CN109837263A - The culture medium of mix bacterium agent and preparation method thereof for cellulase-producing - Google Patents
The culture medium of mix bacterium agent and preparation method thereof for cellulase-producing Download PDFInfo
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- CN109837263A CN109837263A CN201910213724.7A CN201910213724A CN109837263A CN 109837263 A CN109837263 A CN 109837263A CN 201910213724 A CN201910213724 A CN 201910213724A CN 109837263 A CN109837263 A CN 109837263A
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Abstract
The culture medium of mix bacterium agent and preparation method thereof that the invention discloses a kind of for cellulase-producing, the preparation method include: 1) to mix mud with water, then filter to take filtrate, and filtrate sterilizing, concentration are then removed water so that sludge extract is made；2) straw powder, rice bran, maize cob meal, dregs of beans and water are mixed to get mixture, then mixture is subjected to high pressure sealing processing, sucrose, peptone, enuatrol, pH adjusting agent, mashed potatoes, grape juice, sludge extract is then added into system and is mixed so that the culture medium of the mix bacterium agent for cellulase-producing is made；Wherein, the pH of culture medium is 5.8-6.4.The culture medium for being used for the mix bacterium agent of cellulase-producing can fully be decomposed by the mix bacterium agent of cellulase-producing, while the preparation method has the characteristics that process is simply and easy to operate.
Description
Technical field
The present invention relates to culture mediums, and in particular, to a kind of culture medium of the mix bacterium agent for cellulase-producing and its
Preparation method.
Background technique
Main polysaccharide class product of the cellulose as photosynthesis of plant, is the main component of higher plant cell wall, is
Generally acknowledged nature most abundant, most cheap renewable organic substance resource.According to according to a preliminary estimate, the annual crops straw in China
Cellulose total amount is up to 200,000,000 tons or so in stalk (straw, corn stalk, wheat straw etc.).Cellulose decomposition can be Portugal by cellulase
Grape sugar.The enzyme is of great significance in terms of the crises such as the energy, grain, the environmental pollution that solution our times faces.However, so far
Cellulase activity and yield are lower until the present, the production cycle is long, at high cost, all seriously hinder its industrialized application.It is fine
The degradation of dimension element depends on cellulase.Many microorganisms can produce cellulase in nature, and the degradation of cellulose is certainly
The key link of right boundary Carbon cycle.
But it is still very limited to the utilization of cellulose due to the design feature of cellulose, only have 20% fiber at present
Plain substance is exploited, and a large amount of cellulosic material is discarded due to utilization because that can not decompose, and not only results in waste of resources, but also dirty
Contaminate environment.
Summary of the invention
The culture medium of mix bacterium agent and preparation method thereof that the object of the present invention is to provide a kind of for cellulase-producing, should
The culture medium of mix bacterium agent for cellulase-producing can fully be decomposed by the mix bacterium agent of cellulase-producing, while the system
Preparation Method has the characteristics that process is simply and easy to operate.
To achieve the goals above, the present invention provides a kind of systems of the culture medium of mix bacterium agent for cellulase-producing
Preparation Method, comprising:
1) mud is mixed with water, then filters to take filtrate, then go water to extract so that mud is made filtrate sterilizing, concentration
Object；
2) straw powder, rice bran, maize cob meal, dregs of beans and water are mixed to get mixture, mixture is then subjected to high pressure
Then encapsulation process adds sucrose, peptone, enuatrol, pH adjusting agent, mashed potatoes, grape juice, sludge extract into system
It mixes so that the culture medium of the mix bacterium agent for cellulase-producing is made；
Wherein, the pH of culture medium is 5.8-6.4.
The present invention also provides a kind of culture mediums of mix bacterium agent for cellulase-producing, this is used for cellulase-producing
The culture medium of mix bacterium agent is prepared by above-mentioned preparation method.
Through the above technical solutions, sludge extract is made in the present invention first in mud, straw powder, rice are then selected
Chaff, maize cob meal, dregs of beans select sucrose, peptone, enuatrol, pH adjusting agent, mashed potatoes, grape juice, mud to mention as major ingredient
Take object that culture medium is made as auxiliary material, fibre is made in the abundant decomposition which can be conducive to the mix bacterium agent of cellulase-producing
Tie up plain enzyme.
Other features and advantages of the present invention will the following detailed description will be given in the detailed implementation section.
Specific embodiment
Detailed description of the preferred embodiments below.It should be understood that described herein specific
Embodiment is merely to illustrate and explain the present invention, and is not intended to restrict the invention.
The endpoint of disclosed range and any value are not limited to the accurate range or value herein, these ranges or
Value should be understood as comprising the value close to these ranges or value.For numberical range, between the endpoint value of each range, respectively
It can be combined with each other between the endpoint value of a range and individual point value, and individually between point value and obtain one or more
New numberical range, these numberical ranges should be considered as specific open herein.
The present invention provides a kind of preparation methods of the culture medium of mix bacterium agent for cellulase-producing, comprising:
1) mud is mixed with water, then filters to take filtrate, then go water to extract so that mud is made filtrate sterilizing, concentration
Object；
2) straw powder, rice bran, maize cob meal, dregs of beans and water are mixed to get mixture, mixture is then subjected to high pressure
Then encapsulation process adds sucrose, peptone, enuatrol, pH adjusting agent, mashed potatoes, grape juice, sludge extract into system
It mixes so that the culture medium of the mix bacterium agent for cellulase-producing is made；
Wherein, the pH of culture medium is 5.8-6.4.
In step 1) of the invention, the dosage of each material can select in a wide range, but be to be able to higher
Efficient extract sludge extract, it is preferable that in step 1), the weight ratio of mud and water is 1:8-10.
In step 1) of the invention, the mode of sterilizing and concentration can select in a wide range, but in order to
Extract sludge extract to enough higher efficiencies, it is preferable that in step 1), sterilizing is carried out by high-pressure sterilizing pot；Concentration is logical
Cross the progress of rotation concentrating instrument.
In step 2) of the invention, the dosage of each material can select in a wide range, but in order to make culture medium
More cellulase can be decomposited, it is preferable that in step 2), water, straw powder, rice bran, maize cob meal, dregs of beans, sucrose,
Peptone, enuatrol, mashed potatoes, grape juice, sludge extract weight ratio be 100:8-12:3-4:5-8:5-9:1.5-2.5:
0.5-0.9:0.1-0.3:0.4-0.6:0.3-0.6:1-3.
In step 2) of the invention, the type of straw powder can select in a wide range, but in order to make culture medium
More cellulase can be decomposited, it is preferable that in step 2), straw powder is selected from milled powders of cornstalk, kaoliang stalk powder, Wheat Straw
At least one of stalk powder, rice straw powder.
In step 2) of the invention, the type of pH adjusting agent can select in a wide range, but in order to from cost
Upper consideration, it is preferable that in step 2), pH adjusting agent is selected from oxalic acid, acetic acid, citric acid, sodium carbonate, sodium bicarbonate, bicarbonate
At least one of potassium.
In step 2) of the invention, the mode of high pressure sealing processing can select in a wide range, but in order to make
Culture medium can decomposite more cellulase, it is preferable that in step 2), high pressure sealing processing are as follows: in high-pressure sterilizing pot
Middle processing 15-25min.
In the present invention, in order to enable culture medium to decomposite more cellulase, it is preferable that after step 2),
The preparation method further includes that oxygen is passed through into system；It is highly preferred that the ventilatory capacity of oxygen is 0.4- after step 2)
0.6vvm。
In the present invention, the specific type of mix bacterium agent can select in a wide range, but in order to enable culture medium
Enough decomposite more cellulase, it is preferable that mix bacterium agent is by koning trichoderma bacterium, aspergillus niger, Aspergillus glaucus, aspergillus niger, cigarette
At least the two composition in aspergillus.
The present invention also provides a kind of culture mediums of mix bacterium agent for cellulase-producing, this is used for cellulase-producing
The culture medium of mix bacterium agent is prepared by above-mentioned preparation method.
The present invention will be described in detail by way of examples below.In the examples below that, pH adjusting agent be selected from oxalic acid,
At least one of acetic acid, citric acid, sodium carbonate, sodium bicarbonate, saleratus.
Embodiment 1
1) mud is mixed with water according to the weight ratio of 1:9, filtrate is then filtered to take, then by filtrate in high-pressure sterilizing pot
Middle sterilizing removes water by the concentration of spin concentration instrument sludge extract is made；
2) by straw powder (milled powders of cornstalk, kaoliang stalk powder, wheat stalk powder, rice straw powder), rice bran, maize cob meal, beans
The dregs of rice and water are mixed to get mixture, then mixture are handled 20min in high-pressure sterilizing pot, sugarcane is then added into system
Sugar, peptone, enuatrol, pH adjusting agent, mashed potatoes, grape juice, sludge extract mix (water, straw powder, rice bran, corncob
Powder, dregs of beans, sucrose, peptone, enuatrol, mashed potatoes, grape juice, sludge extract weight ratio be 100:10:3.5:7:7:
2:0.8:0.2:0.5:0.4:2) the culture medium A 1 that pH is 6.0 is made.
Embodiment 2
1) mud is mixed with water according to the weight ratio of 1:8, filtrate is then filtered to take, then by filtrate in high-pressure sterilizing pot
Middle sterilizing removes water by the concentration of spin concentration instrument sludge extract is made；
2) by straw powder (milled powders of cornstalk, kaoliang stalk powder, wheat stalk powder, rice straw powder), rice bran, maize cob meal, beans
The dregs of rice and water are mixed to get mixture, then mixture are handled 15min in high-pressure sterilizing pot, sugarcane is then added into system
Sugar, peptone, enuatrol, pH adjusting agent, mashed potatoes, grape juice, sludge extract mix (water, straw powder, rice bran, corncob
Powder, dregs of beans, sucrose, peptone, enuatrol, mashed potatoes, grape juice, sludge extract weight ratio be 100:8:3:5:5:1.5:
0.5:0.1:0.4:0.3:1) the culture medium A 2 that pH is 5.8 is made.
Embodiment 3
1) mud is mixed with water according to the weight ratio of 1:10, filtrate is then filtered to take, then by filtrate in high pressure sterilization
It sterilizes in pot, water is gone so that sludge extract is made by the concentration of spin concentration instrument；
2) by straw powder (milled powders of cornstalk, kaoliang stalk powder, wheat stalk powder, rice straw powder), rice bran, maize cob meal, beans
The dregs of rice and water are mixed to get mixture, then mixture are handled 25min in high-pressure sterilizing pot, sugarcane is then added into system
Sugar, peptone, enuatrol, pH adjusting agent, mashed potatoes, grape juice, sludge extract mix (water, straw powder, rice bran, corncob
Powder, dregs of beans, sucrose, peptone, enuatrol, mashed potatoes, grape juice, sludge extract weight ratio be 100:12:4:8:9:
2.5:0.9:0.3:0.6:0.6:3) the culture medium A 3 that pH is 6.4 is made.
Comparative example 1
Culture medium A 4 is prepared Following the procedure of Example 1, the difference is that sludge extract is not used in step 2).
Application examples 1
Mix bacterium agent (including koning trichoderma bacterium and aspergillus niger) is inoculated in above-mentioned culture medium, inoculum concentration 5%；Then
Oxygen, ventilatory capacity 0.5vvm, stirring rate 360r/min are passed through into above-mentioned culture medium；65h is cultivated at 30 DEG C, finally
It is living to detect culture medium cellulase, concrete outcome is shown in Table 1.
Table 1
| Cellulase activity (U/g) | |
| A1 | 509.0 |
| A2 | 515.3 |
| A3 | 512.6 |
| A4 | 438.8 |
The preferred embodiment of the present invention has been described above in detail, still, during present invention is not limited to the embodiments described above
Detail within the scope of the technical concept of the present invention can be with various simple variants of the technical solution of the present invention are made, this
A little simple variants all belong to the scope of protection of the present invention.
It is further to note that specific technical features described in the above specific embodiments, in not lance
In the case where shield, can be combined in any appropriate way, in order to avoid unnecessary repetition, the present invention to it is various can
No further explanation will be given for the combination of energy.
In addition, various embodiments of the present invention can be combined randomly, as long as it is without prejudice to originally
The thought of invention, it should also be regarded as the disclosure of the present invention.
Claims (10)
1. a kind of preparation method of the culture medium of the mix bacterium agent for cellulase-producing characterized by comprising
1) mud is mixed with water, then filters to take filtrate, water then is removed into filtrate sterilizing, concentration so that the mud is made and extracts
Object；
2) straw powder, rice bran, maize cob meal, dregs of beans and water are mixed to get mixture, the mixture is then subjected to high pressure
Then encapsulation process is added sucrose, peptone, enuatrol, pH adjusting agent, mashed potatoes, grape juice, the mud into system and is mentioned
Object is taken to mix so that the culture medium of the mix bacterium agent for cellulase-producing is made；
Wherein, the pH of the culture medium is 5.8-6.4.
2. preparation method according to claim 1, wherein in step 1), the weight ratio of the mud and water is 1:8-
10。
3. preparation method according to claim 1, wherein in step 1), the sterilizing is carried out by high-pressure sterilizing pot；
It is described to be concentrated by the progress of spin concentration instrument.
4. preparation method according to claim 1, wherein in step 2), the water, straw powder, rice bran, corncob
Powder, dregs of beans, sucrose, peptone, enuatrol, mashed potatoes, grape juice, the sludge extract weight ratio be 100:8-12:3-
4:5-8:5-9:1.5-2.5:0.5-0.9:0.1-0.3:0.4-0.6:0.3-0.6:1-3.
5. preparation method according to claim 1, wherein in step 2), the straw powder is selected from milled powders of cornstalk, sorghum
At least one of bar powder, wheat stalk powder, rice straw powder.
6. preparation method according to claim 1, wherein in step 2), the pH adjusting agent be selected from oxalic acid, acetic acid,
At least one of citric acid, sodium carbonate, sodium bicarbonate, saleratus.
7. preparation method according to claim 1, wherein in step 2), the high pressure sealing processing are as follows: go out in high pressure
15-25min is handled in bacterium pot.
8. preparation method according to claim 1, wherein after step 2), the preparation method further includes to system
In be passed through oxygen.
9. preparation method according to claim 9, wherein after step 2), the ventilatory capacity of oxygen is 0.4-0.6vvm.
10. a kind of culture medium of the mix bacterium agent for cellulase-producing, which is characterized in that described for the mixed of cellulase-producing
The culture medium of combined bacteria agent is prepared by preparation method described in any one of claim 1-9.
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| CN201910213724.7A CN109837263A (en) | 2019-03-20 | 2019-03-20 | The culture medium of mix bacterium agent and preparation method thereof for cellulase-producing |
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3677899A (en) * | 1969-02-03 | 1972-07-18 | Kyowa Hakko Kogyo Kk | Process for producing cellulase |
| CN104561135A (en) * | 2013-10-15 | 2015-04-29 | 丰益(上海)生物技术研发中心有限公司 | Method of using trichoderma reesei to produce aromatic substance |
| CN107384810A (en) * | 2017-09-07 | 2017-11-24 | 芜湖皖江知识产权运营中心有限公司 | A kind of stalk fermentation antimicrobial composition microbial inoculum and preparation method thereof |
-
2019
- 2019-03-20 CN CN201910213724.7A patent/CN109837263A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3677899A (en) * | 1969-02-03 | 1972-07-18 | Kyowa Hakko Kogyo Kk | Process for producing cellulase |
| CN104561135A (en) * | 2013-10-15 | 2015-04-29 | 丰益(上海)生物技术研发中心有限公司 | Method of using trichoderma reesei to produce aromatic substance |
| CN107384810A (en) * | 2017-09-07 | 2017-11-24 | 芜湖皖江知识产权运营中心有限公司 | A kind of stalk fermentation antimicrobial composition microbial inoculum and preparation method thereof |
Non-Patent Citations (1)
| Title |
|---|
| 周峰等: "杏鲍菇栽培原辅材料研究", 《安徽农学通报》 * |
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Application publication date: 20190604 |