CN107245482A - The method of mushroom ferment production cellulase and the application of institute's cellulase-producing - Google Patents
The method of mushroom ferment production cellulase and the application of institute's cellulase-producing Download PDFInfo
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- CN107245482A CN107245482A CN201710363342.3A CN201710363342A CN107245482A CN 107245482 A CN107245482 A CN 107245482A CN 201710363342 A CN201710363342 A CN 201710363342A CN 107245482 A CN107245482 A CN 107245482A
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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
- C12N13/00—Treatment of microorganisms or enzymes with electrical or wave energy, e.g. magnetism, sonic waves
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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
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
- C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
- C12N9/2405—Glucanases
- C12N9/2434—Glucanases acting on beta-1,4-glucosidic bonds
- C12N9/2437—Cellulases (3.2.1.4; 3.2.1.74; 3.2.1.91; 3.2.1.150)
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y302/00—Hydrolases acting on glycosyl compounds, i.e. glycosylases (3.2)
- C12Y302/01—Glycosidases, i.e. enzymes hydrolysing O- and S-glycosyl compounds (3.2.1)
- C12Y302/01004—Cellulase (3.2.1.4), i.e. endo-1,4-beta-glucanase
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Abstract
The invention discloses a kind of method that mushroom ferment produces cellulase, it is related to biological technical field, mushroom strain is inoculated with PDA culture medium and carries out 12 16 d of culture, 60 90 mL fluid nutrient mediums of addition are punched into after 10 mm bacterium piece with aseptic card punch, 24 32 DEG C are placed in, the h of fermented and cultured 96 192 under the conditions of 100 160 r/min shaking table.The configuration such as culture medium of the present invention and the component of feed supplement, concentration is reasonable, work in coordination, pass through rational component and concentration, expression i.e. to mushroom cellulose enzyme gene promotes, induction is carried out in the generation to mushroom cellulase again, will not also it is unreasonable because of component, concentration and caused by enzyme activity suppression, so as to realize the purpose of efficiently production cellulase.The present invention also provides the application of fibrous matter of the cellulase of mushroom ferment production in degraded draft or xylophyta, and a kind of new mode is provided for the fibrous matter in degraded draft or xylophyta.
Description
Technical field
Produce the method for cellulase the present invention relates to biological technical field, more particularly to a kind of mushroom ferment and produce fibre
The application of the plain enzyme of dimension.
Background technology
After cellulase is found in the alimentary canal of snail by scientist, cellulose the problem of microbial degradation on just
By the concern that World Science men are larger.The Natick laboratories of U.S. army then just have studied relevant microbial degradation and existed
Protection question in terms of military cellulosic material, finds cellulose in addition to it can be degraded by microorganisms in later research,
The solid waste problem constantly produced in the raw materials for production produced also very abundant economy, nature is also expected to obtain
Solve.After this, cellulase has obtained more extensive concern.Cellulose be distributed on earth it is most wide, be also content most
Abundant carbohydrate, and the 35%-50% of plant gross dry weight is cellulose.For the mankind, cellulose is even more nature
Key link essential in quantity the hugest recyclability material, the Carbon cycle mechanism of nature is exactly it in boundary
Degradation process.Make full use of cellulose short for solving worldwide significant problem such as world food at present with its conversion condition
The upper tool such as scarce, energy crisis, environmental pollution is of great significance.Cellulase is protected in food processing, Chemical Manufacture, medicine
There is boundless application prospect and ten in the field such as strong, brewing industry, Feed Manufacturing, processing of agriculture product, environmental protection
Divide deep potentiality.However, the enzymatic productivity that cellulase has producing strains is low, and produced enzyme component it is imperfect or
Uneven the problems such as, the mechanism of cellulase effect has not been studied the major reason that must be fully apparent from so far, is also actual
Using and research there is chief reason the most in larger distance.
Mushroom belongs to Tricholomataceae in Agaricales, is that one kind can be medicinal, and the class edible mushroom also liked by the common people turns into
Second largest edible mushroom in the world.Among the people, mushroom taste in culinary art is very delicious, fleshy hypertrophy, fragrance make people mentally refreshing heart spleen, nutrition
Content is extremely enriched again, is sung the praises of by people as " mountain delicacy ", more enjoys " plant queen " good reputation.At present, China is on Lentnus edodes
The yield of 1 year is occupied more than 80% ratio in the total annual production in the world, is in No. 1 in the world, export volume up to 80,000 tons
3.6 ten thousand tons are reached, the first place in the world is also occupy.In cultivating champignon, sawdust, stalk etc. it is this kind of rich in cellulose, hemicellulose,
The material of lignin is the main source of carbon source.Mushroom possess by these big molecular substances resolve into small-molecule substance so as to
The ectoenzyme being enough utilized, this explanation cellulase ties up in mushroom the content for having very abundant.Research mushroom produces polysaccharide at present
It is in the majority, and the research to mushroom ferment cellulase-producing is rarely found, if it will be pole that the blank of this respect, which cannot be filled up,
Big waste.
The content of the invention
Not enough for more than, it is an object of the invention to provide a kind of method that efficient mushroom ferment produces cellulase.
To reach above-mentioned purpose, the present invention is adopted the following technical scheme that：
A kind of method that mushroom ferment produces cellulase, mushroom strain is inoculated with PDA culture medium and carries out culture 12-16 d,
Addition 60-90 mL fluid nutrient mediums after 10 mm bacterium piece are punched into aseptic card punch, 24-32 DEG C, 100-160 r/ is placed in
Fermented and cultured 96-192 h under the conditions of min shaking table.
On the basis of above-mentioned technical proposal, the present invention also has further optimization and improved.
PDA culture medium in the present invention is potato dextrose agar.
Further, the preparation method of the PDA culture medium is：Take the g of potato 200, peeling, stripping and slicing add water 1000
ML, boils, filtered through gauze；It is separately added into the g of sucrose 20, agar 20 g, KH2PO4 3 g、MgSO41.5 g, vitamin B1
0.01 g, heating is dissolved；PH is natural；20 min that sterilize are placed in 121 DEG C of autoclaves, carry out Plate Procedure.
Further, the preparation method of the fluid nutrient medium is：Take the g of potato 200, peeling, stripping and slicing add water 1000
ML, boils, filtered through gauze；It is separately added into sucrose 20 g, KH2PO4 3 g、MgSO41.5 g, the g of vitamin B1 0.01, heating
Dissolve；PH is natural；20 min that sterilize are placed in 121 DEG C of autoclaves, loaded in conical flask.
Further, when adding the fluid nutrient medium, culture nitrogen source or culture carbon source are also added simultaneously；The culture
Nitrogen source is yeast extract, beef extract, maize flour, (NH4)2SO4In one kind, and the culture nitrogen source added amount be 4-6g/
L；The culture carbon source is one kind in sucrose, glucose, carboxymethyl cellulose, bagasse, starch, and the training added
The amount for supporting carbon source is 18-25g/L.
Further, the quantity of the bacterium piece is 3-5 pieces.
Further, when the mushroom strain is cultivated in the PDA culture medium, 15 W, 25 cm ultraviolet lamp bar are placed in
Mutagenic treatment is carried out under part, processing time is 3-8 min.
Further, the fermented incubation time is 144 h.
Further, the shaking speed is 120 r/min.
It is a further object to provide the cellulase of mushroom ferment production in degraded draft or xylophyta
Fibrous matter application.The cellulase that the method for mushroom ferment disclosed by the invention production cellulase is produced can be with
The fibrous matter in degraded draft or xylophyta is efficiently applied to, the degraded of draft or the fibrous matter in xylophyta is improved
Efficiency.
Compared with prior art, the beneficial effects of the invention are as follows：The present invention provides a kind of mushroom ferment production cellulase
Method, by new culture medium and the component of feed supplement, concentration etc., to improve the level that mushroom ferment produces cellulase,
So as to improve the yield of cellulase.The configuration such as new culture medium and the component of feed supplement, concentration is reasonable, work in coordination, and passes through
Rational component and concentration, the i.e. expression to mushroom cellulose enzyme gene promote, and the generation implementation of mushroom cellulase is lured
Lead, will not also it is unreasonable because of component, concentration and caused by enzyme activity suppression, so as to realize the purpose of efficiently production cellulase.This
Invention also provides the application of fibrous matter of the cellulase of mushroom ferment production in degraded draft or xylophyta, for degraded
Fibrous matter in draft or xylophyta provides a kind of new mode.
Embodiment
The technical scheme in the embodiment of the present invention will be clearly and completely described below.Obviously, described implementation
Example only a part of embodiment of the invention, rather than whole embodiments.Based on the embodiment in the present invention, this area is common
The every other embodiment that technical staff is obtained under the premise of creative work is not made, belongs to the model that the present invention is protected
Enclose.
Embodiment 1
The present embodiment provides a kind of method that mushroom ferment produces cellulase, and mushroom strain is inoculated with PDA culture medium and is carried out
14 d are cultivated, are added to after the bacterium piece that 10 mm are punched into aseptic card punch in 75 mL fluid nutrient mediums, bacterium piece is 4, is put
The h of fermented and cultured 144 under the conditions of 28 DEG C, 120 r/min shaking table.
Wherein, the preparation method of PDA culture medium is：Take the g of potato 200, peeling, stripping and slicing, add water 1000 mL, boils,
Filtered through gauze；It is separately added into the g of sucrose 20, agar 20 g, KH2PO4 3 g、MgSO41.5 g, the g of vitamin B1 0.01, heating
Dissolve；PH is natural；20 min that sterilize are placed in 121 DEG C of autoclaves, carry out Plate Procedure.
Wherein, the preparation method of fluid nutrient medium is：Take the g of potato 200, peeling, stripping and slicing, add water 1000 mL, boils,
Filtered through gauze；It is separately added into sucrose 20 g, KH2PO4 3 g、MgSO41.5 g, the g of vitamin B1 0.01, heating are dissolved；PH is certainly
So；20 min that sterilize are placed in 121 DEG C of autoclaves, loaded in conical flask.
Embodiment 2
The present embodiment provides a kind of method that mushroom ferment produces cellulase, and mushroom strain is inoculated with PDA culture medium and is carried out
12 d are cultivated, are added to after the bacterium piece that 10 mm are punched into aseptic card punch in 60 mL fluid nutrient mediums, bacterium piece is 3, is put
The h of fermented and cultured 96 under the conditions of 24 DEG C, 100 r/min shaking table.
When adding fluid nutrient medium, culture carbon source is also added simultaneously, and culture carbon source is sucrose, and the amount of the sucrose of addition is
18g/L。
When mushroom strain is cultivated in PDA culture medium, it is placed under the conditions of 15 W, 25 cm uviol lamp and carries out mutagenic treatment,
Processing time is 3 min.
Wherein, the preparation method of PDA culture medium is：Take the g of potato 200, peeling, stripping and slicing, add water 1000 mL, boils,
Filtered through gauze；It is separately added into the g of sucrose 20, agar 20 g, KH2PO4 3 g、MgSO41.5 g, the g of vitamin B1 0.01, heating
Dissolve；PH is natural；20 min that sterilize are placed in 121 DEG C of autoclaves, carry out Plate Procedure.
Wherein, the preparation method of fluid nutrient medium is：Take the g of potato 200, peeling, stripping and slicing, add water 1000 mL, boils,
Filtered through gauze；It is separately added into sucrose 20 g, KH2PO4 3 g、MgSO41.5 g, the g of vitamin B1 0.01, heating are dissolved；PH is certainly
So；20 min that sterilize are placed in 121 DEG C of autoclaves, loaded in conical flask.
Embodiment 3
The present embodiment provides a kind of method that mushroom ferment produces cellulase, and mushroom strain is inoculated with PDA culture medium and is carried out
16 d are cultivated, are added to after the bacterium piece that 10 mm are punched into aseptic card punch in 90 mL fluid nutrient mediums, bacterium piece is 5, is put
The h of fermented and cultured 192 under the conditions of 32 DEG C, 160 r/min shaking table.
When adding fluid nutrient medium, culture carbon source is also added simultaneously, and culture carbon source is glucose, the glucose of addition
Measure as 25g/L.
When mushroom strain is cultivated in PDA culture medium, it is placed under the conditions of 15 W, 25 cm uviol lamp and carries out mutagenic treatment,
Processing time is 8 min.
Wherein, the preparation method of PDA culture medium is：Take the g of potato 200, peeling, stripping and slicing, add water 1000 mL, boils,
Filtered through gauze；It is separately added into the g of sucrose 20, agar 20 g, KH2PO4 3 g、MgSO41.5 g, the g of vitamin B1 0.01, heating
Dissolve；PH is natural；20 min that sterilize are placed in 121 DEG C of autoclaves, carry out Plate Procedure.
Wherein, the preparation method of fluid nutrient medium is：Take the g of potato 200, peeling, stripping and slicing, add water 1000 mL, boils,
Filtered through gauze；It is separately added into sucrose 20 g, KH2PO4 3 g、MgSO41.5 g, the g of vitamin B1 0.01, heating are dissolved；PH is certainly
So；20 min that sterilize are placed in 121 DEG C of autoclaves, loaded in conical flask.
Embodiment 4
The present embodiment provides a kind of method that mushroom ferment produces cellulase, and mushroom strain is inoculated with PDA culture medium and is carried out
15 d are cultivated, are added to after the bacterium piece that 10 mm are punched into aseptic card punch in 75 mL fluid nutrient mediums, bacterium piece is 4, is put
The h of fermented and cultured 144 under the conditions of 28 DEG C, 120 r/min shaking table.
When adding fluid nutrient medium, culture carbon source is also added simultaneously, and culture carbon source is carboxymethyl cellulose, the carboxylic of addition
The amount of methylcellulose is 20g/L.
When mushroom strain is cultivated in PDA culture medium, it is placed under the conditions of 15 W, 25 cm uviol lamp and carries out mutagenic treatment,
Processing time is 5 min.
Wherein, the preparation method of PDA culture medium is：Take the g of potato 200, peeling, stripping and slicing, add water 1000 mL, boils,
Filtered through gauze；It is separately added into the g of sucrose 20, agar 20 g, KH2PO4 3 g、MgSO41.5 g, the g of vitamin B1 0.01, heating
Dissolve；PH is natural；20 min that sterilize are placed in 121 DEG C of autoclaves, carry out Plate Procedure.
Wherein, the preparation method of fluid nutrient medium is：Take the g of potato 200, peeling, stripping and slicing, add water 1000 mL, boils,
Filtered through gauze；It is separately added into sucrose 20 g, KH2PO4 3 g、MgSO41.5 g, the g of vitamin B1 0.01, heating are dissolved；PH is certainly
So；20 min that sterilize are placed in 121 DEG C of autoclaves, loaded in conical flask.
Embodiment 5
The present embodiment provides a kind of method that mushroom ferment produces cellulase, and mushroom strain is inoculated with PDA culture medium and is carried out
14 d are cultivated, are added to after the bacterium piece that 10 mm are punched into aseptic card punch in 60 mL fluid nutrient mediums, bacterium piece is 3, is put
The h of fermented and cultured 96 under the conditions of 24 DEG C, 100 r/min shaking table.
When adding fluid nutrient medium, culture nitrogen source is also added simultaneously, and culture nitrogen source is yeast extract, the yeast extract of addition
Measure as 18g/L.
When mushroom strain is cultivated in PDA culture medium, it is placed under the conditions of 15 W, 25 cm uviol lamp and carries out mutagenic treatment,
Processing time is 3 min.
Wherein, the preparation method of PDA culture medium is：Take the g of potato 200, peeling, stripping and slicing, add water 1000 mL, boils,
Filtered through gauze；It is separately added into the g of sucrose 20, agar 20 g, KH2PO4 3 g、MgSO41.5 g, the g of vitamin B1 0.01, heating
Dissolve；PH is natural；20 min that sterilize are placed in 121 DEG C of autoclaves, carry out Plate Procedure.
Wherein, the preparation method of fluid nutrient medium is：Take the g of potato 200, peeling, stripping and slicing, add water 1000 mL, boils,
Filtered through gauze；It is separately added into sucrose 20 g, KH2PO4 3 g、MgSO41.5 g, the g of vitamin B1 0.01, heating are dissolved；PH is certainly
So；20 min that sterilize are placed in 121 DEG C of autoclaves, loaded in conical flask.
Embodiment 6
The present embodiment provides a kind of method that mushroom ferment produces cellulase, and mushroom strain is inoculated with PDA culture medium and is carried out
12 d are cultivated, are added to after the bacterium piece that 10 mm are punched into aseptic card punch in 90 mL fluid nutrient mediums, bacterium piece is 5, is put
The h of fermented and cultured 192 under the conditions of 32 DEG C, 150 r/min shaking table.
When adding fluid nutrient medium, culture nitrogen source is also added simultaneously, and culture nitrogen source is beef extract, the beef extract of addition
Measure as 25g/L.
When mushroom strain is cultivated in PDA culture medium, it is placed under the conditions of 15 W, 25 cm uviol lamp and carries out mutagenic treatment,
Processing time is 8 min.
Wherein, the preparation method of PDA culture medium is：Take the g of potato 200, peeling, stripping and slicing, add water 1000 mL, boils,
Filtered through gauze；It is separately added into the g of sucrose 20, agar 20 g, KH2PO4 3 g、MgSO41.5 g, the g of vitamin B1 0.01, heating
Dissolve；PH is natural；20 min that sterilize are placed in 121 DEG C of autoclaves, carry out Plate Procedure.
Wherein, the preparation method of fluid nutrient medium is：Take the g of potato 200, peeling, stripping and slicing, add water 1000 mL, boils,
Filtered through gauze；It is separately added into sucrose 20 g, KH2PO4 3 g、MgSO41.5 g, the g of vitamin B1 0.01, heating are dissolved；PH is certainly
So；20 min that sterilize are placed in 121 DEG C of autoclaves, loaded in conical flask.
Embodiment 7
The present embodiment provides a kind of method that mushroom ferment produces cellulase, and mushroom strain is inoculated with PDA culture medium and is carried out
15 d are cultivated, are added to after the bacterium piece that 10 mm are punched into aseptic card punch in 75 mL fluid nutrient mediums, bacterium piece is 4, is put
The h of fermented and cultured 144 under the conditions of 28 DEG C, 120 r/min shaking table.
When adding fluid nutrient medium, culture nitrogen source is also added simultaneously, and culture nitrogen source is maize flour, the maize flour of addition
Measure as 20g/L.
When mushroom strain is cultivated in PDA culture medium, it is placed under the conditions of 15 W, 25 cm uviol lamp and carries out mutagenic treatment,
Processing time is 5 min.
Wherein, the preparation method of PDA culture medium is：Take the g of potato 200, peeling, stripping and slicing, add water 1000 mL, boils,
Filtered through gauze；It is separately added into the g of sucrose 20, agar 20 g, KH2PO4 3 g、MgSO41.5 g, the g of vitamin B1 0.01, heating
Dissolve；PH is natural；20 min that sterilize are placed in 121 DEG C of autoclaves, carry out Plate Procedure.
Wherein, the preparation method of fluid nutrient medium is：Take the g of potato 200, peeling, stripping and slicing, add water 1000 mL, boils,
Filtered through gauze；It is separately added into sucrose 20 g, KH2PO4 3 g、MgSO41.5 g, the g of vitamin B1 0.01, heating are dissolved；PH is certainly
So；20 min that sterilize are placed in 121 DEG C of autoclaves, loaded in conical flask.
After mushroom ferment culture cellulase has been carried out, producing enzyme efficiency is measured.
Zymotic fluid is obtained after mushroom ferment culture cellulase, zymotic fluid is through 4 layers of filtered through gauze, then by centrifuge 3000
R/min rotating speeds are centrifuged after 15 min, are taken the supernatant of solution, are crude enzyme liquid, now crude enzyme liquid is cellulase solution.Take from
The crude enzyme liquid of the heart is placed in boiling water bath and persistently inactivates 10 min, stand-by.
0.5 mL will be taken after crude enzyme liquid after inactivation suitably dilution, add 2.0 mL substrates（Substrate refers to 0.6g carboxylic first
Base sodium cellulosate is dissolved in the gains of the citric acid solution of 100 mL, pH values 4. 8, and vibration is mixed, in 60 DEG C of conditions
Under, persistently it is incubated after 30 min, after 3, the 5- dinitrosalicylic acid solutions for adding 2.5 mL, puts in boiling water bath after 5 min, horse
It is upper to use originally water cooling.540 nm light absorption value is measured on iMark ELIASAs, the standard curve of glucose is compareed, calculates
The vigor of enzyme.
Technical scheme to embodiment 1 ~ 7 obtains cellulase solution progress enzyme activity calculating, and result of calculation see the table below.
| Scheme | Enzyme activity（mg/mL） |
| Embodiment 1 | 0.23 |
| Embodiment 2 | 0.48 |
| Embodiment 3 | 0.35 |
| Embodiment 4 | 0.33 |
| Embodiment 5 | 0.39 |
| Embodiment 6 | 0.41 |
| Embodiment 7 | 0.45 |
As can be seen from the above table, scheme provided in an embodiment of the present invention has good enzymatic productivity, especially adds
Cultivate carbon source or culture nitrogen source and introduce after ultraviolet source, enzyme activity is obviously improved, and the ability of mushroom cellulase-producing is entered
One step is strengthened, and producing enzyme efficiency greatly improves, the method for illustrating the mushroom ferment production cellulase that the present invention is provided it is simple and
Efficiently, the purpose of the mushroom cellulase-producing of low input high production can be realized.
The foregoing is only a specific embodiment of the invention, but protection scope of the present invention is not limited thereto, any
Those familiar with the art the invention discloses technical scope in, the change or replacement that can be readily occurred in, all should
It is included within the scope of the present invention.Therefore, protection scope of the present invention should using the scope of the claims as
It is accurate.
Claims (9)
1. a kind of method that mushroom ferment produces cellulase, it is characterised in that be inoculated with mushroom strain in PDA culture medium and carry out
12-16 d are cultivated, addition 60-90 mL fluid nutrient mediums after 10 mm bacterium piece are punched into aseptic card punch, 24-32 is placed in
DEG C, fermented and cultured 96-192 h under the conditions of 100-160 r/min shaking table.
2. the method that mushroom ferment according to claim 1 produces cellulase, it is characterised in that：The PDA culture medium
Preparation method be：Take the g of potato 200, peeling, stripping and slicing, add water 1000 mL, boils, filtered through gauze；It is separately added into sucrose 20
G, agar 20 g, KH2PO4 3 g、MgSO41.5 g, the g of vitamin B1 0.01, heating are dissolved；PH is natural；121 DEG C are placed in go out
Sterilize 20 min in bacterium pot, carries out Plate Procedure.
3. the method that mushroom ferment according to claim 1 produces cellulase, it is characterised in that：The fluid nutrient medium
Preparation method be：Take the g of potato 200, peeling, stripping and slicing, add water 1000 mL, boils, filtered through gauze；It is separately added into sucrose 20
g、KH2PO4 3 g、MgSO41.5 g, the g of vitamin B1 0.01, heating are dissolved；PH is natural；It is placed in 121 DEG C of autoclaves and goes out
The min of bacterium 20, loaded in conical flask.
4. the method that mushroom ferment according to claim 1 produces cellulase, it is characterised in that：Adding the liquid
During culture medium, culture nitrogen source or culture carbon source are also added simultaneously；It is described culture nitrogen source be yeast extract, beef extract, maize flour,
(NH4)2SO4In one kind, and the culture nitrogen source added amount be 4-6g/L；The culture carbon source is sucrose, grape
One kind in sugar, carboxymethyl cellulose, bagasse, starch, and the amount of the culture carbon source added is 18-25g/L.
5. the method that mushroom ferment according to claim 1 produces cellulase, it is characterised in that：The quantity of the bacterium piece
For 3-5 pieces.
6. the method that mushroom ferment according to claim 1 produces cellulase, it is characterised in that：The mushroom strain exists
When being cultivated in the PDA culture medium, it is placed under the conditions of 15 W, 25 cm uviol lamp and carries out mutagenic treatment, processing time is 3-8
min。
7. the method that mushroom ferment according to claim 1 produces cellulase, it is characterised in that：During the fermented and cultured
Between be 144 h.
8. the method that mushroom ferment according to claim 1 produces cellulase, it is characterised in that：The shaking speed is
120 r/min。
9. fibre object of the cellulase of any one of claim 1-8 mushroom ferment production in degraded draft or xylophyta
The application of matter.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113662894A (en) * | 2021-08-25 | 2021-11-19 | 云南雅赫生物科技有限公司 | Asiatic pennywort herb enzymatic hydrolysate and preparation method and application thereof |
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2017
- 2017-05-22 CN CN201710363342.3A patent/CN107245482A/en active Pending
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113662894A (en) * | 2021-08-25 | 2021-11-19 | 云南雅赫生物科技有限公司 | Asiatic pennywort herb enzymatic hydrolysate and preparation method and application thereof |
| CN113662894B (en) * | 2021-08-25 | 2023-08-22 | 云南雅赫生物科技有限公司 | Centella enzymolysis fermentation product and preparation method and application thereof |
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