CN108619899B - A kind of biological active filling material comprising Meng Shi pseudomonad - Google Patents

A kind of biological active filling material comprising Meng Shi pseudomonad Download PDF

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CN108619899B
CN108619899B CN201810455474.3A CN201810455474A CN108619899B CN 108619899 B CN108619899 B CN 108619899B CN 201810455474 A CN201810455474 A CN 201810455474A CN 108619899 B CN108619899 B CN 108619899B
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filler
microbial activity
meng shi
parathion
shi pseudomonad
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CN108619899A (en
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任仁
朱利军
柴春燕
晋超
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Dajiang Environment Co ltd
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    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D53/00—Separation of gases or vapours; Recovering vapours of volatile solvents from gases; Chemical or biological purification of waste gases, e.g. engine exhaust gases, smoke, fumes, flue gases, aerosols
    • B01D53/34—Chemical or biological purification of waste gases
    • B01D53/74—General processes for purification of waste gases; Apparatus or devices specially adapted therefor
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    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D53/00—Separation of gases or vapours; Recovering vapours of volatile solvents from gases; Chemical or biological purification of waste gases, e.g. engine exhaust gases, smoke, fumes, flue gases, aerosols
    • B01D53/34—Chemical or biological purification of waste gases
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    • B01D—SEPARATION
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    • B01D53/34—Chemical or biological purification of waste gases
    • B01D53/46—Removing components of defined structure
    • B01D53/68—Halogens or halogen compounds
    • B01D53/70—Organic halogen compounds
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    • 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
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Abstract

The invention belongs to field of environmental improvement, and in particular to a kind of microbial activity filler containing Meng Shi pseudomonad.The Meng Shi pseudomonad is specially Meng Shi pseudomonad (Pseudomonas monteilii) LW-1, deposit number is CGMCC No.13748, the microbial activity filler is adsorbed on filler by Meng Shi pseudomonad bacteria suspension, and filler is seated in wire side hollow sphere and is prepared, the filler be used to handle the generation of agricultural chemicals waste water reservoir containing the organic sulphur waste gas, gas flow 100-5000m3/ h, residence time 1.0-5.0min, 35-55 DEG C of temperature, malathion, parathion, parathion-methyl total concentration be 20-200mg/m3, malathion, parathion, parathion-methyl degradation efficiency can achieve 90% or more.

Description

A kind of biological active filling material comprising Meng Shi pseudomonad
Technical field:
The invention belongs to field of environmental improvement, and in particular to one kind includes Meng Shi pseudomonad (Pseudomonas Monteilii) the biological active filling material of LW-1.
Background technique:
Waste water in pesticide producing contains a large amount of organosulfur pesticide and organo-chlorine pesticide intermediate, complicated component, concentration High, strong toxicity.The product or in which mesosome of part organosulfur pesticide and organo-chlorine pesticide such as parathion, malathion, methyl pair Sulphur phosphorus, trichloroacetaldehyde, phosphorus trichloride etc., have a volatility, and most of organic sulfur and organo-chlorine pesticide have it is pungent Peculiar smell.
Parathion (C10H14NO5It PS) is colourless oil liquid, solubility is 24mg/L (25 DEG C) in water.It is slightly soluble in petroleum ether And kerosene, there can be garlic odour with most immiscible organic solvents.It is promptly hydrolyzed in alkaline medium；In neutral or slightly acidic solution In it is relatively stable；It is all unstable to ultraviolet light and air.There are mutagenicity and animal teratogenesis to microorganism.Belong to hypertoxic type, tool There are acute toxicity, irritation, mutagenicity.Human lymphocyte Sister chromatid exohange is influenced when 200 μ g/L；10μmol/ Human fibroblasts unscheduled DNA synthesis is influenced when L；When 10 μm of ol/L, rat lymphocyte DNA damage is made in 16 hours, IARC carcinogenicity is evaluated as G3 grades；The minimum toxic dose of rat oral (TDLo): 360 μ g/kg (pregnant 2~22d/ postpartum 15d), Influence newborn rat biochemistry and metabolism；The minimum toxic dose of subcutaneous rat (TDLo): 9800 μ g/kg (pregnant 7~13d), lethal tire. Parathion can enter human body by alimentary canal, respiratory tract and complete skin and mucous membrane.Parathion in environment can also pass through Biological concentration occurs for food chain.
Malathion (C10H19O6PS2) it is colourless or pale yellow oily liquid, there is garlic smell.Chance open fire, high fever is flammable；With Strong oxidizer reacts；Phosphorus, sulfur oxide gas are generated by thermal decomposition；It is toxic to animal, it is irritating to eyes, skin Property；Active in 5.0 or less pH, pH7.0 or more is easy hydrolysis failure.Female mice acute oral LD50 is 1751.5mg/kg, male Rat acute LD50 is 1634. 5mg/kg, and the percutaneous LD50 of rat acute is 4000~6150mg/kg.Poisoning symptom is head Bitterly, dizziness, nausea, inability, hidrosis, vomiting, salivation, eye-blurred, myosis, spasm, stupor, flesh fibrillation, pulmonary edema etc.. Generally morbidity is very fast after contact in a short time (oral, sucking, skin, mucous membrane) a large amount of organophosphorus pesticides.Malathion poisoning is dived The volt phase is longer.It absorbs through skin, falls ill in 12h, poisoning symptom shows as appetite stimulator, Nausea and vomiting, abdominal pain, hidrosis, thing Object is fuzzy, myosis, respiratory secretions increase, bronchial spasm expiratory dyspnea, pulmonary edema；Sample Symptoms are muscle bundle shake It quivers, muscle spasmus, myoparalysis；Central nervous system symptom shows as headache, dizzy, insomnia drowsiness, out of strength, irritated, twitch living, dusk Fan, brain edema.The state of an illness is heavier when oral ingestion, and gastrointestinal symptom becomes apparent from.Some cases can have cardiac muscle, liver, renal damage.
Parathion-methyl (C8H10NO5PS): colorless crystalline powder, industrial goods are brown or yellow liquid or solid, are met bright Fire, high fever is flammable, by poisonous gas such as thermal decomposition release phosphorus, sulfur oxides, can decompose rapidly in lye.To big white mouse urgency Property LD50 of passing through mouth be 14~24mg/kg, to rabbit acute percutaneous LD50 be 300~400mg/kg, belong to high toxic pesticide.Methyl pair Sulphur phosphorus tool is tagged and stomach poison function, can inhibit the vigor of cholinesterase in pest nervous system and lethal, and insecticidal spectrum is wide, Chang Jiagong It is used at missible oil or pulvis (see the formulations of pesticide), is mainly used for prevention and treatment various agricultural pest.There is certain cause to microorganism Mutability, such as 667 μ g/ ware of salmonella typhimurium；Escherichia coli: 10mmol/L.Gaseous state, mist state, aerosol state or powdery Easily for respiratory tract absorb, liquid can be through eye mucous membrane, skin or gastrointestinal tract.Into after in vivo, distribution is each to whole body rapidly Organ-tissue, and be firmly combined with histone.
Organo-chlorine pesticide is one of current most popular insecticide, is occupied an important position in pesticide market.So And in organo-chlorine pesticide production and agricultural chemicals waste water treatment process, there is volatile Pesticidal products or intermediate, waved from liquid phase Hair, spills into air, forms air pollution.
Volatile pesticide and intermediate include: metrifonate, DDVP, phosphorus trichloride, dichlobenil, herbicide, thimet, Phoxim, biphenyl, BFPO, alkane and alkane substitute class intermediate, aromatic intermediate etc. (choosing organochlorine), these substances Most of have a pungent smell, some substances such as diethyl phosphite, trichloroacetaldehyde, belong to carcinogenicity, mutagenicity and The cause property substance of the three of teratogenesis.
It is these organic sulfurs with peculiar smell, organic because being aerated, pushing the mechanical disturbances such as water flow in wastewater treatment process Chlorine is spilt into air from water, forms odor pollution.If not being subject to processing appropriate, the organochlorine in air is spilt into, Serious pollution environment, is detrimental to health.
Biotechnology cleans because it has many advantages, such as at low cost, easy to operate, technology, is without secondary pollution, is effective at present Handle one of the important method containing organic compounds.The principle of biologic treating technique is to utilize the biological oxidation of microorganism will Organic sulfur conversion is the process of the harmless or low harmful substances such as sulfate, carbon dioxide, water, as follows:
Malathion → carbon dioxide+sulfate+water；
Parathion-methyl → p-nitrophenol+organic sulfate → sulfate+phosphate+carbon dioxide+water；
Parathion → p-nitrophenol+organic sulfate → sulfate+phosphate+carbon dioxide+water.
The principle of biologic treating technique is to convert carbon dioxide, salt for organochlorine using the biological oxidation of microorganism Hydrochlorate and water etc. be harmless or the process (formula 1) of low harmful substance.
C2HCl3O+O2→CO2+H2O+HCl (formula 1)
Existing research shows that the facultative chemoheterotrophy bacterium in natural environment belongs to pseudomonad (Pseudomonas Sp.) X2 (201310551914.2), Sphingobacterium (Sphingobacterium griseoflavus) SCU-B140 (201510715812.9), Acidithiobacillus ferrooxidans strain GF (Acidithiobacillus ferrooxidans), thermophilic iron hook end The mixed culture of spirillum (Leptospirillum ferriphilum) and Ferroplasma acidiphilum (201310745138.X) etc. has degradation to organosulfur pesticide.But not yet discovery is for handling organic sulfur and organic The Meng Shi pseudomonad (Pseudomonas monteilii) of chlorine, and use Meng Shi pseudomonad processing sulfur-bearing, containing chlorine dirt Contaminate the method for object.
Summary of the invention:
In order to solve, volatility organic sulfur, organochlorine treatment effeciency in the prior art be low, microorganism is on mixed carrier surface Load is uneven, starting is slow, the problem of filler collapses is used for a long time, and the present invention provides one plant of Meng Shi pseudomonad (Pseudomonas monteilii), and the bacterial strain is applied to removing containing in organic sulfur, organochlorine exhaust gas.
The Meng Shi pseudomonad, specially Meng Shi pseudomonad (Pseudomonas monteilii) LW-1 is From the agricultural chemicals waste water containing the organochlorines such as the organic sulfurs such as parathion-methyl, malathion, parathion and phosphorus trichloride, trichloroacetaldehyde Domestication, isolated separation bacterium.The bacterial strain is preserved in Chinese microorganism strain preservation conservator on March 10th, 2017 Meeting common micro-organisms center, deposit number are CGMCC No.13748, preservation address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3 Number, Institute of Microorganism, Academia Sinica, postcode 100101.
The bacterial strain feature is as follows:
Meng Shi pseudomonad (Pseudomonas monteilii) LW-1: cell is in the shape of a rod, peritrichous, gram dye Color is negative, and Starch Hydrolysis, VP, indole reaction are feminine gender, and gelatin liquefaction, glucose response are the positive.Bacterium colony is white, round, Surface is smooth, moistens sticky, flat protrusion.In the natural environment, competitiveness is strong, wide adaptation range, can utilize carbon extensively Hydrate, amino acid, organic acid are carbon source, pH wide adaptation range (pH 3.0-7.0), under high temperature (50 DEG C -60 DEG C) part It can all grow, optimum growth temperature is 30-55 DEG C, pH 4.0-6.5.
The present invention also provides a kind of microbial activity filler, the microbial activity filler is by Meng Shi pseudomonad LW-1 Bacteria suspension is adsorbed on filler, and filler is seated in wire side hollow sphere and is prepared, can preferably treatment and purification containing organic The exhaust gas of sulphur and/or organochlorine.
The filler is one of polyurethane foamed blocks, haydite, volcanic rock or molecular sieve or a variety of；
The packing density is 20-80kg/m3, porosity 50-97%, partial size 10-100mm；
The mass ratio of the microbial bacteria suspension and filler is 1-12: 1；
Meng Shi pseudomonad LW-1 cell concentration 1.0 × 10 in the microbial activity filler5 -9.0×108CFU/g(Filler)；
The wire side hollow sphere material is polyethylene, polyvinyl chloride or polypropylene；
The hollow bulb diameter 6cm of the wire side, is made of two hemisphere, and each hemisphere surface is porous, the aperture in each hole 2-8mm；
There is a circle reinforcing ring in the middle part of the wire side hollow sphere, reinforcing ring is equipped with lock, convenient for unlatching and closure two Hemisphere.
Specific preparation process is as follows for the filler:
(1) preparation of Meng Shi pseudomonad LW-1 bacteria suspension
1. being carried out liquid fermentation medium is accessed after Meng Shi pseudomonad LW-1 slant activation by 10-25% inoculum concentration Expand fermented and cultured step by step；
Fermentation culture conditions are as follows: in 35-45 DEG C, ventilation quantity 10-100L/min, dissolved oxygen > 2.0mg/L, tank presses 0.03- Under the conditions of 0.10Mpa, incubation time 45-55h；
The composition of the liquid fermentation medium are as follows: glucose 0.5g, K2HPO40.5g, KH2PO41.5g, (NH4)2SO40.5g, NaCl 0.5g, MgSO40.2g, CaCl20.05g, FeSO40.02g, water 1000mL, pH value 6.2-7.0, 115 DEG C of sterilizing 20min, are cooled to room temperature after sterilizing and add 4.5mg/L parathion and/or trichloroacetaldehyde (for handling sulfur-bearing 4.5mg/L parathion is added when exhaust gas；4.5mg/L trichloroacetaldehyde is added when for handling chlorine contained exhaust gas；For handle sulfur-bearing and 4.5mg/L parathion and 4.5mg/L trichloroacetaldehyde are added when chlorine contained exhaust gas)；
2. after fermented and cultured, fermentation liquid being placed in refrigerated centrifuge in 4 DEG C, 2000-8000rpm, 10- is centrifuged 20min discards supernatant liquid, and precipitating is diluted with sterile nutrient solution, and acquisition cell concentration is 0.30-0.95g/L Meng Shi pseudomonad LW-1 bacteria suspension；
The composition of the sterile nutrient solution are as follows: glucose 0.5g, K2HPO40.5g, KH2PO41.5g, (NH4)2SO4 0.5g, NaCl 0.5g, MgCl20.2g, CaCl20.05g, FeSO40.02g, water 1000mL, pH value 6.8-7.2,121 DEG C Sterilize 20min；
(2) preparation of microbial activity filler:
1. the filler of packet or grain shape is filled into wire side hollow sphere, the mass ratio of filler and wire side hollow sphere For 0.9-3:1；
2. being sprayed the Meng Shi pseudomonad LW-1 bacteria suspension of step (1) preparation by the hole of wire side hollow sphere to packet Or the surface of the filler of grain shape；
It is mixed 3. the wire side hollow sphere that load has filler is put into rotate in impeller；
4. again to the wire side hollow sphere surface spraying Meng Shi pseudomonad LW-1 bacteria suspension that filler is housed；
It repeats step 1. -4. 2-3 time, equal appendix on filler is made in hollow ball surface and hole to have bacterium cell, acquisition Meng Shi Pseudomonad LW-1 cell concentration reaches 1.0 × 105-9.0×108CFU/g(Filler)Microbial activity filler, 4 DEG C of preservations are standby With.
The side containing organic sulfur and/or organochlorine exhaust gas is handled using mentioned microorganism active filler the present invention also provides a kind of Method, the specific steps are as follows:
(1) there is the microbial activity filler of Meng Shi pseudomonad LW-1 to be placed in a container above-mentioned appendix, be passed through containing pair Sulphur phosphorus gas (when processing sulphur-containing exhaust gas), using parathion as phosphorus source, sulphur source, activates microbial activity filler；Activation time is 2- 15 days；
The concentration of parathion in the gas containing parathion being passed through is 50-200mg/m3；
It is passed through the gas containing trichloroacetaldehyde when handling chlorine contained exhaust gas, microbial activity filler is activated with trichloroacetaldehyde；It is living Changing the time is 2-15 days；
The concentration of the trichloroacetaldehyde is 50-200mg/m3；
It is passed through the gas containing parathion and trichloroacetaldehyde when processing while sulfur-bearing and chlorine contained exhaust gas, activates microbial activity Filler；Activation time is 2-15 days；The concentration of parathion is 50-200mg/m3, the concentration of trichloroacetaldehyde is 50-200mg/m3；
(2) the microbial activity filler after activation is placed in bioreactor, is passed through useless containing organic sulfur and/or organochlorine Gas, with the organic sulfur and/or organochlorine in Meng Shi pseudomonad LW-1 purification exhaust gas, microbial activity packing volume is 0.1- 1000m3, the total concentration of organic sulfur is 20-250mg/m3And/or the total concentration of organochlorine is 5-100mg/m3, gas flow rate is 0.05-10000m3/ h, residence time 1.5-10.0min, purification temperature are 30-65 DEG C；Periodically to microbial activity filler surface Spraying nutrient solution maintains the activity of microbial activity filler, and spray frequency is to spray 1 time for 1-3 days, and each spray rate is 1.0- 200L/min, each spray time are 10-90min；
The composition of the nutrient solution are as follows: glucose 0.5g, K2HPO40.5g, KH2PO41.5g, (NH4)2SO4 0.5g, NaCl 0.5g, MgSO40.2g, CaCl20.05g, FeSO40.02g, water 1000ml, pH value 6.2-7.0.
The utility model has the advantages that
1. Meng Shi pseudomonad (Pseudomonas monteilii) LW-1 degradation organic sulfur with higher of the invention With the efficiency of organochlorine, to the degradation efficiency of organic sulfur up to 90-100%, to the degradation efficiency of organochlorine up to 75% or more； And the laundering period of dominant bacteria is short, and inoculation can quickly start degradation reaction in 1 week, applies and is purifying the exhaust gas life containing organic sulfur In object reactor, the starting period can be shortened.
2. using the effect of a variety of mechanics such as sedimentation, attachment and centrifugation, by the Meng Shi pseudomonad (Pseudomonas that degrades Monteilii) LW-1 appendix forms microbial activity filler, bacterium cell on the light porous material of wire side hollow ball inside It can not only be uniformly adhered to filler surface, and be able to enter in the hole inside filler, the bacterium of microbial activity filler is made Cell appendix amount is high, is not easy to be lost from bioreactor.
3. Meng Shi pseudomonad (Pseudomonas monteilii) LW-1 has, distribution is wide, quantity is more, nutritional need is simple Feature single, breeding is fast, competition colonization power is strong.The pseudomonad inhibits other microorganisms, containing biting iron by secretion antibiotic Element, the siderophore that can be secreted have a clear superiority in the competition of iron.
4. Meng Shi pseudomonad (Pseudomonas monteilii) LW-1 is chemoorganotrophy type, there is catalase, It can utilize and decomposing volatile organic sulfur/organochlorine.
5. Meng Shi pseudomonad (Pseudomonas monteilii) LW-1 has antiviral activity, to TMV (Tobacco mosaic Virus), the viruses such as PVY (marmor upsilon) have inhibiting effect, use is safer.
6. the rough surface, porous of the carrier material for microbial activity filler is suitable for bacterium cell and adheres to；Specific surface area Greatly, be conducive to the volatility such as bacterium cell and parathion, trichloroacetaldehyde organic sulfur, organochlorine contacts；Load the wire side of light filler Hollow sphere hole is more, and good permeability, resistance is small, and mass-transfer efficiency is high；Microbial activity filler weight is light, intensity is high, the pressure loss It is small；It is high temperature resistant, corrosion-resistant；Not perishable deformation, can be used for a long time and adapt to the physical condition of Various Complex.
Detailed description of the invention:
Fig. 1 Meng Shi pseudomonad (Pseudomonas monteilii) LW-1 phylogenetic tree.
Specific embodiment:
In order to which the objects, technical solutions and advantages of this patent are more clearly understood, below in conjunction with specific embodiment, to this Patent is further elaborated.It should be appreciated that specific embodiment described herein is only to explain this patent, and do not have to It is of the invention in limiting.
Strain idenfication:
DNA is extracted: using Full-automatic magnetic beads instrument for extracting nucleic acid Smart LabAssist-16 and respective environment sample DNA The bacterial strain DNA that extraction purification kit (Taiwan dot nanometer technical concern Co., Ltd) screens acquisition respectively extract and Purifying, selects bacterial universal primers F16S-27 and R16S-1492 to carry out PCR amplification, and primer sequence is respectively as follows:
F16S-27(5`-AGAGTTTGATCCTGGCTCAG-3`)
R16S-1492(5`-CGGTTACCTTGTTACGACTTC-3`)
PCR reaction condition setting are as follows: 94 DEG C of initial denaturation 5min；Subsequent 94 DEG C of denaturation 30s, 56 DEG C of annealing 30s, 72 DEG C are prolonged 90s is stretched, 25 periods are recycled；Then 72 DEG C of extension 10min；Last 4 DEG C of holdings 10min.By being purified to PCR product, quality inspection PCR purified product is connected on PMD-18T carrier afterwards, is then transformed into bacillus coli DH 5 alpha competent cell.It will conversion 50 μ L of competent cell afterwards is coated on the LB culture medium containing isopropylthiogalactoside, X-gal and ammonia benzyl.37 DEG C of perseverances Warm incubator is incubated overnight, and carries out LB liquid medium culture from each plate picking 25 white single bacterium spots.Culture 2 hours Afterwards, culture solution is subjected to sequencing analysis.The sequence measured uses DNAMAN software to carry out similitude merger first, and similitude is big It is an operating unit in 97% sequence merger, chooses representative sequence in each operating unit and utilize BLAST and GenBank 16SrDNA gene order in database carries out homology analysis, chooses 1400bp or more length and is compared (Clustelx1.83), Phylogenetic Analysis (MEGA3.1) is carried out using ortho position connection (Neighbour Joining) method, system Development tree is as shown in Figure 1.It is compared by DNA sequence dna, which is Meng Shi pseudomonad (Pseudomonas monteilii), It is named as Meng Shi pseudomonad (Pseudomonas monteilii) LW-1.
Embodiment 1: a kind of microbial activity filler of the removing containing the organic sulphur waste gas and its preparation
A kind of microbial activity filler is to be adsorbed in polyurethane foamed blocks filler by Meng Shi pseudomonad LW-1 bacteria suspension On, and filler is seated in wire side hollow sphere and is prepared；
Packing density is 20kg/m3, porosity 90%, partial size 10mm；The mass ratio of microbial bacteria suspension and filler It is 1: 1；Meng Shi pseudomonad LW-1 cell concentration 1.0 × 10 in microbial activity filler5 CFU/g(filler)；
The wire side hollow sphere material is polyethylene；The hollow bulb diameter 6cm of wire side, is made of, Mei Geban two hemisphere Ball surface is porous, the aperture 2mm in each hole；There is a circle reinforcing ring in the middle part of wire side hollow sphere, reinforcing ring is equipped with lock, convenient for opening Open and be closed two hemisphere.
Specific preparation process is as follows for the filler:
(1) preparation of Meng Shi pseudomonad LW-1 bacteria suspension
1. being carried out liquid fermentation medium is accessed after Meng Shi pseudomonad LW-1 slant activation step by step by 10% inoculum concentration Expand fermented and cultured；
Fermentation culture conditions are as follows: in 35 DEG C, ventilation quantity 10L/min, dissolved oxygen > 2.0mg/L, tank presses 0.03-0.10Mpa Under the conditions of, incubation time 55h；
Slant medium composition are as follows: glucose 0.5g, K2HPO40.5g, KH2PO41.5g, (NH4)2SO40.5g, NaCl 0.5g, MgSO40.2g, CaCl20.05g, FeSO40.02g, water 1000ml, agar 20g, pH value 6.2-7.0,115 DEG C go out Bacterium 20min；
The composition of the liquid fermentation medium are as follows: glucose 0.5g, K2HPO40.5g, KH2PO41.5g, (NH4)2SO40.5g, NaCl 0.5g, MgSO40.2g, CaCl20.05g, FeSO40.02g, water 1000mL, pH value 6.2-7.0, 115 DEG C of sterilizing 20min, are cooled to room temperature after sterilizing and add 4.5mg/L parathion；
2. after fermented and cultured, fermentation liquid being placed in refrigerated centrifuge in 4 DEG C, 2000rpm, 10min is centrifuged, is abandoned Supernatant is removed, precipitating is diluted with sterile nutrient solution, and acquisition cell concentration is 0.30g/L Meng Shi pseudomonad LW-1 bacteria suspension；
The composition of the sterile nutrient solution are as follows: glucose 0.5g, K2HPO40.5g, KH2PO41.5g, (NH4)2SO4 0.5g, NaCl 0.5g, MgCl20.2g, CaCl20.05g, FeSO40.02g, water 1000mL, pH value 6.8-7.2,121 DEG C Sterilize 20min；
(2) preparation of microbial activity filler:
1. packet filler is filled into wire side hollow sphere, the mass ratio of filler and wire side hollow sphere is 0.9:1；
2. being sprayed the Meng Shi pseudomonad LW-1 bacteria suspension of step (1) preparation by the hole of wire side hollow sphere to packet The surface of the filler of shape；
It is mixed 3. the wire side hollow sphere that load has filler is put into rotate in impeller；
4. again to the wire side hollow sphere surface spraying Meng Shi pseudomonad LW-1 bacteria suspension that filler is housed；
1. -4. 2 time it repeats step, makes in hollow ball surface and hole that equal appendix has bacterium cell on filler, obtain Meng Shi vacation Monad LW-1 cell concentration reaches 1.0 × 105CFU/g(filler)Microbial activity filler, 4 DEG C save backup.
Embodiment 2: a kind of microbe stuffing of the removing containing the organic sulphur waste gas and its preparation
A kind of microbial activity filler is to be adsorbed on ceramics filling by Meng Shi pseudomonad LW-1 bacteria suspension, and will fill out Material, which is seated in wire side hollow sphere, to be prepared；Packing density is 80kg/m3, porosity 50%, partial size 100mm；Micro- life The mass ratio of object bacteria suspension and filler is 12: 1；Meng Shi pseudomonad LW-1 cell concentration 9.0 in microbial activity filler × 108CFU/g(filler)；
The wire side hollow sphere material is polyvinyl chloride；The hollow bulb diameter 6cm of wire side, is made of, each two hemisphere Hemisphere surface is porous, the aperture 8mm in each hole；There is a circle reinforcing ring in the middle part of wire side hollow sphere, reinforcing ring is equipped with lock, is convenient for Open and be closed two hemisphere.
Specific preparation process is as follows for the filler:
(1) preparation of Meng Shi pseudomonad LW-1 bacteria suspension
1. being carried out liquid fermentation medium is accessed after Meng Shi pseudomonad LW-1 slant activation step by step by 25% inoculum concentration Expand fermented and cultured；
Fermentation culture conditions are as follows: in 45 DEG C, ventilation quantity 100L/min, dissolved oxygen > 2.0mg/L, tank presses 0.10Mpa condition Under, incubation time 45h；
Slant medium composition are as follows: glucose 0.5g, K2HPO40.5g, KH2PO41.5g, (NH4)2SO40.5g, NaCl 0.5g, MgSO40.2g, CaCl20.05g, FeSO40.02g, water 1000ml, agar 20g, pH value 6.2-7.0,115 DEG C go out Bacterium 20min；
The composition of the liquid fermentation medium are as follows: glucose 0.5g, K2HPO40.5g, KH2PO41.5g, (NH4)2SO40.5g, NaCl 0.5g, MgSO40.2g, CaCl20.05g, FeSO40.02g, water 1000mL, pH value 6.2-7.0,115 DEG C sterilizing 20min, be cooled to room temperature after sterilizing and add 4.5mg/L parathion；
2. after fermented and cultured, fermentation liquid being placed in refrigerated centrifuge in 4 DEG C, 8000rpm, 20min is centrifuged, is abandoned Supernatant is removed, precipitating is diluted with sterile nutrient solution, and acquisition cell concentration is 0.95g/L Meng Shi pseudomonad LW-1 bacteria suspension；
The composition of the sterile nutrient solution are as follows: glucose 0.5g, K2HPO40.5g, KH2PO41.5g, (NH4)2SO4 0.5g, NaCl 0.5g, MgCl20.2g, CaCl20.05g, FeSO40.02g, water 1000mL, pH value 6.8-7.2,121 DEG C Sterilize 20min；
(2) preparation of microbial activity filler:
1. the filler of grain shape is filled into wire side hollow sphere, the mass ratio of filler and wire side hollow sphere is 2:1；
2. being sprayed the Meng Shi pseudomonad LW-1 bacteria suspension of step (1) preparation by the hole of wire side hollow sphere to granulated The surface of the filler of shape；
It is mixed 3. the wire side hollow sphere that load has filler is put into rotate in impeller；
4. again to the wire side hollow sphere surface spraying Meng Shi pseudomonad LW-1 bacteria suspension that filler is housed；
1. -4. 3 time it repeats step, makes in hollow ball surface and hole that equal appendix has bacterium cell on filler, obtain Meng Shi vacation Monad LW-1 cell concentration reaches 9.0 × 108CFU/g(filler)Microbial activity filler, 4 DEG C save backup.
Embodiment 3: a kind of microbe stuffing of the removing containing the organic sulphur waste gas and its preparation
A kind of microbial activity filler is to be adsorbed on volcanic filling material by Meng Shi pseudomonad LW-1 bacteria suspension, and incite somebody to action Filler is seated in wire side hollow sphere and is prepared；Packing density is 50kg/m3, porosity 75%, partial size 50mm；It is micro- The mass ratio of biological bacteria suspension and filler is 8: 1；Meng Shi pseudomonad LW-1 cell concentration 3.0 in microbial activity filler × 107CFU/g(filler)；
The wire side hollow sphere material is polypropylene；The hollow bulb diameter 6cm of wire side, is made of, Mei Geban two hemisphere Ball surface is porous, the aperture 5mm in each hole；There is a circle reinforcing ring in the middle part of wire side hollow sphere, reinforcing ring is equipped with lock, convenient for opening Open and be closed two hemisphere.
Specific preparation process is as follows for the filler:
(1) preparation of Meng Shi pseudomonad LW-1 bacteria suspension
1. being carried out liquid fermentation medium is accessed after Meng Shi pseudomonad LW-1 slant activation step by step by 17% inoculum concentration Expand fermented and cultured；
Fermentation culture conditions are as follows: in 40 DEG C, ventilation quantity 50L/min, dissolved oxygen > 2.0mg/L, tank presses 0.03-0.10Mpa Under the conditions of, incubation time 50h；
Slant medium composition are as follows: glucose 0.5g, K2HPO40.5g, KH2PO41.5g, (NH4)2SO40.5g, NaCl 0.5g, MgSO40.2g, CaCl20.05g, FeSO40.02g, water 1000ml, agar 20g, pH value 6.2-7.0,115 DEG C go out Bacterium 20min；
The composition of the liquid fermentation medium are as follows: glucose 0.5g, K2HPO40.5g, KH2PO41.5g, (NH4)2SO40.5g, NaCl 0.5g, MgSO40.2g, CaCl20.05g, FeSO40.02g, water 1000mL, pH value 6.2-7.0, 115 DEG C of sterilizing 20min, are cooled to room temperature after sterilizing and add 4.5mg/L parathion；
2. after fermented and cultured, fermentation liquid is placed in refrigerated centrifuge in 4 DEG C, 5000rpm, 10-20min is centrifuged, Liquid is discarded supernatant, precipitating is diluted with sterile nutrient solution, and acquisition cell concentration is 0.65g/L Meng Shi pseudomonad LW-1 bacteria suspension；
The composition of the sterile nutrient solution are as follows: glucose 0.5g, K2HPO40.5g, KH2PO41.5g, (NH4)2SO4 0.5g, NaCl 0.5g, MgCl20.2g, CaCl20.05g, FeSO40.02g, water 1000mL, pH value 6.8-7.2,121 DEG C Sterilize 20min；
(2) preparation of microbial activity filler:
1. packet filler is filled into wire side hollow sphere, the mass ratio of filler and wire side hollow sphere is 3:1；
2. being sprayed the Meng Shi pseudomonad LW-1 bacteria suspension of step (1) preparation by the hole of wire side hollow sphere to box-shaped The surface of the filler of shape；
It is mixed 3. the wire side hollow sphere that load has filler is put into rotate in impeller；
4. again to the wire side hollow sphere surface spraying Meng Shi pseudomonad LW-1 bacteria suspension that filler is housed；
1. -4. 3 time it repeats step, makes in hollow ball surface and hole that equal appendix has bacterium cell on filler, obtain Meng Shi vacation Monad LW-1 cell concentration reaches 3.0 × 107CFU/g(filler)Microbial activity filler, 4 DEG C save backup.
A kind of microbe stuffing for removing the exhaust gas containing organochlorine of embodiment 1 ' and its preparation
Parathion in preparation step step (1), is only replaced with the trichlorine of equivalent by filler and the preparation method is the same as that of Example 1 Acetaldehyde.
A kind of microbe stuffing for removing the exhaust gas containing organochlorine of embodiment 2 ' and its preparation
Parathion in preparation step step (1) is only replaced with the trichlorine of equivalent with embodiment 2 by filler and preparation method Acetaldehyde.
A kind of microbe stuffing for removing the exhaust gas containing organochlorine of embodiment 3 ' and its preparation
Parathion in preparation step step (1) is only replaced with the trichlorine of equivalent with embodiment 3 by filler and preparation method Acetaldehyde.
A kind of embodiment 4: method of the microbial activity filler processing containing the organic sulphur waste gas
The method containing the organic sulphur waste gas is handled using microbial activity filler described in embodiment 1 the present invention also provides a kind of, Specific step is as follows:
(1) there is the microbial activity filler of Meng Shi pseudomonad LW-1 to be placed in a container 1 appendix of embodiment, be passed through and contain The gas of parathion activates microbial activity filler using parathion as phosphorus source, sulphur source；Activation time is 15 days；
The concentration of parathion in the gas containing parathion being passed through is 50mg/m3；
(2) the microbial activity filler after activation is placed in bioreactor, is passed through containing the organic sulphur waste gas, with Meng Shi vacation Monad LW-1 purifies the organic sulfur in exhaust gas, and microbial activity packing volume is 0.1m3, malathion and parathion in air inlet Concentration be respectively 17.3mg/m3And 46.7mg/m3, gas flow rate 0.05m3/ h, residence time 1.5min, purification temperature are 30℃；The activity of microbial activity filler is periodically maintained to microbial activity filler surface spraying nutrient solution, spray frequency is 3 days Spray 1 time, each spray rate are 1.0L/min, and each spray time is 10min；Gas outlet malathion and parathion Concentration is reduced to 2.2 mg/m3And 4.3mg/m3, removal rate respectively reaches 87.3% and 90.8%.
The composition of the nutrient solution are as follows: glucose 0.5g, K2HPO40.5g, KH2PO41.5g, (NH4)2SO4 0.5g, NaCl 0.5g, MgSO40.2g, CaCl20.05g, FeSO40.02g, water 1000ml, pH value 6.2-7.0.
A kind of embodiment 5: method of the microbial activity filler processing containing the organic sulphur waste gas
The method containing the organic sulphur waste gas is handled using microbial activity filler described in embodiment 2 the present invention also provides a kind of, Specific step is as follows:
(1) there is the microbial activity filler of Meng Shi pseudomonad LW-1 to be placed in a container 2 gained appendix of embodiment, lead to Enter the gas containing parathion, using parathion as phosphorus source, sulphur source, activates microbial activity filler；Activation time is 2 days；
The concentration of parathion in the gas containing parathion being passed through is 200mg/m3；
(2) the microbial activity filler after activation is placed in bioreactor, is passed through containing the organic sulphur waste gas, with Meng Shi vacation Monad LW-1 purifies the organic sulfur in exhaust gas, and microbial activity packing volume is 1000m3, in air inlet, the concentration of parathion is 162.7mg/m3, the concentration of parathion-methyl is 32.1mg/m3, gas flow rate 10000m3/ h, residence time 10.0min, only Changing temperature is 65 DEG C；The activity of microbial activity filler, spray are periodically maintained to microbial activity filler surface spraying nutrient solution Frequency is to spray 1 time for 1 day, and each spray rate is 200L/min, and each spray time is 90min；Gas outlet, parathion it is dense Degree is reduced to 11.3 mg/m3, the concentration of parathion-methyl is reduced to 2.8mg/m3.Their removal rate respectively reaches 93.1% (parathion) and 91.3% (parathion-methyl).
The composition of the nutrient solution are as follows: glucose 0.5g, K2HPO40.5g, KH2PO41.5g, (NH4)2SO4 0.5g, NaCl 0.5g, MgSO40.2g, CaCl20.05g, FeSO40.02g, water 1000ml, pH value 6.2-7.0.
A kind of embodiment 6: method of the microbial activity filler processing containing the organic sulphur waste gas
The method containing the organic sulphur waste gas is handled using microbial activity filler described in embodiment 3 the present invention also provides a kind of, Specific step is as follows:
(1) there is the microbial activity filler of Meng Shi pseudomonad LW-1 to be placed in a container 3 gained appendix of embodiment, lead to Enter the gas containing parathion, using parathion as phosphorus source, sulphur source, activates microbial activity filler；Activation time is 7 days；
The concentration of parathion in the gas containing parathion being passed through is 100mg/m3；
(2) the microbial activity filler after activation is placed in bioreactor, is passed through containing the organic sulphur waste gas, with Meng Shi vacation Monad LW-1 purifies the organic sulfur in exhaust gas, and microbial activity packing volume is 500m3, in air inlet, the concentration of malathion For 179.2mg/m3, the concentration of parathion-methyl is 56.3mg/m3, gas flow rate 5000m3/ h, residence time 5.0min, only Changing temperature is 50 DEG C；The activity of microbial activity filler, spray are periodically maintained to microbial activity filler surface spraying nutrient solution Frequency is to spray 1 time for 2 days, and each spray rate is 100L/min, and each spray time is the gas outlet 50min., malathion Concentration is reduced to 13.9mg/m3, the concentration of parathion-methyl is reduced to 4.1mg/m3.The removal of malathion and parathion-methyl Rate respectively reaches 92.2% and 92.7%.
The composition of the nutrient solution are as follows: glucose 0.5g, K2HPO40.5g, KH2PO41.5g, (NH4)2SO4 0.5g, NaCl 0.5g, MgSO40.2g, CaCl20.05g, FeSO40.02g, water 1000ml, pH value 6.2-7.0.
A kind of method of microbial activity filler processing exhaust gas containing organochlorine of embodiment 4 '
The present invention also provides a kind of method using the microbial activity filler processing of embodiment 1 ' exhaust gas containing organochlorine, Specific steps will only be replaced for activating the gas of microbial activity filler by gas containing parathion with embodiment 4 in step (1) For the gas containing trichloroacetaldehyde, concentration is constant；To be passed through in step (2) and replace with concentration containing the organic sulphur waste gas is to be containing concentration 37.4mg/m3The exhaust gas of trichloroacetaldehyde.
Gas outlet, trichloroacetaldehyde concentration are reduced to 6.2mg/m3, the removal rate of trichloroacetaldehyde reaches 83.4%.
A kind of method of microbial activity filler processing exhaust gas containing organochlorine of embodiment 5 '
The present invention also provides a kind of method using the microbial activity filler processing of embodiment 2 ' exhaust gas containing organochlorine, Specific steps will only be replaced for activating the gas of microbial activity filler by gas containing parathion with embodiment 5 in step (1) For the gas containing trichloroacetaldehyde, concentration is constant；To be passed through in step (2) and replace with concentration containing the organic sulphur waste gas is to be containing concentration 97.6mg/m3The exhaust gas of trichloroacetaldehyde.
Gas outlet, trichloroacetaldehyde concentration are reduced to 21.3mg/m3, the removal rate of trichloroacetaldehyde reaches 78.2%.
A kind of method of microbial activity filler processing exhaust gas containing organochlorine of embodiment 6 '
The present invention also provides a kind of method using the microbial activity filler processing of embodiment 3 ' exhaust gas containing organochlorine, Specific steps will only be replaced for activating the gas of microbial activity filler by gas containing parathion with embodiment 6 in step (1) For the gas containing trichloroacetaldehyde, concentration is constant；To be passed through in step (2) and replace with concentration containing the organic sulphur waste gas is to be containing concentration 13.5mg/m3The exhaust gas of trichloroacetaldehyde.
Gas outlet, trichloroacetaldehyde concentration are reduced to 2.7mg/m3, the removal rate of trichloroacetaldehyde reaches 80%.
Embodiment 7: experiment effect
3 gained filler of embodiment is used for the processing of following gases:
(1) exhaust gas containing parathion, gas flow 300m are handled3/ h, residence time 100s, purification temperature 30 ℃.Bioreactor continuous operation 2 months, pH 4-7, the mean air entry concentration of parathion are as follows: 25mg/m3, out gas concentration be 2mg/m3, removal rate reaches 92%.
Contrast and experiment is shown: when ferro element is not added in nutrient solution, after reactor is run 2 months, and Meng Shi vacation unit cell Bacterium LW-1 quantity is from 3.0 × 107CFU/g increases to 7.0 × 108CFU/g.It adds when to bioreactor containing 0.02g/L iron member After the nutrient solution of element, after reactor is run 2 months, Meng Shi pseudomonad LW-1 quantity is by 3.0 × 107CFU/g increases to 1.0 ×109CFU/g increases significantly, bioreactor is enable effectively to remove the organic sulfur in exhaust gas.
The composition of the nutrient solution are as follows: glucose 0.5g, K2HPO40.5g, KH2PO41.5g, (NH4)2SO4 0.5g, NaCl 0.5g, MgSO40.2g, CaCl20.05g, FeSO40.02g, water 1000ml, pH value 6.2-7.0；Spray frequency Rate is to spray 1 time for 2 days, and each spray rate is 200L/min, and each spray time is 50min.
(2) for handling the exhaust gas containing malathion of agricultural chemicals waste water treatment process generation, the concentration of malathion is 10mg/ m3, gas flow 3000m3/ h, residence time 95s, 30 DEG C of gas temperature, regular replenishment contains the battalion of 0.02g/L ferro element Nutrient solution (ibid), the removal rate of malathion is close to 100%.
Equipment is run more than 3 months, pH 4-6, Meng Shi pseudomonad (Pseudomonas monteilii) LW-1 quantity Maintain 2.0 × 107-2.5×108CFU/g, while reaching purification purpose, equipment can be run steadily in the long term.
(3) exhaust gas containing methyl mercaptan is handled, gas flow 10L/min, residence time 90s are operated under the conditions of 30 DEG C. Bioreactor continuous operation October, pH4-6, the inlet gas concentration of methyl mercaptan are as follows: 2-5mg/m3, gas concentration is lower than 0.1mg/m out3, Average removal rate: 98%.Reactor Nei Mengshi pseudomonad (Pseudomonas monteilii) LW-1 quantity be 2.0 × 107-2.5×108CFU/g, bioreactor are stable.
In above-mentioned gas treatment process, the starting period of bioreactor is one week.And under equal conditions, it will be in filler After strain replaces with the activated sludge of processing agricultural chemicals waste water, the starting period is 2 weeks-January.
Embodiment 7 ': experiment effect
3 ' gained filler of embodiment is used for the processing of following gases:
Handle the exhaust gas containing trichloroacetaldehyde, gas flow 300m3/ h, residence time 100s, purification temperature are 30 DEG C. Bioreactor continuous operation August, pH 4-6, the mean air entry concentration of trichloroacetaldehyde are as follows: 30mg/m3, out gas concentration be 4.5mg/m3, removal rate reaches 85%.
Contrast and experiment is shown: when ferro element is not added in nutrient solution, after reactor is run 1 month, and Meng Shi vacation unit cell Bacterium LW-1 quantity is from 3.0 × 107CFU/g increases to 7.0 × 107CFU/g.It adds when to bioreactor containing 0.02g/L iron member After the nutrient solution of element, after reactor is run 2 months, Meng Shi pseudomonad LW-1 quantity is by 3.0 × 107CFU/g increases to 3.0 ×108CFU/g increases the organochlorine significantly enable in the permanently effective removing exhaust gas of bioreactor.
The composition of the nutrient solution are as follows: glucose 0.5g, K2HPO40.5g, KH2PO41.5g, (NH4)2SO4 0.5g, NaCl 0.5g, MgSO40.2g, CaCl20.05g, FeSO40.02g, water 1000ml, pH value 6.2-7.0；Spray frequency Rate is to spray 1 time for 2 days, and each spray rate is 200L/min, and each spray time is 50min.
(2) for handling the exhaust gas containing trichloroacetaldehyde of agricultural chemicals waste water treatment process generation, the concentration of trichloroacetaldehyde is 10mg/ m3, gas flow 3000m3/ h, residence time 95s, 30 DEG C of gas temperature, regular replenishment contains the battalion of 0.02g/L ferro element Nutrient solution (ibid), the removal rate 90% of trichloroacetaldehyde, discharge gas reach the atmosphere pollutants emission standards of country's formulation.
Equipment is run more than 3 months, pH 4-6, Meng Shi pseudomonad (Pseudomonas monteilii) LW-1 quantity Maintain 7.0 × 106-7.5×107CFU/g, while reaching purification purpose, equipment can be run steadily in the long term.
In above-mentioned gas treatment process, the starting period of bioreactor is one week.And under equal conditions, it will be in filler After strain replaces with the activated sludge of processing agricultural chemicals waste water, the starting period is 2 weeks-January.
The processing of 8 mixed gas of embodiment
(fermentation medium is only added 4.5mg/L pairs using the same method of embodiment 3 preparation microbial activity filler Sulphur phosphorus replaces with while adding 4.5mg/L parathion and 4.5mg/L trichloroacetaldehyde) method of the processing containing organic sulfur, chlorine exhaust gas, tool Steps are as follows for body:
(1) there is the microbial activity filler of Meng Shi pseudomonad LW-1 to be placed in a container appendix, be passed through containing parathion, The gas of trichloroacetaldehyde activates microbial activity filler；Activation time is 7 days；
The concentration of parathion in the gas containing parathion being passed through is 100mg/m3, the concentration of trichloroacetaldehyde is 100mg/m3；
(2) the microbial activity filler after activation is placed in bioreactor, is passed through containing organic sulfur, chlorine exhaust gas, with illiteracy Family name pseudomonad LW-1 purifies organic sulfur, chlorine in exhaust gas, and microbial activity packing volume is 500 m3, in air inlet, horse traction sulphur Phosphorus 37.7mg/m3, parathion 16.1mg/m39.8 mg/m of trichloroacetaldehyde3；Gas flow rate is 5000m3/ h, residence time 5.0min, purification temperature are 50 DEG C；Periodically microbial activity filler is maintained to microbial activity filler surface spraying nutrient solution Activity, spray frequency are to spray 1 time for 2 days, and each spray rate is 100L/min, and each spray time is the gas outlet 50min., Malathion 3.2mg/m3, parathion 1.3mg/m3Trichloroacetaldehyde 2.3mg/m3.Malathion removal rate 91.5%, parathion 92% trichloroacetaldehyde removal rate 76.5% of removal rate.
The composition of the nutrient solution are as follows: glucose 0.5g, K2HPO40.5g, KH2PO41.5g, (NH4)2SO4 0.5g, NaCl 0.5g, MgSO40.2g, CaCl20.05g, FeSO40.02g, water 1000ml, pH value 6.2-7.0.

Claims (8)

1. a kind of microbial activity filler, which is characterized in that the microbial activity filler is by Meng Shi pseudomonad bacteria suspension It is adsorbed on filler, and filler is seated in wire side hollow sphere and is prepared；
The Meng Shi pseudomonad is specially Meng Shi pseudomonad (Pseudomonas monteilii) LW-1, and deposit number is CGMCC No.13748。
2. a kind of microbial activity filler as described in claim 1, which is characterized in that the filler be polyurethane foamed blocks, One of haydite, volcanic rock or molecular sieve are a variety of；
The packing density is 20-80kg/m3, porosity 50-97%, partial size 10-100mm.
3. a kind of microbial activity filler as described in claim 1, which is characterized in that the microbial bacteria suspension and filler Mass ratio be 1-12: 1.
4. a kind of microbial activity filler as described in claim 1, which is characterized in that covered in the microbial activity filler Family name pseudomonad LW-1 cell concentration 1.0 × 105-9.0×108CFU/g(filler)。
5. a kind of microbial activity filler as described in claim 1, which is characterized in that the matter of the filler and wire side hollow sphere Amount is than being 0.9-3:1.
6. a kind of microbial activity filler as described in claim 1, which is characterized in that the wire side hollow sphere material is poly- Ethylene, polyvinyl chloride or polypropylene.
7. a kind of microbial activity filler as described in claim 1, which is characterized in that the hollow bulb diameter 6cm of the wire side, It is made of two hemisphere, each hemisphere surface is porous, the aperture 2-8mm in each hole；
There is a circle reinforcing ring in the middle part of the wire side hollow sphere, reinforcing ring is equipped with lock, convenient for two hemisphere of unlatching and closure.
8. a kind of microbial activity filler as described in claim 1, which is characterized in that the Meng Shi pseudomonad bacteria suspension The preparation method is as follows:
1. being carried out liquid fermentation medium is accessed after Meng Shi pseudomonad LW-1 slant activation step by step by 10-25% inoculum concentration Expand fermented and cultured；
Fermentation culture conditions are as follows: in 35-45 DEG C, ventilation quantity 10-100L/min, dissolved oxygen > 2.0mg/L, tank presses 0.03- Under the conditions of 0.10Mpa, incubation time 45-55h；
The composition of the liquid fermentation medium are as follows: glucose 0.5g, K2HPO40.5g, KH2PO41.5g, (NH4)2SO4 0.5g, NaCl 0.5g, MgSO40.2g, CaCl20.05g, FeSO40.02g, water 1000mL, pH value 6.2-7.0,115 DEG C go out Bacterium 20min is cooled to room temperature after sterilizing and adds 4.5mg/L parathion and/or 4.5mg/L trichloroacetaldehyde；
2. after fermented and cultured, fermentation liquid is placed in refrigerated centrifuge in 4 DEG C, 2000-8000rpm, 10-20min is centrifuged, Liquid is discarded supernatant, precipitating is diluted with sterile nutrient solution, and acquisition cell concentration is 0.30-0.95g/L Meng Shi pseudomonad LW-1 bacterium Suspension；
The composition of the sterile nutrient solution are as follows: glucose 0.5g, K2HPO40.5g, KH2PO41.5g, (NH4)2SO40.5g, NaCl 0.5g, MgCl20.2g, CaCl20.05g, FeSO40.02g, water 1000mL, pH value 6.8-7.2,121 DEG C of sterilizings 20min。
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