CN108018239A - A kind of salt tolerant cellulose degradation microbial inoculum and preparation method and application - Google Patents
A kind of salt tolerant cellulose degradation microbial inoculum and preparation method and application Download PDFInfo
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- 239000002068 microbial inoculum Substances 0.000 title claims abstract description 66
- 239000001913 cellulose Substances 0.000 title claims abstract description 62
- 229920002678 cellulose Polymers 0.000 title claims abstract description 62
- 150000003839 salts Chemical class 0.000 title claims abstract description 43
- 230000015556 catabolic process Effects 0.000 title claims abstract description 30
- 238000006731 degradation reaction Methods 0.000 title claims abstract description 30
- 238000002360 preparation method Methods 0.000 title abstract description 6
- 241000193755 Bacillus cereus Species 0.000 claims abstract description 38
- 239000010902 straw Substances 0.000 claims abstract description 36
- 244000063299 Bacillus subtilis Species 0.000 claims abstract description 30
- 235000014469 Bacillus subtilis Nutrition 0.000 claims abstract description 30
- 238000000855 fermentation Methods 0.000 claims abstract description 20
- 230000004151 fermentation Effects 0.000 claims abstract description 20
- 239000004480 active ingredient Substances 0.000 claims abstract description 12
- 239000002002 slurry Substances 0.000 claims abstract description 7
- 230000000593 degrading effect Effects 0.000 claims description 52
- 241000589614 Pseudomonas stutzeri Species 0.000 claims description 37
- 240000008042 Zea mays Species 0.000 claims description 19
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 claims description 19
- 235000002017 Zea mays subsp mays Nutrition 0.000 claims description 19
- 235000005822 corn Nutrition 0.000 claims description 19
- 239000007788 liquid Substances 0.000 claims description 19
- 230000001580 bacterial effect Effects 0.000 claims description 18
- 239000003795 chemical substances by application Substances 0.000 claims description 13
- 241000894006 Bacteria Species 0.000 claims description 12
- 239000001963 growth medium Substances 0.000 claims description 11
- 238000000034 method Methods 0.000 claims description 9
- 239000000843 powder Substances 0.000 claims description 7
- 239000007787 solid Substances 0.000 claims description 7
- 230000004913 activation Effects 0.000 claims description 5
- 238000012258 culturing Methods 0.000 claims description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 5
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 4
- 239000008367 deionised water Substances 0.000 claims description 4
- 229910021641 deionized water Inorganic materials 0.000 claims description 4
- 239000002609 medium Substances 0.000 claims description 3
- 229920001817 Agar Polymers 0.000 claims description 2
- 229920000742 Cotton Polymers 0.000 claims description 2
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 2
- 244000068988 Glycine max Species 0.000 claims description 2
- 235000010469 Glycine max Nutrition 0.000 claims description 2
- 239000007836 KH2PO4 Substances 0.000 claims description 2
- 239000001888 Peptone Substances 0.000 claims description 2
- 108010080698 Peptones Proteins 0.000 claims description 2
- 240000006394 Sorghum bicolor Species 0.000 claims description 2
- 235000011684 Sorghum saccharatum Nutrition 0.000 claims description 2
- 235000021307 Triticum Nutrition 0.000 claims description 2
- 244000098338 Triticum aestivum Species 0.000 claims description 2
- 239000008272 agar Substances 0.000 claims description 2
- 235000015278 beef Nutrition 0.000 claims description 2
- 239000001110 calcium chloride Substances 0.000 claims description 2
- 229910001628 calcium chloride Inorganic materials 0.000 claims description 2
- 229940041514 candida albicans extract Drugs 0.000 claims description 2
- 229910052564 epsomite Inorganic materials 0.000 claims description 2
- 239000000284 extract Substances 0.000 claims description 2
- 239000008103 glucose Substances 0.000 claims description 2
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 2
- 235000019319 peptone Nutrition 0.000 claims description 2
- 239000011780 sodium chloride Substances 0.000 claims description 2
- VWDWKYIASSYTQR-UHFFFAOYSA-N sodium nitrate Inorganic materials [Na+].[O-][N+]([O-])=O VWDWKYIASSYTQR-UHFFFAOYSA-N 0.000 claims description 2
- 241000894007 species Species 0.000 claims description 2
- 239000012138 yeast extract Substances 0.000 claims description 2
- 239000010907 stover Substances 0.000 claims 4
- VNWKTOKETHGBQD-UHFFFAOYSA-N methane Chemical compound C VNWKTOKETHGBQD-UHFFFAOYSA-N 0.000 abstract description 20
- 238000011160 research Methods 0.000 abstract description 3
- 239000000654 additive Substances 0.000 abstract 2
- 230000000996 additive effect Effects 0.000 abstract 2
- 230000003413 degradative effect Effects 0.000 abstract 2
- 241000589516 Pseudomonas Species 0.000 abstract 1
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- 239000003513 alkali Substances 0.000 description 6
- 230000000052 comparative effect Effects 0.000 description 4
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- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
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- 150000001875 compounds Chemical class 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 239000002689 soil Substances 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
- 230000002195 synergetic effect Effects 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P39/00—Processes involving microorganisms of different genera in the same process, simultaneously
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P5/00—Preparation of hydrocarbons or halogenated hydrocarbons
- C12P5/02—Preparation of hydrocarbons or halogenated hydrocarbons acyclic
- C12P5/023—Methane
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
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- Y02E—REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
- Y02E50/00—Technologies for the production of fuel of non-fossil origin
- Y02E50/30—Fuel from waste, e.g. synthetic alcohol or diesel
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Abstract
The present invention relates to a kind of salt tolerant cellulose degradation microbial inoculum and preparation method and application.The active ingredient of the salt tolerant cellulose degradation microbial inoculum includes bacillus cereus, Si Shi pseudomonas, any two or three kinds in bacillus subtilis.Salt tolerant cellulose degradation microbial inoculum of the present invention can be applied to the biogas slurry of salt content 0 3%, and can express efficient degradation.Research is found, in marsh gas fermentation processes, when stalk additive amount is 100 300g, the salt tolerant cellulose degradation microbial inoculum additive amount is 10 30g, and degrade 15d, and straw degradative rate is 40% 61%.Complex microbial inoculum of the present invention adapts to northern high salinity area, can be applied under higher 0 3% environment of salinity, has the features such as straw degradative rate is high, and methane gas producing amount is high.
Description
Technical Field
The invention belongs to the field of microorganisms and the field of environmental protection. Specifically, the invention relates to a salt-resistant cellulose degrading microbial inoculum, and a method and application thereof.
Background
Because one third of land in Xinjiang is saline-alkali soil, the salt content of water sources such as canal water and the like is very high, and high-concentration saline-alkali ions have strong inhibiting effect on the activity of conventional cellulose degrading bacteria in saline-alkali environment, the saccharification efficiency in the corn straw biogas fermentation is greatly reduced, the gas yield is low, and therefore, the screening of the high-efficiency salt-resistant cellulose degrading bacteria suitable for saline-alkali areas is very necessary. Therefore, aiming at the problems of serious waste of corn straws, high salinity of canal water source, low yield of straw biogas and the like in northwest regions, the invention takes biogas slurry as a sample, adopts methods such as biotechnology and the like to screen salt-tolerant high-efficiency cellulose degrading strains of the corn straws, and utilizes the cellulose degrading strains to be added into a straw fermentation tank to improve the straw fermentation efficiency.
Disclosure of Invention
The invention provides a composite microbial straw degrading microbial inoculum suitable for a methane tank in a saline-alkali area, aiming at the problems of difficult degradation and long degradation time of crop straws such as corn and the like in the methane tank in the saline-alkali environment, and the composite microbial straw degrading microbial inoculum can promote the rapid degradation of the straws and can improve the fermentation efficiency.
The invention aims to provide a salt-resistant cellulose degradation microbial inoculum.
The active ingredients of the salt-tolerant cellulose degrading microbial inoculum comprise any two or three of Bacillus cereus, Pseudomonas stutzeri and Bacillus subtilis; preferably at least two species of Bacillus cereus and Pseudomonas stutzeri; more preferably, the above three bacteria are included at the same time.
Furthermore, the weight ratio of the bacillus cereus, the pseudomonas stutzeri and the bacillus subtilis in the salt-resistant cellulose degrading microbial inoculum is (1-3) to (0-2).
Further, the total effective viable count of the salt-resistant cellulose degrading bacterial agent is 1.8 multiplied by 109-7×109CFU/g, preferably 2X 109-6×109CFU/g。
In a specific embodiment of the invention, the active ingredients of the salt-tolerant cellulose degrading microbial inoculum are bacillus cereus and pseudomonas stutzeri, and the weight ratio of the bacillus cereus to the pseudomonas stutzeri is 1: 1; and the total effective viable count of the salt-resistant cellulose degrading microbial inoculum is 2.5 multiplied by 109-5×109CFU/g。
In another specific embodiment of the invention, the active ingredients of the salt-tolerant cellulose degrading microbial inoculum are bacillus cereus, pseudomonas stutzeri and bacillus subtilis, and the weight ratio of the bacillus cereus to the pseudomonas stutzeri to the bacillus subtilis is 2:2: 1; and the total effective viable count in the salt-resistant cellulose degrading microbial inoculum is 3 multiplied by 109-6×109CFU/g。
In another specific embodiment of the invention, the active ingredients of the salt-tolerant cellulose degrading microbial inoculum are bacillus cereus, pseudomonas stutzeri and bacillus subtilis, and the weight ratio of the bacillus cereus to the pseudomonas stutzeri to the bacillus subtilis is 3:3: 2; and the total effective viable count in the salt-resistant cellulose degrading microbial inoculum is 2 multiplied by 109-4×109CFU/g。
The salt-resistant cellulose degrading microbial inoculum can be applied to biogas slurry with the salt content of 0-3%, and can express high-efficiency degradability. Researches show that in the process of biogas fermentation, when the adding amount of the straws is 100-300g, the adding amount of the salt-resistant cellulose degrading microbial inoculum is 10-30g, the straws are degraded for 15d, and the straw degradation rate is 40-61%.
The bacillus cereus, pseudomonas stutzeri and bacillus subtilis used in the invention are all known strains and can be purchased from the market.
The second purpose of the invention is to provide a preparation method of the salt-tolerant microbial agent, which comprises the steps of taking bacillus cereus, pseudomonas stutzeri and bacillus subtilis microbial agents, and preparing the bacillus cereus, pseudomonas stutzeri and bacillus subtilis microbial agents according to a conventional method in the field.
Preferably, the preparation method of the salt-tolerant microbial agent comprises the following steps:
1) respectively inoculating Bacillus cereus, Pseudomonas stutzeri and Bacillus subtilis to an NA solid culture medium for activation;
2) respectively inoculating the activated bacterial colonies on the flat plate into corresponding liquid fermentation culture media by using an inoculation loop, and performing shake culture for 24-72 hours to logarithmic phase under the conditions of the temperature of 20-40 ℃ and 180-220rpm to obtain liquid microbial inoculum seed liquid;
3) and (3) amplifying and culturing the seed solution to obtain a bacterial solution with a proper concentration, centrifuging at a high speed, collecting bacteria to prepare a dry powder microbial inoculum, and mixing according to a required proportion to obtain the salt-resistant cellulose degrading microbial inoculum.
Wherein the NA solid culture medium comprises the following components in percentage by weight: 3g of beef extract, 5g of peptone, 2g of yeast extract, 8g of glucose, 16g of agar powder and 1L of deionized water, wherein the pH value is 7.0; the formula of the liquid fermentation medium is as follows: NaCl 0.1g, MgSO4·7H2O 0.3g，NaNO32.5g，KH2PO41.0g，CaCl20.1g, cellulose 20 g.L-1Deionized water was added to 1L, pH 7.0.
The third purpose of the invention is to provide the application of the salt-resistant cellulose degrading microbial inoculum in the aspects of degrading cellulose and the like. The cellulose source comprises corn straw, soybean straw, cotton straw, wheat straw, sorghum straw and the like.
Researches find that the salt-resistant cellulose degrading microbial inoculum is particularly suitable for degrading corn straws by biogas slurry, and can ensure that the degradation rate of the corn straws reaches 61%.
The starting materials used in the present invention are commercially available or may be prepared by methods conventional in the art.
On the basis of the common knowledge in the field, the above preferred conditions can be combined with each other to obtain the preferred embodiments of the invention.
Compared with the prior art, the invention has the beneficial effects that:
the invention provides a salt-resistant cellulose degrading microbial inoculum which can rapidly decompose cellulose and hemicellulose under the high-salt condition, so that the straw degradation rate and the gas production rate can be further accurately calculated. The salt-resistant cellulose degradation microbial inoculum provided by the invention has strong synergistic capability, so that the straw degradation efficiency of a high-salt methane tank is improved, and a key pretreatment technology is provided for straw methane fermentation. The microbial compound inoculant is suitable for northern high-salinity and high-salinity areas, can be applied in the environment with higher salinity of 0-3%, and has the characteristics of high straw degradation rate, high biogas yield and the like.
Detailed Description
The following examples are intended to illustrate the invention but are not intended to limit the scope of the invention. The examples do not show the specific techniques or conditions, according to the technical or conditions described in the literature in the field, or according to the product specifications. The reagents or instruments used are conventional products available from regular distributors, not indicated by the manufacturer.
Example 1
Respectively inoculating Bacillus cereus, Pseudomonas stutzeri and Bacillus subtilis to an NA solid culture medium for activation; respectively inoculating bacterial colonies on the flat plate into corresponding liquid fermentation culture media by using an inoculating ring, and performing shake culture for 24 hours to logarithmic phase at the temperature of 20 ℃ and the speed of 180rpm to obtain liquid microbial inoculum seed liquid; and (3) amplifying and culturing the seed solution to obtain a bacterial solution with higher concentration, centrifuging at a high speed, collecting bacteria, preparing into a dry powder microbial inoculum, and respectively preparing the bacillus cereus microbial inoculum, the pseudomonas stutzeri microbial inoculum and the bacillus subtilis microbial inoculum.
The embodiment also provides a salt-tolerant cellulose degrading microbial inoculum, which comprises the active ingredients of bacillus cereus and pseudomonas stutzeri, wherein the weight ratio of the bacillus cereus to the pseudomonas stutzeri is 1: 1; and the total effective viable count of the salt-resistant cellulose degrading microbial inoculum is 2.5 multiplied by 109-5×109CFU/g。
The salt-resistant cellulose degrading microbial inoculum is applied to methane fermentation, and the salt content of the methane liquid is 3%. When the adding amount of the corn straws is 300g, the adding amount of the salt-resistant cellulose degrading microbial inoculum is 10g, the degradation lasts 15d, and the degradation rate of the corn straws is 40%; when the adding amount of the corn straws is 200g, the adding amount of the salt-resistant cellulose degrading microbial inoculum is 20g, the degradation time is 15d, and the degradation rate of the corn straws is 47%.
Example 2
Respectively inoculating Bacillus cereus, Pseudomonas stutzeri and Bacillus subtilis to an NA solid culture medium for activation; respectively inoculating bacterial colonies on the flat plate into corresponding liquid fermentation culture media by using an inoculating ring, and performing shake culture for 48 hours to a logarithmic phase under the conditions of the temperature of 30 ℃ and the rpm of 200 to obtain liquid microbial inoculum seed liquid; and (3) amplifying and culturing the seed solution to obtain a bacterial solution with higher concentration, centrifuging at a high speed, collecting bacteria, preparing into a dry powder microbial inoculum, and respectively preparing the bacillus cereus microbial inoculum, the pseudomonas stutzeri microbial inoculum and the bacillus subtilis microbial inoculum.
The embodiment also provides a salt-tolerant cellulose degrading microbial inoculum, which comprises the active ingredients of bacillus cereus, pseudomonas stutzeri and bacillus subtilis, wherein the weight ratio of the bacillus cereus to the pseudomonas stutzeri to the bacillus subtilis is 2:2: 1; and total effective viable bacteria in the salt-resistant cellulose degrading microbial inoculumNumber 3X 109-6×109CFU/g。
The salt-resistant cellulose degrading microbial inoculum is applied to biogas fermentation, the salt content of biogas slurry is 3%, the adding amount of corn stalks is 200g, the adding amount of the salt-resistant cellulose degrading microbial inoculum is 20g, the degradation time is 15d, and the degradation rate of the corn stalks is 61%.
Example 3
Respectively inoculating Bacillus cereus, Pseudomonas stutzeri and Bacillus subtilis to an NA solid culture medium for activation; respectively inoculating bacterial colonies on the flat plate into corresponding liquid fermentation culture media by using an inoculating ring, and performing shake culture for 72 hours to a logarithmic phase at the temperature of 40 ℃ and the speed of 220rpm to obtain liquid microbial inoculum seed liquid; and (3) amplifying and culturing the seed solution to obtain a bacterial solution with higher concentration, centrifuging at a high speed, collecting bacteria, preparing into a dry powder microbial inoculum, and respectively preparing the bacillus cereus microbial inoculum, the pseudomonas stutzeri microbial inoculum and the bacillus subtilis microbial inoculum.
The embodiment also provides a salt-tolerant cellulose degrading microbial inoculum, which comprises the active ingredients of bacillus cereus, pseudomonas stutzeri and bacillus subtilis, wherein the weight ratio of the bacillus cereus to the pseudomonas stutzeri to the bacillus subtilis is 3:3: 2; and the total effective viable count in the salt-resistant cellulose degrading microbial inoculum is 2 multiplied by 109-4×109CFU/g。
The salt-resistant cellulose degrading microbial inoculum is applied to methane fermentation, and the salt content of the methane liquid is 3%. When the adding amount of the corn straws is 100g, the adding amount of the salt-resistant cellulose degrading microbial inoculum is 30g, the degradation lasts 15d, and the degradation rate of the corn straws is 58%; when the adding amount of the corn straws is 200g, the adding amount of the salt-resistant cellulose degrading microbial inoculum is 20g, the degradation time is 15d, and the degradation rate of the corn straws is 53%.
Comparative example 1
The effective viable count of the bacillus cereus is 4 multiplied by 109-8×109CFU/g。
Comparative example 2
Pseudomonas stutzeri with an effective viable count of 1X 109-2×109CFU/g。
Comparative example 3
The effective viable count of the bacillus subtilis preparation is 4 multiplied by 109-8×109CFU/g。
Experimental example 1
The microbial inoculum of examples 1-3 and comparative examples 1-3 were used for biogas fermentation, respectively, and no microbial inoculum was added as a blank control, the salt content of the biogas slurry was 3%, the degradation time was 15 days, and the corn stalk degradation rate and gas yield were as shown in table 1 below.
TABLE 1
Although the invention has been described in detail hereinabove with respect to a general description and specific embodiments thereof, it will be apparent to those skilled in the art that modifications or improvements may be made thereto based on the invention. Accordingly, such modifications and improvements are intended to be within the scope of the invention as claimed.
Claims (10)
1. The salt-tolerant cellulose degrading microbial inoculum is characterized in that the active ingredients comprise any two or three of Bacillus cereus (Bacillus cereus), Pseudomonas stutzeri (Pseudomonas stutzeri) and Bacillus subtilis (Bacillus subtilis); preferably at least two species of Bacillus cereus and Pseudomonas stutzeri; more preferably, the above three bacteria are included at the same time.
2. The salt-tolerant cellulose degrading bacterial agent of claim 1, wherein the weight ratio of bacillus cereus, pseudomonas stutzeri and bacillus subtilis in the salt-tolerant cellulose degrading bacterial agent is (1-3) to (0-2).
3. The salt-tolerant cellulose-degrading bacterial agent of claim 1 or 2, wherein the total effective viable count of the salt-tolerant cellulose-degrading bacterial agent is 1.8 x 109-7×109CFU/g, preferably 2X 109-6×109CFU/g。
4. The salt-tolerant cellulose degrading bacterial agent as claimed in claim 1, wherein the active ingredients of the salt-tolerant cellulose degrading bacterial agent are bacillus cereus and pseudomonas stutzeri, and the weight ratio of the bacillus cereus to the pseudomonas stutzeri is 1: 1; and the total effective viable count of the salt-resistant cellulose degrading microbial inoculum is 2.5 multiplied by 109-5×109CFU/g; or,
the active ingredients of the salt-tolerant cellulose degrading microbial inoculum are bacillus cereus, pseudomonas stutzeri and bacillus subtilis, and the weight ratio of the bacillus cereus to the pseudomonas stutzeri to the bacillus subtilis is 2:2: 1; and the total effective viable count in the salt-resistant cellulose degrading microbial inoculum is 3 multiplied by 109-6×109CFU/g; or,
the active ingredients of the salt-tolerant cellulose degrading microbial inoculum are bacillus cereus, pseudomonas stutzeri and bacillus subtilis, and the weight ratio of the bacillus cereus to the pseudomonas stutzeri to the bacillus subtilis is 3:3: 2; and the total effective viable count in the salt-resistant cellulose degrading microbial inoculum is 2 multiplied by 109-4×109CFU/g。
5. The method for preparing the salt-tolerant cellulose degrading bacterial agent of any one of claims 1 to 4, which is characterized by comprising the following steps:
1) respectively inoculating Bacillus cereus, Pseudomonas stutzeri and Bacillus subtilis to an NA solid culture medium for activation;
2) respectively inoculating the activated colonies into corresponding liquid fermentation culture media, and performing shake culture at 20-40 ℃ and 180-220rpm for 24-72 hours to logarithmic phase to obtain liquid microbial inoculum seed liquid;
3) and (3) amplifying and culturing the seed solution to obtain a bacterial solution with a proper concentration, centrifuging at a high speed, collecting bacteria to prepare a dry powder microbial inoculum, and mixing according to a required proportion.
6. The method of claim 5, wherein the NA solid medium is formulated as: 3g of beef extract, 5g of peptone, 2g of yeast extract, 8g of glucose, 16g of agar powder and 1L of deionized water, wherein the pH value is 7.0; and/or the presence of a gas in the gas,
the formula of the liquid fermentation medium is as follows: NaCl 0.1g, MgSO4·7H2O 0.3g，NaNO32.5g，KH2PO41.0g，CaCl20.1g, cellulose 20 g.L-1Deionized water was added to 1L, pH 7.0.
7. The use of a salt-tolerant cellulose-degrading bacterial agent according to any one of claims 1 to 4 or prepared by the method of claim 5 or 6 for degrading cellulose; preferably, the source of cellulose comprises corn stover, soybean stover, cotton stover, wheat straw, sorghum stover.
8. The use according to claim 7, characterized in that in biogas fermentation, the salt content of the biogas slurry is 0-3%.
9. The application of the salt-tolerant cellulose degrading bacteria as claimed in claim 7 or 8, wherein the salt-tolerant cellulose degrading bacteria is added in an amount of 10-30g when the straw is added in an amount of 100-300g in the process of biogas fermentation.
10. Use according to claim 9, wherein the degradation time is 15d or more.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109593677A (en) * | 2018-12-24 | 2019-04-09 | 沈阳农业大学 | Low temperature resistant degraded cellulose composite bacteria agent and its preparation method and application |
| CN112080450A (en) * | 2020-09-30 | 2020-12-15 | 内蒙古农业大学 | Straw degrading bacteria and separation screening method |
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