CN107653295A - Using the method for aspergillus cristatus and fermentation by saccharomyces cerevisiae tea dust production tobacco aromaticss - Google Patents
Using the method for aspergillus cristatus and fermentation by saccharomyces cerevisiae tea dust production tobacco aromaticss Download PDFInfo
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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
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- A24—TOBACCO; CIGARS; CIGARETTES; SIMULATED SMOKING DEVICES; SMOKERS' REQUISITES
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- A24B15/00—Chemical features or treatment of tobacco; Tobacco substitutes, e.g. in liquid form
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Abstract
The invention discloses a kind of method for producing tobacco aromaticss using aspergillus cristatus and fermentation by saccharomyces cerevisiae tea dust, it is that first aspergillus cristatus strain is seeded on CZG flat boards, 25 29 DEG C of 4 7d of culture, aspergillus cristatus ascospore is inoculated on CZG flat boards, 7 10d are cultivated at 25 29 DEG C, sterilized water dilutes, and obtains 10 ˇ3‑10ˇ6/ mL spore suspension；Saccharomyces cerevisiae strain is inoculated on YPD flat boards again, 35 45h is cultivated at 26 30 DEG C, S. cervisiae is inoculated in YPD fluid nutrient mediums, in 26 30 DEG C, the 45h of 100 120rpm concussion and cultivates 35, sterilized water dilutes to obtain 10 ˇ6‑10ˇ9/ mL brewing yeast cell suspension；The Saccharomyces cerevisiae cells suspension of 1 2% parts by weight aspergillus cristatus spore suspension and 2 4% percentage by weights is finally inoculated with tea dust fermentation substrate, ferment 3 5d at 26 29 DEG C, the sterile water filtration for adding 30 40 percentage by weights afterwards is extracted, by extract solution from, supernatant is taken, produces tobacco aromaticss.Spices produced by the present invention also creates aroma in addition to containing tealeaves tea perfume in itself, and the sweet distinctive salubrious bacterium of fragrant and aspergillus cristatus is fragrant.
Description
Technical field
The present invention relates to a kind of method for producing tobacco aromaticss, particularly one kind to use aspergillus cristatus and fermentation by saccharomyces cerevisiae
The method that tea dust produces tobacco aromaticss.
Background technology
The technique for applying and formula of essence spice for cigarette are the core technologies of tobacco industry, with enterprise's cigarette style and brand
Develop closely related.Spices is generally divided into two classes, and one kind is natural perfume material, has been currently known kind more than 3000, a kind of in addition to close
Planted into spices, up to more than 7000, and new kind also continues to bring out with the development of scientific research.Although synthetic perfume exists
It is more than natural perfume material in type and quantity, but due to the raising of concept of health, nowadays consumer increasingly tends to buy day
Right non-harmful product, therefore natural perfume material is also more favourable, and synthetic perfume is significantly larger than in price.Produced for natural perfume material,
US and European provides can only be by the method for the method or microorganisms/enzymes of physics.
The principle of micro-organisms tobacco aromaticss is：Microorganism is by some in fermentation substrate such as starch, protein, fiber
The organic macromolecule materials such as element, pectin and lignin change into flavor matter or aroma component precursor, then special using some
Technique these fragrance matters are extracted, then make an addition to tobacco product.It is more in tobacco microorganism flavouring research at home and abroad
Using the microbial strains from tobacco leaf in itself or from food separation, carried out in a manner of single strain or mixed fermentation.
But now current tobacco aromaticss preparation technology is complicated, cost is high, and fragrance is single, and spice liquid property is not steady enough
It is fixed, it is easily thin out that lower fragrance is deposited for a long time.
The content of the invention
It is an object of the present invention to provide one kind using aspergillus cristatus and fermentation by saccharomyces cerevisiae tea dust production tobacco aromaticss
Method.The characteristics of present invention has simple production process, and cost is relatively low, and the fragrance of spices enriches, and spice liquid property is stable.
Technical scheme：A kind of side that tobacco aromaticss are produced using aspergillus cristatus and fermentation by saccharomyces cerevisiae tea dust
Method, comprise the following steps：
1）Aspergillus cristatus strain is seeded on CZG flat boards, 25-29 DEG C of constant temperature is inverted culture 4-7d, obtains A product；
2）Choose the aspergillus cristatus ascospore coating activated in A product to be inoculated on CZG flat boards, cultivate 7-10d at 25-29 DEG C, treat
After aspergillus cristatus forms ascospore, aspergillus cristatus spore is scraped from flat board with the glass bar of sterilizing, sterilized water dilution, is used
Counting method of blood cell is counted, until obtaining 10 ˇ3-10ˇ6/ mL aspergillus cristatus spore suspension；
3）Saccharomyces cerevisiae strain is inoculated on YPD flat boards, culture 35-45h is inverted in 26-30 DEG C of constant temperature, obtains B product；
4）The S. cervisiae activated in picking B product is inoculated in YPD fluid nutrient mediums, in 26-30 DEG C, 100-120rpm concussion trainings
35-45h is supported, takes out shaking flask, more than 4h is stood and treats yeast cell pellets to shaking flask bottom, incline nutrient solution, and acquisition, which is deposited in, shakes
The brewing yeast cell in bottom of bottle portion, is diluted with sterilized water, is counted using counting method of blood cell, until obtaining 10 ˇ6-10ˇ9/mL
Brewing yeast cell suspension；
5）1-2% percentage by weight aspergillus cristatus spore suspensions and 2-4% percentage by weights are inoculated with tea dust fermentation substrate
Brewing yeast cell suspension, ferment 3-5d at 26-29 DEG C, obtains C product；
6）The sterilized water of 30-40% percentage by weights is added in C product, filters and is extracted after mixing, by extract solution in 5000-
3-7min is centrifuged under the conditions of 6000r/min, takes supernatant, produces tobacco aromaticss.
The foregoing method that tobacco aromaticss are produced using aspergillus cristatus and fermentation by saccharomyces cerevisiae tea dust, the step 1）
In, aspergillus cristatus strain is to be inverted culture 5d in 28 DEG C of constant temperature.
The foregoing method that tobacco aromaticss are produced using aspergillus cristatus and fermentation by saccharomyces cerevisiae tea dust, the step 2）
In, the aspergillus cristatus spore activated in A product is to cultivate 7d at 28 DEG C, and spore suspension concentration is 10 ˇ3/mL。
The foregoing method that tobacco aromaticss are produced using aspergillus cristatus and fermentation by saccharomyces cerevisiae tea dust, the step 3）
In, saccharomyces cerevisiae strain is to be inverted culture 40h in 28 DEG C of constant temperature, obtains B product；
The foregoing method that tobacco aromaticss are produced using aspergillus cristatus and fermentation by saccharomyces cerevisiae tea dust, the step 4）In, B
It in 28 DEG C, 110rpm concussion and cultivate 40h, brewing yeast cell concentration is 10 ˇ that the S. cervisiae activated in product, which is,6/mL。
The foregoing method that tobacco aromaticss are produced using aspergillus cristatus and fermentation by saccharomyces cerevisiae tea dust, the step 5）
In, it is the 4d that fermented at 28 DEG C.
The foregoing method that tobacco aromaticss are produced using aspergillus cristatus and fermentation by saccharomyces cerevisiae tea dust, the step 6）
In, it is the sterilized water that 35 parts by weight are added in C product, carries out extruding filtering extraction with four layers of gauze after mixing, extract solution is existed
5min is centrifuged under the conditions of 6000r/min, takes supernatant, produces tobacco aromaticss.
The foregoing method that tobacco aromaticss are produced using aspergillus cristatus and fermentation by saccharomyces cerevisiae tea dust, by weight,
The tea dust fermentation substrate includes tea leaf powder 34-36 parts, glucose 3.5-9 parts and water 55-61.5 parts.
The foregoing method that tobacco aromaticss are produced using aspergillus cristatus and fermentation by saccharomyces cerevisiae tea dust, by weight,
The tea dust fermentation substrate includes 55 parts of 36 parts of tea leaf powder, 9 parts of glucose and water.
Beneficial effects of the present invention：
One of fermentation strain that the present invention uses is from the separating obtained S. cervisiae of Maotai vinasse（Saccharomyces cerevisiae）, saccharomyces cerevisiae be at present by people understand at most and human society is produced the maximum microorganism of contribution it
One, can by substantial amounts of glucose, maltose, galactolipin and sucrose etc. it is saccharide converted be ethanol, frequently as production beer, fruit wine,
The food drinks such as Spirit and industrial fermentation produce the fermented bacterium of ethanol, in addition protein content in saccharomyces cerevisiae thalline
Height, containing nutriments such as abundant amino acid, vitamin, trace element and enzymes, widely it is applied in production
The fields such as feed, medical treatment.
Another bacterial strain of fermentation that the present invention uses is separating obtained aspergillus cristatus bacterium from Fu-brick tea（Aspergillus cristatus）, aspergillus cristatus category Aspergillus glaucus category is preced with, is the dominant bacteria during Fu-brick tea production is grown dim, can be by tealeaves
The substance decompositions such as starch, tannin convert, and to reduce the bitter taste of tealeaves, make the mellow sweetness of tealeaves flavour, and impart Fu-brick tea
The greasy healthcare function of the steatolysis that disappears, therefore Fu-brick tea is referred to as the tea of life by the border area herdsman such as Xinjiang, Qinghai, Gansu, Ningxia.
Tea dust refers to screen broken, incomplete tealeaves from tealeaves, belongs to a kind of tealeaves low value by-product
Product, typically sell at a low price and be used for deodorization, dehumidifying or as fertilizers for potted flowers to consumer, although the price of tea dust is well below tea
Leaf, but the nutritional ingredient contained by it accounts for the 1/3 of gross dry weight, caffeine 2%-5 % with tealeaves and indifference, wherein Polyphenols,
The % of amino acid 2% -4, and rich in vitamin and potassium (K), iron (F e), selenium (S e) such as dimension A, dimension B, Vc, dimension E, dimension K
With the mineral element such as fluorine (F).
Simple production process of the present invention, and cost is relatively low, and obtained spices is in addition to containing tealeaves tea perfume in itself, also
Aroma is generated, the sweet distinctive salubrious bacterium of fragrant and aspergillus cristatus is fragrant, and fragrance is more abundant, meanwhile, the spice liquid produced
Property is stable, and characteristic chicken flavor still keeps good after being placed in natural conditions next year.
Embodiment
With reference to embodiment, the present invention is further illustrated, but is not intended as the foundation limited the present invention.
Embodiments of the invention
Embodiment 1：A kind of method for producing tobacco aromaticss using aspergillus cristatus and fermentation by saccharomyces cerevisiae tea dust, step are as follows：
1）Aspergillus cristatus strain is seeded on CZG flat boards, 25 DEG C of constant temperature are inverted culture 7d, obtain A product；
2）Choose the aspergillus cristatus ascospore activated in A product to be inoculated on CZG flat boards, cultivate 10d at 25 DEG C, treat aspergillus cristatus shape
Into after ascospore, aspergillus cristatus spore, sterilized water dilution, using counting method of blood cell are scraped from flat board with the glass bar of sterilizing
Counted, until obtaining 10 ˇ3/ mL spore suspension；
3）Saccharomyces cerevisiae strain is inoculated on YPD flat boards, culture 45h is inverted in 26 DEG C of constant temperature, obtains B product；
4）The S. cervisiae activated in picking B product is inoculated in YPD fluid nutrient mediums, in 26 DEG C, 100rpm concussion and cultivate 45h,
Shaking flask is taken out, 4h is stood and treats yeast cell pellets to shaking flask bottom, incline nutrient solution, obtains the wine brewing ferment for being deposited in shaking flask bottom
Mother cell, diluted with sterilized water, counted using counting method of blood cell, until obtaining 10 ˇ6/ mL brewing yeast cell suspension；
5）The wine brewing ferment of 2% percentage by weight aspergillus cristatus seed liquor and 4% percentage by weight is inoculated with tea dust fermentation substrate
Female bacterium seed liquor, ferment 5d at 26 DEG C, obtains C product；
6）The sterilized water of 35% percentage by weight is added in C product, filters and is extracted after mixing, by extract solution in 6000r/min
Under the conditions of centrifuge 5min, take supernatant, produce tobacco aromaticss.
The tea dust fermentation substrate includes 60 parts of 34 parts of tea leaf powder, 6 parts of glucose and water.
The CZG culture medium prescriptions are：Sucrose 20g, sodium chloride 23g, dipotassium hydrogen phosphate lg, nitric acid ammonia 3g, magnesium sulfate
0.5g, agar 20g.
The YPD culture medium prescriptions are：Glucose 20g, yeast extract 5g, peptone 10g, agar 20g.
The YPD Liquid Cultures based formulas is：Glucose 20g, yeast extract 5g, peptone 10g.
Embodiment 2：A kind of method for producing tobacco aromaticss using aspergillus cristatus and fermentation by saccharomyces cerevisiae tea dust, step is such as
Under：
1）Aspergillus cristatus strain is seeded on CZG flat boards, 28 DEG C of constant temperature are inverted culture 5d, obtain A product；
2）Choose the aspergillus cristatus ascospore coating activated in A product and be inoculated in CZG fluid nutrient mediums, cultivate 8d at 28 DEG C, wait to be preced with
After prominent aspergillus forms ascospore, aspergillus cristatus spore, sterilized water dilution, using blood are scraped from flat board with the glass bar of sterilizing
Ball count method is counted, until obtaining 10 ˇ5/ mL spore suspension；
3）Saccharomyces cerevisiae strain is inoculated on YPD flat boards, culture 40h is inverted in 28 DEG C of constant temperature, obtains B product；
4）The S. cervisiae activated in picking B product is inoculated in YPD fluid nutrient mediums, in 28 DEG C, 110rpm concussion and cultivate 40h,
Shaking flask is taken out, 5h is stood and treats yeast cell pellets to shaking flask bottom, incline nutrient solution, obtains the wine brewing ferment for being deposited in shaking flask bottom
Mother cell, diluted with sterilized water, counted using counting method of blood cell, until obtaining 10 ˇ8/ mL brewing yeast cell suspension；
5）The wine brewing ferment of 2 percentage by weight aspergillus cristatus spore suspensions and 4 percentage by weights is inoculated with tea dust fermentation substrate
Mother cell suspension, ferment 4d at 28 DEG C, obtains C product；
6）The sterilized water of 30 percentage by weights is added in C product, filters and is extracted after mixing, by extract solution in 5000r/min
Under the conditions of centrifuge 7min, take supernatant, produce tobacco aromaticss.
The tea dust fermentation substrate includes 61.5 parts of 35 parts of tea leaf powder, 3.5 parts of glucose and water.
The CZG culture mediums and YPD culture medium prescriptions are consistent with embodiment 1.
Embodiment 3：A kind of method for producing tobacco aromaticss using aspergillus cristatus and fermentation by saccharomyces cerevisiae tea dust, step is such as
Under：
1）Aspergillus cristatus strain is seeded on CZG flat boards, 29 DEG C of constant temperature are inverted culture 4d, obtain A product；
2）Choose the aspergillus cristatus ascospore activated in A product and be inoculated in CZG fluid nutrient mediums, cultivate 7d at 29 DEG C, treat coronoid process
After aspergillus forms ascospore, aspergillus cristatus spore, sterilized water dilution, using blood cell are scraped from flat board with the glass bar of sterilizing
Counting method is counted, until obtaining 10 ˇ6/ mL spore suspension；
3）Saccharomyces cerevisiae strain is inoculated on YPD flat boards, culture 35h is inverted in 30 DEG C of constant temperature, obtains B product；
4）The S. cervisiae activated in picking B product is inoculated in YPD fluid nutrient mediums, in 30 DEG C, 120rpm concussion and cultivate 35h,
Shaking flask is taken out, 8h is stood and treats yeast cell pellets to shaking flask bottom, incline nutrient solution, obtains the wine brewing ferment for being deposited in shaking flask bottom
Mother cell, diluted with sterilized water, counted using counting method of blood cell, until obtaining 10 ˇ9/ mL brewing yeast cell suspension；
5）The wine brewing ferment of 1 percentage by weight aspergillus cristatus spore suspension and 2 percentage by weights is inoculated with tea dust fermentation substrate
Mother cell suspension, ferment 3d at 29 DEG C, obtains C product；
6）The sterilized water of 40 percentage by weights is added in C product, filters and is extracted after mixing, by extract solution in 6000r/min
Under the conditions of centrifuge 3min, take supernatant, produce tobacco aromaticss.
The tea dust fermentation substrate includes 55 parts of 36 parts of tea leaf powder, 9 parts of glucose and water.
The CZG culture mediums and YPD culture medium prescriptions are consistent with embodiment 1.
As it can be seen from table 1 the overall organoleptic of the cigarette made through spices produced by the present invention smokes panel test, scoring is higher than control
The scoring of group.
Claims (9)
- A kind of 1. method that tobacco aromaticss are produced using aspergillus cristatus and fermentation by saccharomyces cerevisiae tea dust, it is characterised in that including Following steps：1）Aspergillus cristatus strain is seeded on CZG flat boards, 25-29 DEG C of constant temperature is inverted culture 4-7d, obtains A product；2）Choose the aspergillus cristatus ascospore coating activated in A product to be inoculated on CZG flat boards, cultivate 7-10d at 25-29 DEG C, treat After aspergillus cristatus forms ascospore, aspergillus cristatus spore is scraped from flat board with the glass bar of sterilizing, sterilized water dilution, is used Counting method of blood cell is counted, until obtaining 10 ˇ3-10ˇ6/ mL aspergillus cristatus spore suspension；3）Saccharomyces cerevisiae strain is inoculated on YPD flat boards, culture 35-45h is inverted in 26-30 DEG C of constant temperature, obtains B product；4）The S. cervisiae activated in picking B product is inoculated in YPD fluid nutrient mediums, in 26-30 DEG C, 100-120rpm concussion trainings 35-45h is supported, takes out shaking flask, more than 4h is stood and treats yeast cell pellets to shaking flask bottom, incline nutrient solution, and acquisition, which is deposited in, shakes The brewing yeast cell in bottom of bottle portion, is diluted with sterilized water, is counted using counting method of blood cell, until obtaining 10 ˇ6-10ˇ9/mL Brewing yeast cell suspension；5）1-2% percentage by weight aspergillus cristatus spore suspensions and 2-4% percentage by weights are inoculated with tea dust fermentation substrate Brewing yeast cell suspension, ferment 3-5d at 26-29 DEG C, obtains C product；6）The sterilized water of 30-40% percentage by weights is added in C product, filters and is extracted after mixing, by extract solution in 5000- 3-7min is centrifuged under the conditions of 6000r/min, takes supernatant, produces tobacco aromaticss.
- 2. the method according to claim 1 that tobacco aromaticss are produced using aspergillus cristatus and fermentation by saccharomyces cerevisiae tea dust, It is characterized in that：The step 1）In, aspergillus cristatus strain is to be inverted culture 5d in 28 DEG C of constant temperature.
- 3. the method according to claim 1 that tobacco aromaticss are produced using aspergillus cristatus and fermentation by saccharomyces cerevisiae tea dust, It is characterized in that：The step 2）In, the aspergillus cristatus spore activated in A product is to cultivate 7d at 28 DEG C, and spore suspension concentration is 10ˇ3/mL。
- 4. the method according to claim 1 that tobacco aromaticss are produced using aspergillus cristatus and fermentation by saccharomyces cerevisiae tea dust, It is characterized in that：The step 3）In, saccharomyces cerevisiae strain is to be inverted culture 40h in 28 DEG C of constant temperature, obtains B product.
- 5. the method according to claim 1 that tobacco aromaticss are produced using aspergillus cristatus and fermentation by saccharomyces cerevisiae tea dust, It is characterized in that：The step 4）In, the S. cervisiae activated in B product is in 28 DEG C, 110rpm concussion and cultivate 40h, wine brewing Barm cell concentration is 10 ˇ6/mL。
- 6. the method according to claim 1 that tobacco aromaticss are produced using aspergillus cristatus and fermentation by saccharomyces cerevisiae tea dust, It is characterized in that：The step 5）In, it is the 4d that fermented at 28 DEG C.
- 7. the method according to claim 1 that tobacco aromaticss are produced using aspergillus cristatus and fermentation by saccharomyces cerevisiae tea dust, It is characterized in that：The step 6）In, it is the sterilized water that 35 parts by weight are added in C product, is squeezed after mixing with four layers of gauze Press filtration is extracted, and extract solution is centrifuged into 5min under the conditions of 6000r/min, takes supernatant, produces tobacco aromaticss.
- 8. the method according to claim 1 that tobacco aromaticss are produced using aspergillus cristatus and fermentation by saccharomyces cerevisiae tea dust, It is characterized in that：By weight, the tea dust fermentation substrate includes tea leaf powder 34-36 parts, glucose 3.5-9 parts and water 55-61.5 parts.
- 9. the method according to claim 8 that tobacco aromaticss are produced using aspergillus cristatus and fermentation by saccharomyces cerevisiae tea dust, It is characterized in that：By weight, the tea dust fermentation substrate includes 55 parts of 36 parts of tea leaf powder, 9 parts of glucose and water.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109258370A (en) * | 2018-07-29 | 2019-01-25 | 四川省烟草公司宜宾市公司 | A kind of tobacco planting method improving the sweet fragrant breeze lattice of tobacco leaf |
| CN111011600A (en) * | 2019-12-26 | 2020-04-17 | 江苏省微生物研究所有限责任公司 | Compound microbial feed additive and preparation method thereof |
| CN115851841A (en) * | 2023-01-09 | 2023-03-28 | 福建中烟工业有限责任公司 | Extract of Tieguanyin enzyme-deactivating substance, preparation method of extract, tobacco leaf composition and cigarette |
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