CN106119215A - A kind of fungus fermentation lacquer producing enzyme liquid culture medium and its preparation method and application - Google Patents
A kind of fungus fermentation lacquer producing enzyme liquid culture medium and its preparation method and application Download PDFInfo
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- C12N9/0055—Oxidoreductases (1.) acting on diphenols and related substances as donors (1.10)
- C12N9/0057—Oxidoreductases (1.) acting on diphenols and related substances as donors (1.10) with oxygen as acceptor (1.10.3)
- C12N9/0061—Laccase (1.10.3.2)
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- C12Y110/03—Oxidoreductases acting on diphenols and related substances as donors (1.10) with an oxygen as acceptor (1.10.3)
- C12Y110/03002—Laccase (1.10.3.2)
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Abstract
The invention discloses a kind of fungus fermentation lacquer producing enzyme liquid culture medium and its preparation method and application, fluid medium described in every L includes following component: shaddock peel powder 10～40g, wheatfeed 5～10g, one of bean cake, bean dregs or its mixture 3～10g, ammonium chloride 3～10g, copper sulphate pentahydrate 0.1～0.5g, one of glucose, maltose or its mixture 5～15g, sodium chloride 5～15g, potassium dihydrogen phosphate 1～2g, calcium chloride 0.3～0.8g.The fungus fermentation lacquer producing enzyme liquid culture medium of the present invention, with agricultural by-products primary raw materials such as Pericarpium Citri grandiss, can be obviously improved the laccase fermentation yield of multiple fungus；Culture medium has that preparation is simple, raw material is easy to get and lower-price characteristic, reduces the industrially prepared cost of laccase, and the comprehensive utilization for Pericarpium Citri grandis simultaneously provides a method that.
Description
Technical field
The invention belongs to technical field of bioengineering, be specifically related to a kind of fungus fermentation lacquer producing enzyme liquid culture medium and system thereof
Preparation Method and application.
Background technology
Laccase (EC 1.10.3.2) is a kind of blue blue multicopper oxidase, by Japanese scholars in 1883 first in a kind of Japan
Toxicodendron verniciflnum (Stokes) F. A. Barkley (Rhus verniciflua Stokes) finds, is a currently the only lignin-degrading enzymes realizing industrialized production and application.In addition to plant, research finds
Laccase exists among antibacterial, fungus and insecticide.Especially one class has the Laccase from White Rot Fungus of Putrefaction and produces trees
Measure the highest, be also current topmost laccase production strain simultaneously.Laccase is a kind of one-electron oxidation enzyme, it is possible to using oxygen as
Electron acceptor forms Oxidation to substrate while generating water.
Owing to laccase has relatively low substrate-function specificity, and multiple substrate is had Oxidation, the most study
Find that the substrate specificity of laccase reaches kind more than 200, mainly phenolic compound and derivant thereof or phenolic compound analog
And derivant.Owing to laccase has Degradation to multiple native compound or artificial-synthetic compound so that laccase is not only
Bioenergy, papermaking and textile industry are used widely research, simultaneously at dye wastewater treatment, biological restoration, disregard message
The numerous areas such as paper deinking, Novel Biosensor, biological fuel cell and food inspection also receive much concern.
The principal element that restriction laccase is commonly used at present is that the fermentation preparation cost of laccase is too high.Prepared by the fermentation of laccase
Cost is mainly determined by three factors, and one is the product enzyme level producing bacterial strain, and two is fermentation medium price, and three is fermentation week
Phase；And the composition of fermentation medium the most directly determines the cost of culture medium, simultaneously to producing enzyme level and the change of fermentation period
Change and also there is stronger impact.
Chinese patent CN 102719410 B provides a kind of culture medium prescription being exclusively used in laccase and preparation method thereof, should
The culture medium used in patent mainly contains the compositions such as sweet potato juice, mannose, yeast extract, peptone, casein and maltose, profit
With laccase activity the most about 0.34U/mL in 14 days after fermentation liquid of this culture medium fermentation culture, there is culture medium cost high, fermentation
Time is long, produces the feature that enzyme activity is low.
Chinese patent CN 102399704 B provides the growth of a kind of applicable white rot fungi and produces the fluid medium of laccase
And application, the culture medium used in this patent mainly contains wheat bran, a great number of elements, trace element and price costly
The compositions such as acid hydrolysis casein and vitamin B1, utilizing the laccase activity in this culture medium culturing 13d after fermentation liquid is 50U/mL,
There is culture medium and make complicated, relatively costly and that fermentation efficiency is low feature.
A kind of method that Chinese patent CN 102181410 B provides fermentative production of laccase, the cultivation used in this patent
Base mainly contains glucose, ammonium tartrate, a great number of elements, trace element and price 2,2-azino-two (3-second costly
Base-benzothiazole-6-sulfonic acid) composition such as di-ammonium salts and vitamin B1, reach 56.45U/ after utilizing the fermented 6d of peristome bacterium of the same colour
ML, has culture medium and makes complicated and relatively costly feature.
The fermentation medium of the main producing bacterial strain fungus of laccase usually contains substantial amounts of yeast extract, peptone, dimension at present
The expensive nutrient substance such as raw element, casein or its hydrolysate, and it is typically every several to tens units to produce enzyme level
Milliliter, and fermentation time is generally at about two weeks, even up to one month as long as, causes the fermentation preparation cost of laccase to occupy
Height is lower and becomes restriction laccase industrialized production and the Main Bottleneck problem of application.
China's Shaddock resource enriches, and is one of major country of production and the country of consumption of Fructus Citri grandis.Pericarpium Citri grandis accounts for whole Fructus Citri grandis weight
40%～50%, typically directly process as garbage after grapefruit flesh is processed or edible, cause the very big wave of resource
Take, and discarded Pericarpium Citri grandis due to mouldy, spoiled and to environment.In Pericarpium Citri grandis containing pectin, lignin, cellulose, polysaccharide,
Naringin, esters, phenols, flavonoid and several mineral materials, wherein lignin, naringin, phenols and flavonoid
The materials such as thing are also the substrate specificity of laccase, and therefore Pericarpium Citri grandis is a kind of preferably laccase fermentation raw material or inducer.
Summary of the invention
According to above the deficiencies in the prior art, the technical problem to be solved is to propose a kind of fungi fermentation to produce paint
Enzyme liquid culture medium and its preparation method and application, it is therefore an objective to reduce the industrially prepared cost of laccase, it is simple to the refuse profit of Pericarpium Citri grandis
With.
In order to solve above-mentioned technical problem, the technical solution used in the present invention is:
A kind of fungus fermentation lacquer producing enzyme liquid culture medium, fluid medium described in every L includes following component: shaddock peel powder 10～
40g, wheatfeed 5～10g, one of bean cake, bean dregs or its mixture 3～10g, ammonium chloride 3～10g, copper sulphate pentahydrate 0.1～
0.5g, one of glucose, maltose or its mixture 5～15g, sodium chloride 5～15g, potassium dihydrogen phosphate 1～2g, calcium chloride 0.3
～0.8g.
Described laccase produces strain and includes bolt bacterium (Trametes sp.) LS-10C, Trametes trogii (Trametes hirsute)
LS-7,2 years residual pore fungi (Abortiporus biennis) LS-4 or white rake teeth bacterium (Irpex lacteus) LS-6.Wherein, bolt
Bacterium LS-10C has been preserved in China typical culture collection center, and deposit number is CCTCC NO:M2015191, and preservation date is
On 04 03rd, 2015, preservation address was Wuhan, China Wuhan University.The relevant preservation information of bolt bacterium LS-10C is in application
Number it is 201510391964.8, the patent application of entitled bolt bacterium and application thereof is submitted to.
The preparation method of described fungus fermentation lacquer producing enzyme liquid culture medium, described preparation method comprises the steps,
A, the preparation of shaddock peel powder: collect Pericarpium Citri grandis drying, pulverize, shaddock peel powder of sieving to obtain, wherein, dried Pericarpium Citri grandis is aqueous
Amount is between 5%～10%；
B, the preparation of wheatfeed: size-reduced for wheat bran sieving is obtained；
C, the preparation of fermentation medium:
Component (1): one of glucose, maltose or its mixture 5～15g are dissolved in 100ml water, high temperature sterilize processes
After to be cooled to room temperature standby；
Component (2): be dissolved in 100ml water by potassium dihydrogen phosphate 1～2g, it is standby that high temperature sterilize is cooled to room temperature after processing；
Component (3): successively by sodium chloride 5～15g, calcium chloride 0.3～0.8g, ammonium chloride 3～10g and copper sulphate pentahydrate
0.1～0.5g is dissolved in 100ml water, and it is standby that high temperature sterilize is cooled to room temperature after processing；
Component (4): successively by shaddock peel powder 10～40g, wheatfeed 5～10g, one of bean cake, bean dregs or its mixture 3～
10g is dissolved in 700ml water, and it is standby that high temperature sterilize is cooled to room temperature after processing；
Component (1), component (2), component (3), component (4) being mixed in the ratio of 1:1:1:7, regulation pH value exists
Between 4.0～7.0, i.e. prepare fungus fermentation lacquer producing enzyme fermentation medium.
It is preferred that the temperature of high temperature sterilize is 105～110 DEG C in component (1), the temperature of high temperature sterilize in component (2)
Being 110～115 DEG C, in component (3), the temperature of high temperature sterilize is 115～121 DEG C, and in component (4), the temperature of high temperature sterilize is 121
～123 DEG C.
In step A, the baking temperature of Pericarpium Citri grandis is 50～60 DEG C, pulverizes 80～120 mesh sieve, the Fructus Citri grandis after the process of this condition
Corium farinosum has through the fungus fermentation lacquer producing enzyme fermentation medium that subsequent treatment obtains preferably produces laccase activity value.
Preferably, step B was pulverized 60～120 mesh sieve.
Described regulation pH value uses ammonia or dilute sulfuric acid regulation.
Described ammonia is the ammonia of mass fraction 25%～28%, and the concentration of described dilute sulfuric acid is 0.2mmol/L.
The application in utilizing agricultural wastes Pericarpium Citri grandis of the described fungus fermentation lacquer producing enzyme liquid culture medium.
The medicine have the advantages that
Culture medium the most of the present invention uses agricultural wastes Pericarpium Citri grandis as main culture matrix, reduce laccase industrially prepared
While cost, the comprehensive utilization for Pericarpium Citri grandis provides a method that；
Culture medium the most of the present invention uses the agricultural by-products such as wheat bran, bean dregs or bean cake as aid nutrition composition, and not
Containing expensive raw materials such as ABTS, casein or vitamin B1s, culture medium preparation is simple, raw material is easy to get, cheap；
Culture medium the most of the present invention can be obviously improved the laccase fermentation vigor of multiple fungus, has that fermentation time is short and laccase produces
Measure high feature, multiple fungus fermentation lacquer producing enzyme is had universality.
Accompanying drawing explanation
Below the content expressed by this specification accompanying drawing and the labelling in figure are briefly described:
Fig. 1 be embodiments of the invention 1 produce 4 kinds of fungal culture of laccase inoculation of medium produce after 9 days laccase activity signal
Figure；
Fig. 2 be embodiments of the invention 2 produce 4 kinds of fungal culture of laccase inoculation of medium produce after 9 days laccase activity signal
Figure；
Fig. 3 be embodiments of the invention 3 produce 4 kinds of fungal culture of laccase inoculation of medium produce after 9 days laccase activity signal
Figure.
Detailed description of the invention
Below against accompanying drawing, by the description to embodiment, the present invention is further detailed explanation, to help ability
Field technique personnel have more complete, accurate and deep understanding to inventive concept, the technical scheme of the present invention.
The preparation process of the fungus fermentation lacquer producing enzyme liquid culture medium of the present invention includes:
A, the preparation of shaddock peel powder
Collect fresh Pericarpium Citri grandis and be dried at 50～60 DEG C, making moisture content between 5%～10%, then use pulverizing
Dry Pericarpium Citri grandis is pulverized by machine, crosses 80～120 mesh sieve, i.e. obtain shaddock peel powder after pulverizing.
B, the preparation of wheatfeed
The wheat bran of fresh purchase uses pulverizer cross 60～120 mesh sieve after pulverizing further, i.e. obtains wheatfeed.
C, the preparation of fermentation medium
The preparation of each nutrition composition
Component (1): add one of glucose, maltose or its mixture 5～15g in 100mL tap water, stir and fill
Divide and dissolve, standby.
Component (2): add potassium dihydrogen phosphate 1～2g in 100mL tap water, stirring is also fully dissolved, standby.
Component (3): be sequentially added into sodium chloride 5～15g in 100mL tap water, calcium chloride 0.3～0.8g, ammonium chloride 3～
10g and copper sulphate pentahydrate 0.1～0.5g, stirring is also fully dissolved, standby.
Component (4): be sequentially added into shaddock peel powder 10～40g in 700mL tap water, wheatfeed 5～10g, bean cake, bean dregs it
One or its mixture 3～10g, standby after room temperature places 1～2h after stirring.
The sterilizing of each nutrition composition
High-pressure sterilizing pot is used said components (1) to be taken out after sterilizing 10～20min at 105～110 DEG C and be cooled to
Room temperature, standby.
High-pressure sterilizing pot is used said components (2) to be taken out after sterilizing 15～25min at 110～115 DEG C and be cooled to
Room temperature, standby.
High-pressure sterilizing pot is used said components (3) to be taken out after sterilizing 20～30min at 115～121 DEG C and be cooled to
Room temperature, standby.
High-pressure sterilizing pot is used said components (4) to be taken out after sterilizing 30～40min at 121～123 DEG C and be cooled to
Room temperature, standby.
The mixing of each component of culture medium
In superclean bench, component (1), component (2), component (3) and the component (4) after above-mentioned 4 kinds of sterilizings is pressed 1:1:
The ratio of 1:7 mixes, and uses ammonia (25%～28%) or dilute sulfuric acid (0.2mmol/L) regulation pH, make pH value after mixing
Between 4.0～7.0, i.e. prepare and produce laccase fermentation culture medium.
It is specifically described below by preferred embodiment:
Embodiment 1
By the material ratio in component (1), component (2), component (3) and component (4) needed for adjustment preparation fermentation medium
Example, in making every L fermentation medium contains shaddock peel powder 10g after the mixing of 1:1:1:7 ratio, wheatfeed 10g, bean cake 1g, bean dregs 2g,
Ammonium chloride 3g, copper sulphate pentahydrate 0.2g, glucose 10g, sodium chloride 5g, potassium dihydrogen phosphate 1g, calcium chloride 0.3g, regulation pH is extremely
After 5.0, aseptically subpackage 45mL culture medium, to 250mL triangular flask, is i.e. worth producing laccase fermentation culture medium respectively.
Shaddock peel powder is that fresh Pericarpium Citri grandis is the most size-reduced at 50 DEG C, crosses 100 mesh sieve and prepare, and its moisture content is 5%.
Wheatfeed is that wheat bran is size-reduced, crosses 80 mesh sieve and prepare.
Component (1) sterilising conditions is: 105 DEG C, 15min.
Component (2) sterilising conditions is: 115 DEG C, 20min.
Component (3) sterilising conditions is: 121 DEG C, 20min.
Component (4) sterilising conditions is: 121 DEG C, 30min.
It is seeded to described fermentation medium by 10% inoculum concentration and cultivates bolt bacterium (Trametes sp.) LS-10C, Trametes trogii
(Trametes hirsute) LS-7,2 years residual pore fungi (Abortiporus biennis) LS-4 or white rake teeth bacterium (Irpex
Lacteus) LS-6, the laccase activity cultivated in 9 days after fermentation liquid is up to 707.2U/mL (Fig. 1).
The assay method of laccase activity is: ABTS solution and enzyme liquid are placed in 40 DEG C of thermostat water baths preheating 5min respectively
After, draw 1.5mL enzyme liquid and add in 1.5mL substrate solution, the most fully mix and be separately recorded in 0 to 30 second, 30 to 60 seconds,
Reactant liquor change of light absorption value at 420nm in 60 seconds to 90 seconds, with the meansigma methods note of the light absorption value changing value that above-mentioned 3 times measure
Changing value for reaction system average absorbance value.
The definition of laccase activity is: under above-mentioned enzymatic reaction condition, required for definition 1min internal oxidition 1 μm ol ABTS
Enzyme amount be a laccase activity unit (U), and laccase fermentation vigor is expressed as U/mL.
Embodiment 2
By the material ratio in component (1), component (2), component (3) and component (4) needed for adjustment preparation fermentation medium
Example, makes after mixing in 1:1:1:7 ratio in every L fermentation medium containing shaddock peel powder 30g, wheatfeed 8g, bean cake 5g, ammonium chloride
5g, copper sulphate pentahydrate 0.3g, Portugal maltose 10g, sodium chloride 10g, potassium dihydrogen phosphate 1.5g, calcium chloride 0.5g, regulate pH to 4.0
After aseptically distinguish subpackage 45mL culture medium to 250mL triangular flask, i.e. be worth produce laccase fermentation culture medium.
Shaddock peel powder is that fresh Pericarpium Citri grandis is the most size-reduced at 50 DEG C, crosses 80 mesh sieve and prepare, and its moisture content is 10%.
Wheatfeed is that wheat bran is size-reduced, crosses 80 mesh sieve and prepare.
Component (1) sterilising conditions is: 105 DEG C, 15min.
Component (2) sterilising conditions is: 115 DEG C, 15min.
Component (3) sterilising conditions is: 121 DEG C, 20min.
Component (4) sterilising conditions is: 121 DEG C, 20min.
It is seeded to described product laccase fermentation culture medium culturing bolt bacterium (Trametes sp.) LS-10C, hair by 10% inoculum concentration
Bolt bacterium (Trametes hirsute) LS-7,2 years residual pore fungi (Abortiporus biennis) LS-4 or white rake teeth bacterium
(Irpex lacteus) LS-6, the laccase activity cultivated in 9 days after fermentation liquid is up to 875.9U/mL (Fig. 2).
Embodiment 3
By the material ratio in component (1), component (2), component (3) and component (4) needed for adjustment preparation fermentation medium
Example, makes after mixing in 1:1:1:7 ratio in every L fermentation medium containing shaddock peel powder 30g, wheatfeed 8g, bean cake 5g, ammonium chloride
5g, copper sulphate pentahydrate 0.3g, Portugal maltose 10g, sodium chloride 10g, potassium dihydrogen phosphate 1.5g, calcium chloride 0.5g, regulate pH to 7.0
After aseptically distinguish subpackage 45mL culture medium to 250mL triangular flask, i.e. be worth produce laccase fermentation culture medium.
Shaddock peel powder is that fresh Pericarpium Citri grandis is the most size-reduced at 60 DEG C, crosses 120 mesh sieve and prepare, and its moisture content is 5%.
Wheatfeed is that wheat bran is size-reduced, crosses 100 mesh sieve and prepare.
Component (1) sterilising conditions is: 105 DEG C, 20min.
Component (2) sterilising conditions is: 115 DEG C, 20min.
Component (3) sterilising conditions is: 121 DEG C, 20min.
Component (4) sterilising conditions is: 123 DEG C, 40min.
It is seeded to described product laccase fermentation culture medium culturing bolt bacterium (Trametes sp.) LS-10C, hair by 10% inoculum concentration
Bolt bacterium (Trametes hirsute) LS-7,2 years residual pore fungi (Abortiporus biennis) LS-4 or white rake teeth bacterium
(Irpex lacteus) LS-6, the laccase activity cultivated in 9 days after fermentation liquid is up to 645.7U/mL (Fig. 3).
Embodiment 4
By the material ratio in component (1), component (2), component (3) and component (4) needed for adjustment preparation fermentation medium
Example, in making every L fermentation medium contains shaddock peel powder 30g after the mixing of 1:1:1:7 ratio, wheatfeed 10g, bean cake 5g, bean dregs 5g,
Ammonium chloride 5g, copper sulphate pentahydrate 0.3g, glucose 10g, sodium chloride 5g, potassium dihydrogen phosphate 1g, calcium chloride 0.5g, regulation pH is extremely
After 5.0, aseptically subpackage 45mL culture medium, to 250mL triangular flask, is i.e. worth producing laccase fermentation culture medium respectively.
Shaddock peel powder is that fresh Pericarpium Citri grandis is the most size-reduced at 60 DEG C, crosses 120 mesh sieve and prepare, and its moisture content is 5%.
Wheatfeed is that wheat bran is size-reduced, crosses 120 mesh sieve and prepare.
Component (1) sterilising conditions is: 105 DEG C, 20min.
Component (2) sterilising conditions is: 110 DEG C, 20min.
Component (3) sterilising conditions is: 115 DEG C, 30min.
Component (4) sterilising conditions is: 123 DEG C, 40min.
It is seeded to described product laccase fermentation culture medium culturing bolt bacterium (Trametes sp.) LS-10C, hair by 10% inoculum concentration
Bolt bacterium (Trametes hirsute) LS-7,2 years residual pore fungi (Abortiporus biennis) LS-4 or white rake teeth bacterium
(Irpex lacteus) LS-6, cultivates the laccase activity in 9 days after fermentation liquid and is up to 1011.4U/mL (table 1).
It is respectively adopted PDA (every L contains 200g Rhizoma Solani tuber osi diffusion juice and 20g glucose) above-mentioned with liquid fermentation culture medium culturing
4 kinds of funguses, in the 9 days after fermentation liquid that ferments, laccase activity is up to only 8.4U/mL (table 1).
14 kinds of funguses of table utilize different culture media fermentation lacquer producing enzyme vigor (U/mL)
Above in conjunction with accompanying drawing, the present invention is exemplarily described, it is clear that the present invention implements not by aforesaid way
Restriction, as long as have employed the method design of the present invention and the improvement of various unsubstantialities that technical scheme is carried out, or without changing
Enter and design and the technical scheme of the present invention are directly applied to other occasion, all within protection scope of the present invention.This
Bright protection domain should be as the criterion with the protection domain that claims are limited.
Claims (8)
1. a fungus fermentation lacquer producing enzyme liquid culture medium, it is characterised in that fluid medium described in every L includes following component:
Shaddock peel powder 10～40g, wheatfeed 5～10g, one of bean cake, bean dregs or its mixture 3～10g, ammonium chloride 3～10g, five water sulfur
Acid copper 0.1～0.5g, one of glucose, maltose or its mixture 5～15g, sodium chloride 5～15g, potassium dihydrogen phosphate 1～2g,
Calcium chloride 0.3～0.8g.
The preparation method of fungus fermentation lacquer producing enzyme liquid culture medium the most according to claim 1, it is characterised in that described preparation
Method comprises the steps,
A, the preparation of shaddock peel powder: collect Pericarpium Citri grandis drying, pulverize, shaddock peel powder of sieving to obtain, wherein, dried Pericarpium Citri grandis water content exists
Between 5%～10%；
B, the preparation of wheatfeed: size-reduced for wheat bran sieving is obtained；
C, the preparation of fermentation medium:
Component (1): one of glucose, maltose or its mixture 5～15g are dissolved in 100ml water, high temperature sterilize is cold after processing
But standby to room temperature；
Component (2): be dissolved in 100ml water by potassium dihydrogen phosphate 1～2g, it is standby that high temperature sterilize is cooled to room temperature after processing；
Component (3): successively by sodium chloride 5～15g, calcium chloride 0.3～0.8g, ammonium chloride 3～10g and copper sulphate pentahydrate 0.1～
0.5g is dissolved in 100ml water, and it is standby that high temperature sterilize is cooled to room temperature after processing；
Component (4): successively by shaddock peel powder 10～40g, wheatfeed 5～10g, one of bean cake, bean dregs or its mixture 3～10g are molten
In 700ml water, it is standby that high temperature sterilize is cooled to room temperature after processing；
Component (1), component (2), component (3), component (4) are mixed in the ratio of 1:1:1:7, regulation pH value 4.0～
Between 7.0, i.e. prepare fungus fermentation lacquer producing enzyme fermentation medium.
The preparation method of fungus fermentation lacquer producing enzyme liquid culture medium the most according to claim 2, it is characterised in that component (1)
The temperature of middle high temperature sterilize is 105～110 DEG C, and in component (2), the temperature of high temperature sterilize is 110～115 DEG C, high in component (3)
The temperature of temperature sterilizing is 115～121 DEG C, and in component (4), the temperature of high temperature sterilize is 121～123 DEG C.
The preparation method of fungus fermentation lacquer producing enzyme liquid culture medium the most according to claim 2, it is characterised in that in step A
The baking temperature of Pericarpium Citri grandis is 50～60 DEG C, pulverizes 80～120 mesh sieve.
The preparation method of fungus fermentation lacquer producing enzyme liquid culture medium the most according to claim 2, it is characterised in that in step B
Pulverized 60～120 mesh sieve.
The preparation method of fungus fermentation lacquer producing enzyme liquid culture medium the most according to claim 2, it is characterised in that described regulation
PH value uses ammonia or dilute sulfuric acid regulation.
The preparation method of fungus fermentation lacquer producing enzyme liquid culture medium the most according to claim 6, it is characterised in that described ammonia
For the ammonia of mass fraction 25%～28%, the concentration of described dilute sulfuric acid is 0.2mmol/L.
8. fungus fermentation lacquer producing enzyme liquid culture medium application in utilizing agricultural wastes Pericarpium Citri grandis described in claim 1.
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| CN108265008A (en) * | 2018-02-12 | 2018-07-10 | 江南大学 | 2 years residual pore fungis of one plant of production laccase |
| CN112385835A (en) * | 2020-11-16 | 2021-02-23 | 浙江一鸣食品股份有限公司 | Bitter-taste-free tangerine enzyme and preparation method thereof |
| CN113564058A (en) * | 2021-09-08 | 2021-10-29 | 南昌大学 | Method for promoting production of laccase from Inonotus hirsutus by using food waste |
| CN116024101A (en) * | 2022-11-17 | 2023-04-28 | 中国科学院遗传与发育生物学研究所农业资源研究中心 | Food residue culture medium, and method and application for propagating lignocellulose degrading bacteria |
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| CN108265008A (en) * | 2018-02-12 | 2018-07-10 | 江南大学 | 2 years residual pore fungis of one plant of production laccase |
| CN112385835A (en) * | 2020-11-16 | 2021-02-23 | 浙江一鸣食品股份有限公司 | Bitter-taste-free tangerine enzyme and preparation method thereof |
| CN113564058A (en) * | 2021-09-08 | 2021-10-29 | 南昌大学 | Method for promoting production of laccase from Inonotus hirsutus by using food waste |
| CN113564058B (en) * | 2021-09-08 | 2024-02-09 | 南昌大学 | Method for promoting phellinus lanuginosus to produce laccase by using food waste |
| CN116024101A (en) * | 2022-11-17 | 2023-04-28 | 中国科学院遗传与发育生物学研究所农业资源研究中心 | Food residue culture medium, and method and application for propagating lignocellulose degrading bacteria |
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