CN101831391A - Effective phenylamine degrading strain as well as use and use method thereof - Google Patents
Effective phenylamine degrading strain as well as use and use method thereof Download PDFInfo
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- CN101831391A CN101831391A CN200910058557A CN200910058557A CN101831391A CN 101831391 A CN101831391 A CN 101831391A CN 200910058557 A CN200910058557 A CN 200910058557A CN 200910058557 A CN200910058557 A CN 200910058557A CN 101831391 A CN101831391 A CN 101831391A
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
- Y02W10/00—Technologies for wastewater treatment
- Y02W10/10—Biological treatment of water, waste water, or sewage
Abstract
The invention belongs to the technical field of environment protection, in particular an effective phenylamine degrading strain as well as use and use method thereof. In the invention, a code number of the effective phenylamine degrading bacteria is AN-P1; and the effective phenylamine degrading bacteria is authenticated to be rhodococcus sp, which is preserved in China committee for culture preservation of general microorganism centre with the preservation number of CGMCC No.2783 in DEC.4th, 2008. The effective phenylamine degrading strain is applied to high-concentration phenylamine waste water treatment or phenylamine pollution incident emergency treatment. The effective phenylamine degrading strain has the characteristics of strong endurance capability of high-concentration phenylamine, high degradation speed, no secondary pollution, safe use, no bad influence on microorganism in an original processing system and the like.
Description
Technical field
The invention belongs to environmental protection technical field, be specifically related to a strain efficient degrading bacteria and the application in biological treatment of high density aniline waste water and aniline happen suddenly the pollution incident emergency processing thereof of aniline degradation fast.
Background technology
Aniline is the main noxious pollutant in the industrial discharge waste water such as oil refining, dyestuff, agricultural chemicals, medicine, is modal water pollutant.Aniline is poisonous and harmful " three cause " material, is detrimental to health.Physico-chemical process and biological process are mainly adopted in the processing of aniline waste water at present, and to exist processing efficiency low but physico-chemical process is handled aniline, problems such as cost height, secondary pollution are serious, complicated operation; Biological treatment system is as the waste water treatment process that is most widely used, have Pollutant Treatment speed fast, the efficient height, cost is low, treatment capacity is big, characteristics such as non-secondary pollution, but because aniline is highly toxic substance, in case entering biological treatment system, will suppress high density aniline, damage wherein microorganism system, thereby make biological process handle aniline waste water and be subjected to very big restriction, in recent years, many work have also been carried out both at home and abroad at efficient microphenomenon of degradation of aniline of screening and processing aniline waste water, as: 40 hours degradable 1000mg/L of AN4 bacterial strain, 2000mg/L aniline degraded 12.5% after 44 hours.The 1000mg/L aniline waste water was degraded more than 80% in 48h hour, fell when aniline concentration is higher than 1000mg/L to be subjected to press down.Taiwan patent microbial inoculum H.S.B is by holding concurrently oxygen 30～40h, aerobic 20～30h can processing up to standard less than the aniline waste water of 500mg/L.(Zhang Yifei, Gu Ting, kingdom is auspicious, Deng. the isolation identification of a strain microphenomenon of degradation of aniline and the .2008 of degradation characteristic Environmental Pollution and Control [J] thereof, 30 (02): 12～15. Wei seawards, appoint the source, Wu Chaofei, Deng. the dynamics research [J] of obligate aerobic bacteria aniline degradation waste water. Research of Environmental Sciences, 1999,12 (4): 15～18. chapters are strong, Liu Qing is the microbiological deterioration of aniline [J]. Agricultural University Of Anhui's journal, give birth in 1997,24 (3): 283～286. yellow capital. the full biochemical processing method [P] of nitrobenzene waste water or aniline waste water or its composite waste. and Chinese patent: 200410065840.2,2005-07-27).Aniline waste water biological treatment at present exists low, the problems such as degradation rate is slow, difficulty up to standard of aniline tolerance concentration.
On the other hand, the production technique of aniline industry and various artificial origin make it become the high issuing industry of environmental accident.The aniline blast and the leakage accident that only take place in the period of nineteen ninety-five to 2006 according to incompletely statistics just reach more than 20, the wherein foremost aniline explosive incident that comprises PetroChina Company Limited.'s Jilin Petrochemical and Lanzhou petrochemical industry.Pollute blast and leakage accident at burst, country stipulates that clearly No. [2006] 10, danger (safety supervision always) chemical enterprise must be provided with under the accidental states such as accident pool or Buffer Pool collection, the Disposal Measures of " peace and quiet descend water ".Yet, after aniline burst pollution incident took place, high density aniline waste liquid still need carry out treatment and disposal fast and safely after effectively being isolated and tackling, adopt methods such as charcoal absorption or materialization processing at present mostly, face and handle inconvenience, secondary pollution problems.Therefore, how to utilize the huge removing pollutant of pollution incident spot plant effluent treatment station or sewage work to handle capacity and ability, strengthen the high-concentration waste liquid that quick emergency processing pollution incident produces, become the emphasis of pollution incident urgent technique research and development.
Summary of the invention
The objective of the invention is to take place frequently, and aniline toxicity is big, degraded is difficult, the slow problem of degradation rate at the aniline pollution incident, provide a strain fast degrading high concentration aniline efficient degrading bacteria and be applied to handle the technology of burst pollution incident.
Black Liquor with Efficient Bacteria AN-P1 provided by the present invention is from the sewage active sludge, through enrichment screening, purifying gained.This bacterial strain can be sole carbon source, nitrogenous source and energy growth with aniline.This identification of strains is: rhodococcus Rhodoccocus sp., on December 4th, 2008 in the common micro-organisms center preservation of Chinese culture presevation management committee, preserving number is: CGMCC No.2783.This bacterial strain bacterium colony is incarnadine, circle, neat in edge, convex protuberance, thickness, and colony diameter is about 1.5mm; Gram-positive, oxidase negative, hydrogen peroxide enzyme positive; Can utilize salts such as Citrate trianion, succinate to be carbon source; The nitrate reduction positive.
The AN-P1 bacterium 2000mg/L aniline 32h that degrades under the pure culture condition can make water outlet reach " integrated wastewater discharge standard " (GB 8978-1996) primary standard, the high density aniline of 500～2000mg/L capable of being fast degraded can be used for the biological reinforced processing of high density aniline waste water.In conjunction with sewage disposal conventional activated sludge system, with AN-P1 is function stem, it is made an addition to Sludge System by 0.1 ‰～0.3 ‰ (dry cell weight) inoculum size, 500～2000mg/L aniline that emergency processing aniline burst pollution incident produces, the emergent treatment system that AN-P1 strengthens can reach the relevant emission standards of national regulation at the efficient aniline of removing in the conventional sewage disposal hydraulic detention time 8～35 hours.By 16S r DNA, V3 district fragment PCR-DGGE collection of illustrative plates check and analysis result shows that this bacterial strain can effectively protect the Sewage treatment systems microbial ecological to avoid high density aniline influence.
The present invention has the following advantages:
1, function stem AN-P1 is strong to high density aniline tolerance, degradation speed is fast, and the aniline of 2000mg/L is still had good degradation effect;
2, the high density aniline waste liquid that produces at aniline burst pollution incident, make full use of the huge removing pollutant of pollution incident spot plant effluent treatment station or sewage work and handle capacity and ability, add aniline efficient degrading bacteria emergency processing pollution incident, have easy and simple to handle, fast and safely, feature such as non-secondary pollution.
3, aniline efficient degrading bacteria fermentation culture mode is simple, the required hydraulic detention time of degraded 500～2000mg/L aniline is 10～32 hours, this and sanitary sewage, the trade effluent biological treatment residence time are identical substantially, and it is safe in utilization, can not cause detrimentally affect to microorganism and treatment effect in original treatment system.
Embodiment
Embodiment 1: the screening and separating of microphenomenon of degradation of aniline Rhodoccocus sp.AN-P1 and processing aniline waste water
Gather Chengdu first sewage work's Aerobic Pond active sludge, mud sample is by 10% inoculum size, 30 ℃, 160r/min, progressively improves the aniline concentration enrichment and cultivates.Containing line separation on the 1000mg/L aniline MSB agar plate then, select the bacterium colony of different shape feature, again be forwarded in the MSB substratum of 1000mg/L aniline, it is its degrading aniline ability that the same terms is cultivated with checking down, is further purified until the pure culture bacterial strain that obtains degradation capability.
Synthetic medium (MSB): every 1L contains Na
2HPO
42g, KH
2PO
40.5g, MgSO
47H
2O 0.3g, trace element solution 5mL, aniline 1g.The agar powder that adds 15g/L in the solid medium.
Obtain an effective phenylamine degrading strain as well as AN-P1 through taming final separation and purification repeatedly.Bacterium colony is incarnadine, circle, neat in edge, convex protuberance, thickness, and colony diameter is about 1.5mm; Gram-positive, oxidase negative, hydrogen peroxide enzyme positive; Can utilize salts such as Citrate trianion, succinate to be carbon source; The nitrate reduction positive.It reaches 98% with the likelihood of having a liking for the 16S rDNA sequence of pyridine rhodococcus (Rhodococcus pyridinivorans) analysis of 16S rDNA, is that rhodococcus (Rhodoccocus sp.) is numbered AN-P1. through identifying this bacterium
This bacterium is contained the aniline simulated wastewater (adding aniline in the MSB substratum) of aniline 500mg/L, 1000mg/L, 2000mg/L at pH=7 in 0.3 ‰ inoculum sizes (with dry cell weight) access, 30 ℃, 160rpm cultivates, every 3h sampling and measuring aniline content (as shown in table 1) and COD, measurement result shows respectively by 18h, 24h, 32h degraded, the aniline clearance is higher than 99.9%, water outlet COD difference 47.4,85.5,24.2mg/L, effluent quality satisfy " integrated wastewater discharge standard " (GB8978-1996) primary standard.
Table 1AN-P1 is to the degradation effect of different concns aniline simulated wastewater
Embodiment 2:AN-P1 simulation emergency processing aniline burst pollution incident waste liquid
The aniline that adds a certain amount of (500mg/L, 1000mg/L, 2000mg/L) in sanitary sewage, aniline enters network of drains impact sewage biological treatment system after the simulation burst pollution incident.Experimental system for simulating divides two groups, and one group is contradistinction system, i.e. conventional activated sludge treatment system (MLSS=2000 ± 100mg/L); Second group is the biological reinforced emergent treatment system (MLSS=2000 ± 100mg/L) of 0.3 ‰ AN-P1 bacterial strains for adding with inoculum size.30 ℃, natural pH, (dissolved oxygen is controlled at 3～5mg/L) to 160rpm shaking table cultivation simulation process.The 500mg/L group is every 1h sampling and measuring aniline and COD degradation effect; 1000mg/L, 2000mg/L group is every 4h sampling and measuring aniline and COD degradation effect, shown in result table 2, table 3, the table 4.Simultaneously, to different period sample 16S r DNA in 1000mg/L, the 2000mg/L aniline burst pollution incident simulated wastewater treating processes, V3 district fragment is carried out PCR-DGGE and is analyzed.
Table 2, table 3, table 4 result of study shows, to 500,1000, the aniline pollution incident waste liquid of 2000mg/L, the conventional activated sludge system does not have removal substantially to aniline, and as function stem the conventional activated sludge system is strengthened emergency processing burst pollution incident aniline waste liquid with AN-P1, contain 500mg/L, 1000mg/L, the pollution incident waste liquid of 2000mg/L aniline is handled 10h respectively, 20h, behind the 32h, aniline content and COD reach " integrated wastewater discharge standard " (GB8978-1996) primary standard in the treat effluent, compare with the bacterial strain of having reported, improved the processing speed of biological treatment system greatly aniline tolerance concentration and high density aniline waste water.
Table 2 function yeast AN-P1 emergency processing 500mg/L aniline pollution incident waste liquid effect
Table 3 function yeast AN-P1 emergency processing 1000mg/L aniline pollution incident waste liquid effect
Table 4 function yeast AN-P1 emergency processing 2000mg/L aniline pollution incident waste liquid effect
Microorganism PCR-DGGE analytical results shows in the emergency processing process, process relatively before and after emergent treatment system and contradistinction system were handled the aniline waste liquid, there are A, B, three obvious characteristics bands of C, A is the characteristic strip of AN-P1 function yeast, can both detect characteristic strip clearly in emergent treatment system.Band B band in the conventional activated sludge system is darker, and its band B becomes clear bright behind the consolidation system processing aniline co simulate waste liquid, becomes dominant bacteria.Band C handles band brightness deepening behind the aniline co simulate waste liquid in the conventional activated sludge system, the bacterial number of its representative reduces, and its brightness reduces more obvious when handling high density (2000mg/L) aniline.And band C brightness increases on the contrary to some extent handle aniline in emergent treatment system after, shows that the corresponding bacterial number of band C increases.Swimming lane band number analytical results shows, 1000mg/L, the emergency processing of 2000mg/L aniline liquid waste treatment system 0 hour, contradistinction system bar algebraically is 16, and emergent treatment system bar algebraically is 17 (having more the characteristic strip of an AN-P1 function yeast than contradistinction system); Emergency processing 20,32 hours, contradistinction system bar algebraically is 14, emergent treatment system bar algebraically is 18.Band difference shows before and after handling, when using conventional Sludge System emergency processing high density aniline waste water, swimming lane band reduced number shows that the microorganism in the system can be subjected to the murder by poisoning of aniline, damage, thereby cause microbe population, kind to reduce the system handles degradation.And strengthen the system of emergency processing aniline co simulate waste liquid with the AN-P1 bacterial strain, and can protect the microorganism in the treatment system to avoid the aniline influence, make system aspect bacterial number, kind, all can keep stablizing.PCR-DGGE result can effectively protect the microorganism in original ecosystem to avoid the aniline influence after showing reinforcement.It is safe and feasible that proof AN-P1 is applied to emergency processing aniline burst pollution incident.
Embodiment 3: based on the aniline pollution incident waste liquid emergency processing of SBR technology
Get domestic sewage factory mud and sewage and make up the SBR process system, reactor is numbered (conventional SBR system) No. 1, No. 2 (emergent SBR system), useful volume 4.5L, MLSS=2000 ± 100mg/L, 32: 1 (dissolved oxygens 3～5mg/L) of steam-water ratio, room temperature, normal operation period HRT is 8h, adds 2000mg/L aniline to 1, No. 2 reactors behind the steady running 7d, by 0.3 ‰ inoculation AN-P1 bacterium to 2 reactors, every four hours sampling and measuring aniline content and COD.Measurement result is as shown in table 2.After emergency processing finishes, recover SBR technology and normally move the processing sanitary sewage, detect COD, ammonia nitrogen effluent quality index, detected result is as shown in table 3.
As shown in table 4, aniline and COD detected result show by the water outlet of No. 2 SBR emergent treatment systems of 36h degraded can arrive " integrated wastewater discharge standard " (GB 8978-1996) primary standard.Aniline content and COD are respectively 0.67mg/L, 97.7mg/L.And conventional SBR system aniline clearance only is 1.7%, and COD is 6083.3mg/L.
Table 4 is based on the aniline pollution incident waste liquid emergency processing effect of SBR technology
As shown in table 5, behind the emergency processing aniline pollution incident waste liquid, conventional SBR system normally moves 3 all after dates with emergent SBR system to the removal effect of COD can recover qualified discharge, the normal operation 7 all after dates of emergent SBR system ammonia nitrogen removal effect recover up to standard, and remove efficient and day by day improve, conventional SBR system 14 all after dates can not recover yet.Show with the AN-P1 bacterium strengthen behind the emergent treatment system can not only be rapidly and efficiently removal aniline, and can effectively ensure the purifying property of treatment system to pollutent.
Table 5 based on the aniline pollution incident waste liquid emergency processing of SBR technology after the normal operating performance of system investigate
The foregoing description is a preferred implementation of the present invention; but embodiments of the present invention are not restricted to the described embodiments; other any do not deviate from change, the modification made under spirit of the present invention and the principle, substitutes, combination, simplify; all should be the substitute mode of equivalence, be included within protection scope of the present invention.
Claims (4)
1. an effective phenylamine degrading strain as well as, code name is AN-P1, through being accredited as rhodococcus Rhodoccocussp., on December 4th, 2008 in the common micro-organisms center preservation of Chinese culture presevation management committee, preserving number is: CGMCC No.2783; This mycetocyte form is spherical or rod-short, Gram-positive, and atrichia does not move, and does not produce gemma; Bacterium colony is incarnadine, circle, neat in edge, convex protuberance, thickness; Oxidase negative, hydrogen peroxide enzyme positive; The nitrate reduction positive.
2. the cultural method of the described efficient microphenomenon of degradation of aniline of claim 1 is:
Medium component: every 1L contains Na
2HPO
42g, KH
2PO
40.5g, MgSO
47H
2O 0.3g, trace element solution 5mL, aniline 1g, the agar powder of adding 15g/L in the solid medium;
Cultural method: adopt above-mentioned substratum, bacterial classification is activated, the activation culture condition is 30 ℃, pH 6-8, shaking table 160r/min activation 24 hours; Press the 1-10% inoculum size and enlarge fermentation culture, culture temperature is 25-35 ℃, and shaking speed is 160r/min, ferments 12-16 hour.
3. the purposes of the described efficient microphenomenon of degradation of aniline of claim 1: be applied to that the high density aniline waste water is handled or the emergency processing of aniline burst pollution incident.
4. the using method of the described efficient microphenomenon of degradation of aniline of claim 1 is: when handling the high density aniline waste water, in dry cell weight, add the AN-P1 microbial inoculum in the aniline waste water biological treatment system that has moved by inoculum size 0.1 ‰～0.3 ‰, adjust hydraulic detention time 18～32 hours; During emergency processing aniline burst pollution incident, in dry cell weight, insert the AN-P1 microbial inoculum by inoculum size 0.1 ‰～0.3 ‰ in treatment system, aeration is kept dissolved oxygen 2-6mg/L in the treatment system, adjusts hydraulic detention time 10～32 hours.
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102168037A (en) * | 2010-12-03 | 2011-08-31 | 中国地质大学（武汉） | Application of strain No. 5-1 capable of degrading aniline |
| CN102864107A (en) * | 2012-09-26 | 2013-01-09 | 朱文霞 | Microbial preparation for rehabilitating phenyl amine polluted water body |
| CN105861375A (en) * | 2016-04-29 | 2016-08-17 | 广东工业大学 | Aniline-degrading bacterium and application thereof |
| CN110423709A (en) * | 2019-07-12 | 2019-11-08 | 武汉大学 | One plant height concentration phenol, microphenomenon of degradation of aniline and its application |
| CN110564716A (en) * | 2019-08-05 | 2019-12-13 | 武汉大学 | Bacterium-carrying composite microsphere for synchronously removing phenol and aniline, and preparation method and application thereof |
| CN111908620A (en) * | 2020-07-17 | 2020-11-10 | 中国科学院成都生物研究所 | Method for enhancing degradation of aniline by rhodococcus through environmental stress |
| CN112175878A (en) * | 2020-10-19 | 2021-01-05 | 江苏蓝必盛化工环保股份有限公司 | Aniline efficient degradation microbial inoculum, preparation method thereof and application thereof in chemical wastewater |
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| CN101235358B (en) * | 2008-03-05 | 2010-06-09 | 广东省微生物研究所 | Sludge sinorhizobium with aniline degradation capability and application thereof |
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Cited By (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102168037A (en) * | 2010-12-03 | 2011-08-31 | 中国地质大学（武汉） | Application of strain No. 5-1 capable of degrading aniline |
| CN102168037B (en) * | 2010-12-03 | 2012-12-19 | 中国地质大学（武汉） | Application of strain No. 5-1 capable of degrading aniline |
| CN102864107A (en) * | 2012-09-26 | 2013-01-09 | 朱文霞 | Microbial preparation for rehabilitating phenyl amine polluted water body |
| CN102864107B (en) * | 2012-09-26 | 2014-08-13 | 朱文霞 | Microbial preparation for rehabilitating phenyl amine polluted water body |
| CN105861375A (en) * | 2016-04-29 | 2016-08-17 | 广东工业大学 | Aniline-degrading bacterium and application thereof |
| CN105861375B (en) * | 2016-04-29 | 2019-08-09 | 广东工业大学 | A kind of microphenomenon of degradation of aniline and application thereof |
| CN110423709A (en) * | 2019-07-12 | 2019-11-08 | 武汉大学 | One plant height concentration phenol, microphenomenon of degradation of aniline and its application |
| CN110423709B (en) * | 2019-07-12 | 2021-01-01 | 武汉大学 | High-concentration phenol and aniline degrading bacterium and application thereof |
| CN110564716A (en) * | 2019-08-05 | 2019-12-13 | 武汉大学 | Bacterium-carrying composite microsphere for synchronously removing phenol and aniline, and preparation method and application thereof |
| CN110564716B (en) * | 2019-08-05 | 2021-07-20 | 武汉大学 | Bacterium-carrying composite microsphere for synchronously removing phenol and aniline, and preparation method and application thereof |
| CN111908620A (en) * | 2020-07-17 | 2020-11-10 | 中国科学院成都生物研究所 | Method for enhancing degradation of aniline by rhodococcus through environmental stress |
| CN112175878A (en) * | 2020-10-19 | 2021-01-05 | 江苏蓝必盛化工环保股份有限公司 | Aniline efficient degradation microbial inoculum, preparation method thereof and application thereof in chemical wastewater |
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