CN111690563A - Salt-tolerant staphylococcus lentus and application thereof in industrial NMP wastewater treatment - Google Patents
Salt-tolerant staphylococcus lentus and application thereof in industrial NMP wastewater treatment Download PDFInfo
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- 241000147121 Staphylococcus lentus Species 0.000 title claims abstract description 61
- 238000004065 wastewater treatment Methods 0.000 title claims abstract description 12
- 239000002351 wastewater Substances 0.000 claims abstract description 72
- 230000015556 catabolic process Effects 0.000 claims abstract description 17
- 238000006731 degradation reaction Methods 0.000 claims abstract description 17
- 238000004321 preservation Methods 0.000 claims abstract description 10
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 32
- 238000000855 fermentation Methods 0.000 claims description 32
- 230000004151 fermentation Effects 0.000 claims description 32
- 230000001954 sterilising effect Effects 0.000 claims description 28
- 238000012258 culturing Methods 0.000 claims description 20
- 241001052560 Thallis Species 0.000 claims description 19
- 150000003839 salts Chemical class 0.000 claims description 19
- 241000894006 Bacteria Species 0.000 claims description 18
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- 108010080698 Peptones Proteins 0.000 claims description 18
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 18
- 239000007788 liquid Substances 0.000 claims description 18
- 235000019319 peptone Nutrition 0.000 claims description 18
- 239000011780 sodium chloride Substances 0.000 claims description 16
- 241000191940 Staphylococcus Species 0.000 claims description 15
- 229940041514 candida albicans extract Drugs 0.000 claims description 15
- 239000012138 yeast extract Substances 0.000 claims description 15
- 238000001816 cooling Methods 0.000 claims description 14
- 239000000843 powder Substances 0.000 claims description 14
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- 238000011081 inoculation Methods 0.000 claims description 7
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- 239000008103 glucose Substances 0.000 claims description 6
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- 238000005070 sampling Methods 0.000 claims description 5
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- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 claims description 4
- 230000008859 change Effects 0.000 claims description 4
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- 239000010802 sludge Substances 0.000 abstract description 2
- SECXISVLQFMRJM-UHFFFAOYSA-N N-Methylpyrrolidone Chemical compound CN1CCCC1=O SECXISVLQFMRJM-UHFFFAOYSA-N 0.000 description 52
- 239000002609 medium Substances 0.000 description 18
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- 101100074998 Neurospora crassa (strain ATCC 24698 / 74-OR23-1A / CBS 708.71 / DSM 1257 / FGSC 987) nmp-2 gene Proteins 0.000 description 2
- 230000001580 bacterial effect Effects 0.000 description 2
- 238000006065 biodegradation reaction Methods 0.000 description 2
- 239000001110 calcium chloride Substances 0.000 description 2
- 229910001628 calcium chloride Inorganic materials 0.000 description 2
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- 230000002401 inhibitory effect Effects 0.000 description 1
- 229910017053 inorganic salt Inorganic materials 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
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Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- 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
- C12N1/205—Bacterial isolates
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/44—Staphylococcus
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- 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
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2101/00—Nature of the contaminant
- C02F2101/30—Organic compounds
- C02F2101/34—Organic compounds containing oxygen
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2101/00—Nature of the contaminant
- C02F2101/30—Organic compounds
- C02F2101/38—Organic compounds containing nitrogen
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N2030/022—Column chromatography characterised by the kind of separation mechanism
- G01N2030/027—Liquid chromatography
Abstract
A salt-tolerant staphylococcus lentus and application thereof in industrial NMP wastewater treatment belong to the technical field of industrial wastewater treatment. The staphylococcus lentus CCZU-X is preserved in the China general microbiological culture Collection center of the China Committee for culture Collection of microorganisms with the preservation number of CGMCC No.19016, can be applied to degradation of NMP in industrial NMP salt-containing wastewater, and particularly has the following degradation culture conditions when the NMP is degraded: the temperature is 20-40 ℃, the pH is 5-9, and the concentration of the NMP after treatment is greatly reduced. Compared with the traditional method, the strain and the application method thereof have the advantages of high treatment efficiency, wide application range, simple operation, low requirement on equipment and obviously reduced operation cost, and can solve the problem of low efficiency of the traditional activated sludge method for treating industrial NMP salt-containing wastewater.
Description
Technical Field
The invention relates to the technical field of industrial wastewater treatment, in particular to a salt-tolerant staphylococcus lentus and application thereof in industrial NMP wastewater treatment.
Background
Methyl Pyrrolidone (NMP) is needed in the processes of natural gas recovery, lubricating oil refining, polymer reaction and the like, and NMP remained in waste water pollutes the environment.
At present, in the aspect of NMP wastewater treatment, physical and chemical methods are generally adopted to separate NMP in wastewater. These methods all have the problems of high energy consumption, low efficiency, secondary pollution and the like. Biological treatment is one of the most effective methods at present. The NMP has good microbial degradability, and the content of the NMP in the wastewater is reduced through the degradation of the microbes. However, industrial NMP wastewater contains salts with higher concentration, which has an inhibitory effect on the growth of common microorganisms, so that the conventional activated sludge method cannot effectively treat the wastewater.
The physical and chemical recovery method is usually adopted for treating NMP in the current industrial waste liquid, but the method has low recovery efficiency, incomplete recovery and high cost and needs professional equipment. The biodegradation method is a method which can effectively degrade NMP in wastewater. However, the NMP industrial wastewater is often complex in components and contains more inorganic salt ions, so that common microorganisms cannot survive in the high osmotic pressure environment, and difficulty is brought to biodegradation of the NMP industrial wastewater.
Disclosure of Invention
The technical problem to be solved is as follows: aiming at the problems in the prior art, the invention provides a salt-tolerant staphylococcus lentus strain and application thereof in industrial NMP wastewater treatment. In addition, the strain can keep normal physiological functions in a high-salt environment so as to achieve the aim of degrading residual NMP in high-salt wastewater.
The technical scheme is as follows: salt-tolerant staphylococcus lentusStaphylococcus lentus.) CCZU-X with preservation number of CGMCC No. 19016. The slow staphylococcus (A), (B), (C)Staphylococcus lentus.) CCZU-X colony characteristics: the colony is single, milky white, round, relatively moist and slightly raised. Gram staining was shown to be positive, globular. And the strain can grow in a culture medium with the salt concentration of more than 20%. The strain source is as follows: the salt-tolerant staphylococcus lentus CCZU-X is obtained by screening edible shrimps.
The application of the salt-tolerant staphylococcus lentus CCZU-X in treating industrial salt-containing NMP wastewater.
Preferably, the specific application steps are as follows:
preparing salt-tolerant staphylococcus lentus wet thalli, namely performing activated culture and fermentation culture on salt-tolerant staphylococcus lentus CCZU-X to obtain a fermentation culture solution, centrifuging to obtain salt-tolerant staphylococcus lentus CCZU-X wet thalli, washing the thalli twice by using a phosphate buffer solution with the pH value of 5-9, and storing the thalli at 4 ℃ for later use;
step two, pretreating the wastewater to be treated, carrying out suction filtration for 2-3 times, and adding 1-2 mol/L NaOH into the wastewater to be treated to adjust the pH value to 7.0;
adding salt-tolerant slow staphylococcus CCZU-X wet bacteria into the pretreated wastewater, adding the wet bacteria obtained in the step one into the wastewater treated in the step two, and culturing the mixture in a shaking table at the temperature of 20-50 ℃ and the shaking table rotating speed of 100-250 rpm for 1-72 hours;
and step four, measuring the wastewater treatment effect of the salt-tolerant staphylococcus lentus CCZU-X, namely adding wet bacteria into the wastewater treated in the step three, sampling at regular time, measuring the change value of NMP in the wastewater before and after the salt-tolerant staphylococcus lentus CCZU-X is added, quantitatively measuring the content of NMP in the wastewater by using a high performance liquid chromatography, and calculating the degradation rate of the NMP, wherein the measuring and analyzing method is detailed in QB/T5159-2017.
Preferably, the activation culture step in the first step is as follows: the components of the activation medium were formulated at the following concentrations: 5-100 g/L of yeast extract powder, 10-100 g/L of peptone and 10-100 g/L of NaCl, natural pH, sterilizing at 121 ℃ for 20min, cooling after sterilization, inoculating the slope seeds with the salt-tolerant staphylococcus lentus CCZU-X, and culturing for 1-3 d at 20-40 ℃ under the condition that the rotating speed of a shaking table is 100-250 rpm to serve as activated culture seed liquid.
Preferably, the fermentation culture step in the step one is as follows: the components of the fermentation medium are prepared according to the following concentrations: 5-100 g/L of yeast extract powder or glucose or sucrose, 10-100 g/L of peptone, 5-9 pH, sterilizing at 121 ℃ for 20min, cooling after sterilization, and culturing activated culture seed liquid according to the inoculation volume ratio of 3-10% under the conditions of the temperature of 20-40 ℃ and the rotating speed of a shaker of 100-250 rpm for 1-3 d to obtain a fermentation culture solution.
Preferably, the addition amount of wet bacteria in the wastewater pretreated in the third step is 5-15 g/L.
Has the advantages that: the invention obtains the staphylococcus lentus by screening from edible shrimpsStaphylococcus lentus.) CCZU-X, washing wet thalli by phosphate buffer solution, adding the wet thalli into high-salt NMP wastewater to treat wastewater, reducing the NMP in the wastewater to below 140 mg/L from 2000 mg/L before treatment, adding 0.5-20.0 g/L of wet thalli into the wastewater, treating the wastewater automatically, having low cost and low requirement on equipment, and being a new method for effectively degrading the high-salt NMP wastewater industrially. The salt-tolerant microorganisms can grow well in the salt-containing wastewater, so that the microorganisms can show good competitive advantages and application potential in the treatment of the salt-containing industrial NMP wastewater, and the method has the advantages of low treatment cost, strong adaptability and the like. Compared with the prior art, the maximum degradation rate of the strain CCZU-X can reach 93%, the strain is recovered by a physical method in the prior research, a water collecting tank, a pH adjusting tank, a coagulation reaction tank, a flocculation reaction tank, an efficient sedimentation tank, an intermediate tank, a selective filter, a recycling water tank and an NMP recovery system are required in the recovery, and the required process steps are complicated.
Preservation information: the strain preservation date of the invention is 11 months and 27 days in 2019, and the preservation number is as follows: CGMCC No.19016, and is classified and named as staphylococcus lentus (Staphylococcus lentus.) CCZU-X, the name of the preservation unit is China general microbiological culture Collection center, the preservation addressIs the microbiological research institute of the national academy of sciences No. 3, Xilu No.1, Beijing area facing the sun, the postal code: 100101.
Detailed Description
The invention is further described with reference to specific examples. It should be understood that these examples are for illustrative purposes only and should not be construed as limiting the technical aspects of the present invention.
Example 1: salt-tolerant staphylococcus lentus (A)Staphylococcus lentus.) Separation and domestication of CCZU-X
The salt-tolerant staphylococcus lentus (A) of the inventionStaphylococcus lentus.) The separation and domestication process of CCZU-X is as follows:
1) a sample, an edible shrimp living in nature in high salt seawater;
2) culture medium
Primary screening of culture medium: LB medium. LB medium (g/L): peptone 10, yeast extract 5, sodium chloride 10, agar 20, pH 7.0.
Re-screening the culture medium: LB selective medium with higher salt concentration. And a selection culture medium (g/L) which comprises 10 parts of peptone, 5 parts of yeast extract powder, 10-200 parts of sodium chloride and 7.0 parts of pH.
3) Primary screening, secondary screening and fermentation culture operation
Primary screening: putting 5g of edible sea shrimps living in high-salt seawater in nature into 100 mL of enrichment medium, carrying out enrichment culture at 37 ℃ for 72 h at 180 r/min, carrying out gradient dilution by using sterile water, coating the diluted solution on a primary screening solid medium, carrying out culture at 37 ℃ for 48h, picking bacterial colonies with good growth, different shapes and different colors, carrying out streak separation, and carrying out purification by seven streaks.
Re-screening: and (3) selecting a purified colony in a primary screening solid culture medium by using a sterilized inoculating loop, inoculating the colony on a selective culture medium, culturing for 24 hours at 37 ℃, and observing a strain which can grow under high salt concentration and is numbered as CCZU-X.
The morphological identification and molecular identification of the obtained strain are carried out, and the results are as follows:
salt-tolerant staphylococcus lentus (A)Staphylococcus lentus.) CCZU-X colony characteristics: the colony is single, milky white, round, relatively moist and slightly raised. Gram staining was shown to be positive, globular. And the strain can grow in a culture medium with high salt concentration.
Example 2: salt-tolerant staphylococcus lentus (A)Staphylococcus lentus.) Preparation of wet cells of CCZU-X
The components of the activation medium were formulated at the following concentrations: 5g/L yeast extract powder, 10g/L peptone, 10g/L sodium chloride, natural pH, sterilizing at 121 ℃ for 20 minutes, sterilizing, cooling, inoculating slant seeds (the salt-tolerant slow staphylococcus lentus CCZU-X prepared in example 1), and culturing at 37 ℃ and 180 r/min of shaking table rotation speed for 1 day as an activated culture seed solution.
The components of the fermentation medium are prepared according to the following concentrations: 20 g/L of sucrose, 10g/L of peptone, 10g/L of sodium chloride, pH7.0, sterilizing for 20 minutes at 121 ℃, cooling after sterilization, inoculating activated culture seed liquid according to the inoculation volume ratio of 10 percent, and culturing for 1 day at the temperature of 37 ℃ and the rotating speed of a shaker of 180 r/min to obtain a fermentation culture solution. Centrifuging for 10min (8000 r/min), collecting thallus, washing thallus with phosphate buffer (0.1M, pH 7.0) twice, and storing at 4 deg.C, wherein the wet thallus amount in the fermentation liquid is about 10 g/L.
Example 3: salt-tolerant staphylococcus lentus (A)Staphylococcus lentus.) Preparation of wet cells of CCZU-X
The components of the activation medium were formulated at the following concentrations: 10g/L yeast extract powder, 10g/L peptone, 10g/L sodium chloride, natural pH, sterilizing at 121 ℃ for 20 minutes, sterilizing, cooling, inoculating slant seeds (salt-tolerant slow staphylococcus lentus CCZU-X prepared in example 1), and culturing at 37 ℃ and 180 r/min of shaking table rotation speed for 1 day as an activated culture seed solution.
The components of the fermentation medium are prepared according to the following concentrations: 5g/L of yeast extract powder, 10g/L of peptone, 10g/L of sodium chloride, pH7.0, sterilizing for 20 minutes at 121 ℃, cooling after sterilization, inoculating activated culture seed liquid according to the inoculation volume ratio of 10%, and culturing for 1 day at 37 ℃ and the rotating speed of a shaking table of 180 r/min to obtain a fermentation culture liquid. Centrifuging for 10min (8000 r/min), collecting thallus, washing thallus with phosphate buffer (0.1M, pH 7.0) twice, and storing at 4 deg.C, wherein the wet thallus amount in the fermentation liquid is about 5 g/L.
Example 4: salt-tolerant staphylococcus lentus (A)Staphylococcus lentus.) Preparation of wet cells of CCZU-X
The components of the activation medium were formulated at the following concentrations: 100g/L yeast extract powder, 100g/L peptone, 100g/L sodium chloride, natural pH, sterilizing at 121 ℃ for 20 minutes, sterilizing, cooling, inoculating slant seeds (the salt-tolerant slow staphylococcus lentus CCZU-X prepared in example 1), and culturing at 30 ℃ and 180 r/min of shaking table rotation speed for 1 day to obtain an activated culture seed solution.
The components of the fermentation medium are prepared according to the following concentrations: 100g/L glucose, 100g/L peptone, 100g/L sodium chloride, pH7.0, sterilizing for 20 minutes at 121 ℃, cooling after sterilizing, inoculating activated culture seed liquid according to the inoculation volume ratio of 10%, and culturing for 1 day at 37 ℃ and the rotating speed of a shaking table of 180 r/min to obtain a fermentation culture liquid. Centrifuging for 10min (8000 r/min), collecting thallus, washing thallus with phosphate buffer (0.1M, pH 7.0) twice, and storing at 4 deg.C, wherein the wet thallus amount in the fermentation liquid is about 5 g/L.
By comparing the examples 2, 3 and 4, and combining the growth condition of the strain, the degradation condition of the fermentation liquid and economic factors, the most suitable activation medium is 100g/L of yeast extract powder, 100g/L of peptone, 100g/L of sodium chloride and natural pH, and the most suitable fermentation medium is 100g/L of glucose, 100g/L of peptone, 100g/L of sodium chloride and pH 7.0.
Example 5 salt-tolerant Staphylococcus lentus at different cell concentrations: (Staphylococcus lentus.) Effect of CCZU-X in degrading waste water
In the embodiment, the wastewater to be treated is simulated wastewater which contains 20 g/L of NaCl and CaCl240 g/L，NMP 2 g/L。
Pretreating the wastewater to be treated, carrying out suction filtration for 3 times, and adding 2 mol/L NaOH into the wastewater to be treated to adjust the pH value to 7.0.
Salt-tolerant staphylococcus lentus (b) at different cell concentrationsStaphylococcus lentus.) The effect of CCZU-X in degrading waste water. After the salt-tolerant staphylococcus lentus CCZU-X was activated for 1 day and fermented for 1 day according to the method of example 2, wet cells with a concentration of 5g/L, 7.5 g/L, and 10g/L were weighed and added to a 250 mL conical flask containing 100 mL of NMP pretreated wastewater with a salt content of 100g/L, and were subjected to shake cultivation at 37 ℃ and 180 r/min for 6 days, and NMP was sampled and measured, and the results are shown in Table 1.
Table 1: treating for 6 days under different thallus concentrations, detecting the NMP concentration before and after the wastewater is degraded, and calculating the degradation rate of the NMP
| Bacterial cell amount/g.L-1 | Initial NMP content/mg. L-1 | NMP content/mg. L-1 | NMP degradation rate/%) |
| 5 | 2000 | 793 | 60.35 |
| 7.5 | 2000 | 472.8 | 76.36 |
| 10 | 2000 | 296 | 85.20 |
As can be seen from the table, the degradation rate of NMP increases with the increase of the amount of the bacteria under the same conditions for the same time, indicating that the strain has an effect on the treatment of NMP in wastewater, and the treatment effect is positively correlated with the growth of the strain.
Example 6 salt-tolerant Staphylococcus lentus with a cell concentration of 10g/L (Staphylococcus lentus.) CCZU-X culture effect of degrading wastewater at different times
In the embodiment, the wastewater to be treated is simulated wastewater which contains 20 g/L of NaCl and CaCl240 g/L，NMP 2 g/L。
Pretreating the wastewater to be treated, carrying out suction filtration for 2 times, and adding 1 mol/L NaOH into the wastewater to be treated to adjust the pH value to 7.0.
Salt-tolerant staphylococcus lentus (A), (B) and (C)Staphylococcus lentus.) CCZU-X was cultured for 20 hours in the same manner as in example 2, and then wet cells at a concentration of 10g/L were weighed out and added to a 250 mL Erlenmeyer flask containing 100 mL of NMP-pretreated wastewater having a salt content of 10g/L, followed by shaking culture at 37 ℃ and 180 r/min for 72 hours, during which time NMP content was measured by sampling, and the results are shown in Table 2.
When the salt concentration is 1%, the bacteria are used for treating the wastewater NMP content and the NMP degradation rate in different time (0 h, 12 h, 24 h, 36 h, 48h, 60 h and 72 h).
| Treatment time/h | 0 | 12 | 24 | 36 | 48 | 60 | 72 |
| NMP content/mg. L-1 | 2000 | 1154 | 838 | 634 | 396 | 190 | 140 |
| NMP degradation rate/%) | - | 42.3 | 58.1 | 68.3 | 80.2 | 90.5 | 93.0 |
As can be seen from the table, the degradation rate of the strain with respect to NMP increased with the increase of the treatment time, but the increase of the degradation rate gradually decreased, which is related to the batch process employed in this example.
By utilizing the culture condition and the biotransformation process of the strain, the NMP content in the high-salt NMP wastewater can be reduced. The lower the salt concentration, the better the degradation. Therefore, it is feasible to degrade NMP in industrial high-salinity wastewater.
Example 7
Salt-tolerant staphylococcus lentusStaphylococcus lentus.) CCZU-X with the preservation number of CGMCCNo.19016. The slow staphylococcus (A), (B), (C)Staphylococcus lentus.) CCZU-X colony characteristics: the colony is single, milky white, round, relatively moist and slightly raised. Gram staining was shown to be positive, globular. And the strain can grow in a culture medium with the salt concentration of more than 20%. The strain source is as follows: the salt-tolerant staphylococcus lentus CCZU-X is obtained by screening edible shrimps.
The application of the salt-tolerant staphylococcus lentus CCZU-X in treating industrial salt-containing NMP wastewater. The specific application steps are as follows:
the preparation method comprises the following steps of firstly, preparing salt-tolerant staphylococcus lentus wet thalli, namely performing activated culture and fermentation culture on the salt-tolerant staphylococcus lentus CCZU-X to obtain fermentation culture solution, centrifuging to obtain the salt-tolerant staphylococcus lentus CCZU-X wet thalli, washing the thalli twice by using phosphate buffer solution with the pH of 5, and storing the thalli at 4 ℃ for later use, wherein the activated culture step is as follows: the components of the activation medium were formulated at the following concentrations: 5g/L yeast extract powder, 10g/L peptone and 10g/L NaCl, natural pH, sterilizing at 121 ℃ for 20min, cooling after sterilization, inoculating the slope seeds to the salt-tolerant slow staphylococcus CCZU-X, and culturing for 3 d at 20 ℃ and at the rotating speed of a shaking table of 100 rpm to obtain activated culture seed liquid; the fermentation culture steps are as follows: the components of the fermentation medium are prepared according to the following concentrations: 5g/L of yeast extract powder or glucose or sucrose, 10g/L of peptone, pH5, sterilizing for 20min at 121 ℃, cooling after sterilizing, and culturing the activated culture seed liquid according to the inoculation volume ratio of 3% for 3 d under the conditions of 20 ℃ of temperature and 100 rpm of shaking table rotation speed to obtain a fermentation culture solution;
step two, pretreating the wastewater to be treated, performing suction filtration for 2 times, and adding 1 mol/L NaOH into the wastewater to be treated to adjust the pH value to 7.0;
adding salt-tolerant slow staphylococcus CCZU-X wet bacteria into the pretreated wastewater, adding the wet bacteria in the step one into the wastewater treated in the step two, putting the wastewater into a shaking table for culturing at the temperature of 20 ℃ and the shaking table rotating speed of 100 rpm for 72 hours, wherein the addition amount of the wet bacteria in the pretreated wastewater is 5 g/L;
and step four, measuring the wastewater treatment effect of the salt-tolerant staphylococcus lentus CCZU-X, namely, after the wet bacteria prepared in the step one is added in the step three, sampling at regular time, measuring the change value of NMP in the wastewater before and after the salt-tolerant staphylococcus lentus CCZU-X is added, quantitatively measuring the content of NMP in the wastewater through a high performance liquid chromatography, and calculating the degradation rate of the NMP, wherein the measuring and analyzing method is detailed in QB/T5159-2017.
Example 8
Salt-tolerant staphylococcus lentusStaphylococcus lentus.) CCZU-X with the preservation number of CGMCCNo.19016. The slow staphylococcus (A), (B), (C)Staphylococcus lentus.) CCZU-X colony characteristics: the colony is single, milky white, round, relatively moist and slightly raised. Gram staining was shown to be positive, globular. And the strain can grow in a culture medium with the salt concentration of more than 20%. The strain source is as follows: the salt-tolerant staphylococcus lentus CCZU-X is obtained by screening edible shrimps.
The application of the salt-tolerant staphylococcus lentus CCZU-X in treating industrial salt-containing NMP wastewater. The specific application steps are as follows:
step one, preparing salt-tolerant staphylococcus lentus wet thalli, namely performing activated culture and fermentation culture on salt-tolerant staphylococcus lentus CCZU-X to obtain fermentation culture solution, centrifuging to obtain salt-tolerant staphylococcus lentus CCZU-X wet thalli, washing the thalli twice by using phosphate buffer solution with pH 9, and storing the thalli at 4 ℃ for later use, wherein the activated culture step is as follows: the components of the activation medium were formulated at the following concentrations: 100g/L yeast extract powder, 100g/L peptone and 100g/L NaCl, natural pH, sterilizing at 121 ℃ for 20min, cooling after sterilization, inoculating the slope seeds to the salt-tolerant slow staphylococcus CCZU-X, and culturing for 1 d at 40 ℃ and the rotating speed of a shaking table of 250 rpm to obtain activated culture seed liquid; the fermentation culture steps are as follows: the components of the fermentation medium are prepared according to the following concentrations: 100g/L yeast extract powder or glucose or sucrose, 100g/L peptone, pH 9, sterilizing at 121 ℃ for 20min, cooling after sterilization, culturing the activated culture seed liquid according to the inoculation amount volume ratio of 10% for 1 d under the conditions of 40 ℃ temperature and 250 rpm of shaking table rotation speed to obtain a fermentation culture solution;
step two, pretreating the wastewater to be treated, carrying out suction filtration for 3 times, and adding 2 mol/L NaOH into the wastewater to be treated to adjust the pH value to 7.0;
adding salt-tolerant slow staphylococcus CCZU-X wet bacteria into the pretreated wastewater, adding the wet bacteria in the step one into the wastewater treated in the step two, putting the wastewater into a shaking table for culturing at the temperature of 50 ℃ and the shaking table rotating speed of 250 rpm for 1 h, wherein the addition amount of the wet bacteria in the pretreated wastewater is 15 g/L;
and step four, measuring the wastewater treatment effect of the salt-tolerant staphylococcus lentus CCZU-X, namely, after the wet bacteria prepared in the step one is added in the step three, sampling at regular time, measuring the change value of NMP in the wastewater before and after the salt-tolerant staphylococcus lentus CCZU-X is added, quantitatively measuring the content of NMP in the wastewater through a high performance liquid chromatography, and calculating the degradation rate of the NMP, wherein the measuring and analyzing method is detailed in QB/T5159-2017.
In light of the foregoing description of the preferred embodiment of the present invention, many modifications and variations will be apparent to those skilled in the art without departing from the spirit and scope of the invention. The technical scope of the present invention is not limited to the content of the specification, and must be determined according to the scope of the claims.
Claims (6)
1. Salt-tolerant staphylococcus lentusStaphylococcus lentus.) CCZU-X with the preservation number of CGMCCNo.19016.
2. The use of the salt tolerant staphylococcus lentus CCZU-X of claim 1 for the treatment of industrial salt-containing NMP wastewater.
3. The application of claim 2, wherein the specific application steps are as follows:
preparing salt-tolerant staphylococcus lentus wet thalli, namely performing activated culture and fermentation culture on salt-tolerant staphylococcus lentus CCZU-X to obtain a fermentation culture solution, centrifuging to obtain salt-tolerant staphylococcus lentus CCZU-X wet thalli, washing the thalli twice by using a phosphate buffer solution with the pH value of 5-9, and storing the thalli at 4 ℃ for later use;
step two, pretreating the wastewater to be treated, carrying out suction filtration for 2-3 times, and adding 1-2 mol/L NaOH into the wastewater to be treated to adjust the pH value to 7.0;
adding salt-tolerant slow staphylococcus CCZU-X wet bacteria into the pretreated wastewater, adding the wet bacteria prepared in the step one into the wastewater treated in the step two, and culturing the mixture in a shaking table at the temperature of 20-50 ℃ and the shaking table rotating speed of 100-250 rpm for 1-72 hours;
and step four, measuring the effect of the salt-tolerant slow staphylococcus CCZU-X on wastewater treatment, namely adding wet bacteria into the wastewater treated in the step three, sampling at regular time to measure the change value of NMP in the wastewater before and after the salt-tolerant slow staphylococcus CCZU-X is added, quantitatively measuring the content of the NMP in the wastewater through a high performance liquid chromatography, and calculating the degradation rate of the wastewater.
4. The use of claim 3, wherein the activation culturing step in the first step is as follows: the components of the activation medium were formulated at the following concentrations: 5-100 g/L of yeast extract powder, 10-100 g/L of peptone and 10-100 g/L of NaCl, natural pH, sterilizing at 121 ℃ for 20min, cooling after sterilization, inoculating the slope seeds with the salt-tolerant staphylococcus lentus CCZU-X, and culturing for 1-3 d at 20-40 ℃ under the condition that the rotating speed of a shaking table is 100-250 rpm to serve as activated culture seed liquid.
5. The use of claim 4, wherein the fermentation culture step in the first step is as follows: the components of the fermentation medium are prepared according to the following concentrations: 5-100 g/L of yeast extract powder or glucose or sucrose, 10-100 g/L of peptone, 5-9 pH, sterilizing at 121 ℃ for 20min, cooling after sterilization, and culturing activated culture seed liquid according to the inoculation volume ratio of 3-10% under the conditions of the temperature of 20-40 ℃ and the rotating speed of a shaker of 100-250 rpm for 1-3 d to obtain a fermentation culture solution.
6. The application of claim 3, wherein the addition amount of wet bacteria in the wastewater pretreated in the third step is 5-15 g/L.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2022121718A1 (en) * | 2020-12-07 | 2022-06-16 | 山东晨彰生物科技有限公司 | Application of staphylococcus nepalensis to degradation of residual sugar in fermentation industrial organic wastewater |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105132323A (en) * | 2015-09-08 | 2015-12-09 | 常州大学 | Salt-tolerance bacillus and application thereof in high-salinity wastewater treatment |
-
2020
- 2020-06-16 CN CN202010549899.8A patent/CN111690563A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105132323A (en) * | 2015-09-08 | 2015-12-09 | 常州大学 | Salt-tolerance bacillus and application thereof in high-salinity wastewater treatment |
Non-Patent Citations (2)
| Title |
|---|
| GUORUI ZHANG ET AL.: "Degradation of N-methylpyrrolidone in High Salinity Wastewater by a Halotolerant Microorganism", 《JOURNAL OF BIOLOGY AND LIFE SCIENCE》 * |
| 王丽娟等: "一株耐盐菌的分离及其降解特性", 《化工进展》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2022121718A1 (en) * | 2020-12-07 | 2022-06-16 | 山东晨彰生物科技有限公司 | Application of staphylococcus nepalensis to degradation of residual sugar in fermentation industrial organic wastewater |
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