CN114854620A - N-methylpyrrolidone degrading strain and application thereof - Google Patents
N-methylpyrrolidone degrading strain and application thereof Download PDFInfo
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- SECXISVLQFMRJM-UHFFFAOYSA-N N-Methylpyrrolidone Chemical compound CN1CCCC1=O SECXISVLQFMRJM-UHFFFAOYSA-N 0.000 title claims abstract description 87
- 230000000593 degrading effect Effects 0.000 title claims abstract description 18
- 230000015556 catabolic process Effects 0.000 claims abstract description 25
- 238000006731 degradation reaction Methods 0.000 claims abstract description 25
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- 241000194108 Bacillus licheniformis Species 0.000 claims abstract description 16
- 238000004321 preservation Methods 0.000 claims abstract description 13
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- 235000007079 manganese sulphate Nutrition 0.000 claims description 3
- 239000011702 manganese sulphate Substances 0.000 claims description 3
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 claims description 3
- 235000019319 peptone Nutrition 0.000 claims description 3
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- 239000002351 wastewater Substances 0.000 abstract description 17
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- 230000001580 bacterial effect Effects 0.000 abstract description 4
- 238000001514 detection method Methods 0.000 abstract description 4
- HNJBEVLQSNELDL-UHFFFAOYSA-N pyrrolidin-2-one Chemical compound O=C1CCCN1 HNJBEVLQSNELDL-UHFFFAOYSA-N 0.000 abstract 1
- 241000193830 Bacillus <bacterium> Species 0.000 description 8
- 229910017053 inorganic salt Inorganic materials 0.000 description 5
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 4
- 229920006231 aramid fiber Polymers 0.000 description 4
- 238000005516 engineering process Methods 0.000 description 4
- 108010055527 nuclear matrix protein 2 Proteins 0.000 description 4
- 239000000243 solution Substances 0.000 description 4
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- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 3
- XKMRRTOUMJRJIA-UHFFFAOYSA-N ammonia nh3 Chemical compound N.N XKMRRTOUMJRJIA-UHFFFAOYSA-N 0.000 description 3
- 230000007613 environmental effect Effects 0.000 description 3
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- WHXSMMKQMYFTQS-UHFFFAOYSA-N Lithium Chemical compound [Li] WHXSMMKQMYFTQS-UHFFFAOYSA-N 0.000 description 1
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
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- C12N1/20—Bacteria; Culture media therefor
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- C02F2101/38—Organic compounds containing nitrogen
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- Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
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Abstract
The invention belongs to the technical field of bioengineering, and provides a degrading strain of N-methylpyrrolidone and an application thereof, wherein the strain is Bacillus licheniformis (Bacillus licheniformis), the strain code is YJY22-06, and the preservation number is CGMCC No. 24315; the microbial inoculum obtained by fermenting the strain is directly put into an aerobic tank, the existing treatment process is adopted to show a very obvious degradation effect on high-concentration N-methylpyrrolidone, and the initial concentration of the microbial inoculum is 20000mg L after 12h detection ‑1 The degradation rate of the N-methyl pyrrolidone reaches 100 percent, and the N-methyl pyrrolidone can be applied to 50000mg L in 40h ‑1 The degradation rate of the N-methyl pyrrolidone reaches more than 80 percent, and the bacterial strain and the method can greatly improve the high-concentration N-methylpyridineThe biochemical treatment efficiency of the pyrrolidone waste water.
Description
Technical Field
The invention belongs to the technical field of bioengineering, and provides a degrading strain of N-methylpyrrolidone and application thereof.
Background
N-Methyl-2-pyrrolidone (NMP) is often used as a typical organic polar solvent with strong selectivity and good stability due to its characteristics of low volatility, high polarity, non-corrosiveness, good water solubility and the like, and is an important chemical raw material. N-methylpyrrolidone is widely used in various manufacturing industries such as lithium batteries, dyes, medicines, detergents, adhesives, and the like. The N-methyl pyrrolidone is easy to be discharged into the environment along with wastewater due to the characteristics of good water miscibility, but the N-methyl pyrrolidone is stably transferred in soil and underground water and is not easy to be decomposed under natural conditions due to the characteristics of structural stability, poor biodegradability, strong biological toxicity and the like of the N-methyl pyrrolidone, and the N-methyl pyrrolidone which does not reach the discharge index is released into the environment to cause irreparable harm to an ecosystem and a water area environment, so how to efficiently and quickly eliminate the environmental pollution caused by the N-methyl pyrrolidone becomes an environmental problem to be solved urgently.
At present, the treatment method of the wastewater containing the N-methyl pyrrolidone is mainly a physical and chemical method, and the method has high cost and is easy to cause secondary environmental pollution; in addition, the biological treatment technology has the characteristics of high efficiency, economy, no secondary pollution and the like, can realize non-toxic and harmless environmental management, and is an environmental treatment technology which is widely applied at present. Therefore, screening effective strains capable of eliminating N-methyl pyrrolidone in the environment is the most direct and effective method for solving the problem of environmental pollution of N-methyl pyrrolidone.
Through the search of the inventor, the application of Chinese patent application No. 201610071081.3 discloses a Bacillus subtilis N-methylpyrrolidone-degrading NMP-2 and application thereof, wherein the strain is Bacillus subtilis, and the concentration of N-methylpyrrolidone degrading is 500mg L -1 The degradation rate of the N-methyl pyrrolidone reaches 98 percent in 72 hours; application No. 201910209582.7 discloses Enterobacter for degrading N-methylpyrrolidone and application thereof in wastewater treatment, wherein the degradation concentration of the strain is 2800- -1 ，3000mg L -1 The N-methyl pyrrolidone can be completely degraded in 55 hours; the application No. 202011483934.7 discloses N-methyl pyrrolidone degrading bacteria and application thereof in wastewater treatment, wherein the degrading concentration of the degrading strain bacillus is 4.7-13.6mM, and 12mM of N-methyl pyrrolidone can be completely removed within 23 h; the strains can degrade N-methylpyrrolidone, but the degradation concentration is lower (500-3000mg L) -1 ) And the time is longer, namely 23-72h, and the lower efficiency is difficult to realize wide industrial application.
Therefore, whether better N-methylpyrrolidone degrading strains can be obtained becomes one of the important tasks for researchers.
Disclosure of Invention
Aiming at the defects of the technology, the invention provides a degrading strain of N-methylpyrrolidone and an application thereof, wherein the strain is Bacillus licheniformis (Bacillus licheniformis), the strain code is YJY22-06, and the preservation number is CGMCC No. 24315; the microbial inoculum obtained by fermenting the strain is directly put into an aerobic tank, the existing treatment process is adopted to generate very obvious degradation effect on high-concentration N-methylpyrrolidone, and the initial concentration of the microbial inoculum is 20000mg L after 12h of detection -1 The degradation rate of the N-methyl pyrrolidone reaches 100 percent, and the biochemical treatment efficiency of the high-concentration N-methyl pyrrolidone wastewater can be greatly improved by utilizing the strain and the method.
The specific technical scheme provided by the invention is as follows:
firstly, a new Bacillus licheniformis (Bacillus licheniformis) is obtained and named as YJY22-06, and the strain has the morphological characteristics that: the bacterial colony is small and loose, and the edge is irregular; the 16S rDNA nucleotide complete sequence is shown as Seq ID No:1, the 16S rDNA sequence is subjected to BLAST comparison, and the result shows that the nucleotide sequence of the 16S rDNA of the strain has more than 99 percent of homology with the nucleotide sequence of different strains of Bacillus (Bacillus) and has 100 percent of homology with the strain in which the specific mark is Bacillus licheniformis (Bacillus licheniformis).
The inventor carries out biological preservation on the strain in China general microbiological culture Collection center with the preservation number of CGMCC No.24315, and the strain is detected to be in a survival state.
The inventor conducts experiments on the functions of the strain, and the result shows that compared with the prior disclosed N-methyl pyrrolidone treatment strain, the concentration of pollutants treated by the strain is improved by 5-40 times, the time is shortened by 2-10 times, the strain has better tolerance to wastewater with different concentrations, and the high-efficiency low-cost treatment of high-concentration N-methyl pyrrolidone wastewater in industries such as aramid fiber and the like is realized.
Furthermore, the inventor provides a production method of the microbial inoculum which can be applied industrially, and the specific steps are as follows:
(1) activation of strains: taking out the preserved strain, and activating at room temperature for 2-4 h; the strain is YJY 22-06;
(2) preparing a seed solution: in a clean bench, selecting strains in a test tube inclined plane, directly inoculating the strains into 100ml of sterile LB liquid culture medium, culturing at 30-37 ℃ for 16-20h at 180-200rpm to prepare seed liquid;
(3) fermentation: adding a fermentation medium into a fermentation tank, sterilizing at 121 ℃ for 0.5 hour, inoculating seed liquid according to the volume ratio of 0.2%, controlling the temperature to be 30-37 ℃ in the fermentation process, controlling the tank pressure to be 0.04-0.05MPa, starting rotating speed to be 180 and 200rpm, dissolved oxygen being larger than or equal to 20%, and gas-liquid ratio to be 1: 1; when the dissolved oxygen is reduced to be below 20 percent, the pH value is increased to 8.8, and the microbial inoculum is obtained after the fermentation is finished;
the formula of the fermentation medium comprises the following components in percentage by weight: 1.0-1.5% of glucose, 0.3-0.8% of corn starch, 3.0-4.2% of soybean meal, 0.3-0.6% of peptone, 0.3-0.6% of ammonium sulfate, 0.3-0.7% of yeast powder, 0.02-0.04% of manganese sulfate and the balance of deionized water.
When in specific application, the microbial inoculum obtained by fermentation is added into an aerobic tank for biochemical treatment according to the liquid volume ratio of 0.1-3% in the aerobic tank.
The aerobic tank is generally referred to asA 2 The O aerobic tank in the O treatment system can also be applied to aerobic tanks in other N-methylpyrrolidone-containing sewage treatment systems, and the main reason is that the strain provided by the application is an aerobic strain and has a good treatment effect in the aerobic tanks.
Through field experiment determination, after the raw materials are added into an aerobic tank according to the dosage, the raw materials are treated for 12 hours, and the initial concentration is 20000mg L -1 The degradation rate of the N-methyl pyrrolidone reaches 100 percent, and the N-methyl pyrrolidone can be applied to 50000mg L after being treated for 40 hours -1 The degradation rate of the N-methyl pyrrolidone reaches more than 80 percent.
In conclusion, the microbial inoculum prepared by the strain provided by the invention is applied to the biological treatment of aramid paper wastewater, the strain has very good degradation property on N-methylpyrrolidone, and the initial concentration of 20000mg L in 12h -1 The degradation rate of the N-methylpyrrolidone reaches 100 percent, and the biochemical treatment efficiency of the high-concentration N-methylpyrrolidone wastewater can be greatly improved by utilizing the strain and the method.
Preservation information
Preservation time: 1 month and 17 days 2022
The name of the depository: china general microbiological culture Collection center
The preservation number is: CGMCC No.24315
The address of the depository: microbial research institute of western road 1 institute No. 3 of China academy of sciences, Beijing, Chaoyang
Classification nomenclature Bacillus licheniformis (Bacillus licheniformis)
Drawings
FIG. 1 is a schematic diagram showing the degradation effect of seed liquid of the strain in example 3 on N-methylpyrrolidone;
FIG. 2 is a graph showing the effect of seed liquid of the strain of example 4 on the degradation of N-methylpyrrolidone at higher concentration;
fig. 3 is a schematic diagram of the degradation effect of the microbial inoculum in the aramid paper production wastewater in example 5 on N-methylpyrrolidone.
Detailed Description
The present invention will be described in further detail with reference to the following examples, but it should not be construed that the scope of the above subject matter is limited to the following examples. All the technologies realized based on the above contents of the present invention belong to the scope of the present invention, except for the specific description, the following embodiments are all completed by the conventional prior art, and all the strains adopted in the following embodiments are the strains with the preservation number of CGMCC No. 24315.
The media used in the following examples consisted of:
LB medium composition: tryptone 10g L -1 Yeast powder 5g L -1 Sodium chloride 10g L -1 pH 7.2; the LB solid medium was supplemented with 1.5 wt% agar based on the LB medium described above.
The inorganic salt culture medium comprises the following components: k 2 HPO 4 0.6g L -1 ，KH 2 PO 4 0.5g L -1 ，NaCl 0.1g L -1 ， MgSO 4 0.06g L -1 ，CaCl 2 0.08g L -1 ，FeSO 4 0.008g L -1 ，MnSO 4 0.008g L -1 ， pH 7.2。
If N-methylpyrrolidone is required to be added, it can be added according to specific requirements, and the content is generally controlled to be 20000-50000mg L -1 。
After the preparation of the culture medium, the culture medium is sterilized in a high-pressure steam sterilization pot at the temperature of 121 ℃ for 20min, wherein the addition of N-methyl pyrrolidone is carried out on the inorganic salt culture medium after the sterilization is finished.
Example 1 obtaining of the Strain
Sampling 1 g of a sample from a factory wastewater biochemical treatment device of a new material company Limited in Shandong, adding the sample into 100mL of physiological saline sterilized at 121 ℃ for 20min, uniformly mixing the sample with 180rpm of a shaking table, standing the mixture for 2h after 30min, sucking supernatant, and diluting the supernatant to 10 ℃ by using sterile water in a gradient manner -3 -10 -10 And (3) coating the mixture in LB solid culture medium added with N-methyl pyrrolidone, and culturing the mixture in a biochemical incubator at 35 ℃ for 24 hours.
Selecting single colonies with obvious differences on a culture dish, performing purification culture by adopting a plate streaking separation method, continuously purifying for 3 times to obtain 6 single strains in total, and storing the single strains on a slant and a glycerol tube; the subsequent N-methyl pyrrolidone degradation experiment research is respectively carried out on 6 strains to be selected in an inorganic salt liquid culture medium containing N-methyl pyrrolidone, and finally a strain which has high N-methyl pyrrolidone degradation efficiency, easy culture and stable passage characteristic is screened out, and the morphological characteristics are as follows: the bacterial colony is small, loose and irregular in edge, and is named as YJY 22-06.
The inventors performed 16S rDNA sequencing, the nucleotide sequence of which is shown in Seq ID No:1, and BLAST alignment of the 16S rDNA sequence, and the results showed that the nucleotide sequence of the 16S rDNA of the strain has more than 99% homology with the nucleotide sequence of different strains of Bacillus (Bacillus), and 100% homology with the strain in which the specific marker is Bacillus licheniformis (Bacillus licheniformis). And the biological preservation is carried out in the common microorganism center of China Committee for culture Collection of microorganisms, the preservation number is CGMCC No.24315, and the preservation is detected to be in a survival state.
EXAMPLE 2 fermentation of the Strain
(1) Activating strains: taking out the strain test tube stored in the slant nutrient agar culture medium in a refrigerator at 4 ℃, and activating for 2-4h at room temperature.
(2) Preparing liquid seeds: in an ultra-clean workbench, selecting strains in a test tube inclined plane, directly inoculating the strains into 100ml of sterile LB liquid culture medium, and culturing at 180rpm and 32 ℃ for 16h to prepare seed liquid;
(3) fermentation: adding a fermentation culture medium which accounts for 1/2-2/3 of the volume of the fermentation tank into the fermentation tank, sterilizing, inoculating seed liquid according to the volume ratio of 0.2%, controlling the temperature to be 35 ℃, the tank pressure to be 0.05MPa, the initial rotation speed to be 180rpm, the dissolved oxygen to be not less than 20% and the gas-liquid ratio to be 1:1 in the fermentation process; when the dissolved oxygen is reduced to 20 percent and the pH value is increased to 8.8, the microbial inoculum is obtained after the fermentation is finished, and the viable count of the obtained microbial inoculum is about 10 9 cfu/ml；
The sterilization mode is preferably sterilization at 121 ℃ for 0.5 hour;
the formula of the fermentation medium is as follows: 1.5% of glucose, 0.5% of corn starch, 4.0% of soybean meal, 0.5% of peptone, 0.4% of ammonium sulfate, 0.5% of yeast powder, 0.03% of manganese sulfate and the balance of deionized water.
Example 3 verification of the Effect of the strains
The YJY22-06 seed liquid obtained in the step (2) of the example 2 is directly added into the seed liquid containing 20000mg L -1 In an inorganic salt culture medium of N-methylpyrrolidone, the inoculation amount is 1 percent by volume, the culture is carried out under the conditions of 35 ℃ and 180rpm, a blank control which is not inoculated with YJY22-06 is established, when the measurement is carried out for 12h, the N-methylpyrrolidone in the culture medium is degraded by 100 percent, and the residual amount of the N-methylpyrrolidone in a blank sample is basically unchanged (figure 1).
EXAMPLE 4 degradation Effect of the Strain on high concentration of N-methylpyrrolidone
The YJY22-06 seed solution obtained in the step (2) of example 2 was directly added to a solution containing 50000mg of L -1 In an inorganic salt culture medium of N-methylpyrrolidone, the inoculation amount is 1 percent by volume, the culture is carried out under the conditions of 34 ℃ and 180rpm, a blank control which is not inoculated with YJY22-06 is set up, tracking detection is carried out, and the N-methylpyrrolidone in the culture medium is 9539.58mg L when the determination is carried out for 40h -1 The degradation rate is 80.92%, and the residual quantity of N-methylpyrrolidone in the blank sample is basically not changed (figure 2), which shows that the strain YJY22-06 provided by the invention is applied to 50000mg L -1 The degradation rate of the N-methyl pyrrolidone can reach 80 percent in 40 hours.
Example 5 treatment of aramid fiber paper production wastewater with the bacterial strain
The biochemical treatment device for the waste water generated in the production of the aramid fiber paper is subjected to high-content N-methyl pyrrolidone (the initial content is 18543.65mg L) -1 ) The invention artificially verifies the treatment effect of the strain, namely adding the microbial inoculum prepared in the step (3) in the embodiment 2 into an aerobic pool according to the proportion of 0.2 percent of the volume ratio of liquid in the aerobic pool to verify the degradation effect of the N-methyl pyrrolidone, wherein the temperature of the aerobic pool is 32 ℃, and the dissolved oxygen is 4mg L -1 The pH value is 7.55, the hydraulic retention time is 12 hours, and the content of N-methyl pyrrolidone in a detection system is tracked;
the results show that the N-methylpyrrolidone content in the system is from the initial 18543.65mg L -1 Degraded into0mg L -1 (figure 3), the effluent index of the system is recovered stably, and the YJY22-06 strain greatly relieves the impact of N-methylpyrrolidone on a sewage treatment system.
Comparative example 1
When the microbial inoculum is not added to a certain aramid fiber paper production wastewater biochemical treatment device in embodiment 5, the treatment concentration is 500mg L -1 The N-methyl pyrrolidone can normally run, the hydraulic retention time is 12h, and the N-methyl pyrrolidone in the effluent is 0mg L -1 0.53mg L of ammonia nitrogen -1 The effluent index is qualified; when receiving the wastewater with higher concentration of N-methyl pyrrolidone (800- -1 ) In time, the ammonia nitrogen in the effluent is 263.2mg L after the hydraulic retention time is 12h -1 Serious standard exceeding;
in order to verify the function of the microbial inoculum, the wastewater (800- -1 ) When the biological treatment is carried out, the YJY22-06 microbial inoculum prepared in the step (3) in the embodiment 2 is added into an aerobic tank for biochemical treatment according to the liquid volume ratio of 0.2 percent in the aerobic tank, the temperature of the aerobic tank is 33 ℃, and the dissolved oxygen is 4mg L -1 pH 7.55; after 2h of treatment, the N-methylpyrrolidone in the effluent of the system is reduced to 0mg L -1 0.26mg L of ammonia nitrogen -1 And the effluent index is qualified.
Therefore, after the wastewater with high concentration of N-methylpyrrolidone is impacted, the YJY22-06 strain can better maintain the treatment effect than the original treatment strain in the aerobic pool, can degrade the N-methylpyrrolidone in a short time, cannot influence the treatment effect on other pollutants, and can realize qualified effluent indexes in the shortest time.
Comparative example 2
The application of Chinese patent application No. 201610071081.3 discloses N-methylpyrrolidone degrading bacillus NMP-2 and application thereof, wherein the concentration of N-methylpyrrolidone degraded by bacillus is 500mg L -1 The degradation rate of the N-methyl pyrrolidone reaches 98 percent in 72 hours;
the YJY22-06 of the invention can regulate the concentration to 20000mg L -1 The degradation of N-methylpyrrolidone is completed in 12h, and the N-methylpyrrolidone reacts with the bacillusCompared with the strain NMP-2, the degradation concentration of the YJY22-06 strain to N-methylpyrrolidone is improved by 40 times;
can be applied to the solution containing N-methyl pyrrolidone with the concentration of 1000mg L within 2h -1 The following pollutants are completely degraded, and the time is shortened by 36 times on the contrary under the condition that the concentration of N-methylpyrrolidone is doubled compared with that of the bacillus NMP-2 treatment, so that the effect is obviously improved.
According to the comparison, the strain and the method can greatly improve the biochemical treatment efficiency of the high-concentration N-methylpyrrolidone wastewater.
Sequence listing
<110> Jingbo chemical research institute of yellow river delta Ltd
<120> N-methylpyrrolidone degrading strain and application thereof
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1140
<212> DNA
<213> Bacillus licheniformis (Bacillus licheniformis)
<400> 1
cgatcacttc ggcggctggc tcctaaaagg ttacctcacc gacttcgggt gttacaaact 60
ctcgtggtgt gacgggcggt gtgtacaagg cccgggaacg tattcaccgc ggcatgctga 120
tccgcgatta ctagcgattc cagcttcacg cagtcgagtt gcagactgcg atccgaactg 180
agaacagatt tgtgggattg gcttaacctc gcggtttcgc tgccctttgt tctgtccatt 240
gtagcacgtg tgtagcccag gtcataaggg gcatgatgat ttgacgtcat ccccaccttc 300
ctccggtttg tcaccggcag tcaccttaga gtgcccaact gaatgctggc aactaagatc 360
aagggttgcg ctcgttgcgg gacttaaccc aacatctcac gacacgagct gacgacaacc 420
atgcaccacc tgtcactctg cccccgaagg ggacgtccta tctctaggat tgtcagagga 480
tgtcaagacc tggtaaggtt cttcgcgttg cttcgaatta aaccacatgc tccaccgctt 540
gtgcgggccc ccgtcaattc ctttgagttt cagtcttgcg accgtactcc ccaggcggag 600
tgcttaatgc gttagctgca gcactaaggg gcggaaaccc cctaacactt agcactcatc 660
gtttacggcg tggactacca gggtatctaa tcctgttcgc tccccacgct ttcgctcctc 720
agcgtcagtt acagaccaga gagtcgcctt cgccactggt gttcctccac atctctacgc 780
atttcaccgc tacacgtgga attccactct cctcttctgc actcaagttc cccagtttcc 840
aatgaccctc cccggttgag ccgggggctt tcacatcaga cttaagaaac cgcctgcgag 900
ccctttacgc ccaataattc cggacaacgc ttgccaccta cgtattaccg cagcctgctg 960
gcccgtagtt agccgtggcc tttctggatt aggtaccgtc aggtatcggc cctattcgaa 1020
cggtacctgg tcttccctta tcacagagcc cttacgatcc gaaaaccttc atcacctcac 1080
gcggacgtgc ctacgctgaa cttttcgtcc aattgcgtag aatcccctac gatctgtcct 1140
Claims (4)
- A degrading strain of N-methylpyrrolidone, characterized in that: the strain belongs to Bacillus licheniformis (Bacillus licheniformis), the code of the strain is YJY22-06, the preservation number is CGMCC No.24315, and the 16S rDNA nucleotide complete sequence is shown as Seq ID No: 1.
- 2. A production method of a degrading microbial inoculum of N-methylpyrrolidone is characterized by comprising the following steps: the method comprises the following specific steps:(1) activation of strains: taking out the preserved strain, and activating at room temperature for 2-4 h; the strain is YJY22-06 with the preservation number of CGMCC No. 24315;(2) preparing a seed solution: in a clean bench, selecting strains in a test tube inclined plane, directly inoculating the strains into 100ml of sterile LB liquid culture medium, culturing at 30-37 ℃ for 16-20h at 180-200rpm to prepare seed liquid;(3) fermentation: adding a fermentation culture medium into a fermentation tank, sterilizing at 121 ℃ for 0.5 hour, inoculating seed liquid according to the volume ratio of 0.2%, controlling the temperature at 30-37 ℃ in the fermentation process, controlling the tank pressure at 0.04-0.05MPa, controlling the initial rotation speed at 180-200rpm, controlling the dissolved oxygen content to be equal to or greater than 20%, and controlling the gas-liquid ratio to be 1: 1; when the dissolved oxygen is reduced to be below 20 percent, the pH value is increased to 8.8, and the microbial inoculum is obtained after the fermentation is finished;the formula of the fermentation medium comprises the following components in percentage by weight: 1.0-1.5% of glucose, 0.3-0.8% of corn starch, 3.0-4.2% of soybean meal, 0.3-0.6% of peptone, 0.3-0.6% of ammonium sulfate, 0.3-0.7% of yeast powder, 0.02-0.04% of manganese sulfate and the balance of deionized water.
- 3. Use of the degrading strain of N-methylpyrrolidone according to claim 1 for degrading N-methylpyrrolidone.
- 4. The application of the N-methylpyrrolidone degrading microbial inoculum according to claim 3 in N-methylpyrrolidone degradation, which is characterized in that:the method comprises the step of directly adding the microbial inoculum according to claim 2 into an aerobic tank in the N-methylpyrrolidone-containing sewage treatment system, wherein the adding amount of the microbial inoculum is 0.1-3% of the volume of liquid in the aerobic tank.
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