CN104560798A - Preparation method of halophilic bacteria agent for degrading phenols - Google Patents
Preparation method of halophilic bacteria agent for degrading phenols Download PDFInfo
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- CN104560798A CN104560798A CN201410812036.XA CN201410812036A CN104560798A CN 104560798 A CN104560798 A CN 104560798A CN 201410812036 A CN201410812036 A CN 201410812036A CN 104560798 A CN104560798 A CN 104560798A
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- halophilic bacterium
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- 241000894006 Bacteria Species 0.000 title claims abstract description 140
- 238000002360 preparation method Methods 0.000 title claims abstract description 20
- 230000000593 degrading effect Effects 0.000 title abstract description 7
- 150000002989 phenols Chemical class 0.000 title abstract 2
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N phenol group Chemical group C1(=CC=CC=C1)O ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 claims abstract description 102
- 239000002351 wastewater Substances 0.000 claims abstract description 45
- 238000012216 screening Methods 0.000 claims abstract description 35
- 239000000725 suspension Substances 0.000 claims abstract description 25
- 239000007788 liquid Substances 0.000 claims abstract description 21
- 239000000126 substance Substances 0.000 claims abstract description 15
- 239000010802 sludge Substances 0.000 claims abstract description 12
- 230000001580 bacterial effect Effects 0.000 claims abstract description 8
- 239000001963 growth medium Substances 0.000 claims abstract description 7
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 40
- 238000000926 separation method Methods 0.000 claims description 32
- 239000002068 microbial inoculum Substances 0.000 claims description 24
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 claims description 18
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 14
- 238000001914 filtration Methods 0.000 claims description 14
- 230000001954 sterilising effect Effects 0.000 claims description 14
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 12
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims description 12
- 239000001888 Peptone Substances 0.000 claims description 12
- 108010080698 Peptones Proteins 0.000 claims description 12
- 230000015556 catabolic process Effects 0.000 claims description 12
- 238000006731 degradation reaction Methods 0.000 claims description 12
- 239000000463 material Substances 0.000 claims description 12
- 235000019319 peptone Nutrition 0.000 claims description 12
- 230000001681 protective effect Effects 0.000 claims description 12
- 239000006228 supernatant Substances 0.000 claims description 12
- 238000012360 testing method Methods 0.000 claims description 12
- 239000000203 mixture Substances 0.000 claims description 9
- 239000012528 membrane Substances 0.000 claims description 8
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 6
- 229910021536 Zeolite Inorganic materials 0.000 claims description 6
- 229940041514 candida albicans extract Drugs 0.000 claims description 6
- 238000005119 centrifugation Methods 0.000 claims description 6
- 230000009514 concussion Effects 0.000 claims description 6
- HNPSIPDUKPIQMN-UHFFFAOYSA-N dioxosilane;oxo(oxoalumanyloxy)alumane Chemical compound O=[Si]=O.O=[Al]O[Al]=O HNPSIPDUKPIQMN-UHFFFAOYSA-N 0.000 claims description 6
- 239000012153 distilled water Substances 0.000 claims description 6
- 235000011187 glycerol Nutrition 0.000 claims description 6
- 239000000843 powder Substances 0.000 claims description 6
- 235000019600 saltiness Nutrition 0.000 claims description 6
- 238000012546 transfer Methods 0.000 claims description 6
- 239000012138 yeast extract Substances 0.000 claims description 6
- 239000010457 zeolite Substances 0.000 claims description 6
- JEGUKCSWCFPDGT-UHFFFAOYSA-N h2o hydrate Chemical compound O.O JEGUKCSWCFPDGT-UHFFFAOYSA-N 0.000 claims description 2
- 238000000034 method Methods 0.000 abstract description 20
- 150000003839 salts Chemical class 0.000 abstract description 5
- 230000000694 effects Effects 0.000 abstract description 4
- 239000003795 chemical substances by application Substances 0.000 abstract 5
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 8
- 230000008569 process Effects 0.000 description 7
- 210000002421 cell wall Anatomy 0.000 description 6
- 210000004027 cell Anatomy 0.000 description 4
- 244000005700 microbiome Species 0.000 description 4
- 230000003204 osmotic effect Effects 0.000 description 4
- 108010074858 plaferon Proteins 0.000 description 4
- 239000010865 sewage Substances 0.000 description 4
- 239000011780 sodium chloride Substances 0.000 description 4
- 241000251468 Actinopterygii Species 0.000 description 3
- 230000003647 oxidation Effects 0.000 description 3
- 238000007254 oxidation reaction Methods 0.000 description 3
- 108090000623 proteins and genes Proteins 0.000 description 3
- 102000004169 proteins and genes Human genes 0.000 description 3
- 108090000790 Enzymes Proteins 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 230000004913 activation Effects 0.000 description 2
- 238000003889 chemical engineering Methods 0.000 description 2
- 239000002894 chemical waste Substances 0.000 description 2
- 238000004939 coking Methods 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 239000003344 environmental pollutant Substances 0.000 description 2
- 231100000719 pollutant Toxicity 0.000 description 2
- ISPYQTSUDJAMAB-UHFFFAOYSA-N 2-chlorophenol Chemical compound OC1=CC=CC=C1Cl ISPYQTSUDJAMAB-UHFFFAOYSA-N 0.000 description 1
- GAKLFAZBKQGUBO-UHFFFAOYSA-N 2-methyl-3-nitrophenol Chemical compound CC1=C(O)C=CC=C1[N+]([O-])=O GAKLFAZBKQGUBO-UHFFFAOYSA-N 0.000 description 1
- FRXSZNDVFUDTIR-UHFFFAOYSA-N 6-methoxy-1,2,3,4-tetrahydroquinoline Chemical compound N1CCCC2=CC(OC)=CC=C21 FRXSZNDVFUDTIR-UHFFFAOYSA-N 0.000 description 1
- ZAMOUSCENKQFHK-UHFFFAOYSA-N Chlorine atom Chemical compound [Cl] ZAMOUSCENKQFHK-UHFFFAOYSA-N 0.000 description 1
- VGGSQFUCUMXWEO-UHFFFAOYSA-N Ethene Chemical compound C=C VGGSQFUCUMXWEO-UHFFFAOYSA-N 0.000 description 1
- 108090001030 Lipoproteins Proteins 0.000 description 1
- 102000004895 Lipoproteins Human genes 0.000 description 1
- MSFSPUZXLOGKHJ-UHFFFAOYSA-N Muraminsaeure Natural products OC(=O)C(C)OC1C(N)C(O)OC(CO)C1O MSFSPUZXLOGKHJ-UHFFFAOYSA-N 0.000 description 1
- 241000254809 Natrinema altunense Species 0.000 description 1
- 108010013639 Peptidoglycan Proteins 0.000 description 1
- 108010001267 Protein Subunits Proteins 0.000 description 1
- 102000002067 Protein Subunits Human genes 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000000460 chlorine Substances 0.000 description 1
- 229910052801 chlorine Inorganic materials 0.000 description 1
- 230000001112 coagulating effect Effects 0.000 description 1
- 150000001896 cresols Chemical class 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 239000000645 desinfectant Substances 0.000 description 1
- 238000007599 discharging Methods 0.000 description 1
- 239000000975 dye Substances 0.000 description 1
- 235000013601 eggs Nutrition 0.000 description 1
- DNJIEGIFACGWOD-UHFFFAOYSA-N ethanethiol Chemical compound CCS DNJIEGIFACGWOD-UHFFFAOYSA-N 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 230000008595 infiltration Effects 0.000 description 1
- 238000001764 infiltration Methods 0.000 description 1
- 229910017053 inorganic salt Inorganic materials 0.000 description 1
- 239000011810 insulating material Substances 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 230000005012 migration Effects 0.000 description 1
- 238000013508 migration Methods 0.000 description 1
- 239000000178 monomer Substances 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000000575 pesticide Substances 0.000 description 1
- 239000003208 petroleum Substances 0.000 description 1
- 238000005120 petroleum cracking Methods 0.000 description 1
- 231100000614 poison Toxicity 0.000 description 1
- 230000007096 poisonous effect Effects 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 229920003987 resole Polymers 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
- 239000008399 tap water Substances 0.000 description 1
- 235000020679 tap water Nutrition 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 125000002256 xylenyl group Chemical class C1(C(C=CC=C1)C)(C)* 0.000 description 1
Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2101/00—Nature of the contaminant
- C02F2101/30—Organic compounds
- C02F2101/34—Organic compounds containing oxygen
- C02F2101/345—Phenols
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- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Microbiology (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Genetics & Genomics (AREA)
- Biotechnology (AREA)
- Water Supply & Treatment (AREA)
- Tropical Medicine & Parasitology (AREA)
- Virology (AREA)
- Medicinal Chemistry (AREA)
- Biomedical Technology (AREA)
- Environmental & Geological Engineering (AREA)
- Hydrology & Water Resources (AREA)
- Biodiversity & Conservation Biology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Purification Treatments By Anaerobic Or Anaerobic And Aerobic Bacteria Or Animals (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses a preparation method of a halophilic bacteria agent for degrading phenolic wastewater, and belongs to the field of preparation of halophilic bacteria agents. The method comprises the following steps: selecting activated sludge of high-salinity wastewater with a salt content of 3-15 percent discharged by a chemical plant in an industrial park to prepare a bacterial suspension; taking the bacterial suspension, and marking, separating and screening halophilic bacteria on a flat plate prepared by a halophilic bacteria separating and screening culture medium to obtain halophilic bacteria liquid; separating and screening to obtain the halophilic bacteria agent for degrading phenols. The prepared halophilic bacteria agent is used for degrading phenolic substances and can be used for treating the wastewater with over-high phenolic substance content to obtain a good treatment effect. The halophilic bacteria agent is applied to treatment of the wastewater with higher salt content and higher phenol content, can specially solve the problem of special wastewater, is economic and effective and has important significance and wide application prospect.
Description
Technical field
The present invention relates to microbial inoculum preparation field, particularly relate to the preparation method of a kind of degraded containing the halophilic bacterium microbial inoculum of phenol.
Background technology
In industrial park, waste water has quantity discharged greatly, and pollution range is wide, and discharging modes are complicated; Pollutant kind is various, and fluctuation of concentration amplitude is large; Pollution substance strong toxicity, harm is large, and after pollutant emission, migration and variation rule difference is large; Recover the feature such as more difficult.
In industrial park waste water, phenolic wastewater is mainly from industrial sector and petroleum cracking ethene, synthesizing phenol, tynex, synthetic dyestuff, organic pesticide and resol production processes such as coke-oven plant, producer gas plant, petroleum chemical plant, insulating material factorys.Main containing phenol-based compounds in phenolic wastewater, as phenol, cresols, xylenol and nitrocresol etc.Phenol-based compounds is a kind of protoplasma poisonous substance, can make protein coagulating.When in water, the mass concentration of phenol reaches 0.1 ~ 0.2mg/L, namely the flesh of fish has peculiar smell, can not eat; Mass concentration is increased to 1mg/L, can affect fish and lay eggs, and containing phenol 5 ~ 10mg/L, fish will mortality.Can HUMAN HEALTH be affected containing phenol in tap water, even if only have 0.002mg/L containing phenol mass concentration in water, also can produce chlorophenol stench with chlorine disinfectant.Usually be the phenolic wastewater of 1000mg/L by mass concentration. be called high-concentration phenolic wastewater, after this waste water must reclaim phenol, then process.Mass concentration is less than the phenolic wastewater of 1000mg/L, is called lower concentration phenolic wastewater.Usually this kind of waste water circulation is used, by the aftertreatment of phenol concentration and recovery.The method reclaiming phenol has solvent extration, steam blow-off method, absorption method, closed circulation method etc.Discharge after the method such as the oxidation of waste water applicable biological, chemical oxidation, physical chemistry oxidation of below 300mg/L processes containing phenol mass concentration or reclaim.
Biologic treating technique is widely used in sewage disposal because it has the feature of economical and efficient, but the infiltration of microorganism growth to environment is pressed with certain requirement, osmotic pressure in environment should in the pressure range that microorganism cells plasma membrane can bear, and the size of osmotic pressure is directly proportional to strength of solution, in solution, inorganic salt concentration is higher, then osmotic pressure is higher.Microorganism is in Hyposmolality solution (NaCl massfraction 0.01%), and in solution, water molecules infiltrates in microbe in a large number, and microorganism cells is expanded, and severe patient breaks; And in high osmotic pressure environment, in microbe, water molecules is seeped in vitro in a large number, result causes cell generation plasmolysis that growth is suppressed, and severe patient is dead.
There is in the cell of halophilic bacterium, particularly Natrinema altunense sp quite high ionic concn, and cell wall components is special, containing peptidoglycan and based on lipoprotein.The complete of this cell wall structure is maintained by ionic linkage, and high density, for the combination between its cell wall protein subunit, keeps cyto-architectural integrity to be required.When the salt concn in environment is lower, cell wall protein depolymerization is protein monomers on the one hand, cell wall is lost complete; On the other hand, intraor extracellular ionic concn balance is broken, and cell water-swelling, finally causes cell wall to break, the complete self-dissolving of thalline.The generation of halophilic bacterium enzyme, stable and activation plays all need high salt concentration.The generation of organism endoenzyme, stable and activation plays all need certain physico chemical factor.The enzyme of halophilic bacterium is halophilism.Its generation, stable and play activity and need high salt concentration condition.And do not prepare halophilic bacterium very well at present, particularly degraded is containing the method for the halophilic bacterium of phenol.
Summary of the invention
Based on the problem existing for above-mentioned prior art, the invention provides a kind of degraded containing the preparation method of the halophilic bacterium microbial inoculum of phenol, can utilize the active sludge of the high slat-containing wastewater of chemical plant emission in industrial park, the degraded for the preparation of water treatment contains the halophilic bacterium of phenol.
For solving the problems of the technologies described above, the invention provides a kind of degraded containing the preparation method of the halophilic bacterium microbial inoculum of phenol, it is characterized in that, comprising:
(1) bacteria suspension is made:
The saltiness choosing chemical plant emission in industrial park is that the active sludge of the high slat-containing wastewater of 3% ~ 15% makes bacteria suspension;
(2) separation screening halophilic bacterium:
Get the flat lining out separation screening halophilic bacterium that described bacteria suspension makes at halophilic bacterium separation screening substratum and obtain halophilic bacterium bacterium liquid;
(3) separation screening degraded is containing the halophilic bacterium of phenol:
By peptone 5 ~ 15g, yeast extract paste 1 ~ 10g, NaCl5 ~ 15g, after phenol 0.1 ~ 0.2g, distilled water 1000ml mixing, be adjusted to 7.0 ~ 7.2 by pH value, within 20 minutes, be mixed with LB phenol substratum in 121 DEG C of sterilizings;
Oxford cup is added in described LB phenol culture medium flat plate; the described halophilic bacterium bacterium liquid that step 2 obtains is added in the cup of Oxford; with not adding the Oxford cup of bacterium liquid as contrast; 37 DEG C; cultivate 24 ~ 36h; the bacterium liquid choosing large more than the 5mm of transparent circle is compared with the control the halophilic bacterium of degradation of phenol, adds stablizer and protective material and namely complete the preparation of degraded containing the halophilic bacterium microbial inoculum of phenol in the halophilic bacterium of described degraded containing phenol.
In aforesaid method, described stablizer adopt in glycerine, ethanol, ethylene glycol one or more, add-on is described degraded containing 0.5 ~ 5% of the halophilic bacterium weight of phenol; Described protective material adopt in zeolite powder, diatomite, Powdered Activated Carbon one or more, add-on is described degraded containing 1 ~ 10% of the halophilic bacterium weight of phenol.
In aforesaid method, in described making bacteria suspension step, by as follows for the mode that the active sludge of high slat-containing wastewater makes bacteria suspension:
The waste water water sample gathering 200ml puts into beaker, with filter paper filtering, then hold in centrifuge tube, with 12000r/min centrifugation 30min, pour out supernatant liquor, whole supernatant liquor is filtered by bacterial filter, then takes off filtering membrane, filtering membrane is put into the beaker containing aseptic water sample, filter paper is fully soaked and obtains solution; The described solution taking out 1ml with the transfer pipet of sterilizing puts into aseptic test tube, in this test tube, add 9ml sterilized water, fully after concussion, allows it fully mix, namely makes bacteria suspension.
In aforesaid method, in described separation screening halophilic bacterium step, halophilic bacterium separation screening substratum used is: by extractum carnis 2 ~ 5g, peptone 5 ~ 15g, NaCl5 ~ 15g, Na
2sO
43 ~ 10g, KCl 0.5 ~ 1.5g, NaHCO
30.1 ~ 1g, KBr 0.1 ~ 1.5g, H
3bO
30.01 ~ 0.15g, NaF 0.001 ~ 0.015g, MgCl
21 ~ 15g and CaCl
20.5 ~ 1.5g, adding distil water to 1000ml, after pH value is adjusted to 7.0 ~ 7.2, through the substratum that 121 DEG C of sterilizings 20 minutes are obtained.
Beneficial effect of the present invention is: the halophilic bacterium microbial inoculum suitability of obtained degraded phenolic wastewater is more wide in range, can be used for the process of a series of phenolic wastewater of coking chemical waste water, chemical engineering sewage etc. in industrial park, in conjunction with different treatment process, good treatment effect can be reached.
Embodiment
Be clearly and completely described the technical scheme in the embodiment of the present invention below, obviously, described embodiment is only the present invention's part embodiment, instead of whole embodiments.Based on embodiments of the invention, those of ordinary skill in the art, not making the every other embodiment obtained under creative work prerequisite, belong to protection scope of the present invention.Method:
The embodiment of the present invention provides halophilic bacterium microbial inoculum of a kind of phenolic wastewater of degrading and preparation method thereof, for the biological treating of high-concentration phenolic wastewater, specifically comprises the following steps:
(1) bacteria suspension is prepared in the collection of sample:
Choose the active sludge of the high slat-containing wastewater of chemical plant emission in industrial park as sample, the saltiness of waste water is 3% ~ 15%.
The water sample gathering 200ml puts into beaker, with filter paper filtering, then hold in the middle of centrifuge tube, with 12000r/min centrifugation 30min, pour out supernatant liquor, whole supernatant liquor is filtered by bacterial filter, then takes off filtering membrane, put it in the beaker containing aseptic water sample, filter paper is fully soaked.Take out this solution of 1ml with the transfer pipet of bacterium of having gone out and put into aseptic test tube, in this test tube, add 9ml sterilized water, fully after concussion, allow it fully mix, make bacteria suspension;
(2) separation screening halophilic bacterium:
Get the flat lining out that bacteria suspension is made at halophilic bacterium separation screening substratum, separation screening halophilic bacterium.The composition of halophilic bacterium separation screening substratum has extractum carnis 3g, peptone 7g, NaCl6g, Na
2sO
48g, KCl 1g, NaHCO
30.5g, KBr0.4g, H
3bO
30.12g, NaF 0.008g, MgCl
29g, CaCl
21g, adding distil water is to 1000ml, and pH value is adjusted to 7.0 ~ 7.2.121 DEG C of sterilizing 20min obtain.
3, separation screening degraded is containing the halophilic bacterium of phenol:
Preparation LB phenol substratum.Peptone 10g, yeast extract paste 9g, NaCl 15g, phenol 0.2g, distilled water 1000ml, pH value is adjusted to 7.0 ~ 7.2.121 DEG C of sterilizing 20min obtain.
In LB phenol culture medium flat plate, adding Oxford cup, in the cup of Oxford, add halophilic bacterium bacterium liquid, with not adding the Oxford cup of bacterium liquid as contrast, 37 DEG C, cultivating 24h.The bacterium liquid choosing large more than the 5mm of transparent circle is compared with the control the halophilic bacterium of degradation of phenol.
(4) in degraded is containing the halophilic bacterium of phenol, stablizer and protective material is added.Stablizer adopts glycerine, ethanol, one or more in ethylene glycol, and add-on is degraded containing 0.5 ~ 5% of the halophilic bacterium weight of phenol; Protective material adopts zeolite powder, diatomite, one or more in Powdered Activated Carbon, and add-on is degraded containing 1 ~ 10% of the halophilic bacterium weight of phenol.
Below in conjunction with specific embodiment, the invention will be further described.
Embodiment 1
The present embodiment provides a kind of and prepares the method for degraded containing the halophilic bacterium microbial inoculum of phenol, comprises the following steps:
(1) bacteria suspension is prepared in the collection of sample:
Choose the active sludge of the high slat-containing wastewater of chemical plant emission in industrial park as sample, the saltiness of waste water is 8%;
The water sample gathering 200ml puts into beaker, with filter paper filtering, then hold in the middle of centrifuge tube, with 12000r/min centrifugation 30min, pour out supernatant liquor, whole supernatant liquor is filtered by bacterial filter, then takes off filtering membrane, put it in the beaker containing aseptic water sample, filter paper is fully soaked; Take out this solution of 1ml with the transfer pipet of bacterium of having gone out and put into aseptic test tube, in this test tube, add 9ml sterilized water, fully after concussion, allow it fully be mixed;
(2) separation screening halophilic bacterium:
Get the flat lining out that bacteria suspension is made at halophilic bacterium separation screening substratum, separation screening halophilic bacterium.The composition of halophilic bacterium separation screening substratum has extractum carnis 2 ~ 5g, peptone 5 ~ 15g, NaCl5 ~ 15g, Na
2sO
43 ~ 10g, KCl 0.5 ~ 1.5g, NaHCO
30.1 ~ 1g, KBr 0.1 ~ 1.5g, H
3bO
30.01 ~ 0.15g, NaF 0.001 ~ 0.015g, MgCl
21 ~ 15g, CaCl
20.5 ~ 1.5g, adding distil water is to 1000ml, and pH value is adjusted to 7.0 ~ 7.2.121 DEG C of sterilizing 20min obtain;
(3) separation screening degraded is containing the halophilic bacterium of phenol:
Preparation LB phenol substratum.Peptone 5 ~ 15g, yeast extract paste 1 ~ 10g, NaCl5 ~ 15g, phenol 0.1 ~ 0.2g, distilled water 1000ml, pH value is adjusted to 7.0 ~ 7.2.121 DEG C of sterilizing 20min obtain.
In LB phenol culture medium flat plate, add Oxford cup, add halophilic bacterium bacterium liquid in the cup of Oxford, contrast, 37 DEG C with the Oxford cup not adding bacterium liquid, cultivate 24 ~ 36h, the bacterium liquid choosing large more than the 5mm of transparent circle is compared with the control the halophilic bacterium of degradation of phenol;
(4) in degraded is containing the halophilic bacterium of phenol, stablizer and protective material is added.Stablizer adopts glycerine, ethanol, one or more in ethylene glycol, and add-on is 0.5 ~ 5% of the halophilic bacterium weight of degradation of phenol; Protective material adopts zeolite powder, diatomite, one or more in Powdered Activated Carbon, and add-on is 1 ~ 10% of the halophilic bacterium weight of degradation of phenol.
Embodiment 2
The embodiment of the present invention provides halophilic bacterium microbial inoculum of a kind of phenolic wastewater of degrading and preparation method thereof.This halophilic bacterium microbial inoculum is for high-concentration phenolic wastewater.
The concrete preparation method of the embodiment of the present invention is as follows:
(1) sample collecting makes bacteria suspension:
Choose the active sludge of the high slat-containing wastewater of chemical plant emission in industrial park as sample, the saltiness of waste water is 10%.
The water sample gathering 200ml puts into beaker, with filter paper filtering, then hold in the middle of centrifuge tube, with 12000r/min centrifugation 30min, pour out supernatant liquor, whole supernatant liquor is filtered by bacterial filter, then takes off filtering membrane, put it in the beaker containing aseptic water sample, filter paper is fully soaked.Take out this solution of 1ml with the transfer pipet of bacterium of having gone out and put into aseptic test tube, in this test tube, add 9ml sterilized water, fully after concussion, allow it fully mix, make bacteria suspension;
(2) separation screening halophilic bacterium:
Get the flat lining out that bacteria suspension is made at halophilic bacterium separation screening substratum, separation screening halophilic bacterium.The composition of halophilic bacterium separation screening substratum has extractum carnis 5g, peptone 9g, NaCl8g, Na
2sO
45g, KCl 0.8g, NaHCO
30.9g, KBr0.5g, H
3bO
30.14g, NaF 0.009g, MgCl
210g, CaCl
20.8g, adding distil water is to 1000ml, and pH value is adjusted to 7.0 ~ 7.2, and 121 DEG C of sterilizing 20min obtain;
(3) separation screening degraded is containing the halophilic bacterium of phenol:
Preparation LB phenol substratum.Peptone 10g, yeast extract paste 5g, NaCl 10g, phenol 0.2g, distilled water 1000ml, pH value is adjusted to 7.0 ~ 7.2, and 121 DEG C of sterilizing 20min obtain;
In LB phenol culture medium flat plate, add Oxford cup, add halophilic bacterium bacterium liquid in the cup of Oxford, with not adding the Oxford cup of bacterium liquid as contrast, 37 DEG C, cultivate 24h, the bacterium liquid choosing large more than the 5mm of transparent circle is compared with the control the halophilic bacterium of degradation of phenol;
(4) in degraded is containing the halophilic bacterium of phenol, stablizer and protective material is added.Stablizer adopts glycerine, ethanol, one or more in ethylene glycol, and add-on is 0.5 ~ 5% of the halophilic bacterium weight of degradation of phenol; Protective material adopts zeolite powder, diatomite, one or more in Powdered Activated Carbon, and add-on is 1 ~ 10% of the halophilic bacterium weight of degradation of phenol.
Embodiment 3
The embodiment of the present invention provides halophilic bacterium microbial inoculum of a kind of phenolic wastewater of degrading and preparation method thereof.This halophilic bacterium microbial inoculum is for high-concentration phenolic wastewater.
The concrete preparation method of the embodiment of the present invention is as follows:
(1) bacteria suspension is made in the collection of sample
Choose the active sludge of the high slat-containing wastewater of chemical plant emission in industrial park as sample, the saltiness of waste water is 15%.
The water sample gathering 200ml puts into beaker, with filter paper filtering, then hold in the middle of centrifuge tube, with 12000r/min centrifugation 30min, pour out supernatant liquor, whole supernatant liquor is filtered by bacterial filter, then takes off filtering membrane, put it in the beaker containing aseptic water sample, filter paper is fully soaked.Take out this solution of 1ml with the transfer pipet of bacterium of having gone out and put into aseptic test tube, in this test tube, add 9ml sterilized water, fully after concussion, allow it fully mix, make bacteria suspension.
(2) separation screening halophilic bacterium:
Get the flat lining out that bacteria suspension is made at halophilic bacterium separation screening substratum, separation screening halophilic bacterium.The composition of halophilic bacterium separation screening substratum has extractum carnis 5g, peptone 5g, NaCl6g, Na
2sO
47g, KCl 0.8g, NaHCO
30.5g, KBr0.5g, H
3bO
30.13g, NaF 0.01g, MgCl
210g, CaCl
21g, adding distil water is to 1000ml, and pH value is adjusted to 7.0 ~ 7.2.121 DEG C of sterilizing 20min obtain.
(3) degraded is containing the separation screening of the halophilic bacterium of phenol
Preparation LB phenol substratum.Peptone 8g, yeast extract paste 7g, NaCl 13g, phenol 0.2g, distilled water 1000ml, pH value is adjusted to 7.0 ~ 7.2.121 DEG C of sterilizing 20min obtain.
In LB phenol culture medium flat plate, adding Oxford cup, in the cup of Oxford, add halophilic bacterium bacterium liquid, with not adding the Oxford cup of bacterium liquid as contrast, 37 DEG C, cultivating 24h.The bacterium liquid choosing large more than the 5mm of transparent circle is compared with the control the halophilic bacterium of degradation of phenol.
(4) in degraded is containing the halophilic bacterium of phenol, stablizer and protective material is added.Stablizer adopts glycerine, ethanol, one or more in ethylene glycol, and add-on is 0.5 ~ 5% of the halophilic bacterium weight of degradation of phenol.Protective material adopts zeolite powder, diatomite, one or more in Powdered Activated Carbon, and add-on is 1 ~ 10% of the halophilic bacterium weight of degradation of phenol.
The halophilic bacterium microbial inoculum of degraded phenolic wastewater the various embodiments described above obtained, is applied in engineering project, measures water inlet phenol and water outlet phenol, specific as follows:
The halophilic bacterium microbial inoculum of the degraded phenolic wastewater obtained by each embodiment drops in the Aerobic Pond of each domestic sewage factory, control temperature at 20 ~ 38 DEG C, pH value 6.0 ~ 9.0, dissolved oxygen at 3 ~ 6mg/L, sludge settling ratio SV
30be 15% ~ 30%, the residence time 3 ~ 6h, influent COD index is 400 ~ 650mg/L, and water outlet COD index is 60 ~ 100mg/L, and water inlet phenol content 100 ~ 200mg/L, return sludge ratio controls at 1:1 ~ 1:4.
Can be found out by above-mentioned contrast, in three engineerings of the halophilic bacterium microbial inoculum of the degraded phenolic wastewater of interpolation various embodiments of the present invention, the clearance of phenol all reaches more than 90%, apparently higher than the engineering of not adding microbial inoculum, final outflow water phenol all can be made to reach country-level A standard.And in operational process, adding of the halophilic bacterium microbial inoculum of degraded phenolic wastewater is more convenient, and capacity of resisting impact load is comparatively strong, and operational management is convenient, and cost is not high.
In sum, the suitability of the halophilic bacterium microbial inoculum of the degraded phenolic wastewater of the embodiment of the present invention is more wide in range, can be used for the process of a series of phenolic wastewater of coking chemical waste water, chemical engineering sewage etc. in industrial park, in conjunction with different treatment process, good treatment effect can be reached.
The above; be only the present invention's preferably embodiment, but protection scope of the present invention is not limited thereto, is anyly familiar with those skilled in the art in the technical scope that the present invention discloses; the change that can expect easily or replacement, all should be encompassed within protection scope of the present invention.Therefore, protection scope of the present invention should be as the criterion with the protection domain of claims.
Claims (4)
1. degraded is containing a preparation method for the halophilic bacterium microbial inoculum of phenol, it is characterized in that, comprising:
(1) bacteria suspension is made:
The saltiness choosing chemical plant emission in industrial park is that the active sludge of the high slat-containing wastewater of 3% ~ 15% makes bacteria suspension;
(2) separation screening halophilic bacterium:
Get the flat lining out separation screening halophilic bacterium that described bacteria suspension makes at halophilic bacterium separation screening substratum and obtain halophilic bacterium bacterium liquid;
(3) separation screening degraded is containing the halophilic bacterium of phenol:
By peptone 5 ~ 15g, yeast extract paste 1 ~ 10g, NaCl5 ~ 15g, after phenol 0.1 ~ 0.2g, distilled water 1000ml mixing, be adjusted to 7.0 ~ 7.2 by pH value, within 20 minutes, be mixed with LB phenol substratum in 121 DEG C of sterilizings;
Oxford cup is added in described LB phenol culture medium flat plate; the described halophilic bacterium bacterium liquid that step 2 obtains is added in the cup of Oxford; with not adding the Oxford cup of bacterium liquid as contrast; 37 DEG C; cultivate 24 ~ 36h; the bacterium liquid choosing large more than the 5mm of transparent circle is compared with the control the halophilic bacterium of degradation of phenol, adds stablizer and protective material and namely complete the preparation of degraded containing the halophilic bacterium microbial inoculum of phenol in the halophilic bacterium of described degraded containing phenol.
2. a kind of degraded according to claim 1 is containing the preparation method of halophilic bacterium microbial inoculum of phenol, it is characterized in that, described stablizer adopt in glycerine, ethanol, ethylene glycol one or more, add-on is described degraded containing 0.5 ~ 5% of the halophilic bacterium weight of phenol; Described protective material adopt in zeolite powder, diatomite, Powdered Activated Carbon one or more, add-on is described degraded containing 1 ~ 10% of the halophilic bacterium weight of phenol.
3. a kind of degraded according to claim 1 and 2 is containing the preparation method of the halophilic bacterium microbial inoculum of phenol, it is characterized in that, in described making bacteria suspension step, by as follows for the mode that the active sludge of high slat-containing wastewater makes bacteria suspension:
The waste water water sample gathering 200ml puts into beaker, with filter paper filtering, then hold in centrifuge tube, with 12000r/min centrifugation 30min, pour out supernatant liquor, whole supernatant liquor is filtered by bacterial filter, then takes off filtering membrane, filtering membrane is put into the beaker containing aseptic water sample, filter paper is fully soaked and obtains solution; The described solution taking out 1ml with the transfer pipet of sterilizing puts into aseptic test tube, in this test tube, add 9ml sterilized water, fully after concussion, allows it fully mix, namely makes bacteria suspension.
4. a kind of degraded according to claim 1 and 2 is containing the preparation method of the halophilic bacterium microbial inoculum of phenol, it is characterized in that, in described separation screening halophilic bacterium step, halophilic bacterium separation screening substratum used is: by extractum carnis 2 ~ 5g, peptone 5 ~ 15g, NaCl5 ~ 15g, Na
2sO
43 ~ 10g, KCl 0.5 ~ 1.5g, NaHCO
30.1 ~ 1g, KBr 0.1 ~ 1.5g, H
3bO
30.01 ~ 0.15g, NaF 0.001 ~ 0.015g, MgCl
21 ~ 15g and CaCl
20.5 ~ 1.5g, adding distil water to 1000ml, after pH value is adjusted to 7.0 ~ 7.2, through the substratum that 121 DEG C of sterilizings 20 minutes are obtained.
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