CN108707562B - Microbial preparation for repairing tri-nitrogen contaminated soil and preparation method and application thereof - Google Patents
Microbial preparation for repairing tri-nitrogen contaminated soil and preparation method and application thereof Download PDFInfo
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Abstract
The invention discloses a microbial preparation for repairing tri-nitrogen contaminated soil, which is characterized by comprising a pseudomonas stutzeri SC221-M microbial inoculum and an aerobic denitrifying bacterium capable of carrying out heterotrophic nitrificationParacoccus pantotrophusATCC 35512 bacterial agent and immobilized glucose oxidase. The invention also discloses a microbial preparation for repairing the tri-nitrogen contaminated soil and an application thereof. The combined use of the two microbial agents is found that the effect in denitrification is superior to the effect of using the two microbial agents independently and jointly, the ammonia nitrogen removal rate reaches over 95 percent, the nitrite nitrogen removal rate reaches over 95 percent, and the nitrate nitrogen removal rate reaches over 96 percent.
Description
Technical scheme
The invention belongs to the technical field of soil remediation, and particularly relates to a microbial preparation for remediating tri-nitrogen contaminated soil, and a preparation method and application thereof.
Background
The soil is the target of the migration, direct current and deposition of pollutants in the atmosphere, water and solid wastes in the environment, and is the long-term environmental pollution, with the accelerated development of social economy and industrialization, certain industrial waste water such as coking waste water, synthetic ammonia fertilizer waste water and the like causes trinitrogen pollution, namely ammonia nitrogen, nitrite nitrogen and nitrate nitrogen pollution, the ammonia nitrogen pollution mainly comes from the decomposition products of nitrogenous organic matters in domestic sewage under the action of microorganisms, the nitrite nitrogen is a carcinogenic substance and has great harm to the health of human bodies, on one hand, nitrate can cause the algification phenomenon of the water body, on the other hand, nitrate can be reduced into nitrite in intestines and stomach to form carcinogenic nitrosamine, so that the life health is harmed, at present, the pollution of underground water nitrate in China is serious, and the leaching loss of the nitrate in farmland soil is the main reason of underground water pollution.
The technology and method for remedying the soil pollution have been studied since the end of 70 s and the beginning of 80 s abroad, and the remediation of the polluted soil is generally carried out by physical and chemical methods in the initial stage, such as a heat treatment method and a chemical leaching method, but the cost is high, and the technology and the method are not suitable for large-area application. In the late 80 s and early 90 s, the research on the bioremediation method of soil pollution by microorganisms and plant catabolism based on physical and chemical processes is started abroad, the bioremediation method is developed very rapidly in recent years, is economical compared with physical and chemical methods, is not easy to generate secondary pollution, and is more suitable for the remediation of large-area soil, the bioremediation technology is started to be used as a main treatment technology for soil remediation and plays an increasingly important role, but the existing remediation technology has the problems of low ammonia nitrogen removal rate, unstable microorganisms, easy inactivation of enzymatic treatment and the like, and therefore, the problem of ammonia nitrogen-polluted soil needs to be solved and remedied comprehensively.
Disclosure of Invention
The purpose of the invention is as follows: the invention aims to provide a microbial preparation for repairing trinitrogen polluted soil, which is mainly used for repairing pollution of ammonia nitrogen, nitrite nitrogen and nitrate nitrogen.
The invention also aims to provide a preparation method and application of the microbial preparation for repairing the soil polluted by the trinitrogen.
The technical scheme is as follows: the invention provides a microbial preparation for repairing trinitrogen-polluted soil, which comprises a Pseudomonas stutzeri SC221-M microbial inoculum, an aerobic denitrifying bacterium Paracoccus pantophus ATCC 35512 microbial inoculum and immobilized glucose oxidase.
Wherein the weight ratio of the Pseudomonas stutzeri SC221-M microbial inoculum to the aerobic denitrifying bacteria Paracoccus pantophus ATCC 35512 microbial inoculum is 0.1-2: 1-5.
Wherein the immobilized glucose oxidase accounts for 1/10-1/5 of the weight of the microbial preparation.
Wherein, in the microbial preparation, the viable bacteria content of the Pseudomonas stutzeri SC221-M microbial inoculum is 5 multiplied by 108～8×1010cfu/g, the viable bacteria content of the heterotrophic nitrification-aerobic denitrification bacterium Paracoccus pantophus ATCC 35512 microbial inoculum is 3 multiplied by 109～5×1010cfu/g。
Wherein, in the microbial preparation, the concentration of the immobilized glucose oxidase is 20-30U/mg.
The invention also relates to a method for preparing the microbial preparation for repairing the soil polluted by the trinitrogen, which comprises the following steps:
1) uniformly mixing a Pseudomonas stutzeri SC221-M microbial inoculum and an aerobic denitrifying bacterium Paracoccus pantophus ATCC 35512 microbial inoculum according to the mass ratio of 0.1-2: 1-5 to obtain a composite microbial inoculum;
2) adding the immobilized glucose oxidase into the composite microbial inoculum prepared in the step 1), and uniformly mixing to obtain the microbial preparation for repairing the trinitrogen polluted soil.
The preparation method of the Pseudomonas stutzeri SC221-M microbial inoculum comprises the following steps: culturing the pseudomonas stutzeri SC221-M to a logarithmic phase, then inoculating strains cultured to the logarithmic phase into a culture medium according to the inoculation amount of 3-10% of the volume of the culture medium, shaking the culture medium at 30-35 ℃ at 180-220 rpm by shaking table, culturing for 40-48 hours to obtain a fermentation liquor of the pseudomonas stutzeri SC221-M, adding an adsorbent into the fermentation liquor, and drying to obtain the pseudomonas stutzeri SC221-M microbial inoculum; the adsorbent is one or more of bentonite and activated carbon.
Wherein the culture medium is prepared by taking wheat straw powder and sweet potato vine powder as carbon sources and taking ammonium sulfate and peptone as nitrogen sources.
The preparation method of the heterotrophic nitrification-aerobic denitrification bacterium Paracoccus pantophus ATCC 35512 microbial inoculum comprises the following steps: culturing anaerobic nitrification-aerobic denitrifying bacteria Paracoccus pantophus ATCC 35512 in an R2A liquid culture medium to logarithmic phase, then inoculating strains cultured to logarithmic phase into an R2A liquid culture medium according to the inoculation amount of 3-6% of the volume of the culture medium, shaking the table at 120-180 rpm at the temperature of 30-40 ℃, culturing for 24-48 hours, adding an adsorbent, and drying to obtain an anaerobic nitrification-aerobic denitrifying bacteria Paracoccus pantophus ATCC 35512 microbial inoculum; the adsorbent is one or more of bentonite or activated carbon.
The invention also comprises the application of the microbial preparation for repairing the soil polluted by the trinitrogen in the remediation of the soil polluted by the trinitrogen.
Has the advantages that: the invention firstly mixes the Pseudomonas stutzeri SC221-M microbial inoculum and the anaerobic nitrification-aerobic denitrifying bacteria Paracoccus pantophus ATCC 35512 microbial inoculum with immobilized glucose oxidase to prepare the microbial preparation, and the microbial preparation utilizes the nitrification and denitrification of the anaerobic nitrification-aerobic denitrifying bacteria Paracoccus pantophus ATCC 35512 microbial inoculum to carry out NH (ammonia-nitrogen) reaction in the same system4 +The inventor discovers that the effect is superior to the effect of respectively using two bactericides singly and jointly in the aspect of denitrification through the combined use of the two microbial preparations, the ammonia nitrogen removal rate reaches more than 95%, the nitrite nitrogen removal rate reaches more than 95%, and the nitrate nitrogen removal rate reaches more than 96%.
Detailed Description
The Pseudomonas stutzeri SC221-M strain is a gift from the institute of microorganisms of the Chinese academy of sciences, and the heterotrophic nitrification-aerobic denitrification bacterium Paracoccus pantophus ATCC 35512 is purchased from American ATCC cell bank. All reagents in the invention are commercially available.
Example 1
Immobilization of glucose oxidase: adding mesoporous molecular sieve material SBA-15 into glucose oxidase at the temperature of 10 ℃; the pH was 4.5; the rotating speed is 180 rpm; immobilization was carried out for 10 hours.
The preparation method of the Pseudomonas stutzeri SC221-M microbial inoculum comprises the following steps: culturing Pseudomonas stutzeri SC221-M to logarithmic phase, wherein the culture medium is a culture medium which takes wheat straw powder and sweet potato vine powder as carbon sources and takes ammonium sulfate and peptone as nitrogen sources, inoculating strains cultured to logarithmic phase into the culture medium according to the inoculation amount of 3% of the volume of the culture medium, culturing for 48 hours at 30 ℃, shaking table shaking 220rpm to obtain fermentation liquor of Pseudomonas stutzeri SC221-M, adding activated carbon into the fermentation liquor, and drying to obtain the Pseudomonas stutzeri SC221-M microbial inoculum. The viable bacteria content of the Pseudomonas stutzeri SC221-M microbial inoculum is 5 multiplied by 108cfu/g。
The preparation method of the bacterium agent of the heterotrophic nitrification-aerobic denitrification bacterium Paracoccus pantophus ATCC 35512 comprises the following steps: culturing the heterotrophic nitrification-aerobic denitrifying bacteria Paracoccus pantophus ATCC 35512 in an R2A liquid culture medium (purchased from Qingdao Haibobo organisms) to logarithmic phase, then inoculating the strain cultured to logarithmic phase into the R2A liquid culture medium according to the inoculation amount of 3% of the volume of the culture medium, shaking the shaking table at 30 ℃, shaking at 180rpm, culturing for 48 hours, adding bentonite, and drying to prepare the heterotrophic nitrification-aerobic denitrifying bacteria Paracoccus pantophus ATCC 35512 microbial inoculum. The viable bacteria content of the heterotrophic nitrification-aerobic denitrification bacterium Paracoccus ATCC 35512 microbial inoculum is 3 multiplied by 109cfu/g。
A method of microbial preparation for remediation of a soil contaminated with trinitrogen, the method comprising the steps of:
1) uniformly mixing a Pseudomonas stutzeri SC221-M microbial inoculum and an aerobic denitrifying bacterium Paracoccus pantophus ATCC 35512 microbial inoculum according to the mass ratio of 0.1: 1 to obtain a composite microbial inoculum;
2) adding the immobilized glucose oxidase into the composite microbial inoculum prepared in the step 1), wherein the immobilized glucose oxidase accounts for 1/10 of the weight of the microbial preparation, the concentration of the immobilized glucose oxidase is 30U/mg, and uniformly mixing to obtain the microbial preparation for repairing the trinitrogen-contaminated soil.
Example 2
Immobilization of glucose oxidase: adding mesoporous molecular sieve material SBA-15 into glucose oxidase at the temperature of 8 ℃; the pH was 3.5; the rotating speed is 180 rpm; immobilization was carried out for 10 hours.
The preparation method of the Pseudomonas stutzeri SC221-M microbial inoculum comprises the following steps: culturing Pseudomonas stutzeri SC221-M to logarithmic phase, wherein the culture medium is a culture medium which takes wheat straw powder and sweet potato vine powder as carbon sources and takes ammonium sulfate and peptone as nitrogen sources, inoculating the strain cultured to logarithmic phase into the culture medium according to the inoculation amount of 10% of the volume of the culture medium, culturing for 24 hours at 35 ℃, shaking table shaking 180rpm, and obtaining Pseudomonas stutzeri SC221-MAdding bentonite into fermentation liquor of the pseudomonas stutzeri SC221-M, and drying to obtain the pseudomonas stutzeri SC221-M microbial inoculum. The viable bacteria content of the Pseudomonas stutzeri SC221-M microbial inoculum is 8 multiplied by 1010cfu/g。
The preparation method of the bacterium agent of the heterotrophic nitrification-aerobic denitrifying bacterium Paracoccus pantographic ATCC 35512 comprises the following steps: culturing the heterotrophic nitrification-aerobic denitrifying bacteria Paracoccus pantophus ATCC 35512 in an R2A liquid culture medium (purchased from Qingdao Haibobo organisms) to logarithmic phase, then inoculating the strain cultured to logarithmic phase into the R2A liquid culture medium according to the inoculation amount of 6% of the volume of the culture medium, shaking the shaking table at 40 ℃, shaking at 180rpm, culturing for 24 hours, adding activated carbon, and drying to prepare the heterotrophic nitrification-aerobic denitrifying bacteria Paracoccus pantophus ATCC 35512 microbial inoculum. The viable bacteria content of the heterotrophic nitrification-aerobic denitrification bacterium Paracoccus pantophus ATCC 35512 microbial inoculum is 5 multiplied by 1010cfu/g。
A method of microbial preparation for remediation of a soil contaminated with trinitrogen, the method comprising the steps of:
1) the pseudomonas stutzeri SC221-M microbial inoculum and the anaerobic nitrification-aerobic denitrification bacterium Paracoccus pantophus ATCC 35512 microbial inoculum are mixed according to the following ratio of 2: 5 to obtain the composite microbial inoculum;
2) adding the immobilized glucose oxidase into the composite microbial inoculum prepared in the step 1), wherein the immobilized glucose oxidase accounts for 1/5 of the weight of the microbial preparation, the concentration of the immobilized glucose oxidase is 20U/mg, and uniformly mixing to obtain the microbial preparation for repairing the trinitrogen-polluted soil.
Example 3
Immobilization of glucose oxidase: adding mesoporous molecular sieve material SBA-15 into glucose oxidase at the temperature of 12 ℃; the pH was 5.5; the rotating speed is 120 rpm; immobilization was carried out for 12 hours.
The preparation method of the Pseudomonas stutzeri SC221-M microbial inoculum comprises the following steps: culturing Pseudomonas stutzeri SC221-M to logarithmic phase, wherein the culture medium is prepared from wheat straw powder and sweet potato vine powder as carbon source, ammonium sulfate and peptone as nitrogen source, and the volume of the culture medium is 6%Inoculating the strain cultured to the logarithmic phase into a culture medium, culturing for 44 hours at the temperature of 32 ℃ by shaking a table at 200rpm to obtain the fermentation liquor of the pseudomonas stutzeri SC221-M, adding activated carbon into the fermentation liquor, and drying to obtain the pseudomonas stutzeri SC221-M microbial inoculum. The viable bacteria content of the Pseudomonas stutzeri SC221-M microbial inoculum is 4 multiplied by 1010cfu/g。
The preparation method of the bacterium agent of the heterotrophic nitrification-aerobic denitrification bacterium Paracoccus pantophus ATCC 35512 comprises the following steps: culturing the heterotrophic nitrification-aerobic denitrifying bacteria Paracoccus pantophus ATCC 35512 in an R2A liquid culture medium (purchased from Haibobo organisms in Qingdao) to logarithmic phase, then inoculating the strain cultured to logarithmic phase into the R2A liquid culture medium according to the inoculation amount of 4% of the volume of the culture medium, shaking the shaking table at 35 ℃, shaking at 150rpm, culturing for 36 hours, adding activated carbon, and drying to prepare the heterotrophic nitrification-aerobic denitrifying bacteria Paracoccus pantophus ATCC 35512 microbial inoculum. The viable bacteria content of the bacterium agent of the heterotrophic nitrification-aerobic denitrification bacterium Paracoccus pantophus ATCC 35512 is 2.5 multiplied by 1010cfu/g。
A method of microbial preparation for remediation of a soil contaminated with trinitrogen, the method comprising the steps of:
1) uniformly mixing a Pseudomonas stutzeri SC221-M microbial inoculum and an aerobic nitrification-aerobic denitrifying bacterium Paracoccus pantophus ATCC 35512 microbial inoculum according to the mass ratio of 0.1: 5 to obtain a composite microbial inoculum;
2) adding the immobilized glucose oxidase into the composite microbial inoculum prepared in the step 1), wherein the immobilized glucose oxidase accounts for 3/20 of the weight of the microbial preparation, the concentration of the immobilized glucose oxidase is 25U/mg, and uniformly mixing to obtain the microbial preparation for repairing the trinitrogen-polluted soil.
Comparative example 1 composite microbial agent prepared from non-immobilized enzyme preparation and microbial agent
The preparation method of the Pseudomonas stutzeri SC221-M microbial inoculum comprises the following steps: culturing Pseudomonas stutzeri SC221-M to logarithmic phase, wherein the culture medium is prepared by taking wheat straw powder and sweet potato vine powder as carbon sources and taking ammonium sulfate and peptone as nitrogen sources, and then culturing to logarithmic phase according to the inoculation amount of 3% of the volume of the culture mediumInoculating the strain into a culture medium, culturing for 48 hours at 30 ℃ by shaking at 220rpm in a shaking table to obtain a fermentation liquor of the pseudomonas stutzeri SC221-M, adding activated carbon into the fermentation liquor, and drying to obtain the pseudomonas stutzeri SC221-M microbial inoculum. The viable bacteria content of the Pseudomonas stutzeri SC221-M microbial inoculum is 5 multiplied by 108cfu/g。
The preparation method of the bacterium agent of the heterotrophic nitrification-aerobic denitrification bacterium Paracoccus pantophus ATCC 35512 comprises the following steps: culturing the heterotrophic nitrification-aerobic denitrifying bacteria Paracoccus pantophus ATCC 35512 in an R2A liquid culture medium (purchased from Qingdao Haibobo organisms) to logarithmic phase, then inoculating the strain cultured to logarithmic phase into the R2A liquid culture medium according to the inoculation amount of 3% of the volume of the culture medium, shaking the shaking table at 30 ℃, shaking at 180rpm, culturing for 48 hours, adding bentonite, and drying to prepare the heterotrophic nitrification-aerobic denitrifying bacteria Paracoccus pantophus ATCC 35512 microbial inoculum. The viable bacteria content of the heterotrophic nitrification-aerobic denitrification bacterium Paracoccus pantophus ATCC 35512 microbial inoculum is 3 multiplied by 109cfu/g。
A method of microbial preparation for remediation of a soil contaminated with trinitrogen, the method comprising the steps of:
1) uniformly mixing a Pseudomonas stutzeri SC221-M microbial inoculum and an aerobic denitrifying bacterium Paracoccus pantophus ATCC 35512 microbial inoculum according to the mass ratio of 0.1: 1 to obtain a composite microbial inoculum;
2) adding glucose oxidase into the composite microbial inoculum prepared in the step 1), wherein the glucose oxidase accounts for 1/10 of the weight of the microbial preparation, and the concentration of the glucose oxidase is 30U/mg, and uniformly mixing to obtain the microbial preparation for repairing the trinitrogen contaminated soil.
Comparative example 2 Using only Pseudomonas stutzeri SC221-M inoculum and immobilized oxidase
Immobilization of glucose oxidase: adding mesoporous molecular sieve material SBA-15 into glucose oxidase at the temperature of 8 ℃; the pH was 3.5; the rotating speed is 180 rpm; immobilization was carried out for 10 hours.
The preparation method of the Pseudomonas stutzeri SC221-M microbial inoculum comprises the following steps: culturing Pseudomonas stutzeri SC221-M to logarithmic phase in the culture mediumAnd (2) taking wheat straw powder and sweet potato vine powder as a carbon source and ammonium sulfate and peptone as a culture medium of a nitrogen source, inoculating the strain cultured to the logarithmic phase into the culture medium according to the inoculation amount of 10% of the volume of the culture medium, culturing at 35 ℃ and shaking at 180rpm in a shaking table for 24 hours to obtain fermentation liquor of the pseudomonas stutzeri SC221-M, adding activated carbon into the fermentation liquor, and drying to obtain the pseudomonas stutzeri SC221-M microbial inoculum. The viable bacteria content of the Pseudomonas stutzeri SC221-M microbial inoculum is 8 multiplied by 1010cfu/g。
A method of microbial preparation for remediation of a soil contaminated with trinitrogen, the method comprising the steps of:
adding the immobilized glucose oxidase into the pseudomonas stutzeri SC221-M microbial inoculum, wherein the immobilized glucose oxidase accounts for 1/5 of the weight of the pseudomonas stutzeri SC221-M microbial inoculum, and the concentration of the immobilized glucose oxidase is 20U/mg, and uniformly mixing to obtain the microbial preparation for repairing the tri-nitrogen contaminated soil.
Comparative example 3 Using only the bacterium Paracoccus pantophus ATCC 35512 as an aerobic nitrification-aerobic denitrification bacterium and the immobilized oxidase
Immobilization of glucose oxidase: adding mesoporous molecular sieve material SBA-15 into glucose oxidase at the temperature of 12 ℃; the pH was 5.5; the rotating speed is 120 rpm; immobilization was carried out for 12 hours.
The preparation method of the bacterium agent of the heterotrophic nitrification-aerobic denitrification bacterium Paracoccus pantophus ATCC 35512 comprises the following steps: culturing the heterotrophic nitrification-aerobic denitrifying bacteria Paracoccus pantophus ATCC 35512 in an R2A liquid culture medium (purchased from Qingdao Haibobo organisms) to logarithmic phase, then inoculating the strain cultured to logarithmic phase into the R2A liquid culture medium according to the inoculation amount of 4% of the volume of the culture medium, shaking the shaking table at the speed of 150rpm for culturing for 36 hours at the temperature of 35 ℃, adding activated carbon, and drying to prepare the heterotrophic nitrification-aerobic denitrifying bacteria Paracoccus pantophus ATCC 35512 microbial inoculum. The viable bacteria content of the bacterium agent of the heterotrophic nitrification-aerobic denitrification bacterium Paracoccus pantophus ATCC 35512 is 2.5 multiplied by 1010cfu/g。
A method of microbial preparation for remediation of a soil contaminated with trinitrogen, the method comprising the steps of:
adding the immobilized glucose oxidase into an anaerobic nitrification-aerobic denitrification bacterium Paracoccus pantographic ATCC 35512 microbial inoculum, wherein the immobilized glucose oxidase accounts for 3/20 of the weight of the anaerobic nitrification-aerobic denitrification bacterium Paracoccus pantographic ATCC 35512 microbial inoculum, the concentration of the immobilized glucose oxidase is 25U/mg, and the immobilized glucose oxidase is uniformly mixed to obtain the microbial preparation for repairing the three-nitrogen polluted soil.
Example 4
The method is suitable for measuring ammonia nitrogen, nitrite nitrogen and nitrate nitrogen in soil according to the standard of HJ 636 & 2012 & lt & gt solution extraction-spectrophotometry for measuring ammonia nitrogen, nitrite nitrogen and nitrate nitrogen in soil published by the national environmental ministry.
Sample pretreatment: extracting the trinitrogen in the soil by using a potassium chloride solution, centrifugally separating the extracting solution, and taking the supernatant for analysis.
1. And (3) measuring ammonia nitrogen: reacting ammonia ions with phenol → blue indophenol dye → 630nm spectrophotometer;
2. determination of nitrite nitrogen: reacting nitrate nitrogen with sulfanilamide → diazonium salt → coupled reaction with hydrochloric acid N- (1-naphthyl) -ethylenediamine → red dye → 543nm spectrophotometer for determination;
3. determination of nitrate nitrogen: nitrate nitrogen reduction → nitrite nitrogen → colorimetric reaction in the same manner as above → 543nm spectrophotometer measurement (total content of nitrite nitrogen + nitrate) → total content of nitrite nitrogen-total content of nitrate nitrogen
The microbial preparation for repairing the soil polluted by the trinitrogen is used for testing the degradation capability of ammonia nitrogen, nitrite nitrogen and nitrate nitrogen in the soil:
1) selecting soil polluted by ammonia nitrogen, nitrite nitrogen, nitrate nitrogen and the like, averagely dividing the soil into 18 parts, respectively placing the 18 parts in a sterilized container, and dividing the 18 parts of sterilized soil into 6 groups of 3 parts each.
2) Taking the microbial preparations for repairing the soil polluted by the three nitrogen prepared in the examples 1-3 and the comparative examples 1-3, adding the microbial preparation for repairing the soil polluted by the three nitrogen prepared in each example/comparative example into one group of soil correspondingly, and uniformly mixing, wherein the mass of the microbial preparation for repairing the soil polluted by the three nitrogen added into the soil accounts for 0.5% of the mass of the soil.
3) The contents of ammonia nitrogen, nitrite nitrogen and nitrate nitrogen in the treated soil were measured two months later, and the average value of 3 parts of each group was obtained, and the results are shown in table 1.
TABLE 1
Claims (6)
1. A microbial preparation for repairing three-nitrogen contaminated soil is characterized by comprising a Pseudomonas stutzeri SC221-M microbial inoculum and an aerobic denitrifying bacteriumParacoccus pantotrophusATCC 35512 microbial inoculum and immobilized glucose oxidase, the Pseudomonas stutzeri SC221-M microbial inoculum and the heterotrophic nitrification-aerobic denitrification bacteriaParacoccus pantotrophusThe weight ratio of ATCC 35512 microbial inoculum is 0.1-2: 1-5, the immobilized glucose oxidase accounts for 1/10-1/5 of the weight of the microbial preparation, and the viable bacteria content of the SC221-M microbial inoculum of the pseudomonas stutzeri in the microbial preparation is 5 multiplied by 108~8×1010 cfu/g, said heterotrophic nitrification-aerobic denitrification bacteriaParacoccus pantotrophusThe viable bacteria content of ATCC 35512 microbial inoculum is 3 multiplied by 109~5×1010 cfu/g, wherein the concentration of the immobilized glucose oxidase in the microbial preparation is 20-30U/mg.
2. A method of preparing the microbial preparation of claim 1 for remediation of a soil contaminated with trinitrogen, the method comprising the steps of:
1) the pseudomonas stutzeri SC221-M microbial inoculum and the heterotrophic nitrification-aerobic denitrification bacteriaParacoccus pantotrophusThe ATCC 35512 microbial inoculum is uniformly mixed according to the mass ratio of 0.1-2: 1-5 to obtain a composite microbial inoculum;
2) adding the immobilized glucose oxidase into the composite microbial inoculum prepared in the step 1), and uniformly mixing to obtain the microbial preparation for repairing the trinitrogen polluted soil.
3. The method for preparing a microbial preparation for remediating a trinitrogen-contaminated soil according to claim 2, wherein the preparation method of the pseudomonas stutzeri SC221-M microbial inoculum comprises: culturing Pseudomonas stutzeri SC221-M to a logarithmic phase, inoculating strains cultured to the logarithmic phase into a culture medium according to an inoculation amount of 3-10% of the volume of the culture medium, oscillating in a shaking table at 30-35 ℃ at 180-220 rpm, culturing for 40-48 hours to obtain a fermentation broth of Pseudomonas stutzeri SC221-M, adding an adsorbent into the fermentation broth, and drying to obtain a Pseudomonas stutzeri SC221-M microbial inoculum; the adsorbent is one or more of bentonite and activated carbon.
4. The method for preparing a microbial preparation for remediating tri-nitrogen contaminated soil as claimed in claim 3, wherein the culture medium is a culture medium in which wheat straw powder and sweet potato vine powder are used as a carbon source, and ammonium sulfate and peptone are used as a nitrogen source.
5. The method for preparing a microbial preparation for remediating a trinitrogen-contaminated soil as set forth in claim 4, wherein the heterotrophic nitrification-aerobic denitrification bacteriaParacoccus pantotrophusThe preparation method of the ATCC 35512 microbial inoculum comprises the following steps: nitrifying the heterotrophic bacteria and denitrifying the bacteria aerobicallyParacoccus pantotrophusATCC 35512 is cultured in an R2A liquid culture medium to logarithmic phase, then the strain cultured to logarithmic phase is inoculated into the R2A liquid culture medium according to the inoculation amount of 3-6% of the volume of the culture medium, the strain is shaken in a shaking table at 30-40 ℃ at 120-180 rpm and cultured for 24-48 hours, an adsorbent is added, and the obtained product is dried to prepare the heterotrophic nitrification-aerobic denitrification bacteriaParacoccus pantotrophusATCC 35512 microbial inoculum; the adsorbent is one or more of bentonite or activated carbon.
6. Use of the microbial preparation for remediating triaza-contaminated soil as set forth in claim 1 for remediating triaza-contaminated soil.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102876617A (en) * | 2012-10-18 | 2013-01-16 | 浙江大学 | Pseudomonas and purpose of pseudomonas |
| CN105274024A (en) * | 2015-09-28 | 2016-01-27 | 张胜平 | Microbial preparation for processing industrial fermentation wastewater |
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| Title |
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| Analysis of the activation mechanism of Pseudomonas stutzeri cytochrome c peroxidase through an electron transfer chain;de Sousa等;《JOURNAL OF BIOLOGICAL INORGANIC CHEMISTRY》;20110831;第16卷(第6期);第881-888页 * |
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