CN109820239A - A method of strengthening tobacco stem shred fermentation process - Google Patents
A method of strengthening tobacco stem shred fermentation process Download PDFInfo
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- CN109820239A CN109820239A CN201910093125.6A CN201910093125A CN109820239A CN 109820239 A CN109820239 A CN 109820239A CN 201910093125 A CN201910093125 A CN 201910093125A CN 109820239 A CN109820239 A CN 109820239A
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Abstract
The present invention discloses a kind of method for strengthening tobacco stem shred fermentation process, it is handled using bacillus amyloliquefaciens GUHP-86 preparation for the tobacco stem shred of fermentation process, the bacillus amyloliquefaciens GUHP-86 preparation includes bacillus amyloliquefaciens GUHP-86, for the bacillus amyloliquefaciens GUHP-86 on January 4th, 2016 in China typical culture collection center preservation, deposit number is CCTCC M 2016003.Bacillus amyloliquefaciens GUHP-86 preparation of the present invention, can strengthen tobacco stem shred fermentation process, have and increase cigarette perfume, soft flue gas, increase exquisiteness, reduce irritation, hence it is evident that improve flue gas matter and improve the effect of the practical safety of buccal cigarette.Meanwhile the invention also discloses a kind of methods for improving protease and amylase activity in tobacco stem shred fermentation process.
Description
Technical field
The invention belongs to microorganisms and technical field of tobacco processing, relate in particular to a kind of tobacco stem shred hair of reinforcing
The method of ferment process.
Background technique
When using combustion process does not occur for smoke-free tobacco product (Smokeless tobacco products, STPs),
Harmful pyrolysis product will not be generated, smokeless generates, can use in public places, be low harmful environment-friendly type new tobacco products
A kind of important composition form.
Buccal cigarette is the staple product form of STPs, is the fastest-rising product category of existing market share, by the world
The concern of each main tobacco enterprise, and a large amount of fund and the relevant R&D work of energy progress are put into the field.
In the raw materials for production for sucking tobacco product, offal is a kind of important adjunct ingredient, usually with tobacco stem shred powder
Form use in the oral cavity, it is tobacco stem shred as auxiliary material have harmful components it is low, it is at low cost, tobacco waste effective use etc. it is excellent
Gesture.Tobacco stem shred use can be substantially reduced the price of mouth containing smoke product.But stem is used also as auxiliary material in buccal cigarette
The unfavorable factor for bringing perfume quantity to reduce, therefore the use of the quality that biofermentation method promotes stem is to determine mouth containing smoke product wind
Lattice feature and the key factor for reducing price, to influence consumption market to the acceptance level of product.
Microbial fermentation is an effective way for improving the tobacco stem shred material quality of buccal cigarette.Research table both domestic and external
It is bright, stem perfume quantity and alcoholization time can be effectively improved by microbial fermentation, hence it is evident that promote the jealous of stem, while may be used also
The xylon gas of stem are reduced, the aroma quality and fragrance quantitative change when using stem are good.In fermentation process, microorganism carries out metabolism work
The dynamic protein consumed in stem, cellulose, starch and some undesirable constituents etc., while microorganism pass through the one of metabolism generation
A little substances can increase aromatic odor to stem again, and stem various composition ratio is made to tend to coordinate, thus improve the perfume quantity of stem,
The qualities such as fragrance make miscellaneous gas, irritation decline, have preferably edible comfort.
In addition, buccal cigarette is using the tobacco stem shred smokeless tobacco stem product as filling auxiliary material, wherein fermentation alcoholization
Can be generated in the process containing potentially harmful Components Chemical ingredient, thus it is tobacco stem shred in some intrinsic harmful components by serious shadow
Ring the edible safety of buccal cigarette.TSNAs (tobacco specific nitrosamines) is potentially harmful ingredient important in tobacco product.Mouthful
Containing tobacco product, TSNAs forms the alkaloid being widely considered to be by tobacco after everfermentation refines and nitrous reactant salt is formed,
Therefore nitrite is one of the precursor substance for generating nitrosamine, then reducing content of nitrite in tobacco leaf will be helpful to reduce
Content of nitrosamines improves the edible safety of buccal cigarette.The study found that the presence of microorganism can generate the content of nitrite
Very big influence, nitrite can drop in many types in microorganism, be then seeded into tobacco leaf by screening drop nitrate microorganism
Content of nitrosamines in stem can be reduced by carrying out fermentation.
Meanwhile both effectiveness ingredient is often generated in microbial metabolism, such as generate a variety of enzymes, some enzymes itself are to mentioning
The fragrance of high tobacco leaf is helpful, and the protein in tobacco leaf can be hydrolyzed such as protease；Some enzymes are the enzymes beneficial to human health,
Such as Nattokinase (nattokinase abbreviation NK), blood viscosity can be reduced with thrombus, improve blood circulation, softening and increase
Blood vessel elasticity, amylase play critically important effect to digestive system, and a series of problems will be brought to body by lacking this enzyme, such as
Skin erythema feels depressed and the psychological relevant issues such as allergy.The generation of these biologically active functional enzymes can be more
The edible quality of buccal cigarette is improved well.
However it is tobacco stem shred and be different from tobacco leaf, tobacco leaf itself with regard to softer, and offal itself is harder, is unfavorable for sending out
Ferment process, so for promoting the microorganism of Tobacco Fermentation Process that might not can promote tobacco stem shred fermentation process.
Accordingly it is desirable to find the microorganism of other New raxas, with for tobacco stem shred fermentation process, and realizes and subtract
Evil, flavouring and the effect for improving bioactive functions enzymatic activity.
Summary of the invention
First aspect present invention is related to a kind of method for strengthening tobacco stem shred fermentation process, uses bacillus amyloliquefaciens
GUHP-86 preparation is handled for the tobacco stem shred of fermentation process, the bacillus amyloliquefaciens GUHP-86 preparation
Preparation method the following steps are included:
(1) tryptone 0.5-2g, yeast extract 0.5-1.5g, NaCl 6-8g, glucose 0.5-2 g are weighed, is supplemented
Distilled water accesses the solution starch gemma bar to 1000mL, and in 110-130 DEG C of high pressure steam sterilization 10-30min after cooling
Bacterium GUHP-86 carries out culture 18h ± 2h in 30-40 DEG C, 170-190r/min, obtains seed liquor；
(2) seed liquor described in 1000mL is centrifuged under 2-5 DEG C, 7000-9000r 10-20min, collects precipitating, and use 6-
The sterile saline of 9g/L washs the precipitating, and the sterile saline of 1000 mL is then added into the precipitating, shakes
It is even, obtain the bacillus amyloliquefaciens GUHP-86 preparation；
The bacillus amyloliquefaciens GUHP-86 is protected on January 4th, 2016 in China typical culture collection center
Hiding, deposit number are CCTCC M 2016003.
Preferably, the sterile saline need to be in 121 DEG C of high pressure steam sterilization 20min.
Preferably, tobacco stem shred fermentation step is as follows: weighing the tobacco stem shred hermetic bag in 14cm × 20cm specification of 20g
It is interior, the bacterium solution (i.e. 10%-30% inoculum concentration) in 2-6mL centrifuge tube is drawn in tobacco stem shred, and 4mL has sugared salting liquid in tobacco
It in stem, mixes well, is cultivated in 37 DEG C of incubators, periodically taken a breath daily, and add one when humidity reduces
Surely there is sugared salting liquid.
Preferably, described to have sugared salting liquid preparation step as follows: to weigh MgCl25g、KCl 0.5g、CaCl20.5g、 NaCl
0.5g, glucose 2g, moisturizing to 1000mL, dissolution have obtained sugared salting liquid.
Second aspect of the present invention provides a kind of for improving protease and amylase activity in tobacco stem shred fermentation process
Method, handled using the bacillus amyloliquefaciens GUHP-86 preparation for the tobacco stem shred of fermentation process.
Compared with the existing technology, the invention has the following advantages:
(1) bacillus amyloliquefaciens GUHP-86 preparation of the invention can strengthen the fermentation process of tobacco.It is solved through the present invention
Bacillus amyloliquefaciens GUHP-86 preparation (20% inoculum concentration) treated it is tobacco stem shred fermentation 15 days after, to tobacco stem shred sense
Official evaluation compared with without bacillus amyloliquefaciens GUHP-86 preparation handle it is tobacco stem shred increase 19.10%, i.e., solution of the invention
Bacillus amyloliquefaciens GUHP-86 preparation, which has, increases cigarette perfume, soft flue gas, increases exquisiteness, reduces irritation, hence it is evident that improves cigarette
The effect of makings improves tobacco stem shred quality.Meanwhile through bacillus amyloliquefaciens GUHP-86 preparation of the present invention (20% inoculation
Amount) after tobacco stem shred fermentation that treated 21 days, content of nitrite has dropped 66.39% compared with Nicotiana tabacum stem, therefore, adopts
The tobacco stem shred practical safety that can also improve buccal cigarette is handled with bacillus amyloliquefaciens GUHP-86 preparation.
(2) bacillus amyloliquefaciens GUHP-86 preparation of the invention can be improved natto in tobacco stem shred fermentation process and swash
The activity of enzyme, protease and amylase.After bacillus amyloliquefaciens GUHP-86 preparation (20% inoculum concentration) of the present invention processing
Tobacco stem shred fermentation 15 days after, tobacco stem shred middle Nattokinase enzyme activity and proteinase activity be respectively 10.48FU/g and
1308.32IU/g.Meanwhile through bacillus amyloliquefaciens GUHP-86 preparation (20% inoculum concentration) of the present invention treated tobacco stems
After silk fermentation 15 days, it is tobacco stem shred in obviously detect the presence of amylase, but without bacillus amyloliquefaciens GUHP- of the present invention
86 preparations processing it is tobacco stem shred it is fermented after and be not present amylase.And Nattokinase, protease and amylase not only itself
The enzyme helpful or beneficial to human health to the fragrance that raising is tobacco stem shred, these biologically active functional enzymes
Generation can preferably improve the edible quality of buccal cigarette.
Specific embodiment
Below with reference to embodiment, the present invention is described in further detail, but is not limitation of the present invention, based on this
Any transformation or improvement that invention training centre is done, each fall within protection scope of the present invention.Be not specified in embodiment particular technique or
Condition person described technology or conditions or carries out according to the literature in the art according to product description.Agents useful for same or
Production firm person is not specified in instrument, is the conventional products that can be obtained by purchase.
Embodiment 1
The present embodiment is the preparation of bacillus amyloliquefaciens GUHP-86 preparation, specifically includes the following steps:
(1) tryptone 1.25g, yeast extract 0.625g, NaCl 6.25g, glucose 0.625g are weighed, supplement is steamed
Distilled water accesses the bacillus amyloliquefaciens GUHP-86 to 1000mL, and in 121 DEG C of high pressure steam sterilization 20min after cooling
Culture 18h ± 2h is carried out in 37 DEG C, 180r/min, obtains seed liquor；
(2) seed liquor described in 1000mL is centrifuged under 4 DEG C, 8000r 10min, collects precipitating, and with the sterilizing of 8.5g/L
It is precipitated described in brine, the sterile saline of 1000mL is then added into the precipitating, shakes up, obtains the solution
Bacillus amyloliquefaciens GUHP-86 preparation.
The bacillus amyloliquefaciens GUHP-86 is protected on January 4th, 2016 in China typical culture collection center
Hiding, deposit number are CCTCC M 2016003.
The sterile saline need to be in 121 DEG C of high pressure steam sterilization 20min.
Embodiment 2
It is tobacco stem shred in the hermetic bag of 14cm × 20cm specification that 20g is weighed in the present embodiment, draws Xie Dian described in 2mL
Afnyloliquefaciens GUHP-86 preparation (i.e. 10% inoculum concentration) is in tobacco stem shred, and 4mL has sugared salting liquid in tobacco stem shred, sufficiently
It mixes, obtains sample 1.
It is described to have sugared salting liquid preparation step as follows: to weigh MgCl25g、KCl 0.5g、CaCl20.5g, NaCl 0.5g, Portugal
Grape sugar 2g, moisturizing to 1000mL, dissolution have obtained sugared salting liquid.
It is tobacco stem shred in the hermetic bag of 14cm × 20cm specification to weigh 20g, absorption 2mL sterile saline, 4mL have sugar
Salting liquid is mixed well, obtains comparative sample 1 in tobacco stem shred.
Sample 1 and comparative sample 1 are cultivated in 37 DEG C of incubators simultaneously, periodically taken a breath daily, and work as humidity
Adding when reduction centainly has sugared salting liquid.
Steps are as follows for subjective appreciation: more than 10 position this field sensory test personnel being asked to carry out sense organ to fermented tobacco stem sample
Evaluation.Method of smokeing panel test is that buccal cigarette is placed directly between upper lip and upper row's tooth, the pipe tobacco granular mass being put into lip every time
For 0.2g, standard is set to 10min using the time.It is uniform to the entrance mouthfeel of tobacco stem shred fermented sample, fragrance, strength, release
Property it is 15 points each, irritation, is grown compliance (below standard quasi- spued using the time of buccal cigarette edible time is considered as poor compliance)
Each 10 points of taste, pleasant impression, total score 100 is divided, and is evaluated.
To carry out 121 DEG C, 5min pasteurize after fermentation, subjective appreciation is carried out.
Embodiment 3
It is tobacco stem shred in the hermetic bag of 14cm × 20cm specification that 20g is weighed in the present embodiment, draws Xie Dian described in 4mL
Afnyloliquefaciens GUHP-86 preparation (i.e. 20% inoculum concentration) is in tobacco stem shred, and 4mL has sugared salting liquid in tobacco stem shred, sufficiently
It mixes, obtains sample 2.
It is described to have sugared salting liquid preparation step as follows: to weigh MgCl25g、KCl 0.5g、CaCl20.5g, NaCl 0.5g, Portugal
Grape sugar 2g, moisturizing to 1000mL, dissolution have obtained sugared salting liquid.
It is tobacco stem shred in the hermetic bag of 14cm × 20cm specification to weigh 20g, absorption 4mL sterile saline, 4mL have sugar
Salting liquid is mixed well, obtains comparative sample 2 in tobacco stem shred.
Sample 2 and comparative sample 2 are cultivated in 37 DEG C of incubators simultaneously, periodically taken a breath daily, and work as humidity
Adding when reduction centainly has sugared salting liquid.
Please more than 10 position this field sensory test personnel to fermented tobacco stem sample carry out subjective appreciation.Method of smokeing panel test is will
Buccal cigarette is placed directly between upper lip and upper row's tooth, and the pipe tobacco granular mass being put into lip every time is 0.2g, when standard uses
Between be set to 10min.It is irritation, suitable to the entrance mouthfeel of tobacco stem shred fermented sample, fragrance, strength, each 15 points of uniformity of release
Answering property (below standard quasi- spued using the time of buccal cigarette edible time is considered as poor compliance), flavour, each 10 points of pleasant impression, total score
It 100 points, is evaluated.
To carry out 121 DEG C, 5min pasteurize after fermentation, subjective appreciation is carried out.
Embodiment 4
It is tobacco stem shred in the hermetic bag of 14cm × 20cm specification that 20g is weighed in the present embodiment, draws Xie Dian described in 6mL
Afnyloliquefaciens GUHP-86 preparation (i.e. 30% inoculum concentration) is in tobacco stem shred, and 4mL has sugared salting liquid in tobacco stem shred, sufficiently
It mixes, obtains sample 3.
It is described to have sugared salting liquid preparation step as follows: to weigh MgCl25g、KCl 0.5g、CaCl20.5g, NaCl 0.5g, Portugal
Grape sugar 2g, moisturizing to 1000mL, dissolution have obtained sugared salting liquid.
It is tobacco stem shred in the hermetic bag of 14cm × 20cm specification to weigh 20g, absorption 6mL sterile saline, 4mL have sugar
Salting liquid is mixed well, obtains comparative sample 3 in tobacco stem shred.
Sample 3 and comparative sample 3 are cultivated in 37 DEG C of incubators simultaneously, periodically taken a breath daily, and work as humidity
Adding when reduction centainly has sugared salting liquid.
Steps are as follows for subjective appreciation: more than 10 position this field sensory test personnel being asked to carry out sense organ to fermented tobacco stem sample
Evaluation.Method of smokeing panel test is that buccal cigarette is placed directly between upper lip and upper row's tooth, the pipe tobacco granular mass being put into lip every time
For 0.2g, standard is set to 10min using the time.It is uniform to the entrance mouthfeel of tobacco stem shred fermented sample, fragrance, strength, release
Property it is 15 points each, irritation, is grown compliance (below standard quasi- spued using the time of buccal cigarette edible time is considered as poor compliance)
Each 10 points of taste, pleasant impression, total score 100 is divided, and is evaluated.
To carry out 121 DEG C, 5min pasteurize after fermentation, subjective appreciation is carried out.
Embodiment 3, embodiment 4, the sensory evaluation score value of different samples during the fermentation is shown in Table 1 in embodiment 5.
By result in table 1 it is found that bacillus amyloliquefaciens GUHP-86 preparation of the invention can strengthen the fermentation of tobacco
Journey.It is right after through bacillus amyloliquefaciens GUHP-86 preparation (20% inoculum concentration) of the present invention, treated tobacco stem shred fermentation 15 days
Tobacco stem shred sensory evaluation tobacco stem shred is increased compared with what is handled without bacillus amyloliquefaciens GUHP-86 preparation
19.10%, i.e., bacillus amyloliquefaciens GUHP-86 preparation of the invention, which has, increases cigarette perfume, soft flue gas, increases exquisiteness,
Reduce irritation, hence it is evident that the effect for improving flue gas matter improves tobacco stem shred quality.Meanwhile inoculum concentration be 20% when, sense organ is commented
Valence scoring highest, ferment effect is best, and bigger to tobacco stem shred improvement, obtained fermentation tobacco quality is more preferable.
The sensory evaluation score value of the different samples of table 1 during the fermentation is shown in Table 1
Embodiment 5
By the sample 2 in embodiment 3, the fermented tobacco stem after comparative sample 2 fermented 15 days be named as fermented sample 2,
Ferment comparative sample 2, respectively measure fermented sample 2, fermentation comparative sample 2 in natto kinase activity.
Crude enzyme liquid extracts: weighing fermented sample 1g, is accurate to 0.0002g.For 24 hours with the extraction of 25mL phosphate buffer solution, so
It is filtered afterwards with qualitative filter paper at a slow speed, filtrate is stand-by.
1.4mLTris-HCl (50mmol/L, pH7.8) buffer and 0.4mL fibrinogen solution are added into test tube
0.1mL fibrin ferment (20U/mL) is added after 37 DEG C of incubation 5min in (7.2mg/mL), then 37 DEG C of incubation 10min form artificial blood
Bolt, is added 0.1mL sample crude enzyme liquid, and it is whole that 2mL trichloroacetic acid (0.2moL/L) solution left standstill 20min is added in 37 DEG C of incubation 60min
It only reacts, 13000r/min is centrifuged 10min, and supernatant is taken to measure absorbance at 275nm wavelength.Enzyme activity definition: per minute
Enzyme amount required for absorbance increases by 0.01 at 275nm is defined as the fibrin degradation enzyme activity (FU) of 1 unit.
Different fermentations sample Nattokinase vitality test the results are shown in Table 2.
From Table 2, it can be seen that fermentation 2 Nattokinase enzyme activity of comparative sample is 0.29FU/g, 2 Nattokinase of fermented sample
Enzyme activity is 10.48FU/g, shows that bacillus amyloliquefaciens GUHP-86 preparation can be improved natto in tobacco stem shred fermentation process and swash
Enzymatic activity.
2 different fermentations sample Nattokinase vitality test result of table
Embodiment 6
By the sample 2 in embodiment 4, the fermented tobacco stem after comparative sample 2 fermented 15 days be named as fermented sample 2,
Fermentation comparative sample 2 measures proteinase activity in fermented sample 2 and fermentation comparative sample 2 respectively.
Crude enzyme liquid extracts: weighing fermented sample 1g, is accurate to 0.0002g.Then it is extracted with 25mL phosphate buffer solution
For 24 hours, it is then filtered with qualitative filter paper at a slow speed, filtrate is stand-by.
Enzymatic activity is measured using casein plate method.Trypsase commercial enzyme is configured to 10IU/mL, 25IU/ respectively
ML, 50IU/mL, 75IU/mL, 100IU/mL, 125IU/mL concentration respectively take 10uL point sample in the casein plate hole newly prepared
In, it is inverted into 37 DEG C of incubators after placement 10min and is taken out after 16h naturally, measurement dissolution loop diameter calculates each dissolution circle
Area, to measure dissolution circle area (x, mm2) and trypsase vigor (y, IU/mL) relationship.
Measure sample enzyme activity.It takes fermented tobacco stem crude enzyme liquid 10uL point sample on casein plate, so places 10min
After be inverted into 37 DEG C of incubators and taken out after 16h, measurement dissolution loop diameter calculates each dissolution and encloses area, according to dissolving circle face
Product (x, mm2) and prolease activity (y, IU/mL) relationship calculate sample protein enzyme enzyme activity.Measuring fermentation 2 enzyme activity of comparative sample is
137.86IU/g, 2 proteinase activity of fermented sample are 1308.32IU/g, show that bacillus amyloliquefaciens GUHP-86 preparation exists
High enzyme protease living can be generated in tobacco stem shred fermentation process.
Embodiment 7
By the sample 2 in embodiment 4, the fermented tobacco stem after comparative sample 2 fermented 15 days be named as fermented sample 2,
Fermentation comparative sample 2 measures in fermented sample 2 and fermentation comparative sample 2 whether produce amylase respectively.
LB plate added with soluble starch is punched, water is then accessed and extracts the resulting extracting solution of fermented sample 2,
Discovery has transparent circle after iodine dye, shows that it produces amylase.
LB plate added with soluble starch is punched, water is then accessed and extracts the extracting solution that fermentation comparative sample 2 obtains,
It does not find significantly to change after iodine dye, shows that it does not produce amylase substantially.
Embodiment 8
Content of nitrite in 2 fermentation process of sample 2 and comparative sample in embodiment 4 is measured.
Content of nitrite is measured using naphthodiamide hydrochloric acid method.It weighs 3.0g to clay into power the tobacco stem shred of shape, be put in
In 250mL conical flask, 10mL is then added and is saturated borax soln, stirs evenly, distilled water 150mL is added, in 70 degree of water-baths
Heating 30min simultaneously takes out.It is filtered.Then filtrate is poured into 250mL volumetric flask, it is ferrous that 5mL is added while rotating
Potassium cyanide solution shakes up and 5mL zinc acetate solution is added, with precipitating proteins.Water is added into scale, is shaken up, is filtered with filter paper.
Above-mentioned processed filtrate 50mL learn from else's experience in the volumetric flask of 100mL, 2mL is added and is saturated basic lead acetate solution,
Add water to scale, shake up, then filters.It takes filter liquor 50mL in 100mL volume bottle, 0.1mL sulfuric acid solution is added, water is added to arrive
Scale shakes up, and then refilters, and filtrate is spare.
20mL treated filtrate is measured, 2mL p-aminobenzene sulfonic acid (4g/L) is added in 50mL colorimetric cylinder, stirring is put
It sets 3 minutes~5 minutes, 1mL hydrochloride naphthodiamide solution is then added, adds water to scale, 15 minutes is stood after stirring, with 2 lis
Rice colorimetric cylinder, Yu Bochang 545nm place survey absorbance, according to absorbance calculating content of nitrite.
Content of nitrite measurement result is shown in Table 3 in different sample fermentation process.
Content of nitrite measurement result in the different sample fermentation process of table 3
As can be seen from Table 3, through bacillus amyloliquefaciens GUHP-86 preparation (20% inoculum concentration) of the present invention treated cigarette
After careless stem ferments 21 days, content of nitrite compared with without bacillus amyloliquefaciens GUHP-86 preparation handle it is tobacco stem shred under
Dropped 66.39%, therefore, using bacillus amyloliquefaciens GUHP-86 preparation handle it is tobacco stem shred not only make tobacco stem shred flavouring,
The functional enzyme of multiple beneficial is generated, and greatly reduces tobacco stem shred middle content of nitrite, this will improve the food of buccal cigarette
Use safety.
Claims (2)
1. a kind of method for strengthening tobacco stem shred fermentation process, which is characterized in that use bacillus amyloliquefaciens GUHP-86 preparation
It handles for the tobacco stem shred of fermentation process, the preparation method of the bacillus amyloliquefaciens GUHP-86 preparation includes
Following steps:
(1) tryptone 0.5-2g, yeast extract 0.5-1.5g, NaCl 6-8g, glucose 0.5-2g, supplement distillation are weighed
Water accesses the bacillus amyloliquefaciens to 1000mL, and in 110-130 DEG C of high pressure steam sterilization 10-30min after cooling
GUHP-86 carries out culture 18h ± 2h in 30-40 DEG C, 170-190r/min, obtains seed liquor；
(2) seed liquor described in 1000mL is centrifuged under 2-5 DEG C, 7000-9000r 10-20min, collects precipitating, and use 6-9g/L
Sterile saline wash the precipitating, the sterile saline of 1000mL is then added into the precipitating, shakes up, obtains
The bacillus amyloliquefaciens GUHP-86 preparation；
The bacillus amyloliquefaciens GUHP-86 is protected on January 4th, 2016 in China typical culture collection center preservation
Hiding number is CCTCC M 2016003.
2. a kind of method for improving protease and amylase activity in tobacco stem shred fermentation process, which is characterized in that make
It is handled with the bacillus amyloliquefaciens GUHP-86 preparation for the tobacco stem shred of fermentation process.
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| CN201910093125.6A CN109820239A (en) | 2019-01-30 | 2019-01-30 | A method of strengthening tobacco stem shred fermentation process |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910093125.6A CN109820239A (en) | 2019-01-30 | 2019-01-30 | A method of strengthening tobacco stem shred fermentation process |
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| CN109820239A true CN109820239A (en) | 2019-05-31 |
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Application publication date: 20190531 |