, ornithine decarboxylase activity,
Acid can be produced, make milk liquefy；Can be with N-Acetyl-D-glucosamine, Mannitol, glucose, salicin, D- 6-(.alpha.-D-galactosido)-D-glucose .s, D-ribose, flesh
Alcohol, sucrose, maltose, D-glucitol, L-arabinose, DL-LACTIC ACID salt, L-Alanine glycogen, L-PROLINE histidine, Fructus Citri Limoniae
16S rRNA of the hydrochlorate for growth, its 16S rRNA sequence and Bacillus methylotrophicus in the culture medium of carbon source
Sequence alignment similarity reaches 100%.
Microorganism can produce huge high activity enzyme system, such as polysaccharide hydrolysis enzyme, protease, esterification in breeding
Enzyme, redox enzymes and lyases etc., the collaboration of complicated metabolism in enzymatic catalysis, chemical action and microbial body are made
With under, the effect such as decomposed as raw material with leaf tobacco extract, degraded, being aoxidized, being reduced, being polymerized, being coupled, being converted forms complicated
Low molecular compound, including various perfume compounds, such as alcohols, aldehydes, ketone, acids, lipid, phenols, pyran,
Pyridines and terpenes etc., these aroma substances are also undoubtedly the aroma component of Nicotiana tabacum L. or the additive for Nicotiana tabacum L..
It is an advantage of the invention that：
1st, the present invention is using the mixed extract of Nicotiana tabacum L. extracting solution, Grifola frondosa extracting solution and litchi nucleus extract fluid composition as carbon
Source, nitrogen source and substrate, are Medicated cigarette origin material, easily allow microbial enzyme system to utilize come product cigarette perfume of fermenting, its fermenting tobacco leaf
Extract has the harmless advantage of natural environmental-protective.
2nd, using the raw Raoul bacterium Raoultella planticola VP4-4CCTCC NO of plant:M 2012005 and food first
Alcohol bacillus methylotrophicus VJ4-1 carry out fermenting and producing jointly has carried recombination tobacco leaf flue gas matter
Rise, flue gas is fuller, sweet sense is preferable, reduces the fermenting tobacco leaf extract that flue gas stimulates, improving recombination tobacco leaf stimulates big, mouth
The uncomfortable suction taste defect of sense, can be used with other essence spice for cigarette mixing preparations.
3rd, the inventive method technical process is simple, and raw material is cigarette product origin Nicotiana tabacum L., has prospect useful industrially.
Specific embodiment
A part of embodiment is set forth below to be described in further detail the relevant technical problem of the present invention, it is necessary to
This points out that specific examples below is simply further illustrated to the present invention, does not represent limiting the scope of the invention.Its
Other people still fall within protection scope of the present invention according to some nonessential modifications and adjustment that the present invention makes.
It is following to plant raw Raoul bacterium, the entitled Raoultella planticola VP4-4 of the bacterial strain, in January 11 in 2012
It is preserved in Wuhan University's China typical culture collection center day, preserving number is CCTCC NO:M 2012005.
Following food methanol bacillus cereuss, the entitled Bacillus methylotrophicus VJ4-1 of the bacterial strain, in 2012
January 11 was preserved in Wuhan University's China typical culture collection center, deposit number CCTCC NO:M 2012004.
Embodiment 1
A kind of preparation method of the fermenting tobacco leaf extract for reducing stimulating, it comprises the following steps：
1) Nicotiana tabacum L. is crushed to into 80 mesh, then will be filtered after offal and water 1: 50 reflux, extract, 3h in mass ratio, be repeated 1 times back
Stream extracts filtrate, and merging filtrate obtains final product Nicotiana tabacum L. extracting solution, and sterilizes；
2) Semen Litchi for weighing 80 parts of Nicotiana tabacum L. extracting solution, 20 parts of Grifola frondosa extracting solution and 10 parts by ratio of weight and the number of copies is carried
Liquid is taken, mix homogeneously obtains mixed extract；
3) actication of culture：Raw Raoul bacterium Raoultella planticola VP4-4 CCTCC NO will be planted:M
2012005 are inoculated on the raw Raoul bacterium slant medium of plant, at 38 DEG C, obtain activated spawn, be placed in 4 after quiescent culture 24h
In DEG C refrigerator；Wherein, planting raw Raoul bacterium slant medium is：Beerwort：120mL, sucrose：20g/L, agar：15g/L.
4) prepare and plant raw Raoul bacterium seed culture fluid：By step 3) in gained activated spawn, be inoculated into plant Sheng Lawu
That bacterium seed culture medium, under the conditions of 25 DEG C, 100r/min shaken cultivation 72h, continuous switching 3 times are obtained and plant life Raoul bacterium
Seed culture fluid；Wherein, plant the sub- liquid culture medium of raw Raoul strain to consist of：Maltose：50g/L, peptone：10g/L, chlorination
Sodium：5g/L.
5) eat methanol bacillus cereuss slant culture：Will food methanol bacillus methylotrophicus
VJ4-1 CCTCC NO:M 2012004 is inoculated in potato dextrose agar slant culture medium, under the conditions of 30 DEG C, pH7,
Quiescent culture 48h；Wherein, potato glucose slant medium：Rhizoma Solani tuber osi immersion 200mL/L, glucose 20g/L, agar
13g/L；
6) prepare food methanol bacillus cereuss seed culture fluid：With step 5) obtained by slant culture, be inoculated into food methanol
Bacillus cereuss seed culture medium, under the conditions of 30 DEG C, 100rpm shaken cultivation 48h, continuous switching 2 times are obtained food methanol spore
Bacillus seed culture fluid；Wherein, eat methanol bacillus cereuss seed liquid culture medium to consist of：Maltose 50g/L, peptone 10g/L,
Sodium Chloride 5g/L；
7) raw Raoul bacterium seed culture fluid and food methanol bacillus cereuss seed culture fluid will be planted by weight 1: 1 mixing,
Obtain mixed bacteria liquid；
8) ferment：Mixed bacteria liquid 2% is inoculated into into above-mentioned mixed extract by volume, postvaccinal mixed extract
Tinning amount be 25%, pH5,40 DEG C of temperature, rotating speed 100r/min, lucifuge condition bottom fermentation 7 days, do not obtain fermentation liquid；
9) it is 90% ethanol by volume 1: 2 by fermentation liquid and volume parts, precipitates overnight, 10000r/min centrifugations
10min, is evaporated to the extractum that relative density is 1.0633 after filtration, obtain final product the fermenting tobacco leaf extract 1 for reducing stimulating.
Embodiment 2
A kind of preparation method of the fermenting tobacco leaf extract 2 for reducing stimulating, it comprises the following steps：
1) Nicotiana tabacum L. is crushed to into 60 mesh, then will be filtered after offal and water 1: 40 reflux, extract, 3h in mass ratio, be repeated 3 times back
Stream extracts filtrate, and merging filtrate obtains final product Nicotiana tabacum L. extracting solution, and sterilizes；
2) Semen Litchi for weighing 60 parts of Nicotiana tabacum L. extracting solution, 30 parts of Grifola frondosa extracting solution and 10 parts by ratio of weight and the number of copies is carried
Liquid is taken, mix homogeneously obtains mixed extract；
3) actication of culture：Raw Raoul bacterium Raoultella planticola VP4-4 CCTCC NO will be planted:M
2012005 are inoculated on the raw Raoul bacterium slant medium of plant, at 30 DEG C, obtain activated spawn, be placed in 4 after quiescent culture 72h
In DEG C refrigerator；Wherein, planting raw Raoul bacterium slant medium is：Beerwort：120mL, sucrose：20g/L, agar：15g/L；
4) prepare and plant raw Raoul bacterium seed culture fluid：By step 3) in gained activated spawn, be inoculated into plant Sheng Lawu
That bacterium seed culture medium, under the conditions of 32 DEG C, 100r/min shaken cultivation 48h, continuous switching 3 times are obtained and plant life Raoul bacterium
Seed culture fluid；Wherein plant the sub- liquid culture medium of raw Raoul strain to consist of：Maltose：50g/L, peptone：10g/L, chlorination
Sodium：5g/L；
5) eat methanol bacillus cereuss slant culture：Will food methanol bacillus methylotrophicus
VJ4-1 CCTCC NO:M 2012004 is inoculated in potato dextrose agar slant culture medium, under the conditions of 25 DEG C, pH9,
Quiescent culture 24h；Wherein, potato glucose slant medium：Rhizoma Solani tuber osi immersion 200mL/L, glucose 20g/L, agar
13g/L；
6) prepare food methanol bacillus cereuss seed culture fluid：With step 5) obtained by slant culture, be inoculated into food methanol
Bacillus cereuss seed culture medium, under the conditions of 25 DEG C, 100rpm shaken cultivation 72h, continuous switching 4 times are obtained food methanol spore
Bacillus seed culture fluid；Wherein, eat methanol bacillus cereuss seed liquid culture medium to consist of：Maltose 50g/L, peptone 10g/L,
Sodium Chloride 5g/L；
7) raw Raoul bacterium seed culture fluid and food methanol bacillus cereuss seed culture fluid will be planted by weight 1: 3 mixing,
Obtain mixed bacteria liquid；
8) ferment：Mixed bacteria liquid 0.5% is inoculated into into above-mentioned mixed extract, postvaccinal mixed extract by volume
Tinning amount be 30%, pH7,36 DEG C of temperature, rotating speed 100r/min, lucifuge condition bottom fermentation 7 days, do not obtain fermentation liquid；
9) it is 80% ethanol by volume 1: 2 by fermentation liquid and volume parts, precipitates overnight, 10000r/min centrifugations
10min, is evaporated to the extractum that relative density is 1.0722 after filtration, obtain final product the fermenting tobacco leaf extract 2 for reducing stimulating.
Embodiment 3
A kind of preparation method of the fermenting tobacco leaf extract 3 for reducing stimulating, it comprises the following steps：
1) Nicotiana tabacum L. is crushed to into 50 mesh, then will be filtered after offal and water 1: 20 reflux, extract, 1h in mass ratio, be repeated 1 times back
Stream extracts filtrate, and merging filtrate obtains final product Nicotiana tabacum L. extracting solution, and sterilizes；
2) Semen Litchi for weighing 70 parts of Nicotiana tabacum L. extracting solution, 35 parts of Grifola frondosa extracting solution and 15 parts by ratio of weight and the number of copies is carried
Liquid is taken, mix homogeneously obtains mixed extract；
3) actication of culture：Raw Raoul bacterium Raoultella planticola VP4-4 CCTCC NO will be planted:M
2012005 are inoculated on slant medium, at 25 DEG C, obtain activated spawn, be placed in refrigerator after quiescent culture 24h；Wherein, tiltedly
Face culture medium is：Beerwort：120mL, sucrose：20g/L, agar：15g/L.
4) prepare and plant raw Raoul bacterium seed culture fluid：By step 3) in gained activated spawn, be inoculated into seed culture
Base, under the conditions of 25 DEG C, 100r/min shaken cultivation 72h, continuous switching 1 time are obtained and plant raw Raoul bacterium seed culture fluid；Its
In, plant the sub- liquid culture medium of raw Raoul strain and consist of：Maltose：50g/L, peptone：10g/L, Sodium Chloride：5g/L.
5) eat methanol bacillus cereuss slant culture：Will food methanol bacillus methylotrophicus
VJ4-1 CCTCC NO:M 2012004 is inoculated in potato dextrose agar slant culture medium, under the conditions of 35 DEG C, pH5,
Quiescent culture 24h；Wherein, potato glucose slant medium：Rhizoma Solani tuber osi immersion 200mL/L, glucose 20g/L, agar
13g/L；
6) prepare food methanol bacillus cereuss seed culture fluid：With step 5) obtained by slant culture, be inoculated into food methanol
Bacillus cereuss seed culture medium, under the conditions of 35 DEG C, 150rpm shaken cultivation 24h, continuous switching 2 times are obtained food methanol spore
Bacillus seed culture fluid；Wherein, eat methanol bacillus cereuss seed liquid culture medium to consist of：Maltose 50g/L, peptone 10g/L,
Sodium Chloride 5g/L；
7) raw Raoul bacterium seed culture fluid and food methanol bacillus cereuss seed culture fluid will be planted by weight 1: 2 mixing,
Obtain mixed bacteria liquid；
8) ferment：Seed culture fluid 2% is inoculated into into above-mentioned mixed extract, postvaccinal mixed extract by volume
Tinning amount be 40%, pH8,30 DEG C of temperature, rotating speed 100r/min, lucifuge condition bottom fermentation 3 days, do not obtain fermentation liquid；
9) by fermentation liquid and 95% ethanol by volume 1: 6, precipitates overnight, 4000r/min centrifugation 30min subtract after filtration
Pressure is concentrated into the extractum that relative density is 1.0112, obtains final product the fermenting tobacco leaf extract 3 for reducing stimulating.
Embodiment 4
A kind of preparation method of the fermenting tobacco leaf extract 4 for reducing stimulating, it comprises the following steps：
1) Nicotiana tabacum L. is crushed to into 50 mesh, then will be filtered after offal and water 1: 50 reflux, extract, 2h in mass ratio, be repeated 2 times back
Stream extracts filtrate, and merging filtrate obtains final product Nicotiana tabacum L. extracting solution, and sterilizes；
2) Semen Litchi for weighing 50 parts of Nicotiana tabacum L. extracting solution, 25 parts of Grifola frondosa extracting solution and 18 parts by ratio of weight and the number of copies is carried
Liquid is taken, mix homogeneously obtains mixed extract；
3) actication of culture：Raw Raoul bacterium Raoultella planticola VP4-4 CCTCC NO will be planted:M
2012005 are inoculated on slant medium, at 30 DEG C, obtain activated spawn after quiescent culture 48h, are placed in 4 DEG C of refrigerators；Its
In, slant medium is：Beerwort：120mL, sucrose：20g/L, agar：15g/L.
4) prepare and plant raw Raoul bacterium seed culture fluid：By step 3) in gained activated spawn, be inoculated into plant Sheng Lawu
That bacterium seed culture medium, under the conditions of 38 DEG C, 150r/min shaken cultivation 72h, continuous switching 2 times are obtained and plant life Raoul bacterium
Seed culture fluid；Wherein, seed liquid culture medium is consisted of：Maltose：50g/L, peptone：10g/L, Sodium Chloride：5g/L.
5) eat methanol bacillus cereuss slant culture：Will food methanol bacillus methylotrophicus
VJ4-1 CCTCC NO:M 2012004 is inoculated in potato dextrose agar slant culture medium, under the conditions of 30 DEG C, pH6,
Quiescent culture 48h；Wherein, potato glucose slant medium：Rhizoma Solani tuber osi immersion 200mL/L, glucose 20g/L, agar
13g/L；
6) prepare food methanol bacillus cereuss seed culture fluid：With step 5) obtained by slant culture, be inoculated into food methanol
Bacillus cereuss seed culture medium, under the conditions of 30 DEG C, 120rpm shaken cultivation 48h, continuous switching 3 times are obtained food methanol spore
Bacillus seed culture fluid；Wherein, eat methanol bacillus cereuss seed liquid culture medium to consist of：Maltose 50g/L, peptone 10g/L,
Sodium Chloride 5g/L；
7) raw Raoul bacterium seed culture fluid and food methanol bacillus cereuss seed culture fluid will be planted by weight 1: 1 mixing,
Obtain mixed bacteria liquid；
8) ferment：Seed culture fluid 15% is inoculated into into above-mentioned mixed extract, postvaccinal mixed extraction by volume
The tinning amount of liquid be 20%, pH6,35 DEG C of temperature, rotating speed 150r/min, lucifuge condition bottom fermentation 5 days, do not obtain fermentation liquid；
9) by fermentation liquid and 95% ethanol by volume 1: 3, precipitates overnight, 8000r/min centrifugation 20min subtract after filtration
Pressure is concentrated into the extractum that relative density is 1.0522, obtains final product the fermenting tobacco leaf extract for reducing stimulating.
Embodiment 5
The application of tobacco fermentation extract
Above-mentioned fermenting tobacco leaf extract is pressed recombination tobacco leaf amount 0.08%, 0.10%, 0.12% by inventor, is respectively added to
On recombination tobacco leaf.Recombination tobacco leaf is loaded in hermetic bag, 80 DEG C of constant temperature 30min in baking oven are placed, it is suitable to recombination tobacco leaf moisture,
Roll into cigarette, be put into 22 DEG C+1, relative humidity be 60%+2% climatic chambers in 48h, ask the expert group of smokeing panel test to be smoked panel test.It is right
According to the Medicated cigarette that the recombination tobacco leaf for only spray plus equal amount leaf tobacco extract is rolled.Jing expert group of smokeing panel test smokes panel test repeatedly, it is believed that：This
Fermenting tobacco leaf extract, with recombination tobacco leaf amount ratio be 0.08%, 0.10% when, with make recombination tobacco leaf flue gas matter lifted, cigarette
Gas is fuller, and sweet sense is more preferable, reduces the advantage that flue gas stimulates, and the suction taste that improving recombination tobacco leaf stimulates big, mouthfeel uncomfortable lacks
Fall into.
1. fermenting tobacco leaf extract of table is added to the sensory evaluating smoking in recombination tobacco leaf
Other unspecified parts are prior art.Although above-described embodiment is made that to the present invention retouching in detail
State, but it be only a part of embodiment of the invention, rather than whole embodiments, people can with according to the present embodiment without
Other embodiment is obtained under the premise of creativeness, these embodiments belong to the scope of the present invention.
Claims (8)
1. it is a kind of reduce stimulate fermenting tobacco leaf extract preparation method, it comprises the following steps：
1) Nicotiana tabacum L. is crushed to into 60～80 mesh, then will be filtered after offal and water 1: 30～50 1～3h of reflux, extract, in mass ratio, weight
Multiple 1～3 reflux, extract, filtrate, merging filtrate obtain final product Nicotiana tabacum L. extracting solution, and sterilize；
2) 40～80 parts of Nicotiana tabacum L. extracting solution, 20～40 parts of Grifola frondosa extracting solution and 5～20 parts are weighed by ratio of weight and the number of copies
Litchi nucleus extract fluid, mix homogeneously obtain mixed extract；
3) plant raw Raoul bacterium actication of culture：Raw Raoul bacterium Raoultella planticola VP4-4CCTCC NO will be planted:
M 2012005 is inoculated on the raw Raoul bacterium slant medium of plant, at 25～38 DEG C, is activated after 24～72h of quiescent culture
Strain, is placed in refrigerator；
4) prepare and plant raw Raoul bacterium seed culture fluid：By step 3) in gained activated spawn, be inoculated into the raw Raoul bacterium of plant
Seed culture medium, under the conditions of 25～38 DEG C, 100～150r/min shaken cultivation 24h～72h is continuously transferred 1～3 time, is obtained
Plant raw Raoul bacterium seed culture fluid；
5) eat methanol bacillus cereuss slant culture：Will food methanol bacillus methylotrophicus VJ4-
1CCTCC NO:M 2012004 is inoculated in potato dextrose agar slant culture medium, in 25～35 DEG C, pH5～9 condition
Under, 24～72h of quiescent culture；
6) prepare food methanol bacillus cereuss seed culture fluid：With step 5) obtained by slant culture, be inoculated into food methanol spore
Bacillus seed culture medium, under the conditions of 25～35 DEG C, 100～150rpm shaken cultivation 24h～72h is continuously transferred 1～4 time, system
Methanol bacillus cereuss seed culture fluid must be eaten；
7) raw Raoul bacterium seed culture fluid and food methanol bacillus cereuss seed culture fluid will be planted by weight 1: 1～3 mixing, is obtained
To mixed bacteria liquid；
8) ferment：Mixed bacteria liquid 0.5～2% is inoculated in above-mentioned mixed extract by volume, postvaccinal mixed extraction
The tinning amount of liquid is 25～45%, pH5～8,30～40 DEG C of temperature, 100～200r/min of rotating speed, not lucifuge condition bottom fermentation 3
～7 days, obtain fermentation liquid；
9) it is 80～90% ethanol by volume 1: 2～6 by fermentation liquid and volume parts, precipitates overnight, 4000～10000r/
Min is centrifuged 10～30min, and the extractum that relative density is 1.0522～1.0833 is evaporated to after filtration, and obtaining final product reduction stimulates
Fermenting tobacco leaf extract.
2. the preparation method of the fermenting tobacco leaf extract of stimulation is reduced according to claim 1, it is characterised in that：The step
2) in, Semen Litchi of the mixed extract by 50～70 parts of Nicotiana tabacum L. extracting solution, 25～35 parts of Grifola frondosa extracting solution and 8～18 parts
Extracting solution is constituted, and mix homogeneously obtains mixed extract.
3. the preparation method of the fermenting tobacco leaf extract of stimulation is reduced according to claim 2, it is characterised in that：The mixing
Extracting solution is made up of the litchi nucleus extract fluid of 60 parts of Nicotiana tabacum L. extracting solution, 30 parts of Grifola frondosa extracting solution and 10 parts.
4. the preparation method of the fermenting tobacco leaf extract of stimulation is reduced according to Claims 2 or 3, it is characterised in that：It is described
Step 3) in, the life Raoul bacterium slant medium of planting is：Beerwort：120mL, sucrose：20g/L, agar：15g/L.
5. the preparation method of the fermenting tobacco leaf extract of stimulation is reduced according to Claims 2 or 3, it is characterised in that：It is described
Step 4) in, the sub- liquid culture medium of raw Raoul strain of planting is consisted of：Maltose：50g/L, peptone：10g/L, Sodium Chloride：
5g/L。
6. the preparation method of the fermenting tobacco leaf extract of stimulation is reduced according to Claims 2 or 3, it is characterised in that：It is described
Step 5) in, potato glucose slant medium：Rhizoma Solani tuber osi immersion 200mL/L, glucose 20g/L, agar 13g/L.
7. the preparation method of the fermenting tobacco leaf extract of stimulation is reduced according to Claims 2 or 3, it is characterised in that：It is described
Step 6) in, food methanol bacillus cereuss seed liquid culture medium is consisted of：Maltose 50g/L, peptone 10g/L, Sodium Chloride 5g/L.
8. a kind of application of the fermenting tobacco leaf extract extractum for being reduced stimulating according to obtained by claim 1, it is characterised in that：It is described
Reduce, in the fermenting tobacco leaf extract extractum addition recombination tobacco leaf for stimulating, adding cigarette fermenting tobacco leaf extract in recombination tobacco leaf
Mass ratio is 0.08～0.12%.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109349678A (en) * | 2018-11-09 | 2019-02-19 | 湖北中烟工业有限责任公司 | A kind of preparation method and applications of compound leaf tobacco extract |
| CN111657533A (en) * | 2020-07-14 | 2020-09-15 | 上海烟草集团有限责任公司 | Recycling method of surplus extracting solution in reconstituted tobacco production process |
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| CN102618475A (en) * | 2012-04-18 | 2012-08-01 | 湖北中烟工业有限责任公司 | Bacillus methylotrophicus and preparation method as well as application of tobacco cimicifugae extract |
| CN102851241A (en) * | 2012-09-05 | 2013-01-02 | 湖北中烟工业有限责任公司 | Preparation method for irritation reducing fermentation tobacco extract, and applications of irritation reducing fermentation tobacco extract in recombinant tobacco |
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2016
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102618475A (en) * | 2012-04-18 | 2012-08-01 | 湖北中烟工业有限责任公司 | Bacillus methylotrophicus and preparation method as well as application of tobacco cimicifugae extract |
| CN102851241A (en) * | 2012-09-05 | 2013-01-02 | 湖北中烟工业有限责任公司 | Preparation method for irritation reducing fermentation tobacco extract, and applications of irritation reducing fermentation tobacco extract in recombinant tobacco |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109349678A (en) * | 2018-11-09 | 2019-02-19 | 湖北中烟工业有限责任公司 | A kind of preparation method and applications of compound leaf tobacco extract |
| CN109349678B (en) * | 2018-11-09 | 2021-04-16 | 湖北中烟工业有限责任公司 | Preparation method and application of composite tobacco leaf extract |
| CN111657533A (en) * | 2020-07-14 | 2020-09-15 | 上海烟草集团有限责任公司 | Recycling method of surplus extracting solution in reconstituted tobacco production process |
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