CN108516621A - A kind of preparation method of the biological active filling material comprising the short shape bacillus of Nie Shi - Google Patents
A kind of preparation method of the biological active filling material comprising the short shape bacillus of Nie Shi Download PDFInfo
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- CN108516621A CN108516621A CN201810455505.5A CN201810455505A CN108516621A CN 108516621 A CN108516621 A CN 108516621A CN 201810455505 A CN201810455505 A CN 201810455505A CN 108516621 A CN108516621 A CN 108516621A
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- filler
- short shape
- shape bacillus
- nie shi
- microbial activity
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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
- 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
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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
- C02F2003/001—Biological treatment of water, waste water, or sewage using granular carriers or supports for the microorganisms
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- Biodiversity & Conservation Biology (AREA)
- Hydrology & Water Resources (AREA)
- Engineering & Computer Science (AREA)
- Environmental & Geological Engineering (AREA)
- Water Supply & Treatment (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Purification Treatments By Anaerobic Or Anaerobic And Aerobic Bacteria Or Animals (AREA)
- Immobilizing And Processing Of Enzymes And Microorganisms (AREA)
Abstract
The invention belongs to field of environmental improvement, and in particular to a kind of preparation method of the microbial activity filler containing the short shape bacillus of Nie Shi.The short shape bacillus of Nie Shi is specially the short shape bacillus of Nie Shi (Brachybacterium nesterenkovii) NY 1, and deposit number is CGMCC No.13747.Microbial activity filler containing the short shape bacillus NY of Nie Shi 1 prepared by the method has the efficiency of higher degradation organochlorine, up to 90 96%；And the laundering period of dominant bacteria is short, can quickly start degradation reaction in 1 week, applies in purifying the exhaust gas bioreactor containing organochlorine, can shorten the starting period.
Description
Technical field:
The invention belongs to field of environmental improvement, and in particular to one kind includes the short shape bacillus (Brachybacterium of Nie Shi
Nesterenkovii) the preparation method of the biological active filling material of NY-1.
Background technology：
Organo-chlorine pesticide be for prevent in the constituent of plant pest contain organic chlorine element organic compound,
It is generally acknowledged critically important a kind of persistence organic pollutant.In organo-chlorine pesticide production and agricultural chemicals waste water processing procedure, tool
There are volatile Pesticidal products or intermediate, volatilize, spilt into air from liquid phase, forms air pollution.Volatile pesticide
And intermediate includes：Metrifonate, DDVP, phosphorus trichloride, DDT, trichloroacetaldehyde urea, sodium trichloroacetate, six six six, Emhorn
Agent, Niran, endrin, heptachlor, toxaphene, hexachloro-benzene, mirex, trichloroacetaldehyde etc., most of these substances have pungent
Smell, some substances such as trichloroacetaldehyde, belong to carcinogenicity, mutagenicity and teratogenesis three cause property substances.
The colourless volatile oily liquids of trichloroacetaldehyde, irritant smell, 97.8 DEG C of boiling point, -57.5 DEG C of fusing point are relatively close
Degree 1.5121, refractive index 1.4580 is dissolved in water, ethyl alcohol, ether and chloroform, trichloroacetaldehyde hydrate is generated with hydrate, with second
Alcohol is the mixture that raw material reacts that generation trichloroacetaldehyde closes trichloroacetaldehyde with alcohol at 80~90 DEG C with chlorine, then anti-with the concentrated sulfuric acid
It answers, is made through distillation, be used as organic synthesis raw material.It is such as used to manufacture DDT, metrifonate, DDVP insecticide and trichlorine
Acetaldehyde carbamide weedicide not only causes to seriously affect to ecological environment, but also has high risks to health.It is chronically at easily
In the organo-chlorine pesticide environment of volatilization, can cause headache, it is dizzy, furious it is congested, shed tears keep in dark place, cough, pharyngalgia, it is weak, perspire,
Salivation, nausea, loss of appetite, insomnia and Head And Face cacesthesia etc., moderate intoxication person also vomits in addition to having with above-mentioned symptom
It spits, abdominal pain, has aches in the limbs, twitches, cyanosis, expiratory dyspnea, tachycardia etc.；Severe intoxication person obviously aggravates except above-mentioned symptom
Outside, still there are high fever, hidrosis, contraction of muscle, epilepsy seizure, or even dead.
Currently, the treatment technology of the exhaust gas of the volatile organic matter containing organochlorine includes absorbing treatment technology, catalysis burning
Treatment technology, liquid absorption treatment technology, biologic treating technique, Treatment by Photocatalysis Oxidation technology, low-temperature plasma treatment technology,
Membrane-base absorption technology etc..Compared with other physical chemistry processing methods, bioremediation has high treating effect, investment and fortune
Row expense is low, safety is good, non-secondary pollution, the advantages that being easily managed.Its principle is the biological oxidation using microorganism
Convert organochlorine to CO2、H2O, chlorate, hydrogen ion and other (formulas 1).
In exhaust-gas treatment bioreactor, certain thickness filler is usually loaded as microorganism carrier, makes micro- life
The biomembrane that object is formed can be attached on filler particles, and can also provide necessary nutrition to microorganism.Biologic packing material
The factors such as material, the size of specific surface area, gas distribution performance, intensity and density directly affect the treatment effeciency and biological respinse of exhaust gas
The operational reliability of device.The type of filler has very much, can be divided into organic filler and inorganic filler.Organic filler include polyurethane,
Polyethylene, composting material etc.；Inorganic filler includes activated carbon, haydite, volcanic rock, perlite etc..Each filler differs from one another.
The materials such as grain activated carbon, haydite, volcanic rock, polyurethane, rough surface, porosity height, large specific surface area, surface has affine
Property, it is easy to microorganism attachment and growth, forms biomembrane, and property is stablized, will not decomposes.Therefore, application is more.But poly- ammonia
Ester material quality is soft, bad mechanical property, is used for a long time and packing layer collapse phenomenon easily occurs；The materials such as haydite, volcanic rock contain greatly
The minerals of amount are corroded by water, exhaust gas and microorganism, and surface forms rickle, block gas circuit, increase packing layer
The pressure loss, make reactor for treatment effect decline, energy consumption increase.
The inoculum for purifying the bioreactor of the exhaust gas containing organochlorine is mostly at organochlorine pesticide pollution soil or sewage
The activated sludge of factory is managed, the microorganism laundering period is long, and bioreactor starts slow.Function bacterium after inoculation domestication enrichment, can contract
The startup time of short bioreactor.But in the microorganism of laboratory's domestication enrichment, competitiveness is weak, in practical applications,
Usually replaced by the strong microorganism of competitiveness.
Invention content：
In order to solve, volatility organochlorine treatment effeciency in the prior art is low, microorganism is uneven in mixed carrier area load
It is even, start it is slow, the problem of filler collapses is used for a long time, the present invention provides the short shape bacillus of one plant of Nie Shi
(Brachybacterium nesterenkovii) NY-1, and the bacterial strain is applied in removing exhaust gas containing organochlorine.
The short shape bacillus of Nie Shi (Brachybacterium nesterenkovii) NY-1 according to the present invention, for from containing
The agricultural chemicals waste water domestication of the substances such as trichloroacetaldehyde, metrifonate, DDVP, phosphorus trichloride, isolated separation bacterium.The bacterial strain is
It is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on March 10th, 2017, deposit number is
CGMCC No.13747, preservation address：Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, Institute of Microorganism, Academia Sinica,
Postcode 100101.
The bacterial strain feature is as follows：
Gram-positive bacteria, form are rod-short, and size is (0.5~0.75) μ m (1.0~2.5) μm, does not produce interior life
Spore, without motion；NY-1 bacterial strains can utilize Pfansteihl, acetic acid, pyruvic acid, α -one base-glutaric acid, D- galactolipins, rough gentian
Carbon source of the various saccharides substance such as disaccharides, alpha-D-glucose and D- xyloses as the bacterial strain can also utilize l-Alanine, L- paddy
The nitrogen source of propylhomoserin, altheine and butanediamine as it；NY-1 bacterial strains can produce catalase, Esterified Enzyme, alcohol dehydrogenase
A variety of enzymes such as enzyme and acetaldehyde dehydrogenase.
Growth temperature is 20-37 DEG C, and optimum growth temperature is 30 DEG C.Growth pH is 6-11, and the pH of optimum growth is 7.
Acid is produced using rhamnose, D- sucrose, D- melibioses etc., remains to survive in the case where pH is 11 strong basicity environment.
The present invention also provides a kind of microbial activity filler, the microbial activity filler is by the short shape bacillus of Nie Shi
(Brachybacterium nesterenkovii) NY-1 bacteria suspensions are adsorbed on filler, and it is hollow that filler is seated in wire side
It is prepared in ball, it can better exhaust gas of the treatment and purification containing organochlorine.
The filler is one or more in polyurethane foamed blocks, haydite, volcanic rock or molecular sieve；
The packing density is 20-900kg/m3, porosity 55-90%, grain size 4-85mm；
The mass ratio of the microbial bacteria suspension and filler is 2-90: 10；
The short shape bacillus NY-1 cell concentrations of Nie Shi are 2.5 × 10 in the microbial activity filler7CFU/g-1.0×
1010CFU/g(filler)；
The wire side hollow ball 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 are a circle reinforcing ring, reinforcing ring to be equipped with lock in the middle part of the wire side hollow ball, convenient for opening and being closed two
Hemisphere.
The specific preparation process of the filler is as follows：
(1) preparation of the short shape bacillus NY-1 bacteria suspensions of Nie Shi
1. the short shape bacillus NY-1 of Nie Shi are accessed into liquid fermentation medium after inclined-plane culture, by 10-25% inoculum concentrations into
Row expands fermented and cultured step by step；
Fermentation culture conditions are：In 36 DEG C, ventilation quantity 10-100L/min, dissolved oxygen>2.0mg/L, tank press 0.03-
Under the conditions of 0.10Mpa, incubation time 48h；
The group of the liquid fermentation medium becomes：Glucose 0.5g, KH2PO42.0g, NaHCO32.0g, KNO3
2.0g, NH4Cl 0.5g, MgCl2·6H2O 0.5g, FeSO4·7H2O 0.01g, water 1000ml, pH value 6.8-7.2,121 DEG C go out
Bacterium 20min, sterilizing postcooling to room temperature add 2.0mg/L trichloroacetaldehydes；
2. after fermented and cultured, zymotic fluid being placed in refrigerated centrifuge in 4 DEG C, 2000-8000rpm, 10- is centrifuged
20min discards supernatant liquid, and precipitation is diluted with sterile nutrient solution, and acquisition cell concentration is the short shape bacillus NY- of 0.5-1.5g/L Nie Shi
1 bacteria suspension；
The group of the sterile nutrient solution becomes：Glucose 0.5g, KH2PO42.0g, NaHCO32.0g, KNO32.0g
NH4Cl 0.5g, MgCl2·6H2O 0.5g, FeCl2·7H2O 0.01g, water 1000ml, pH value 6.8-7.2,121 DEG C of sterilizings
20min；
(2) preparation of microbial activity filler：
1. the filler of packet or grain shape is filled into wire side hollow ball, wire side hollow ball is with packing quality ratio
100：100-450:；
2. the short shape bacillus NY-1 bacteria suspensions of Nie Shi prepared by step (1) are sprayed by the hole of wire side hollow ball to packet
Or the surface of the filler of grain shape；
3. having the wire side hollow ball of filler to be put into impeller load rotates mixing；
4. again to the short shape bacillus NY-1 bacteria suspensions of wire side hollow ball surface spraying Nie Shi equipped with filler；
Step 1. -4. 2-3 time is repeated, makes in hollow ball surface and hole that equal appendix has bacterium cell, acquisition Nie Shi on filler
Short shape bacillus NY-1 cell concentrations reach 2.5 × 107CFU/g-1.0×1010CFU/g(filler)Microbial activity filler, 4 DEG C guarantor
It deposits spare.
It is specific to walk the present invention also provides a kind of method handling the gas containing trichloroacetaldehyde using mentioned microorganism active filler
It is rapid as follows：
(1) above-mentioned appendix is had to micro- life of the short shape bacillus of Nie Shi (Brachybacterium nesterenkovii) NY-1
Object active filler is placed in a container, is passed through the gas containing trichloroacetaldehyde, using trichloroacetaldehyde as carbon source and chlorine source, activates microorganism
Active filler；Soak time is 2-15 days；
A concentration of 10-the 250mg/m of the trichloroacetaldehyde3；
(2) the microbial activity filler after activation is placed in bioreactor, is passed through the gas containing trichloroacetaldehyde, uses alunite
The short shape bacillus of family name (Brachybacterium nesterenkovii) NY-1 purifies the trichloroacetaldehyde in exhaust gas, microbial activity
Packing volume is 0.01-100m3, the total concentration of trichloroacetaldehyde is 10-250mg/m3, gas flow rate 0.05-10000m3/ h, stops
It is 20-35 DEG C to stay time 0.5-5min, purification temperature；Periodically microorganism is maintained to microbial activity filler surface spraying nutrient solution
The activity of active filler, spray frequency are to spray 1 time for 1-3 days, and each spray rate is 1.0-200L/min, each spray time
For 10-90min；
The group of the nutrient solution becomes：Glucose 0.5g, yeast powder 0.2g, KH2PO42.0g, NaHCO32.0g
KNO32.0g, NH4Cl 0.5g, MgCl2·6H2O 0.5g, FeCl2·7H2O 0.01g, water 1000ml, pH 6.5-7.0；
The degradation capability of tracing detection trichloroacetaldehyde, the present invention use gas chromatography tracing detection trichloroacetaldehyde；It is described
Gas chromatography be：Trichloroacetaldehyde concentration, chromatographic column HP-5 are measured using Agilent GC-6890N chromatographics
Capillary column (30m × 0.32mm × 0.25 μm), vaporizing chamber, FPD detectors, column temperature be respectively set to 250 DEG C, 250 DEG C, 100
DEG C, sample size 100 μ L, N2Flow velocity is 45mL/min.The results show that using above-mentioned filler to the discarded progress containing trichloroacetaldehyde
The degradation rate of purified treatment, trichloroacetaldehyde reaches 90% or more.
Advantageous effect：
1. the short shape bacillus NY-1 of Nie Shi of the present invention have the efficiency of higher degradation organochlorine, up to 90-96%；And
The laundering period of dominant bacteria is short, can quickly start degradation reaction in 1 week, applies in exhaust gas bioreactor of the purification containing organochlorine
In, the starting period can be shortened.
2. using the effect of a variety of mechanics such as sedimentation, attachment and centrifugation, the short shape bacillus NY-1 appendix of the Nie Shi that will degrade are in net
On the light porous material of face hollow ball inside, microbial activity filler is formed, bacterium cell can not only be uniformly adhered to fill out
Expect surface, and can enter in the hole inside filler, keeps the bacterium cell appendix amount of microbial activity filler high, be not easy from biology
It is lost in reactor.
3. Nie Shi is short, shape bacillus NY-1 competitiveness is very strong, can pass through using chlorinated organics as carbon source and chlorine source
It is more to grab Pfansteihl, acetic acid, pyruvic acid, α -one base-glutaric acid, D- galactolipins, gentiobiose, alpha-D-glucose and D- xyloses etc.
Kind glucide, generates a variety of enzymes such as catalase, Esterified Enzyme, alcohol dehydrogenase and acetaldehyde dehydrogenase.
It, being capable of the continuously and effectively purification volatilization in alkaline environment 4. the short shape bacillus NY-1 of Nie Shi have very strong alkali resistance
Property organic exhaust gas containing chlorine.
5. 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 organochlorine such as bacterium cell and trichloroacetaldehyde to contact；The wire side hollow ball hole for loading light filler is more, thoroughly
Gas is good, and resistance is small, and mass-transfer efficiency is high；Microbial activity filler weight is light, intensity is high, the pressure loss is small；High temperature resistant, corrosion resistant
Erosion；Not perishable deformation, can be used for a long time and adapt to the physical condition of Various Complex.
Description of the drawings：
The short shape bacillus of Fig. 1 Nie Shi (Brachybacterium nesterenkovii) NY-1 (NCBI GenBank sequences KY
979105) phylogenetic tree.
Specific implementation mode：
In order to make the object, technical solution and advantage of this patent be 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
It is extracted using Full-automatic magnetic beads instrument for extracting nucleic acid Smart LabAssist-16 and MO-BIO PowerSoil DNA
Kit (Taiwan dot nanometer technical concern Co., Ltd) is extracted and the obtained short shape bacillus of bacterial strain Nie Shi of purifying
The DNA of (Brachybacterium nesterenkovii) NY-1.Take 5 μ L DNA samples under the conditions of 120V, 20-25min into
Row electrophoresis detection carries out observation quality inspection under gel-electrophoretic apparatus.
Select bacterial universal primers F16S-27 (5`-AGAGTTTGATCCTGGCTCAG-3`) and R16S-1492 (5`-
CGGTTACCTTGTTACGACTTC-3` PCR amplification) is carried out.PCR reaction conditions are：94 DEG C of pre-degeneration 3min, 94 DEG C of denaturation
30s, 54 DEG C of annealing 30s, 72 DEG C of extension 90s recycle 30 periods；Then 72 DEG C of extension 10min；Last 4 DEG C of holdings.
Successively pass through PCR product purifying and quality inspection, the connection of PCR purified products, E. coli competent it is heat-shock transformed,
And carry out plasmid extraction, sequencing.
Sequence merger using DNAMAN by the similitude measured more than 97% is an OTU (operating unit), is chosen every
Sequence is represented in a OTU using the GenBank databases and BLAST of NCBI compare between sequence (1400bp or more length),
Utilize Phylogenetic Analysis (MEGA3.1) software ortho position connection (Neighbour Joining) method phylogenetic tree construction (such as Fig. 1
It is shown).
A kind of microbial activity filler of the removing exhaust gas containing organochlorine of embodiment 1 and its preparation
A kind of microbial activity filler, the microbial activity filler are by the short shape bacillus (Brachybacterium of Nie Shi
Nesterenkovii) NY-1 bacteria suspensions are adsorbed on filler, and filler is seated in wire side hollow ball and is prepared, can be more
Good exhaust gas of the treatment and purification containing organochlorine.
The filler is volcanic rock；Packing density is 900kg/m3, porosity 55%, grain size 4mm；Microbial bacteria is outstanding
The mass ratio of liquid and filler is 2: 10；The short shape bacillus NY-1 cell concentrations of Nie Shi are 2.5 × 10 in microbial activity filler7CFU/
g(filler)；
The wire side hollow ball 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 ball, reinforcing ring is equipped with lock, convenient for opening
Open and be closed two hemisphere.
The specific preparation process of the filler is as follows：
(1) preparation of the short shape bacillus NY-1 bacteria suspensions of Nie Shi
1. the short shape bacillus NY-1 of Nie Shi are accessed into liquid fermentation medium after inclined-plane culture, by 10% inoculum concentration carry out by
Grade expands fermented and cultured；
Fermentation culture conditions are：In 36 DEG C, ventilation quantity 10-100L/min, dissolved oxygen>2.0mg/L, tank press 0.03-
Under the conditions of 0.10Mpa, incubation time 48h；
The group of the liquid fermentation medium becomes：Glucose 0.5g, KH2PO42.0g, NaHCO32.0g, KNO3
2.0g, NH4Cl 0.5g, MgCl2·6H2O 0.5g, FeSO4·7H2O 0.01g, water 1000ml, pH value 6.8-7.2,121 DEG C go out
Bacterium 20min, sterilizing postcooling to room temperature add 2.0mg/L trichloroacetaldehydes；
2. after fermented and cultured, zymotic fluid being placed in refrigerated centrifuge in 4 DEG C, 2000rpm, 10min is centrifuged, discards
Supernatant, precipitation are diluted with sterile nutrient solution, and acquisition cell concentration is the short shape bacillus NY-1 bacteria suspensions of 0.5g/L Nie Shi；
The group of the sterile nutrient solution becomes：Glucose 0.5g, KH2PO42.0g, NaHCO32.0g, KNO32.0g
NH4Cl 0.5g, MgCl2·6H2O 0.5g, FeCl2·7H2O 0.01g, water 1000ml, pH value 6.8-7.2,121 DEG C of sterilizings
20min；
(2) preparation of microbial activity filler：
1. the filler of square is filled into wire side hollow ball, wire side hollow ball is 100 with packing quality ratio：450；
2. the short shape bacillus NY-1 bacteria suspensions of Nie Shi prepared by step (1) are arrived filler by the hole spray of wire side hollow ball
Surface；
3. having the wire side hollow ball of filler to be put into impeller load rotates mixing；
4. again to the short shape bacillus NY-1 bacteria suspensions of wire side hollow ball surface spraying Nie Shi equipped with filler；
1. -4. 3 time it repeats step, makes in hollow ball surface and hole that equal appendix has bacterium cell, acquisition Nie Shi short on filler
Shape bacillus NY-1 cell concentrations reach 2.5 × 107CFU/g(filler)Microbial activity filler, 4 DEG C save backup.
A kind of microbe stuffing of the removing exhaust gas containing organochlorine of embodiment 2 and its preparation
A kind of microbial activity filler, the microbial activity filler are by the short shape bacillus (Brachybacterium of Nie Shi
Nesterenkovii) NY-1 bacteria suspensions are adsorbed on filler, and filler is seated in wire side hollow ball and is prepared, can be more
Good exhaust gas of the treatment and purification containing organochlorine.
The filler is polyurethane foamed blocks；Packing density is 20kg/m3, porosity 90%, grain size 85mm；Micro- life
The mass ratio of object bacteria suspension and filler is 90: 10；In microbial activity filler the short shape bacillus NY-1 cell concentrations of Nie Shi be 1.0 ×
1010CFU/g(filler)；
The wire side hollow ball material is polypropylene；The hollow bulb diameter 6cm of wire side, is made of, Mei Geban two hemisphere
Ball surface is porous, the aperture 8mm in each hole；There is a circle reinforcing ring in the middle part of wire side hollow ball, reinforcing ring is equipped with lock, convenient for opening
Open and be closed two hemisphere.
The specific preparation process of the filler is as follows：
(1) preparation of the short shape bacillus NY-1 bacteria suspensions of Nie Shi
1. the short shape bacillus NY-1 of Nie Shi are accessed into liquid fermentation medium after inclined-plane culture, by 25% inoculum concentration carry out by
Grade expands fermented and cultured；
Fermentation culture conditions are：In 36 DEG C, ventilation quantity 10-100L/min, dissolved oxygen>2.0mg/L, tank press 0.03-
Under the conditions of 0.10Mpa, incubation time 48h；
The group of the liquid fermentation medium becomes：Glucose 0.5g, KH2PO42.0g, NaHCO32.0g, KNO3
2.0g, NH4Cl 0.5g, MgCl2·6H2O 0.5g, FeSO4·7H2O 0.01g, water 1000ml, pH value 6.8-7.2,121 DEG C go out
Bacterium 20min, sterilizing postcooling to room temperature add 2.0mg/L trichloroacetaldehydes；
2. after fermented and cultured, zymotic fluid being placed in refrigerated centrifuge in 4 DEG C, 8000rpm, 20min is centrifuged, discards
Supernatant, precipitation are diluted with sterile nutrient solution, and acquisition cell concentration is the short shape bacillus NY-1 bacteria suspensions of 1.5g/L Nie Shi；
The group of the sterile nutrient solution becomes：Glucose 0.5g, KH2PO42.0g, NaHCO32.0g, KNO32.0g
NH4Cl 0.5g, MgCl2·6H2O 0.5g, FeCl2·7H2O 0.01g, water 1000ml, pH value 6.8-7.2,121 DEG C of sterilizings
20min；
(2) preparation of microbial activity filler：
1. granular filler is filled into wire side hollow ball, wire side hollow ball is 100 with packing quality ratio：100；
2. the short shape bacillus NY-1 bacteria suspensions of Nie Shi prepared by step (1) are arrived filler by the hole spray of wire side hollow ball
Surface；
3. having the wire side hollow ball of filler to be put into impeller load rotates mixing；
4. again to the short shape bacillus NY-1 bacteria suspensions of wire side hollow ball surface spraying Nie Shi equipped with filler；
1. -4. 2 time it repeats step, makes in hollow ball surface and hole that equal appendix has bacterium cell, acquisition Nie Shi short on filler
Shape bacillus NY-1 cell concentrations reach 1.0 × 1010CFU/g(filler)Microbial activity filler, 4 DEG C save backup.
A kind of microbe stuffing of the removing exhaust gas containing organochlorine of embodiment 3 and its preparation
A kind of microbial activity filler, the microbial activity filler are by the short shape bacillus (Brachybacterium of Nie Shi
Nesterenkovii) NY-1 bacteria suspensions are adsorbed on filler, and filler is seated in wire side hollow ball and is prepared, can be more
Good exhaust gas of the treatment and purification containing organochlorine.
The filler is molecular sieve；Packing density is 500kg/m3, porosity 75%, grain size 50mm；Microbial bacteria
The mass ratio of suspension and filler is 50: 10；In microbial activity filler the short shape bacillus NY-1 cell concentrations of Nie Shi be 6.0 ×
108CFU/g(filler)；
The wire side hollow ball material is polyvinyl chloride；The hollow bulb diameter 6cm of wire side, is made of, each two hemisphere
Hemisphere surface is porous, the aperture 5mm in each hole；There are a circle reinforcing ring, reinforcing ring to be equipped with lock, be convenient in the middle part of wire side hollow ball
Open and be closed two hemisphere.
The specific preparation process of the filler is as follows：
(1) preparation of the short shape bacillus NY-1 bacteria suspensions of Nie Shi
1. the short shape bacillus NY-1 of Nie Shi are accessed into liquid fermentation medium after inclined-plane culture, by 15% inoculum concentration carry out by
Grade expands fermented and cultured；
Fermentation culture conditions are：In 36 DEG C, ventilation quantity 10-100L/min, dissolved oxygen>2.0mg/L, tank press 0.03-
Under the conditions of 0.10Mpa, incubation time 48h；
The group of the liquid fermentation medium becomes：Glucose 0.5g, KH2PO42.0g, NaHCO32.0g, KNO3
2.0g, NH4Cl 0.5g, MgCl2·6H2O 0.5g, FeSO4·7H2O 0.01g, water 1000ml, pH value 6.8-7.2,121 DEG C go out
Bacterium 20min, sterilizing postcooling to room temperature add 2.0mg/L trichloroacetaldehydes；
2. after fermented and cultured, zymotic fluid being placed in refrigerated centrifuge in 4 DEG C, 5000rpm, 15min is centrifuged, discards
Supernatant, precipitation are diluted with sterile nutrient solution, and acquisition cell concentration is the short shape bacillus NY-1 bacteria suspensions of 1.0g/L Nie Shi；
The group of the sterile nutrient solution becomes：Glucose 0.5g, KH2PO42.0g, NaHCO32.0g, KNO32.0g
NH4Cl 0.5g, MgCl2·6H2O 0.5g, FeCl2·7H2O 0.01g, water 1000ml, pH value 6.8-7.2,121 DEG C of sterilizings
20min；
(2) preparation of microbial activity filler：
1. the filler of square is filled into wire side hollow ball, wire side hollow ball is 100 with packing quality ratio：250；
2. the short shape bacillus NY-1 bacteria suspensions of Nie Shi prepared by step (1) are arrived filler by the hole spray of wire side hollow ball
Surface；
3. having the wire side hollow ball of filler to be put into impeller load rotates mixing；
4. again to the short shape bacillus NY-1 bacteria suspensions of wire side hollow ball surface spraying Nie Shi equipped with filler；
1. -4. 3 time it repeats step, makes in hollow ball surface and hole that equal appendix has bacterium cell, acquisition Nie Shi short on filler
Shape bacillus NY-1 cell concentrations reach 6.0 × 108CFU/g(filler)Microbial activity filler, 4 DEG C save backup.
The method that a kind of 4 microbial activity filler of embodiment handles the exhaust gas containing organochlorine
The present invention also provides a kind of sides handling the gas containing trichloroacetaldehyde using 1 gained microbial activity filler of embodiment
Method is as follows：
(1) 1 gained appendix of embodiment there is into the short shape bacillus of Nie Shi (Brachybacterium nesterenkovii) NY-1
Microbial activity filler be placed in a container, be passed through the gas containing trichloroacetaldehyde, using trichloroacetaldehyde as carbon source and chlorine source, activation
Microbial activity filler；Soak time is 15 days；
A concentration of 20mg/m of the trichloroacetaldehyde3；
(2) the microbial activity filler after activation is placed in bioreactor, is passed through the gas containing trichloroacetaldehyde, uses alunite
Trichloroacetaldehyde in the short shape bacillus NY-1 purifications exhaust gas of family name, microbial activity packing volume is 1m3, trichloroacetaldehyde in air inlet
A concentration of 20mg/m3, gas flow rate 100m3/ h, residence time 0.5min, purification temperature are 20 DEG C, pH 10-11；Periodically to
Microbial activity filler surface spraying nutrient solution maintain microbial activity filler activity, spray frequency be 1 day spray 1 time, often
Secondary spray rate is 1.0L/min, and each spray time is 10min；A concentration of 1.0mg/m of gas outlet trichloroacetaldehyde3, removal rate
Reach 95%；
The group of the nutrient solution becomes：Glucose 0.5g, yeast powder 0.2g, KH2PO42.0g, NaHCO32.0g
KNO32.0g, NH4Cl 0.5g, MgCl2·6H2O 0.5g, FeCl2·7H2O 0.01g, water 1000ml, pH 6.5-7.0.
The method that a kind of 5 microbial activity filler of embodiment handles the exhaust gas containing organochlorine
The present invention also provides a kind of sides handling the gas containing trichloroacetaldehyde using 2 gained microbial activity filler of embodiment
Method is as follows：
(1) 2 gained appendix of embodiment there is into the short shape bacillus of Nie Shi (Brachybacterium nesterenkovii) NY-1
Microbial activity filler be placed in a container, be passed through the gas containing trichloroacetaldehyde, using trichloroacetaldehyde as carbon source and chlorine source, activation
Microbial activity filler；Soak time is 2 days；
A concentration of 250mg/m of the trichloroacetaldehyde3；
(2) the microbial activity filler after activation is placed in bioreactor, is passed through the gas containing trichloroacetaldehyde, uses alunite
Trichloroacetaldehyde in the short shape bacillus NY-1 purifications exhaust gas of family name, microbial activity packing volume is 100m3, trichloroacetaldehyde in air inlet
A concentration of 180mg/m3, gas flow rate 10000m3/ h, residence time 5min, purification temperature are 35 DEG C, pH 10-11；Periodically
It is to spray 1 time for 3 days that the activity of microbial activity filler, spray frequency are maintained to microbial activity filler surface spraying nutrient solution,
Each spray rate is 200L/min, and each spray time is 90min；A concentration of 7.8mg/m of gas outlet trichloroacetaldehyde3, removal
Rate reaches 95.7%；
The group of the nutrient solution becomes：Glucose 0.5g, yeast powder 0.2g, KH2PO42.0g, NaHCO32.0g
KNO32.0g, NH4Cl 0.5g, MgCl2·6H2O 0.5g, FeCl2·7H2O 0.01g, water 1000ml, pH 6.5-7.0.
The method that a kind of 6 microbial activity filler of embodiment handles the exhaust gas containing organochlorine
The present invention also provides a kind of sides handling the gas containing trichloroacetaldehyde using 3 gained microbial activity filler of embodiment
Method is as follows：
(1) 3 gained appendix of embodiment there is into the short shape bacillus of Nie Shi (Brachybacterium nesterenkovii) NY-1
Microbial activity filler be placed in a container, be passed through the gas containing trichloroacetaldehyde, using trichloroacetaldehyde as carbon source and chlorine source, activation
Microbial activity filler；Soak time is 5 days；
A concentration of 250mg/m of the trichloroacetaldehyde3；
(2) the microbial activity filler after activation is placed in bioreactor, is passed through the gas containing trichloroacetaldehyde, uses alunite
Trichloroacetaldehyde in the short shape bacillus NY-1 purifications exhaust gas of family name, microbial activity packing volume is 50m3, trichloroacetaldehyde in air inlet
A concentration of 218mg/m3, gas flow rate 5000m3/ h, residence time 3min, purification temperature are 30 DEG C, pH 10-11；Periodically
It is to spray 1 time for 2 days that the activity of microbial activity filler, spray frequency are maintained to microbial activity filler surface spraying nutrient solution,
Each spray rate is 100L/min, and each spray time is 50min；A concentration of 10.3mg/m of gas outlet trichloroacetaldehyde3, removal
Rate reaches 95.3%；
The group of the nutrient solution becomes：Glucose 0.5g, yeast powder 0.2g, KH2PO42.0g, NaHCO32.0g
KNO32.0g, NH4Cl 0.5g, MgCl2·6H2O 0.5g, FeCl2·7H2O 0.01g, water 1000ml, pH 6.5-7.0.
Embodiment 7：Experimental comparison's effect
3 gained filler of embodiment is used for the processing of following gases：
(1) exhaust gas containing trichloroacetaldehyde, gas flow 0.2m are handled3/ h, residence time 8s, purification temperature 25
℃.Bioreactor continuous operation January, the mean air entry of pH 10-11, trichloroacetaldehyde are a concentration of：527mg/m3, go out gas concentration
For 35mg/m3, removal rate reaches 91%.Cell concentration 6.0 × 108CFU/g(filler)。
Contrast and experiment is shown：When pH is 5-6, bioreactor continuous operation January, the short shape bacillus NY-1 quantity of Nie Shi
From 6 × 108CFU/g increases to 9.2 × 108CFU/g.When pH in bioreactor is adjusted to 10-11, reactor is run 1 month
Afterwards, the short shape bacillus NY-1 quantity of Nie Shi is by 6 × 108CFU/g increases to 3.9 × 109CFU/g increases notable.After 3 months, Nie Shi
Short shape bacillus NY-1 quantity maintains 7.1 × 108-5.2×109CFU/g.As it can be seen that the short shape bacillus NY-1 of Nie Shi can keep number for a long time
Amount and activity enable the trichloroacetaldehyde in the permanently effective removing exhaust gas of bioreactor.
(2) it is used to handle the exhaust gas containing trichloroacetaldehyde of agricultural chemicals waste water processing procedure generation, a concentration of 50mg/ of trichloroacetaldehyde
m3, gas flow 5000m3The removal rate of/h, residence time 85s, 25 DEG C of gas temperature, trichloroacetaldehyde are more than 90%.
Equipment is run more than 2 months, and the short shape bacillus NY-1 quantity of pH 7-8.5, Nie Shi maintains 8.04 × 109-1.4×
1010CFU/g, reach purification trichloroacetaldehyde while, equipment can be run steadily in the long term.
In above-mentioned gas processing procedure, the starting period of bioreactor is 3-7 days.And under equal conditions, it will be in filler
Strain replace with agricultural chemicals waste water processing activated sludge after, the starting period be 2 weeks-January.
Claims (7)
1. a kind of preparation method of the microbial activity filler comprising the short shape bacillus of Nie Shi, which is characterized in that the microorganism lives
Property filler be adsorbed on filler by the short shape bacillus NY-1 bacteria suspensions of Nie Shi, and filler is seated in wire side hollow ball prepare and
At；
The short shape bacillus of Nie Shi is specially the short shape bacillus of Nie Shi (Brachybacterium nesterenkovii) NY-1, is protected
It is CGMCC No.13747 to hide number.
The specific preparation process of the filler is as follows：
(1) preparation of the short shape bacillus NY-1 bacteria suspensions of Nie Shi
1. the short shape bacillus NY-1 of Nie Shi are accessed into liquid fermentation medium after inclined-plane culture, by 10-25% inoculum concentrations carry out by
Grade expands fermented and cultured；
2. after fermented and cultured, zymotic fluid is placed in refrigerated centrifuge in 4 DEG C, 2000-8000rpm, 10-20min is centrifuged,
Liquid is discarded supernatant, precipitation is diluted with sterile nutrient solution, and it is outstanding for the short shape bacillus NY-1 bacterium of 0.5-1.5g/L Nie Shi to obtain cell concentration
Liquid；
(2) preparation of microbial activity filler：
1. the filler of packet or grain shape is filled into wire side hollow ball, wire side hollow ball is 100 with packing quality ratio：
100-450:；
2. the short shape bacillus NY-1 bacteria suspensions of Nie Shi prepared by step (1) by the hole of wire side hollow ball spray to packet or
The surface of the filler of particle shape shape；
3. having the wire side hollow ball of filler to be put into impeller load rotates mixing；
4. again to the short shape bacillus NY-1 bacteria suspensions of wire side hollow ball surface spraying Nie Shi equipped with filler；
Step 1. -4. 2-3 time is repeated, makes in hollow ball surface and hole that equal appendix has bacterium cell, the acquisition short shapes of Nie Shi on filler
Bacillus NY-1 cell concentrations reach 2.5 × 107CFU/g-1.0×1010CFU/g(filler)Microbial activity filler, 4 DEG C of preservations are standby
With.
2. a kind of preparation method of the microbial activity filler comprising the short shape bacillus of Nie Shi as described in claim 1, feature
It is, fermentation culture conditions are：In 36 DEG C, ventilation quantity 10-100L/min, dissolved oxygen>2.0mg/L, tank press 0.03-0.10Mpa
Under the conditions of, incubation time 48h；
The group of the liquid fermentation medium becomes：Glucose 0.5g, KH2PO42.0g, NaHCO32.0g, KNO32.0g
NH4Cl 0.5g, MgCl2·6H2O 0.5g, FeSO4·7H2O 0.01g, water 1000ml, pH value 6.8-7.2,121 DEG C of sterilizings
20min, sterilizing postcooling to room temperature add 2.0mg/L trichloroacetaldehydes.
3. a kind of preparation method of the microbial activity filler comprising the short shape bacillus of Nie Shi as described in claim 1, feature
It is, the group of the sterile nutrient solution becomes：Glucose 0.5g, KH2PO42.0g, NaHCO32.0g, KNO32.0g, NH4Cl
0.5g, MgCl2·6H2O 0.5g, FeCl2·7H2O 0.01g, water 1000ml, pH value 6.8-7.2,121 DEG C of sterilizing 20min.
4. a kind of preparation method of the microbial activity filler comprising the short shape bacillus of Nie Shi as described in claim 1, feature
It is, the filler is one or more in polyurethane foamed blocks, haydite, volcanic rock or molecular sieve；
The packing density is 20-900kg/m3, porosity 55-90%, grain size 4-85mm.
5. a kind of preparation method of the microbial activity filler comprising the short shape bacillus of Nie Shi as described in claim 1, feature
It is, the mass ratio of the microbial bacteria suspension and light porous filler is 2-90: 10.
6. a kind of preparation method of the microbial activity filler comprising the short shape bacillus of Nie Shi as described in claim 1, feature
It is, the wire side hollow ball material is polyethylene, polyvinyl chloride or polypropylene.
7. a kind of preparation method of the microbial activity filler comprising the short shape bacillus of Nie Shi as described in claim 1, feature
It is, the hollow bulb diameter 6cm of the wire side is made of two hemisphere, and each hemisphere surface is porous, the aperture 2- in each hole
8mm；
There are a circle reinforcing ring, reinforcing ring to be equipped with lock in the middle part of the wire side hollow ball, convenient for opening and being closed two hemisphere.
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| CN201810455505.5A CN108516621B (en) | 2018-05-14 | 2018-05-14 | Preparation method of bioactive filler containing Netherobacter brevis |
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| CN201810455505.5A CN108516621B (en) | 2018-05-14 | 2018-05-14 | Preparation method of bioactive filler containing Netherobacter brevis |
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101941762A (en) * | 2010-09-08 | 2011-01-12 | 福建微水环保技术有限公司 | Method for treating wastewater by utilizing composite microbial preparation |
| CN103266099A (en) * | 2013-05-29 | 2013-08-28 | 中国科学院生态环境研究中心 | Ochrobactrum anthropi SL1 active filler and preparation method thereof |
| CN103819267A (en) * | 2014-02-07 | 2014-05-28 | 湖南泰谷生物科技股份有限公司 | Bacterial manure for peanut planting and preparation method of bacterial manure |
| CN107930378A (en) * | 2017-11-17 | 2018-04-20 | 北京沃太斯环保科技发展有限公司 | For handling the filtering tower packing of VOCs gases and foul gas |
-
2018
- 2018-05-14 CN CN201810455505.5A patent/CN108516621B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101941762A (en) * | 2010-09-08 | 2011-01-12 | 福建微水环保技术有限公司 | Method for treating wastewater by utilizing composite microbial preparation |
| CN103266099A (en) * | 2013-05-29 | 2013-08-28 | 中国科学院生态环境研究中心 | Ochrobactrum anthropi SL1 active filler and preparation method thereof |
| CN103819267A (en) * | 2014-02-07 | 2014-05-28 | 湖南泰谷生物科技股份有限公司 | Bacterial manure for peanut planting and preparation method of bacterial manure |
| CN107930378A (en) * | 2017-11-17 | 2018-04-20 | 北京沃太斯环保科技发展有限公司 | For handling the filtering tower packing of VOCs gases and foul gas |
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