CN111662897A - Preparation method of modified wheat bran material immobilized microbial agent - Google Patents
Preparation method of modified wheat bran material immobilized microbial agent Download PDFInfo
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- CN111662897A CN111662897A CN202010564139.4A CN202010564139A CN111662897A CN 111662897 A CN111662897 A CN 111662897A CN 202010564139 A CN202010564139 A CN 202010564139A CN 111662897 A CN111662897 A CN 111662897A
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- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
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Abstract
The invention discloses a preparation method of a modified wheat bran material immobilized microbial agent, which comprises the following steps: s1, preparing bacterial liquid for later use; s2, preparation of a carrier: gradually carrying out starch hydrolysis and protein hydrolysis modification on the wheat bran to prepare a carrier for later use, wherein the hydrolysis sequence is not sequential; s3, preparation of microbial inoculum: mixing the bacterial liquid obtained in S1 with the carrier obtained in S2 according to the ratio of 1 mL: (0.5-1) g, and drying to constant weight. The invention modifies common wheat bran to be used as a carrier of a microbial agent, has high immobilization efficiency, creates rich micropore growth environment for microorganisms because the specific surface area of the modified wheat bran is greatly increased, increases the adsorption quantity and the accommodation quantity of the microorganisms, can be used as a novel carrier, and the viable count in the microbial agent prepared by adopting the carrier is far beyond the execution standard.
Description
Technical Field
The invention relates to the technical field of microbial fermentation, in particular to a preparation method of a modified wheat bran material immobilized microbial inoculum.
Background
Along with the rapid development of economy in China, the living standard of people is continuously improved, the scale of the planting industry, the breeding industry and the industry in China is continuously increased, the yield of organic solid wastes such as livestock manure, garden wastes, sludge, kitchen wastes and the like is increased day by day, and the traditional modes of burning, landfill and the like not only easily cause secondary pollution, but also waste resources. In recent years, the technology of treating the garbage by using a biological method is mature day by day, compared with the traditional mode, the garbage treated by using the biological method is thorough in treatment, free of secondary pollution and good in safety, the transportation cost and the labor cost are reduced, and the treated product can be used for preparing organic fertilizer and is beneficial to recycling of resources.
The key of the biological method for treating the garbage lies in the selection and control of a microbial agent, and in order to more conveniently and durably use the microbial agent, the immobilized microbial technology is the key. Immobilization is the physical or chemical means to confine free microorganisms within a specific spatial range, retaining their inherent activity. The immobilization technology mainly comprises physical adsorption, an embedding method, a crosslinking method and the like, wherein the selection of a proper immobilization material is critical. At present, common carriers comprise inorganic carriers (zeolite, kaolin, activated carbon), organic carriers (corn meal, soybean meal powder) and the like, but the materials have the problems of poor adsorbability, easy secondary pollution, difficult mining, non-regeneration, high cost and the like, and are difficult to be used in the actual garbage treatment in large quantities, so that a carrier which is strong in adsorbability, high in stability, low in price and friendly to the environment is needed.
Disclosure of Invention
The purpose of the invention is as follows: in order to overcome the defects of the background art, the invention discloses a preparation method of a modified wheat bran material immobilized microbial agent.
The technical scheme is as follows: the preparation method of the modified wheat bran material immobilized microbial agent comprises the following steps:
s1, preparing bacterial liquid for later use;
s2, preparation of a carrier: gradually carrying out starch hydrolysis and protein hydrolysis modification on the wheat bran to prepare a carrier for later use, wherein the hydrolysis sequence is not sequential;
s3, preparation of microbial inoculum: mixing the bacterial liquid obtained in S1 with the carrier obtained in S2 according to the ratio of 1 mL: (0.5-1) g, and drying to constant weight.
The preparation method of S1 comprises activating the strain stored at-20-80 deg.C, inoculating into prepared LB liquid culture medium, and fermenting at constant temperature for 1-2 days.
Further, in S2:
starch hydrolysis comprises pulverizing wheat bran, weighing, and adding NaH2PO4NaOH buffer solution, preheating in a constant temperature incubator for 30-50min, adding α amylase, adjusting pH to 6-6.5, hydrolyzing at 60-65 deg.C for 3-5h, washing with distilled water, and oven drying to constant weight.
The protein hydrolysis is carried out by pulverizing wheat bran, weighing, adding NaH2PO4NaOH buffer solution, preheating in a constant temperature incubator for 30-50min, adding protease, adjusting pH to 12.5-13, hydrolyzing at 45-50 deg.C for 2-3h, washing with distilled water, and oven drying to constant weight.
Further, the mass ratio of the wheat bran to the alpha amylase is 10: (0.2-0.3);
the mass ratio of the wheat bran to the protease is 10: (2-3).
When the wheat bran is modified, the hydrolysis sequence is not in sequence, but the starch hydrolysis and the protein hydrolysis need to be respectively operated step by step.
Has the advantages that: compared with the prior art, the invention has the advantages that: firstly, common wheat bran is modified and used as a carrier of a microbial agent, the immobilization efficiency is high, and the modified wheat bran has a greatly increased specific surface area, so that a rich micropore growth environment is created for microorganisms, the adsorption capacity and the accommodation capacity of the microorganisms are increased, and the modified wheat bran can be used as a novel carrier; the viable count in the microbial inoculum prepared by the carrier is far beyond the execution standard; and the modified wheat bran has the advantages of environmental protection, no pollution to the environment, no toxicity or harm to organisms, low price of the bran, and capability of greatly reducing the economic cost in production and solving the problem of arrangement of byproducts in flour production, thereby being a way for reasonably utilizing resources.
Detailed Description
The technical solution of the present invention will be further described with reference to the following examples.
Example 1
A preparation method of a modified wheat bran material immobilized microbial agent comprises the following steps:
1. preparing bacterial liquid: activating the strain stored in a low-temperature (-40 ℃) refrigerator, inoculating the activated strain into a prepared 100mL LB liquid culture medium, and placing the LB liquid culture medium in a constant-temperature shaking table for fermentation for 1d for later use.
2. Preparing a carrier:
(1) hydrolysis of starch in wheat bran: pulverizing wheat bran, sieving with 50 mesh sieve, weighing 50g in triangular flask, adding 500mL NaH2PO4And (2) putting the NaOH buffer solution into a constant-temperature incubator, preheating for 50min, adding 1.5g of α amylase (the mass ratio of the wheat bran to the α amylase is 10: 0.3), adjusting the pH value to 6, putting the mixture into an environment at 60 ℃, hydrolyzing for 5h, washing the mixture with distilled water, and drying the mixture until the weight is constant, wherein the enzymolysis rate can reach 95-99%.
(2) Hydrolysis of proteins in wheat bran: 30g of the debranched bran of (1) are weighed into a triangular flask, and 300mL of NaH are added2PO4And (2) putting the NaOH buffer solution into a constant-temperature incubator, preheating for 50min, adding 9g of protease (the mass ratio of the wheat bran to the protease is 10: 3), adjusting the pH value to 13, putting the mixture into a 50 ℃ environment for hydrolysis for 2h, washing the mixture with distilled water, drying the mixture until the weight is constant, and enabling the enzymolysis rate to reach 85% -92%.
TABLE 1 specific surface area of bran under different treatments
3. Preparation of a microbial inoculum: and (3) fully and uniformly mixing the bacterial suspension obtained in the step (1) and the wheat bran from which the starch and the protein are removed in a ratio of 1:1 (mL: g), and placing the mixture in a forced air drying oven to dry the mixture until the weight is constant. The colony number in the microbial inoculum is 3.5 x 109CFU/g, the implementation standard is referred to GB 20287-2006.
TABLE 2 agricultural microbial inoculum standards
Example 2
A preparation method of a modified wheat bran material immobilized microbial agent comprises the following steps:
1. preparing bacterial liquid: activating the strain stored in a low-temperature (-20 ℃) refrigerator, inoculating the activated strain into a prepared 100mL LB liquid culture medium, and placing the LB liquid culture medium in a constant-temperature shaking table for fermentation for 2d for later use.
2. Preparing a carrier:
(1) hydrolysis of starch in wheat bran: pulverizing wheat bran, sieving with 50 mesh sieve, weighing 100g in triangular flask, adding 1L NaH2PO4And (2) preheating the NaOH buffer solution in a constant-temperature incubator for 30min, adding α amylase (the mass ratio of the wheat bran to the α amylase is 10: 0.2), adjusting the pH value to 6.5, hydrolyzing the mixture in an environment at 65 ℃ for 3h, washing the hydrolyzed product with distilled water, and drying the washed product until the weight is constant, wherein the enzymolysis rate can reach more than 95%.
(2) Hydrolysis of proteins in wheat bran: 60g of the de-starched bran of (1) are weighed into a triangular flask and 600ml of NaH is added2PO4And (2) putting the NaOH buffer solution into a constant-temperature incubator, preheating for 30min, adding 12g of protease (the mass ratio of the wheat bran to the protease is 10: 2), adjusting the pH value to 12.5, putting the mixture into a 50 ℃ environment, hydrolyzing for 3h, washing the mixture with distilled water, and drying to constant weight. The enzymolysis rate can reach more than 85%.
3. Preparation of a microbial inoculum: and (3) fully and uniformly mixing the bacterial suspension obtained in the step (1) and the wheat bran from which the starch and the protein are removed in a ratio of 1:0.5 (mL: g), and placing the mixture in a forced air drying oven to dry the mixture to constant weight. The colony number in the microbial inoculum is 3.2 x 109CFU/g, the implementation standard is referred to GB 20287-2006.
Example 3
A preparation method of a modified wheat bran material immobilized microbial agent comprises the following steps:
1. preparing bacterial liquid: activating the strain stored in a low-temperature (-80 ℃) refrigerator, inoculating the activated strain into a prepared 100mL LB liquid culture medium, and placing the LB liquid culture medium in a constant-temperature shaking table for fermentation for 1.5 days for later use.
2. Preparing a carrier:
(1) hydrolysis of starch in wheat bran: pulverizing wheat bran, sieving with 50 mesh sieve, weighing 100g in triangular flask, adding 1L NaH2PO4-NaAnd (2) preheating the OH buffer solution in a constant-temperature incubator for 40min, adding α amylase (the mass ratio of the wheat bran to the α amylase is 10: 0.25), adjusting the pH value to 6.3, hydrolyzing in an environment at 62 ℃ for 4h, washing with distilled water, and drying to constant weight, wherein the enzymolysis rate can reach more than 95%.
(2) Hydrolysis of proteins in wheat bran: 60g of the de-starched bran of (1) are weighed into a triangular flask and 600ml of NaH is added2PO4NaOH buffer solution, preheating in a constant temperature incubator for 40min, adding 15g protease (the mass ratio of wheat bran to protease is 10: 2.5), adjusting pH to 12.8, hydrolyzing in 48 deg.C environment for 2.5h, washing with distilled water, and oven drying to constant weight. The enzymolysis rate can reach more than 85%.
3. Preparation of a microbial inoculum: and (3) fully and uniformly mixing the bacterial suspension obtained in the step (1) and the wheat bran from which the starch and the protein are removed according to the proportion of 1:0.8 (mL: g), and placing the mixture in a forced air drying oven to dry the mixture to constant weight. The colony number in the microbial inoculum is 3.3 x 109CFU/g, the implementation standard is referred to GB 20287-2006.
The number of viable bacteria in the microbial inoculum prepared in the examples 1-3 is equal to or larger than 30 hundred million/g, and far exceeds the execution standard.
Claims (6)
1. A preparation method of a modified wheat bran material immobilized microbial agent is characterized by comprising the following steps:
s1, preparing bacterial liquid for later use;
s2, preparation of a carrier: gradually carrying out starch hydrolysis and protein hydrolysis modification on the wheat bran to prepare a carrier for later use, wherein the hydrolysis sequence is not sequential;
s3, preparation of microbial inoculum: mixing the bacterial liquid obtained in S1 with the carrier obtained in S2 according to the ratio of 1 mL: (0.5-1) g, and drying to constant weight.
2. The method for preparing the immobilized microbial inoculum of modified wheat bran material of claim 1, which is characterized in that: the preparation method of S1 comprises activating the strain stored at-20-80 deg.C, inoculating into prepared LB liquid culture medium, and fermenting at constant temperature for 1-2 days.
3. The method for preparing the immobilized microbial inoculum of modified wheat bran material of claim 1, which is characterized in that: s2 hydrolyzing starch to obtain powder, weighing, and adding NaH2PO4NaOH buffer solution, preheating in a constant temperature incubator for 30-50min, adding α amylase, adjusting pH to 6-6.5, hydrolyzing at 60-65 deg.C for 3-5h, washing with distilled water, and oven drying to constant weight.
4. The method for preparing the immobilized microbial inoculum of modified wheat bran material of claim 3, which is characterized in that: the mass ratio of the wheat bran to the alpha amylase is 10: (0.2-0.3).
5. The method for preparing the immobilized microbial inoculum of modified wheat bran material of claim 1, which is characterized in that: s2 hydrolyzing the protein to obtain powder, weighing, and adding NaH2PO4NaOH buffer solution, preheating in a constant temperature incubator for 30-50min, adding protease, adjusting pH to 12.5-13, hydrolyzing at 45-50 deg.C for 2-3h, washing with distilled water, and oven drying to constant weight.
6. The method for preparing the immobilized microbial inoculum of modified wheat bran material of claim 5, which is characterized in that: the mass ratio of the wheat bran to the protease is 10: (2-3).
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112194513A (en) * | 2020-09-22 | 2021-01-08 | 武汉鸿谷致远科技有限公司 | Biological deodorant |
| CN115025231A (en) * | 2022-06-15 | 2022-09-09 | 江西省药品检验检测研究院 | Dry inoculation method of traditional Chinese medicinal materials |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107006674A (en) * | 2017-02-21 | 2017-08-04 | 范恒 | A kind of extraction of gluten protein and method of modifying |
| CN110643593A (en) * | 2019-11-14 | 2020-01-03 | 中国科学院沈阳应用生态研究所 | Preparation method of modified corn straw material immobilized agricultural microbial agent |
-
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107006674A (en) * | 2017-02-21 | 2017-08-04 | 范恒 | A kind of extraction of gluten protein and method of modifying |
| CN110643593A (en) * | 2019-11-14 | 2020-01-03 | 中国科学院沈阳应用生态研究所 | Preparation method of modified corn straw material immobilized agricultural microbial agent |
Non-Patent Citations (1)
| Title |
|---|
| 杨健等: "小麦麸皮低聚木糖提取液絮凝工艺技术研究", 《粮食与饲料工业》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112194513A (en) * | 2020-09-22 | 2021-01-08 | 武汉鸿谷致远科技有限公司 | Biological deodorant |
| CN115025231A (en) * | 2022-06-15 | 2022-09-09 | 江西省药品检验检测研究院 | Dry inoculation method of traditional Chinese medicinal materials |
| CN115025231B (en) * | 2022-06-15 | 2024-03-19 | 江西省药品检验检测研究院 | Dry inoculation method of traditional Chinese medicinal materials |
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