CN104774785A - Oil paint wastewater degrading bacterium and screening and enriching method thereof - Google Patents
Oil paint wastewater degrading bacterium and screening and enriching method thereof Download PDFInfo
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- CN104774785A CN104774785A CN201510113113.7A CN201510113113A CN104774785A CN 104774785 A CN104774785 A CN 104774785A CN 201510113113 A CN201510113113 A CN 201510113113A CN 104774785 A CN104774785 A CN 104774785A
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- oil
- base paint
- wastewater degradation
- paint wastewater
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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
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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
- 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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- 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
- C12N1/36—Adaptation or attenuation of cells
Abstract
The invention discloses an oil paint wastewater degrading bacterium and a screening and enriching method thereof, and belongs to the technical field of microbe screening. The method comprises the following steps: (1) preparing a bacterium liquid; (2) preparing an oil paint wastewater degrading bacterial system; (3) screening the oil paint wastewater degrading bacterial system; (4) re-screening the purified degrading bacteria; wherein in the step (2), glucose is added into the bacterium liquid obtained in the step (1), then oil paint acidic wastewater is added, the bacterium is inoculated into an inorganic salt culture medium to carry out subculture in the absence of light on a shaking table; in each subculture, the amount of glucose in the culture medium is less than that of previous generation; in each subculture the glucose amount is 7d, the subculture is carried out for at least five generations, and the culture medium does not contain any glucose in the last subculture. The biological method for processing oil paint wastewater has the advantages of low energy consumption, mild reaction conditions, and high efficiency, and the decomposition efficiency of the degrading bacterium strains on oil paint wastewater can reach 80% or more.
Description
Technical field
The invention belongs to microbe to screen technical field, be specifically related to a kind of oil-base paint wastewater degradation bacterium and screening enriching method thereof.
Background technology
Along with domestic automobile industry is flourish, the consumption of automotive paints increases year by year, and the paint waste water thereupon produced also can not be ignored.In automotive paints spraying process, hand spray has at least the over-sprayed paint fog of 40%-60% to disperse, and electrostatic spraying also can produce the over-sprayed paint fog of more than 10%, and over-sprayed paint fog agglomerate and sink is discharged by workshop recirculated water and produced paint waste water.And oil-base paint waste water mainly results from the pretreatment stage of vehicle spray painting, according to statistics by 2010, China's auto output reaches 1,000 ten thousand, and automobile pollution reaches 5,200 ten thousand, if produce oil-base paint waste water 0.5m by producing each automobile
3calculate, 5,000,000 m will be produced every year
3oil-base paint waste water.Oil-base paint waste water complicated components, Pollutant levels are high, environmental hazard is strong, and this makes the process of oil-base paint waste water very difficult.
The treatment process of current oiliness paint waste water mainly contains coagulant sedimentation, pollutent flocculation sediment is made by adding flocculation agent, then remove through sloping plate deposition pool device etc., this method effectively can remove oil, macromolecule resin etc. in oil-base paint waste water, but can not fundamentally pollution degradation material; Ultrafiltrationmembrane process is under the effect of static pressure difference impellent, make the solvent in oil-base paint waste water and little solute particles from high pressure feed liquid side through the filtrate of the high molecular functional ultra-filtration membrane to low-tension side with selective penetrated property, this method obviously can reduce the concentration of oil-base paint waste water, but complex operation, the membrane module life-span is long, cost is higher; Light electrolysis-chemical oxidization method process oil-base paint waste water is mainly under the effect of chemical oxidizing agent, the organism in oil-base paint waste water is made to be degraded to the stable inorganic salts little to natural environment influence further, this technology drops into industrialization at present, but there is the problems such as filler easily lumps, replacement difficulty, reactor easily block.Utilizing Biochemical method oil-base paint waste water to have, energy consumption is low, reaction conditions is gentle and efficiency is higher, and anaerobic process can also reclaim biogas, more meets that national energy-saving reduces discharging, the strategic guideline of Sustainable development.The screening domestication that research has exclusive adapted microbes is cultivated, and for the tolerance and the stability that improve bio-reaction system, improves degradation efficiency significant.
Summary of the invention
Primary and foremost purpose of the present invention is to overcome the shortcoming that exists in prior art with not enough, provides a kind of oil-base paint wastewater degradation bacterium.
Another object of the present invention is to provide the screening enriching method of described oil-base paint wastewater degradation bacterium, the method effectively raises the degradation efficiency of oil-base paint waste water.
Object of the present invention is achieved through the following technical solutions: a kind of oil-base paint wastewater degradation bacterium, and this bacterial strain is microbacterium (microbacterium sp.) XR001, and preservation from date is on December 23rd, 2014, and deposit number is CGMCC No.10237; Depositary institution is China Committee for Culture Collection of Microorganisms's common micro-organisms center (CGMCC); Depositary institution address is No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City.
The screening enriching method of described oil-base paint wastewater degradation bacterium, comprises the steps:
(1) prepare bacterium liquid: add in sterilized water by vehicle coating wastewater sewage draining exit mud, the concentration of vehicle coating wastewater sewage draining exit mud is 0.08g/mL, and shaking table lucifuge cultivates 24h, obtains bacteria suspension; Inoculate bacteria suspension in minimal medium after sterilization, the inoculum size of bacteria suspension is 5% of minimal medium volume, and shaking table lucifuge cultivates 7d, obtains bacterium liquid;
(2) prepare oil-base paint wastewater degradation fungus strain: added by glucose in the bacterium liquid of step (1), by the 5% inoculation oil-base paint waste water that bacteria liquid is long-pending, shaking table lucifuge carries out Secondary Culture, obtains oil-base paint wastewater degradation fungus strain nutrient solution; During each Secondary Culture, in bacterium liquid, the amount of glucose all reduces 1/5 than previous generation, the amount of oil-base paint waste water all increases than previous generation, each increasing amount be bacteria liquid long-pending 5%, each Secondary Culture 7d, in Secondary Culture >=five generation, during last culture, in substratum, no longer add glucose;
(3) oil-base paint wastewater degradation fungus strain is screened: in minimal medium, add agar, after sterilizing, add after minimal medium solidifies and be also coated with evenly in the oil-base paint wastewater degradation fungus strain nutrient solution of the step (2) of gradient dilution, cultivate 7d for 30 DEG C, the bacterium colony of transparent circle is had around picking, repeat inoculation culture and separation of ruling, obtain the oil-base paint wastewater degradation fungus strain of purifying;
(4) carry out multiple sieve to the oil-base paint wastewater degradation fungus strain of purifying: be inoculated in LB substratum by the oil-base paint wastewater degradation fungus strain of step (3) purifying, after growing bacterium colony, being prepared into concentration is OD
600the bacterium liquid of=1, for subsequent use;
In step (1),
Described shaking table lucifuge condition is 30 DEG C, 130r/min;
The component of described minimal medium is: NaCl 0.5g, NH
4nO
30.4g, KH
2pO
41.0g, K
2hPO
41.0g, yeast leaching powder 0.2g, liquid microelement 4mL, distilled water 1000mL, adjust pH is 7.2;
Consisting of of described liquid microelement: MgSO
42.0g, CuSO
40.5g, MnSO
40.5g, FeSO
47H
2o 0.5g, CaCl
20.5g, distilled water 500mL.
In step (2),
The concentration of described oil-base paint waste water is 200mg/L; The weight of described glucose is 0.5% of minimal medium weight;
The condition of described shaking table lucifuge in 30 DEG C, 130r/min cultivates 7 days;
The component of described minimal medium is: NaCl 0.5g, NH
4nO
30.4g, KH
2pO
41.0g, K
2hPO
41.0g, yeast leaching powder 0.2g, liquid microelement 4mL, distilled water 1000mL, adjust pH is 7.2;
Consisting of of described liquid microelement: MgSO
42.0g, CuSO
40.5g, MnSO
40.5g, FeSO
47H
2o 0.5g, CaCl
20.5g, distilled water 500mL.
In step (3),
The weight of described agar is 2% of minimal medium weight;
The volume ratio of the oil-base paint wastewater degradation fungus strain nutrient solution of described minimal medium and step (2) is 100:1;
Described gradient dilution is that the oil-base paint wastewater degradation fungus strain nutrient solution of step (2) is diluted 10 respectively according to volume
3, 10
4, 10
5doubly;
The component of described minimal medium is: NaCl 0.5g, NH
4nO
30.4g, KH
2pO
41.0g, K
2hPO
41.0g, yeast leaching powder 0.2g, liquid microelement 4mL, distilled water 1000mL, adjust pH is 7.2;
Consisting of of described liquid microelement: MgSO
42.0g, CuSO
40.5g, MnSO
40.5g, FeSO
47H
2o 0.5g, CaCl
20.5g, distilled water 500mL.
In step (4),
Consisting of of described LB plate culture medium: peptone 2.5g, extractum carnis 0.75g, NaCl 1.25g, distilled water 250mL.
The present invention has following advantage and effect relative to prior art: the present invention adopts Biochemical method oil-base paint waste water, has that energy consumption is low, reaction conditions is gentle and efficiency is higher.The screening domestication that the present invention has exclusive adapted microbes is cultivated, and degradation bacteria strains can reach more than 80% to the degradation efficiency of oil-base paint waste water, and for improving the tolerance of bio-reaction system and stability, raising degradation efficiency is significant.
Embodiment
Below in conjunction with embodiment, the present invention is described in further detail, but embodiments of the present invention are not limited thereto.
Embodiment 1
(1) collection in bacterium source and the preparation of bacteria suspension: gather Yancheng DONGFENG YUEDA Kia Motors Paint Factory sewage draining exit soil, is placed in 4 DEG C of refrigerators and saves backup by mud; 20g is gathered the sterilized water that the soil of returning is placed in 250mL, and 30 DEG C vibrate 12 hours, make bacteria suspension;
(2) get 5mL bacteria suspension and inject the Erlenmeyer flask filling 100mL minimal medium, be positioned in 30 DEG C of constant-temperature tables, 7d is cultivated in 130r/min concussion, obtains bacterium liquid;
(3) minimal medium: NaCl 0.5g, NH
4nO
30.4g, KH
2pO
41.0g, K
2hPO
41.0g, yeast leaching powder 0.2g, liquid microelement 4mL, distilled water 1000mL, adjust pH is 7.2; Wherein the consisting of of liquid microelement: MgSO
42.0g, CuSO
40.5g, MnSO
40.5g, FeSO
47H
2o 0.5g, CaCl
20.5g, distilled water 500mL;
(4) get the bacterium liquid in 1mL step (2), add in the minimal medium of 100mL, 30 DEG C, 7d is cultivated in 130r/min concussion, and the content wherein adding oil-base paint in minimal medium, to 50mg/L, adds the content of glucose to 200mg/L;
(5) the bacterium liquid in 1mL step (4) is got, add in the minimal medium of 100mL, 30 DEG C, 7d is cultivated in 130r/min concussion, wherein add the content of oil-base paint in minimal medium to 100mg/L, add the content of glucose to 100mg/L;
(6) get the bacterium liquid in 1mL step (5), add in the minimal medium of 100mL, 30 DEG C, 7d is cultivated in 130r/min concussion, and the content wherein adding oil-base paint in minimal medium, to 150mg/L, adds the content of glucose to 50mg/L;
(7) get the bacterium liquid in 1mL step (6), add in the minimal medium of 100mL, 30 DEG C, 7d is cultivated in 130r/min concussion, and the content wherein adding oil-base paint in minimal medium, to 200mg/L, no longer adds glucose;
(8) the bacterium liquid in step (7) is carried out gradient dilution, described gradient dilution is that the oil-base paint solution fungus strain nutrient solution in step (7) is diluted 10 respectively according to volume
3, 10
4, 10
5doubly, coat on solid plate, cultivate 7d for 30 DEG C, have the bacterium colony of transparent circle around picking, repeated inoculation is cultivated and separation of ruling, and obtains the oil-base paint wastewater degradation bacterium of purifying.
(9) multiple sieve is carried out to the oil-base paint wastewater degradation bacterium of purifying: oil-base paint wastewater degradation bacterium step (8) obtained is inoculated in LB plate culture medium, cultivate 7d, after growing bacterium colony, bacterial classification is scraped in sterilized water, bacterium is dispersed as bacterium liquid, the concentration regulating bacterium liquid is OD
600=1, for subsequent use.Consisting of of described LB plate culture medium: peptone 2.5g, extractum carnis 0.75g, NaCl 1.25g, distilled water 250mL.
The oil-base paint wastewater degradation fungus strain finally obtained can with oil-base paint waste water for substrate grows, and aerobic degradation test shows that flora can carry out effective aerobic degradation to oil-base paint waste water; When the concentration of oil-base paint waste water is 200mg/L, after after 7d, degradation rate reaches 79%, 10d, degradation efficiency reaches 83%.
Above-described embodiment is the present invention's preferably embodiment; but embodiments of the present invention are not restricted to the described embodiments; change, the modification done under other any does not deviate from spirit of the present invention and principle, substitute, combine, simplify; all should be the substitute mode of equivalence, be included within protection scope of the present invention.
Claims (9)
1. an oil-base paint wastewater degradation bacterium, is characterized in that: this bacterial strain is microbacterium (microbacterium sp.) XR001, and preservation from date is on December 23rd, 2014, and deposit number is CGMCC No.10237.
2. the screening enriching method of oil-base paint wastewater degradation bacterium according to claim 1, is characterized in that, comprise the steps:
(1) prepare bacterium liquid: add in sterilized water by vehicle coating wastewater sewage draining exit mud, the concentration of vehicle coating wastewater sewage draining exit mud is 0.08g/mL, and shaking table lucifuge cultivates 24h, obtains bacteria suspension; Inoculate bacteria suspension in minimal medium after sterilization, the inoculum size of bacteria suspension is 5% of minimal medium volume, and shaking table lucifuge cultivates 7d, obtains bacterium liquid;
(2) prepare oil-base paint wastewater degradation fungus strain: added by glucose in the bacterium liquid of step (1), by the 5% inoculation oil-base paint waste water that bacteria liquid is long-pending, shaking table lucifuge carries out Secondary Culture, obtains oil-base paint wastewater degradation fungus strain nutrient solution; During each Secondary Culture, in bacterium liquid, the amount of glucose all reduces 1/5 than previous generation, the amount of oil-base paint waste water all increases than previous generation, each increasing amount be bacteria liquid long-pending 5%, each Secondary Culture 7d, in Secondary Culture >=five generation, during last culture, in substratum, no longer add glucose;
(3) oil-base paint wastewater degradation fungus strain is screened: in minimal medium, add agar, after sterilizing, add after minimal medium solidifies and be also coated with evenly in the oil-base paint wastewater degradation fungus strain nutrient solution of the step (2) of gradient dilution, cultivate 7d for 30 DEG C, the bacterium colony of transparent circle is had around picking, repeat inoculation culture and separation of ruling, obtain the oil-base paint wastewater degradation fungus strain of purifying;
(4) carry out multiple sieve to the oil-base paint wastewater degradation fungus strain of purifying: be inoculated in LB substratum by the oil-base paint wastewater degradation fungus strain of step (3) purifying, after growing bacterium colony, being prepared into concentration is OD
600the bacterium liquid of=1, for subsequent use.
3. the screening enriching method of oil-base paint wastewater degradation bacterium according to claim 2, is characterized in that, the shaking table lucifuge condition described in step (1) is 30 DEG C, 130r/min.
4. the screening enriching method of oil-base paint wastewater degradation bacterium according to claim 2, is characterized in that, the concentration of the oil-base paint waste water described in step (2) is 200mg/L; The weight of described glucose is 0.5% of minimal medium weight.
5. the screening enriching method of oil-base paint wastewater degradation bacterium according to claim 2, is characterized in that, the condition of the shaking table lucifuge described in step (2) be in 30 DEG C, 130r/min cultivates 7d.
6. the screening enriching method of oil-base paint wastewater degradation bacterium according to claim 2, is characterized in that, the component of the minimal medium described in step (2) is: NaCl 0.5g, NH
4nO
30.4g, KH
2pO
41.0g, K
2hPO
41.0g, yeast leaching powder 0.2g, liquid microelement 4mL, distilled water 1000mL, adjust pH is 7.2; Consisting of of described liquid microelement: MgSO
42.0g, CuSO
40.5g, MnSO
40.5g, FeSO
47H
2o 0.5g, CaCl
20.5g, distilled water 500mL.
7. the screening enriching method of oil-base paint wastewater degradation bacterium according to claim 2, is characterized in that, the weight of the agar described in step (3) is 2% of minimal medium weight; The volume ratio of the oil-base paint wastewater degradation fungus strain nutrient solution of described minimal medium and step (2) is 100:1.
8. the screening enriching method of oil-base paint wastewater degradation bacterium according to claim 2, is characterized in that, the gradient dilution described in step (3) is that the oil-base paint wastewater degradation fungus strain nutrient solution of step (2) is diluted 10 respectively according to volume
3, 10
4, 10
5doubly.
9. the screening enriching method of oil-base paint wastewater degradation bacterium according to claim 2, is characterized in that, consisting of of the LB plate culture medium described in step (4): peptone 2.5g, extractum carnis 0.75g, NaCl1.25g, distilled water 250mL.
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Cited By (1)
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