CN104774785B - A kind of oil paint wastewater degradation bacterium and its screening enrichment method - Google Patents
A kind of oil paint wastewater degradation bacterium and its screening enrichment method Download PDFInfo
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- CN104774785B CN104774785B CN201510113113.7A CN201510113113A CN104774785B CN 104774785 B CN104774785 B CN 104774785B CN 201510113113 A CN201510113113 A CN 201510113113A CN 104774785 B CN104774785 B CN 104774785B
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- waste water
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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
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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 a kind of oil paint wastewater degradation bacterium and its screening enrichment methods, belong to microbe to screen technical field.This method comprises the following steps: (1) preparing bacterium solution；(2) oil paint wastewater degradation fungus strain is prepared；(3) oil paint wastewater degradation fungus strain is screened；(4) secondary screening is carried out to the degradation bacteria of purifying；Wherein step (2) is that glucose is added in the bacterium solution of step (1), add oiliness urushic acid waste water, it is linked into minimal medium, shaking table is protected from light secondary culture, the amount of glucose is reduced than previous generation in culture medium when each secondary culture, and the amount of oiliness paint waste water increases than previous generation, each secondary culture 7d, secondary culture more than generation, does not add glucose five in culture medium when last is commissioned to train feeding.The present invention uses Biochemical method oiliness paint waste water, has that low energy consumption, reaction condition is mild and efficiency is higher, and degradation bacteria strains can achieve 80% or more to the degradation efficiency of oiliness paint waste water.
Description
Technical field
The invention belongs to microbe to screen technical fields, and in particular to a kind of oil paint wastewater degradation bacterium and its screening enrichment
Method.
Background technique
As domestic automobile industry flourishes, the dosage of automotive paints increases year by year, the paint waste water generated therewith
It can not be ignored.In automotive paints spraying process, the hand spray at least over-sprayed paint fog of 40%-60% disperses, electrostatic spraying
Also 10% or more over-sprayed paint fog can be generated, over-sprayed paint fog agglomerate and sink generates paint waste water by the discharge of workshop recirculated water.And
Oiliness paint waste water occurs mainly with the pretreatment stage of vehicle spray painting, and according to statistics by 2010, China's auto output is up to 10,000,000
, car ownership is up to 52,000,000, if generating oiliness paint waste water 0.5m by each automobile is produced3It calculates, to generate 500 every year
Ten thousand m3Oiliness paint waste water.Oiliness paint waste water complicated components, pollutant concentration are high, environmental hazard is strong, this makes oiliness paint waste water
Processing it is very difficult.
The processing method of current oiliness paint waste water mainly has coagulant sedimentation, keeps pollutant flocculation heavy by adding flocculant
It forms sediment, is then removed through sloping plate deposition pool device etc., this method can effectively remove oil, macromolecule resin in oiliness paint waste water
Deng, but cannot fundamentally pollution degradation substance；Ultrafiltrationmembrane process is under the action of static pressure difference motive force, so that oil paint is useless
Solvent and small solute particles in water penetrate the high molecular functional ultrafiltration membrane with selective penetrated property to low from high pressure feed liquid side
The filtrate of side is pressed, this method can be substantially reduced the concentration of oiliness paint waste water, but cumbersome, the membrane module service life is long, cost
It is higher；Light electrolysis-chemical oxidization method processing oiliness paint waste water mainly under the action of chemical oxidizing agent, makes oiliness paint waste water
In organic matter be further degraded to the stable inorganic salts small to natural environment influence, industry has been put into the technology at present
Change, but there is fillers easily to agglomerate, replacement is difficult, reactor is easy the problems such as blocking.Utilize Biochemical method oiliness
Paint waste water has that low energy consumption, reaction condition is mild and efficiency is higher, and anaerobic processes can also recycle biogas, be more in line with national section
The strategic guideline of energy emission reduction, sustainable development.Studying, there is the screening of exclusive adapted microbes to tame culture, give birth to for improving
The tolerance and stability of object reaction system improve degradation efficiency and are of great significance.
Summary of the invention
The primary purpose of the present invention is that overcoming shortcoming and deficiency existing in the prior art, a kind of oiliness paint waste water is provided
Degradation bacteria.
Another object of the present invention is to provide the screening enrichment method of the oil paint wastewater degradation bacterium, this method has
The degradation efficiency for improving oiliness paint waste water of effect.
The purpose of the invention is achieved by the following technical solution: a kind of oil paint wastewater degradation bacterium, which is microbacterium
(microbacterium sp.) XR001, preservation from date are on December 23rd, 2014, deposit number CGMCC
No.10237；Depositary institution is China Committee for Culture Collection of Microorganisms's common micro-organisms center (CGMCC)；Depositary institution
Address is Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3.
The screening enrichment method of the oil paint wastewater degradation bacterium, includes the following steps:
(1) it prepares bacterium solution: vehicle coating wastewater sewage draining exit sludge is added in sterile water, vehicle coating wastewater sewage draining exit is dirty
The concentration of mud is 0.08g/mL, and shaking table is protected from light culture for 24 hours, obtains bacteria suspension；Bacterium is inoculated in minimal medium after sterilization
Suspension, the inoculum concentration of bacteria suspension are the 5% of minimal medium volume, and shaking table is protected from light culture 7d, obtain bacterium solution；
(2) it prepares oil paint wastewater degradation fungus strain: glucose being added in the bacterium solution of step (1), by the 5% of bacterium solution volume
It is inoculated with oiliness paint waste water, shaking table is carried out in the dark secondary culture, obtains oil paint wastewater degradation fungus strain culture solution；Each secondary culture
When bacterium solution in the amount of glucose reduce 1/5 than previous generation, the amount of oiliness paint waste water increases than previous generation, and each incrementss are
The 5% of bacterium solution volume, each secondary culture 7d, secondary culture >=five generations no longer add Portugal in culture medium when last is commissioned to train feeding
Grape sugar；
(3) it screens oil paint wastewater degradation fungus strain: agar being added into minimal medium, after sterilizing, trained to inorganic salts
It is added in the oil paint wastewater degradation fungus strain culture solution through (2) the step of gradient dilution and is coated with uniformly, 30 DEG C after supporting base solidification
7d is cultivated, has the bacterium colony of transparent circle around picking, repeats inoculated and cultured and separation of crossing, the oiliness paint waste water purified
Degradation bacterium series；
(4) secondary screening is carried out to the oil paint wastewater degradation fungus strain of purifying: the oil paint wastewater degradation bacterium that step (3) are purified
System is inoculated in LB culture medium, and after growing bacterium colony, being prepared into concentration is OD600=1 bacterium solution, it is spare；
In step (1),
It is 30 DEG C, 130r/min that the shaking table, which is protected from light condition,；
The component of the minimal medium are as follows: NaCl 0.5g, NH4NO30.4g, KH2PO41.0g, K2HPO41.0g
Yeast extract 0.2g, liquid microelement 4mL, distilled water 1000mL, adjusting pH value is 7.2；
The composition of the liquid microelement are as follows: MgSO42.0g, CuSO40.5g, MnSO40.5g, FeSO4·7H2O
0.5g, CaCl20.5g, distilled water 500mL.
In step (2),
The concentration of the oiliness paint waste water is 200mg/L；The weight of the glucose is minimal medium weight
0.5%；
The condition that the shaking table is protected from light is to cultivate 7 days in 30 DEG C, 130r/min；
The component of the minimal medium are as follows: NaCl 0.5g, NH4NO30.4g, KH2PO41.0g, K2HPO41.0g
Yeast extract 0.2g, liquid microelement 4mL, distilled water 1000mL, adjusting pH value is 7.2；
The composition of the liquid microelement are as follows: MgSO42.0g, CuSO40.5g, MnSO40.5g, FeSO4·7H2O
0.5g, CaCl20.5g, distilled water 500mL.
In step (3),
The weight of the agar is the 2% of minimal medium weight；
The volume ratio of the oil paint wastewater degradation fungus strain culture solution of the minimal medium and step (2) is 100:1；
The gradient dilution is to dilute the oil paint wastewater degradation fungus strain culture solution of step (2) respectively according to volume
103、104、105Times；
The component of the minimal medium are as follows: NaCl 0.5g, NH4NO30.4g, KH2PO41.0g, K2HPO41.0g
Yeast extract 0.2g, liquid microelement 4mL, distilled water 1000mL, adjusting pH value is 7.2；
The composition of the liquid microelement are as follows: MgSO42.0g, CuSO40.5g, MnSO40.5g, FeSO4·7H2O
0.5g, CaCl20.5g, distilled water 500mL.
In step (4),
The composition of the LB plating medium are as follows: peptone 2.5g, beef extract 0.75g, NaCl 1.25g, distilled water
250mL。
The present invention has the following advantages and effects with respect to the prior art: the present invention is useless using Biochemical method oil paint
Water, with low energy consumption, reaction condition is mild and efficiency is higher.There is the present invention screening of exclusive adapted microbes to tame training
It supports, degradation bacteria strains can achieve 80% or more to the degradation efficiency of oiliness paint waste water, for improving the tolerance of bio-reaction system
Property and stability, improve degradation efficiency be of great significance.
Specific embodiment
Below with reference to embodiment, the present invention is described in further detail, and embodiments of the present invention are not limited thereto.
Embodiment 1
(1) preparation of the acquisition of bacterium source and bacteria suspension: acquisition Yancheng DONGFENG YUEDA Kia Motors Paint Factory sewage draining exit soil,
Sludge is placed in 4 DEG C of refrigerators and is saved backup；The soil of 20g acquisition back is placed in the sterile water of 250mL, 30 DEG C of oscillations 12
Hour, bacteria suspension is made；
(2) it takes the injection of 5mL bacteria suspension to fill in the conical flask of 100mL minimal medium, is placed in 30 DEG C of constant-temperature tables
Interior, 130r/min shake culture 7d obtains bacterium solution；
(3) minimal medium: NaCl 0.5g, NH4NO30.4g, KH2PO41.0g, K2HPO41.0g, yeast extract
0.2g, liquid microelement 4mL, distilled water 1000mL, adjusting pH value is 7.2；The wherein composition of liquid microelement are as follows: MgSO42.0g
CuSO40.5g, MnSO40.5g, FeSO4·7H2O 0.5g, CaCl20.5g, distilled water 500mL；
(4) bacterium solution in 1mL step (2) is taken, is added in the minimal medium of 100mL, 30 DEG C, 130r/min concussion training
7d is supported, the content of oil paint is wherein added in minimal medium to 50mg/L, the content of glucose is added to 200mg/L；
(5) bacterium solution in 1mL step (4) is taken, is added in the minimal medium of 100mL, 30 DEG C, 130r/min concussion training
7d is supported, the content of oil paint is wherein added in minimal medium to 100mg/L, the content of glucose is added to 100mg/L；
(6) bacterium solution in 1mL step (5) is taken, is added in the minimal medium of 100mL, 30 DEG C, 130r/min concussion training
7d is supported, the content of oil paint is wherein added in minimal medium to 150mg/L, the content of glucose is added to 50mg/L；
(7) bacterium solution in 1mL step (6) is taken, is added in the minimal medium of 100mL, 30 DEG C, 130r/min concussion training
7d is supported, the content of oil paint is wherein added in minimal medium to 200mg/L, no longer plus glucose；
(8) bacterium solution in step (7) is subjected to gradient dilution, the gradient dilution is by the oil paint solution in step (7)
Fungus strain culture solution dilutes 10 according to volume respectively3、104、105Times, it is coated on solid plate, 30 DEG C of culture 7d, has around picking
The bacterium colony of transparent circle, repeated inoculation culture and separation of crossing, the oil paint wastewater degradation bacterium purified.
(9) secondary screening is carried out to the oil paint wastewater degradation bacterium of purifying: the oil paint wastewater degradation bacterium that step (8) obtains is connect
In LB plating medium, culture 7d scrapes strain in sterile water after growing bacterium colony kind, and bacterium is dispersed as bacterium solution, adjusts
The concentration of bacterium solution is OD600=1, it is spare.The composition of the LB plating medium are as follows: peptone 2.5g, beef extract 0.75g, NaCl
1.25g, distilled water 250mL.
Finally obtained oil paint wastewater degradation fungus strain can be grown by substrate of oiliness paint waste water, and aerobic degradation tests table
Bright flora can carry out effective aerobic degradation to oiliness paint waste water；When the concentration of oiliness paint waste water is 200mg/L, after 7d
Degradation efficiency reaches 83% after degradation rate reaches 79%, 10d.
The above embodiment is a preferred embodiment of the present invention, but embodiments of the present invention are not by above-described embodiment
Limitation, other any changes, modifications, substitutions, combinations, simplifications made without departing from the spirit and principles of the present invention,
It should be equivalent substitute mode, be included within the scope of the present invention.
Claims (1)
1. a kind of oil paint wastewater degradation bacterium, it is characterised in that: the bacterial strain is microbacterium (microbacterium sp.)
XR001, preservation from date are on December 23rd, 2014, and deposit number is CGMCC No.10237.
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