CN100475950C - Bacillus megaterium and use thereof - Google Patents
Bacillus megaterium and use thereof Download PDFInfo
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- CN100475950C CN100475950C CNB2006100466420A CN200610046642A CN100475950C CN 100475950 C CN100475950 C CN 100475950C CN B2006100466420 A CNB2006100466420 A CN B2006100466420A CN 200610046642 A CN200610046642 A CN 200610046642A CN 100475950 C CN100475950 C CN 100475950C
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- toluene
- megaterium
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- bacillus megaterium
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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
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/20—Air quality improvement or preservation, e.g. vehicle emission control or emission reduction by using catalytic converters
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- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The Bacillus megaterium J1 is prepared by separating out from active sludge in sewage disposal tank of petrochemical plant, purifying and culturing. The preservative number is CGMCC No. 1487. It has excellent efficient degradable performance for toluene and can be used to dispose waste water and waste gas containing toluene, polluted water and soil.
Description
Technical field
The present invention relates to a strain bacillus megaterium and the application in administering toluene concentration thereof.
Background technology
Oil and various oil fuel excessive spills and the seepage of leakage and underground oil storage facility causes underground water to suffer the pollution of aromatic hydrocarbon, and be very serious in the world.Wherein mononuclear aromatics such as benzene and toluene pollutent is paid close attention to by people most, they all are water-soluble best compounds in oil and the various oil fuel, has constituted organic pollutant the most common in the underground water.And benzene and toluene etc. belong to volatile organic compounds, have become a big class atmospheric polluting material that is only second to particulate pollutant, and it is very harmful to atmospheric seriously polluted.Along with developing rapidly of organic synthesis industry and petrochemical industry, the use of this class organic compound is more and more, especially the consumption of toluene increases day by day, not only human body is had hormesis, and the human central nervous system is had the acute hazard effect.Its volatilization gas diffuses in the face of land, current and the settling after entering atmospheric layer, brings very big harm for human body and environment.Thereby improvement contains the organic exhaust gas of toluene and the underground water and the soil of toluene concentration has become an instant job.
Administering in the organic contamination, Production by Catalytic Combustion Process and extraction facture are generally adopted in the past, but these methods are not only consuming time but also expense is extremely expensive.The biological treating technology can be utilized the microbiological deterioration organic pollutant in the physical environment, and can implement original position and administer, and relative with cost cheap and generally accepted with its technical feasibility by people, have development prospect widely.At aspects such as waste gas, waste water, waste residue and soil pollutions, biological treating is being brought into play more and more important effect.
The biological treating technology has obtained significant progress through the development of last a century in environmental improvement, developed many new technologies, and wherein biological reinforcing technology is a hot technology.Biological reinforcing technology results from the mid-1970s in last century, has been able to extensive studies and application since the eighties.This technology is exactly in order to improve the processing power of biological treatment system, and in this system, add from the dominant bacteria of occurring in nature screening or the high-efficiency strain that produces by assortment of genes technology, to remove a certain or a certain class objectionable impurities, use of the reaction of enhanced biological amount to a certain specific environment or specific pollutant.The key condition of implementing biological reinforcing technology is exactly the efficient degrading bacteria that obtains at target contaminant.Therefore, but around screening some more valuable specificity ground degraded toluene and similar organic bacterial strains thereof, and be applied to always be the problem that this area need solve in the improvement of toluene concentration.
CN1450163A discloses warm pseudomonas in the having a liking for of strain degraded toluene, can be only carbon source with toluene, is applicable to the processing of lower concentration toluene waste gas.JP2000224994, US6,395,539 and US6,551,814 all disclose a kind of gene segment of the degraded toluene of encoding, and can be converted into the toluene of degrading in the intestinal bacteria by this gene segment.UP5,079,166 has disclosed a kind of pseudomonas and the gene segment of the toluene list of the toluene of can degrading (adding) oxygenase, can be converted into the toluene of degrading in the intestinal bacteria by this gene segment." China Environmental Science " 1999.19 (6): mention among the 505-509, under intensified anti-nitrated condition, can utilize denitrifying bacterium degraded benzene and toluene.ES2085239 discloses a strain pseudomonas (CECT 4501) and has been used for the biodegradable method of aromatic hydrocarbons.This pseudomonas can be only carbon source with toluene, dimethylbenzene and ethylbenzene, thereby with it removing.This microorganism can be used for removing toluene, dimethylbenzene and the ethylbenzene etc. in trade effluent, contaminated water, soil and the storage tank clout.
According to the record of prior art, existing microorganism is limited to the degradation capability of toluene, and is only applicable to lower concentration toluene, and is not suitable for high density toluene.
Summary of the invention
The purpose of this invention is to provide bacterial isolates of strain degraded toluene and uses thereof.
Bacterial strain provided by the invention is bacillus megaterium (Bacillus megaterium) J1, be designated hereinafter simply as B.megaterium J1, be preserved in China Committee for Culture Collection of Microorganisms common micro-organisms center (being called for short CGMCC) on October 10th, 2005, deposit number is CGMCC No.1487.
The morphological specificity of this B.megaterium J1 bacterial strain is: Gram-positive, and shaft-like; Cell dia＞1 μ m forms gemma, and gemma does not expand, and is non-circular.
The physiological and biochemical property of this B.megaterium J1 bacterial strain sees Table 1:
The physiological and biochemical property of table 1 B.megaterium J1 bacterial strain
| The experimental project result | The experimental project result |
| Gram-positive | Utilize carbohydrate to produce acid |
| Cell shape is shaft-like | Glucose+ |
| Cell dia＞1 μ m+ | Wood sugar- |
| The formation gemma+ | L-arabinose- |
| Gemma expands- | N.F,USP MANNITOL+ |
| The gemma circle- | Utilize the glucose aerogenesis- |
| Catalase+ | Utilize Citrate trianion+ |
| Oxydase- | Nitrate reduction+ |
| Anaerobic growth- | 50 ℃ of growths- |
| The VP experiment- | The PH5.7 growth+ |
| VP＜PH6 + | The 7%NaCl growth+ |
| VP＞PH7 - | The starch hydrolysis+ |
| The methyl red experiment+ | Gelatine liquefication+ |
| The decomposition casein+ |
B.megaterium J1 bacterial strain of the present invention is different with the bacterial strain of other degraded toluene, compares with Pseudomonas sp CGMCC No.0850 with Bacillusbataviensis PBA CGMCC No.1092, and sibship is far away, is not the microorganism of same genus; Compare with B.megaterium No1 CGMCC No.0674 bacterial strain, though belong to of the same race, but the purposes of the two is completely different, B.megaterium No1 bacterial strain is used for tobacco processing course, with the pungency and the assorted gas of reduction cigarette, and B.megaterium J1 bacterial strain of the present invention is used for the biological treating process of toluene concentration.
B.megaterium J1 bacterial strain of the present invention can be simple inorganics (carbonic acid gas, water) and cell integral part through Degradation and Transformation with sole carbon source and the energy of toluene as growth and breeding.Bacterial strain of the present invention can tolerate the toluene of high density, and toluene is had efficient single-minded degradation property, thereby can be widely used in the biological treating of toluene concentration, especially is fit to handle the high density pollution that causes as sudden situations such as leakages.The B.megateriumJ1 bacterial strain can be strengthened general biological treatment, improves processing efficiency, saves the treatment time.
Biomaterial preservation explanation
B.megateriumJ1 bacterial strain provided by the invention is preserved in China Committee for Culture Collection of Microorganisms common micro-organisms center (CGMCC); Address: No. 13, North No.1 Row, Zhongguancun, Haidian District, Beijing City Institute of Microorganism, Academia Sinica; Deposit number: CGMCC No.1487; Preservation date: on October 10th, 2005.
Embodiment
Specify the screening process and the effect of microorganism strains of the present invention below.
The separation and the cultivation of embodiment 1, megaterium J1 bacterial strain
1, the separation of bacterial strain
Gather mud sample in sewage work of Fushun No.3 Petroleum Factory.Get 1 sludge water solution and add in the screening culture medium, add 0.1g toluene again, smear evenly, be placed in the incubator, cultivated 2 days for 30 ℃ with spreader; With transfering loop the bacterial classification that filters out is forwarded in the LB substratum then, places incubator to cultivate 1 day for 30 ℃, the gained bacterial classification is B.megaterium J1 bacterial classification.
Wherein the component of screening culture medium is:
(NH
4)
2SO
4 5g； K
2HPO
4 1g；
KH
2PO
4 4g； NaCl 2g；
MgSO
47H
2O 1g; Agar 15g;
Water 1000mL; PH 7.0.
2, the cultivation of B.megaterium J1 bacterial strain
With transfering loop the bacterial classification that filters out is connected to and shakes in the bottle, liquid amount is 50mL, cultivates in the shaking culture case.Used substratum is that LB substratum or other natural medium all can; Culture condition is: 30 ℃ of temperature, and rotating speed 150 commentaries on classics/min, incubation time 22h is when cell growth reaches logarithmic growth latter stage, through centrifugal or filter and can obtain microbial inoculum.The component of wherein said LB substratum is:
Yeast extract paste extract 5g; Peptone 10g;
NaCl 10g; Agar 15g;
Water 1000mL; PH 7.0.
B.megaterium J1 bacterial strain of the present invention can descend at aerobic condition and separate toluene, therefore can be applicable to contain waste water, waste gas, the water of toluene concentration and the biological treating process of soil of toluene.
Embodiment 2
The cultivation of B.megaterium J1 bacterial strain is identical with embodiment 1.Get 2 and shake bottle, add 50mL sterilized water and 1.5mL toluene respectively.Get one of them and shake bottle as the blank group, another shakes bottle and adds cultured J1 microbial inoculum, and making its OD value is 5.0.Shake bottle with two and be placed on oscillatory reaction on the shaking table, the shaking table condition is: 30 ℃ of temperature, rotating speed 150 commentaries on classics/min.Sampling analysis behind the 10h the results are shown in Table 2.
Table 2B.megaterium J1 handles the result who contains toluene sewage
Embodiment 3
The cultivation of B.megateriumJ1 bacterial strain is identical with embodiment 1.Get 2 and shake bottle, add the 50mL sterilized water respectively, add 4g again by the soil of 1.5mL toluene concentration.Get one of them and shake bottle as the blank group; Another shakes bottle and adds cultured B.megaterium J1 microbial inoculum, and making its OD value is 5.0.Shake bottle with 2 and be placed on oscillatory reaction on the shaking table, the shaking table condition is: 30 ℃ of temperature; Rotating speed 150 commentaries on classics/min.Sampling analysis behind the 10h the results are shown in table 3.
Table 3B.megaterium J1 handles the result of toluene concentration soil
Embodiment 4
The cultivation of B.megateriumJ1 bacterial strain is identical with embodiment 1.Get 1 and shake bottle, dress 1L sterilized water adds the B.megateriumJ1 microbial inoculum again, and making its OD value is 5.0.Add about 150mL slag toward shaking in the bottle, will shake and bottle be placed on the shaking table about shaking cultured cell 4h.The shaking table condition is: 30 ℃ of temperature; Rotating speed 150 commentaries on classics/min.Take out slag after treating on the slag to generate the mycoderm of capacity, (among the φ 2cm * H50cm), be that 30 ℃, inlet toluene concentration are 100.0mg/L in temperature, airshed is under the operational condition of 80L/h to the biological packing tower of packing into, and the average degradation rate of toluene is 95.54%.
Claims (2)
1, a strain bacillus megaterium (Bacillus megaterium) J1, CGMCC No.1487.
2, the described bacterial strain of claim 1 contains application in toluene waste water, waste gas and the soil in improvement.
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|---|---|---|---|
| CNB2006100466420A CN100475950C (en) | 2006-05-19 | 2006-05-19 | Bacillus megaterium and use thereof |
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|---|---|---|---|
| CNB2006100466420A CN100475950C (en) | 2006-05-19 | 2006-05-19 | Bacillus megaterium and use thereof |
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| CN100475950C true CN100475950C (en) | 2009-04-08 |
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Families Citing this family (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101921714B (en) * | 2009-09-10 | 2011-11-09 | 浙江师范大学 | Bacillus megaterium Bm-10 strain for highly producing poly-beta-hydroxybutyrate as well as screening method and application thereof |
| CN102453678A (en) * | 2010-10-26 | 2012-05-16 | 中国科学院生态环境研究中心 | Microorganism composite bacterial agent for restoration of saline alkali soil polluted by petroleum |
| CN102465103B (en) * | 2010-11-04 | 2013-04-10 | 中国石油化工股份有限公司 | Aerobic denitrification methylobacterium phyllosphaerae and application thereof |
| CN107118973B (en) * | 2017-01-20 | 2019-10-22 | 厦门欧米克生物科技有限公司 | A method of utilizing micro-organisms parahydroxyben-zaldehyde |
| CN108504585B (en) * | 2017-08-17 | 2021-05-14 | 北京沃太斯环保科技发展有限公司 | Benzene degrading bacterium for treating atmospheric pollution and preparation method and application thereof |
| CN107699513B (en) * | 2017-09-13 | 2020-08-21 | 博元生态修复（北京）有限公司 | Black and odorous water body degrading bacterium and application thereof |
| CN109182211B (en) * | 2018-10-16 | 2021-09-07 | 中国石油化工股份有限公司 | Composite biological agent for preventing and treating SRB in wide-temperature environment and use method |
| CN113025535B (en) * | 2021-04-26 | 2023-06-13 | 广东工业大学 | Bacillus GDUTAN11 and application thereof in degradation of benzene series |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6057268A (en) * | 1997-12-12 | 2000-05-02 | Organica, Inc. | Solid form compositions for treating natural bodies of water |
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Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6057268A (en) * | 1997-12-12 | 2000-05-02 | Organica, Inc. | Solid form compositions for treating natural bodies of water |
Non-Patent Citations (2)
| Title |
|---|
| 生物过滤塔甲苯降解性能研究. 李国文等.环境科学，第22卷第2期. 2001 |
| 生物过滤塔甲苯降解性能研究. 李国文等.环境科学，第22卷第2期. 2001 * |
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