CN105861319A - Method for preparing tamarind fruit spice for cigarettes through trichoderma aureoviride - Google Patents
Method for preparing tamarind fruit spice for cigarettes through trichoderma aureoviride Download PDFInfo
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- CN105861319A CN105861319A CN201610240447.5A CN201610240447A CN105861319A CN 105861319 A CN105861319 A CN 105861319A CN 201610240447 A CN201610240447 A CN 201610240447A CN 105861319 A CN105861319 A CN 105861319A
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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
- C12N1/145—Fungal isolates
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/645—Fungi ; Processes using fungi
- C12R2001/885—Trichoderma
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- 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
- A24B3/12—Steaming, curing, or flavouring tobacco
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- 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
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- 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
- C12P1/02—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes by using fungi
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Abstract
The invention discloses a method for preparing tamarind fruit spice for cigarettes through trichoderma aureoviride. A bacterial strain is (Trichoderma aureoviride) HLMM-04 CGMCC No.10839, and has the ability of producing cellulose and then promoting the cellulose to be decomposed into oligosaccharide or monosaccharide. The bacterial strain is activated, fermented, filtered and dialyzed to prepare a cellulose crude enzyme solution used for fermenting tamarind fruits, then, extraction, filtration and vacuum concentration are performed, and spice for cigarettes is prepared. The optimal enzyme producing condition of the HLMM-04 bacterial strain is that the initial pH of a culture medium is 6.5, the culture temperature is 28 DEG C, and culture time is 5d. Perfuming results prove that the prepared spice has the effects of increasing cigarette flavor richness, promoting the secretion of saliva or body fluid, comforting the mouth cavity, improving remaining taste and enabling fragrance of cigarettes to be pure and mild.
Description
Technical field
The invention belongs to fermentable and application thereof, be specifically related to a strain trichoderma aureoviride new strains.
Meanwhile, the method that the invention still further relates to utilize this bacterial strain to prepare cigarette Fructus Tamarindi Indicae spice.
Background technology
Flavoring and casing is the key link in cigarette process, is effectively to promote cigarette smoking quality, forms volume
One of effective means of the peculiar style of cigarette.The tobacco aromatics using added in China Nicotiana tabacum L. at present is to use to pass mostly
Gained is allocated in the extraction of system method again, although production operation is simple, but there is also fragrance dullness, improves
The shortcomings such as cigarette smoking DeGrain.Utilize microorganism that perfume material carries out fermentation to increase tradition and carry
Take the content of multiple aroma substance in spice, promote fragrance rich, and then it is abundant, high to produce fragrance
The Medicated cigarette of quality, meets consumer's requirement to cigarette quality.
Fructus Tamarindi Indicae (Tamarindus indica) is also known as tamarind, tamarind etc..Caesalpiniaceae Fructus Tamarindi Indicae belongs to.According to
Compendium of Material Medica is recorded, Fructus Tamarindi Indicae have quench the thirst disappear heat, help digestion, the function such as nourishing the liver to improve visual acuity.It is usually used in controlling
Treat heatstroke, dyspepsia, food stagnation, stomachache, chronic gastritis, constipation, hypertension and iron-deficient heart disease.
Promoting the production of body fluid to quench thirst, disappear pyrolysis summer-heat, refrigerant tasty and refreshing, enjoys endless aftertastes.
Trichoderma (Trichoderma spp.) is distributed widely in nature, residual rotten wood, seed, plant
In body, fertilizer, soil and air especially the most.Trichoderma growing nutrient requires simple, fast growth,
Highly beneficial for industrialized production.Trichoderma aureoviride (Trichoderma aureoviride) is trichoderma
One, as a kind of important bio-control factors, has broad spectrum activity to the antagonism of pathogenic, makes more
Property, moreover it is possible to producing multiple enzyme decomposes the plant and animal residues in soil, increases soil fertility.To Huang
The report of green trichoderma focuses mostly at aspects such as soil improvement, Biological control and bio-feritlizers.In prior art
There is not yet and trichoderma aureoviride is used for Fructus Tamarindi Indicae fermentation prepares the report of tobacco aromatics using.
Summary of the invention
Present invention aims to the deficiencies in the prior art, it is provided that a strain trichoderma aureoviride new strains, should
Bacterial strain has cellulase-producing, and then promotes that cellulose decomposition becomes the ability of oligosaccharide or monosaccharide.
The present invention also aims to provide a kind of side utilizing this strain fermentation Fructus Tamarindi Indicae to prepare tobacco aromatics using
Method.
The purpose of the present invention is achieved by the following technical programs.
Except as otherwise noted, percent of the present invention is mass percent.
A kind of trichoderma aureoviride (Trichoderma aureoviride) HLMM-04 bacterial strain, CGMCC No.
10839, it has cellulase-producing, and then promotes that cellulose decomposition becomes the ability of oligosaccharide or monosaccharide.
The method utilizing described bacterial strain to prepare tobacco aromatics using, comprises the following steps:
(1), after trichoderma aureoviride HLMM-04CGMCC No.10839 bacterial strain being activated twice, it is inoculated in dress
Have in the triangular flask of 200mL PDA fluid medium, at 20～35 DEG C, 180rpm bottom fermentation cultivate 1～
7d；Described PDA fluid medium: Rhizoma Solani tuber osi 200g, glucose 20g, mend distilled water to 1L,
PH5.5～8.5,115 DEG C of autoclaving 15min；
(2) filtering fermentation liquor removes thalline, takes supernatant and is slowly added to ammonium sulphate to 80% saturation, and 4
Standing overnight at DEG C, centrifugal collecting precipitate is also dissolved in 2mL buffer, then carry out with buffer
Analysis, obtains crude enzyme liquid；Described buffer is: 0.02mol/L Na2HPO4-NaH2PO4, pH 7.3；
(3) Fructus Tamarindi Indicae decontamination is cleaned, add the water of double weight, mix homogeneously after crushing, adjust pH extremely
6.0；Add trichoderma aureoviride HLMM-04 crude enzyme liquid, fermentation, crude enzyme liquid addition be Fructus Tamarindi Indicae quality 5%～
20%, fermentation temperature is 30 DEG C, fermentation time 2～5h；Reflux, extract, 3h, filter cleaner, by supernatant
Liquid concentrating under reduced pressure, obtains required tobacco aromatics using.
Further, for preventing spice from going bad, by 5% interpolation propylene glycol of tobacco aromatics using weight.
In step (1), optimum perfume condition of producing is initial pH 6.5, and cultivation temperature 28 DEG C, during cultivation
Between 5d.
Relative to prior art, the invention have the advantages that
(1) trichoderma aureoviride is applied to tobacco aromatics using preparation field by the present invention first.Use trichoderma aureoviride
HLMM-04 bacterial strain fermentation liquor processes Fructus Tamarindi Indicae, compared with untreated control, protects at original Fructus Tamarindi Indicae characteristic perfume
While holding, there is increase cigarette perfume (or spice) rich, promote the production of body fluid, comfortable oral cavity, improve pleasant impression, make cigarette flavor
The effect of alcohol sum.
(2) present invention is with low cost, simple to operate, it is simple to realize large-scale industrial production, application
Have good prospects.
The explanation of preservation biomaterial
The bacterial strain of the present invention, on May 22nd, 2015, is deposited in Chinese microorganism strain preservation pipe
Reason committee's common micro-organisms center (CGMCG)；This centre address: city of the BeiJing, China Chaoyang District North Star
West Road 1 institute 3, Institute of Microorganism, Academia Sinica.The named trichoderma aureoviride of this strain classification
(Trichoderma aureoviride) HLMM-04, preserving number is CGMCC No.10839.
Detailed description of the invention
Below by embodiment, the present invention is described in further detail, but embodiment is not to this
The restriction of bright technical scheme.
Embodiment 1
1, the suitable condition of enzyme production of trichoderma aureoviride HLMM-04 bacterial strain
The 1.1 initial pH of the culture medium impact on HLMM-04 strain enzyme-producing
The pH of fluid medium is transferred to 5.5,6.0,6.5,7.0,7.5,8.0,8.5 respectively, 28
DEG C, shaken cultivation, after 3 days, measures cellulase activity, shows that optimum initial pH is under 120rpm
6.5。
The impact on HLMM-04 strain enzyme-producing of 1.2 cultivation temperature
Postvaccinal liquid fermentation medium is respectively placed in 20 DEG C, 25 DEG C, 28 DEG C, 32 DEG C, at 35 DEG C,
After 120r/min shaken cultivation 3d, measure cellulase activity, show that optimum cultivation temperature is 28 DEG C.
The impact on HLMM-04 strain enzyme-producing of 1.3 incubation times
The fluid medium (pH 6.5) inoculated is placed in climatic chamber, 28 DEG C, 120r/min
Shaken cultivation, every 24h measures primary fiber element enzyme and lives, and cultivates and reaches to cellulase activity when the 5th day
Big value.
2, trichoderma aureoviride HLMM-04 crude enzyme liquid preparation method
According to above optimum results, after trichoderma aureoviride HLMM-04 bacterial strain is activated twice, it is inoculated in dress
Having in the triangular flask of 200mL PDA fluid medium (pH 6.5), at 28 DEG C, 180rpm issues
Ferment cultivates 5d.Filter and remove thalline.Take supernatant and be slowly added to ammonium sulphate to 80% saturation, at 4 DEG C
Standing overnight, centrifugal collecting precipitate is also dissolved in 2mL buffer (0.02mol/L
Na2HPO4-NaH2PO4Buffer, pH 7.3) in, dialyse with same buffer, obtain
Crude enzyme liquid.
3, the method that trichoderma aureoviride HLMM-04 strain fermentation prepares tobacco aromatics using
Fructus Tamarindi Indicae decontamination is cleaned, adds the water of double weight, mix homogeneously after crushing, adjust pH to 6.0.
Add trichoderma aureoviride HLMM-04 crude enzyme liquid, fermentation.Crude enzyme liquid addition is the 15% of Fructus Tamarindi Indicae quality,
Fermentation temperature is 30 DEG C, fermentation time 3h.Reflux, extract, 3h, filter cleaner, reduces pressure supernatant dense
Contracting, obtains Fructus Tamarindi Indicae fermentation tobacco aromatics using.
For preventing from going bad, by 5% interpolation propylene glycol of fermentation spice weight.
4, product feature and physicochemical property
Gained fermentation tobacco aromatics using is rufous stream shape mastic, does not clarifies.Product physicochemical property is shown in Table 1.
The physical and chemical index of table 1 trichoderma aureoviride HLMM-04 fermentation Fructus Tamarindi Indicae spice
5, perfuming cigarette experiment
Take Fructus Tamarindi Indicae fermentation tobacco aromatics using 0.1g, after taking sterilized water dilution 50 times, be uniformly sprayed at 50g and do not add
In Herba Pelargonii Graveolentis group, load in clean sealing bag, after placing 2h at room temperature, by tobacco shred in 120 DEG C of baking ovens
Middle drying, makes moisture reach 11%-13%.Climatic chamber is put in tobacco shred and comparison after processing
In, under the conditions of temperature 22 DEG C, humidity 60%, balance 24h, roll into Medicated cigarette, by the expert that smokes panel test by hand
The group of smokeing panel test of composition carries out sensory evaluating smoking.Not ferment, Fructus Tamarindi Indicae extracts spice and fermentation of bacillus subtilis
Fructus Tamarindi Indicae spice is comparison.Perfuming result shows, Fructus Tamarindi Indicae trichoderma aureoviride HLMM-04 is fermented spice and compares
Difference substantially, has increase cigarette perfume (or spice) rich, promotes the production of body fluid, and comfortable oral cavity is improved pleasant impression, made cigarette flavor
The effect of alcohol sum.Evaluation result is shown in Table 2.
Table 2 Fructus Tamarindi Indicae fermentation spice smoking result
Embodiment 2
Repeat embodiment 1, have following difference: crude enzyme liquid addition is the 5% of Fructus Tamarindi Indicae quality, fermentation
Time 2h.
1, product feature and physicochemical property
Gained fermentation tobacco aromatics using is rufous stream shape mastic, does not clarifies.Product physicochemical property is shown in Table 3.
The physical and chemical index of table 3 trichoderma aureoviride HLMM-04 fermentation Fructus Tamarindi Indicae spice
2, perfuming cigarette experiment
Perfuming result shows, Fructus Tamarindi Indicae trichoderma aureoviride HLMM-04 fermentation spice is obvious with control area, tool
There is increase cigarette perfume (or spice) rich, promote the production of body fluid, reduce the effect stimulated.Evaluation result is shown in Table 4.
Table 4 Fructus Tamarindi Indicae fermentation spice smoking result
Embodiment 3
Repeat embodiment 1, have following difference: crude enzyme liquid addition is the 20% of Fructus Tamarindi Indicae quality, fermentation
Time 5h.
1, product feature and physicochemical property
Gained fermentation tobacco aromatics using is rufous stream shape mastic, does not clarifies.Product physicochemical property is shown in Table 5.
The physical and chemical index of table 5 trichoderma aureoviride HLMM-04 fermentation Fructus Tamarindi Indicae spice
2, perfuming cigarette experiment
Perfuming result shows, Fructus Tamarindi Indicae trichoderma aureoviride HLMM-04 fermentation spice is obvious with control area, tool
There are increase acid perfume, the effect of oral cavity comfortableness of promoting the production of body fluid, improve.Evaluation result is shown in Table 6.
Table 6 Fructus Tamarindi Indicae fermentation spice smoking result
Claims (4)
1. trichoderma aureoviride (Trichoderma aureoviride) HLMM-04 bacterial strain, CGMCC No.
10839, it has cellulase-producing, and then promotes that cellulose decomposition becomes the ability of oligosaccharide or monosaccharide.
2. the method utilizing bacterial strain described in claim 1 to prepare tobacco aromatics using, comprises the following steps:
(1), after trichoderma aureoviride HLMM-04CGMCC No.10839 bacterial strain being activated twice, it is inoculated in dress
Have in the triangular flask of 200mL PDA fluid medium, at 20～35 DEG C, 180rpm bottom fermentation cultivate 1～
7d；Described PDA fluid medium: Rhizoma Solani tuber osi 200g, glucose 20g, mend distilled water to 1L,
PH5.5～8.5,115 DEG C of autoclaving 15min；
(2) filtering fermentation liquor removes thalline, takes supernatant and is slowly added to ammonium sulphate to 80% saturation, and 4
Standing overnight at DEG C, centrifugal collecting precipitate is also dissolved in 2mL buffer, then carry out with buffer
Analysis, obtains crude enzyme liquid；Described buffer is: 0.02mol/L Na2HPO4-NaH2PO4, pH 7.3；
(3) Fructus Tamarindi Indicae decontamination is cleaned, add the water of double weight, mix homogeneously after crushing, adjust pH extremely
6.0；Add trichoderma aureoviride HLMM-04 crude enzyme liquid, fermentation, crude enzyme liquid addition be Fructus Tamarindi Indicae quality 5%～
20%, fermentation temperature is 30 DEG C, fermentation time 2～5h；Reflux, extract, 3h, filter cleaner, by supernatant
Liquid concentrating under reduced pressure, obtains required tobacco aromatics using.
The method preparing tobacco aromatics using the most according to claim 2, it is characterised in that: for preventing perfume (or spice)
Material is rotten, by 5% interpolation propylene glycol of tobacco aromatics using weight.
The method preparing tobacco aromatics using the most according to claim 2, it is characterised in that: step (1)
In optimum fermentation culture conditions be initial pH 6.5, cultivation temperature 28 DEG C, incubation time 5d.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201610240447.5A CN105861319B (en) | 2016-04-18 | 2016-04-18 | A method of cigarette tamarind fragrance is prepared using trichoderma aureoviride fermentation |
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| CN201610240447.5A CN105861319B (en) | 2016-04-18 | 2016-04-18 | A method of cigarette tamarind fragrance is prepared using trichoderma aureoviride fermentation |
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| CN105861319B CN105861319B (en) | 2019-04-05 |
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101731741A (en) * | 2008-11-11 | 2010-06-16 | 安徽中烟工业公司 | Processing material for improving smoking quality of white hairy tobacco |
| CN102010787A (en) * | 2009-09-04 | 2011-04-13 | 湖北中烟工业有限责任公司 | Casing flavor for reconstituted tobacco leaves produced by using paper-making method |
| CN102226129A (en) * | 2011-05-12 | 2011-10-26 | 湖北中烟工业有限责任公司 | Preparation method of composite extract for fermentation type tobacco |
| CN104522879A (en) * | 2014-11-10 | 2015-04-22 | 浙江中烟工业有限责任公司 | Tobacco flavor for improving taste quality of upper tobacco leaf group |
| CN104983054A (en) * | 2015-06-04 | 2015-10-21 | 湖北中烟工业有限责任公司 | Preparation method and application of extract to improving cigarette sour and sweet note |
-
2016
- 2016-04-18 CN CN201610240447.5A patent/CN105861319B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101731741A (en) * | 2008-11-11 | 2010-06-16 | 安徽中烟工业公司 | Processing material for improving smoking quality of white hairy tobacco |
| CN102010787A (en) * | 2009-09-04 | 2011-04-13 | 湖北中烟工业有限责任公司 | Casing flavor for reconstituted tobacco leaves produced by using paper-making method |
| CN102226129A (en) * | 2011-05-12 | 2011-10-26 | 湖北中烟工业有限责任公司 | Preparation method of composite extract for fermentation type tobacco |
| CN104522879A (en) * | 2014-11-10 | 2015-04-22 | 浙江中烟工业有限责任公司 | Tobacco flavor for improving taste quality of upper tobacco leaf group |
| CN104983054A (en) * | 2015-06-04 | 2015-10-21 | 湖北中烟工业有限责任公司 | Preparation method and application of extract to improving cigarette sour and sweet note |
Non-Patent Citations (2)
| Title |
|---|
| POKROVSKY VS 等: "Enzymatic properties and anticancer activity of L-lysine α-oxidase from Trichoderma cf. aureoviride Rifai BKMF-4268D.", 《ANTICANCER DRUGS》 * |
| 宋金柱 等: "黄绿木霉纤维素酶提纯及其性质研究", 《哈尔滨工业大学学报》 * |
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