CN103087948A - Bacillus megatherium capable of degrading polycyclic aromatic hydrocarbon and application thereof - Google Patents
Bacillus megatherium capable of degrading polycyclic aromatic hydrocarbon and application thereof Download PDFInfo
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- CN103087948A CN103087948A CN2013100004898A CN201310000489A CN103087948A CN 103087948 A CN103087948 A CN 103087948A CN 2013100004898 A CN2013100004898 A CN 2013100004898A CN 201310000489 A CN201310000489 A CN 201310000489A CN 103087948 A CN103087948 A CN 103087948A
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- polycyclic aromatic
- aromatic hydrocarbons
- aromatic hydrocarbon
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Abstract
The invention discloses bacillus megatherium capable of degrading polycyclic aromatic hydrocarbon and application thereof. The bacillus megatherium has the collection number of CGMCC 6900 and is preserved in China General Microbiological Culture Collection Center. The bacillus megatherium has high long-term tolerance and degradation capacity in high-concentration polycyclic aromatic hydrocarbon, and the defects of low environmental tolerance in actual lab strain application at present can be solved.
Description
Technical field
The invention belongs to field of environment protection, in particular to a kind of bacillus megaterium and application thereof that can be used in environment protection.
Background technology
Polycyclic aromatic hydrocarbons (PAHs) class organic compound accounts for more than 1/3rd in more than 1000 kinds of environmental carcinogens, and the polycyclic aromatic hydrocarbons in environment is mainly derived from coal combustion, the waste water in petroleum cracking and industrial production.Because natural degradation speed is extremely slow, the semi-invariant of polycyclic aromatic hydrocarbons in environment continues to increase with industrial development, brings great threat for national health.In recent years due to polycyclic aromatic hydrocarbons cause carcinogenic, the teratogenesis case is of common occurrence, the degraded of polycyclic aromatic hydrocarbons be treated as the important issue of current environmental strategies.The characteristic of microbiological deterioration and high-cleanness efficient with it becomes the best approach and the research emphasis of generally acknowledged at present removal Polycyclic Aromatic Hydrocarbonat Existing in Environment.The microbiological deterioration research on mechanism not only helps to improve the degradation efficiency of single bacterium, also can be the difference configuration composite flora according to the microbiological deterioration approach, and carrying out the polycyclic aromatic hydrocarbons combined degradation provides theoretical foundation.On the other hand, the bio-transformation potentiality that the plurality of enzymes that exists in the degrading polycyclic aromatic hydrocarbons approach (as single, double oxygenase etc.) has are also the focuses of recently paying close attention to, understanding to enzyme in degradation pathway is also will provide thinking for utilizing such bacterial strain to carry out bio-transformation.
The research work of polycyclic aromatic hydrocarbon-degrading bacteria at present mainly concentrates on efficient degrading bacteria, mechanism of degradation, degrading genes and degraded influence factor aspect.Luxuriant and rich with fragrance have minimum carinogenicity structural unit--He Wan district, K district for a kind of thrcylic aromatic hydrocarbon is modal a kind of multi-ring aromatic hydrocarbons pollutent in Soil sediment, often is selected as the metabolism substrate of sole carbon source in such research.The PAHs bacterium for degrading that is separated in recent years mainly comprises Rhodopseudomonas, bacillus, mycobacterium, Sphingomonas, genus arthrobacter, Burkholderia genus and Flavobacterium etc.Wherein the degradation pathway of Rhodopseudomonas, mycobacterium, Sphingomonas, genus arthrobacter all is well studied.
Summary of the invention
The object of the invention is to solve problems of the prior art, providing a kind of can have the bacterial strain of long-term tolerance and degradation capability in the high density polycyclic aromatic hydrocarbons, can solve the present laboratory strains practical application aspect drawback poor to environmental resistance.And experiment is also carried out multiple sieve with the organophosphorus pesticide of phosphorous chlorine, is the ability that bacterial strain has had two kinds of organic pollutants of metabolism simultaneously.
The present invention relates to a kind of genus bacillus, it is characterized in that preserving number is CGMCC 6900, be preserved in China Committee for Culture Collection of Microorganisms common micro-organisms culture presevation administrative center.
The present invention also relates to the effect of above-mentioned genus bacillus in degrading polycyclic aromatic hydrocarbons on the other hand, and described polycyclic aromatic hydrocarbons includes but not limited to the polycyclic aromatic hydrocarbons that 2-10 phenyl ring forms; Preferably, described polycyclic aromatic hydrocarbons contains phenanthrene.
In another preferred embodiment of the present invention, described genus bacillus can contain organophosphorus pesticide OP-DDT, PP-DDE, PP-DDD, PP-DDT, at least a in parathion-methyl and thiophos, two kinds or three kinds or whole situations are carried out degrading polycyclic aromatic hydrocarbons.
Microorganism information
Bacillus megaterium of the present invention, Classification And Nomenclature is bacillus megaterium (Bacillus megaterium), be preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, preservation address: Datun Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica, preserving number is CGMCC 6900, and preservation date is on November 30th, 2012.
Embodiment
Extract bacterial strain and cultivate from river silt, the continuous domestication method take the high density Phenanthrene as sole carbon source, domestication continues one month; With six kinds of different organophosphorus pesticide OP-DDT, PP-DDE, PP-DDD, PP-DDT, parathion-methyl and thiophos carry out multiple sieve, select the bacterial strain that organophosphorus pesticide is had height endurability, the bacterial strain that filters out is preserved in China Committee for Culture Collection of Microorganisms common micro-organisms culture presevation administrative center, and deposit number is CGMCC 6900.
Luxuriant and rich with fragrance degradation experiment
Experiment material
1) phenanthrene (Phenanthrene) is available from Fisher company, and purity is 98%; Mud; Mix dried bacterium;
2) HDM substratum: KH
2PO
41.000 g/L, K
2HPO
41.000 g/L, NH
4NO
32.000 g/L, MgSO
47H
2O 0.300 g/L, CaCl
22H
2O 0.001 g/L, FeSO
47H
2O 0.001 g/L, 1.0ml micronutrient element Solution H
3BO
32.90 g/L, MnCl
24H
2O 1.80 g/L, ZnSO
47H
2O 0.20 g/L, Na
2MoO
42H
2O 0.40 g/L, CuSO
45H
2O 0.08 g/L, Co (NO
3)
26H
2O 0.05 g/L;
3) minimal medium: 4g NaNO
3, 1.5g KH
2PO4,0.5g Na
2HPO
4, 0.0011g FeSO
47H
2O, 0.2g MgSO
47H
2O, 0.01g CaCl
2, add water to 1000 mL, 121 ℃ of autoclaving 30 min;
4) TY substratum: glucose 10.0 g, peptone 5.0g, yeast extract paste 5.0 g, NaCl 5.0 g, inorganic salt solution 2.0 mL add water to 1000 mL and transfer 7.5,115 ℃ of sterilization 20 min of pH.
Experimental implementation
1) cone mouth bottle 250ml, sterilization.121 ℃ of conditions, 30min;
2) HDM nutrient solution 2L, 121 ℃, sterilization 30min, standby;
3) 100ml graduated cylinder, 121 ℃, sterilization 30min, standby;
4) phenanthrene, 5g/L.Solvent is normal hexane.Filtration sterilization;
5) in aseptic operating platform, with 2) in the luxuriant and rich with fragrance solution handled well add in sterilized triangular flask, every bottle of 2ml.With aseptic sealed membrane sealing.Volatilization 18h is to the phenanthrene appearance that crystallization is arranged;
6) with 3) in the went out graduated cylinder amount 2 of bacterium) in the HDM nutrient solution, add 5) in contain in luxuriant and rich with fragrance cone mouth bottle, each 100ml.Make the luxuriant and rich with fragrance 100mg/L of arrival;
7) TY liquid nutrient medium, 100ml.115 ℃, sterilization, 30min.Standby;
8) experimental strain adds 7) nutrient solution in, 30 ℃, 200r/min cultivates 16h;
9) get 8) middle 50ml liquid, centrifugal;
10) thalline 9) is assigned to OD=1 with sterilized water;
11) with 10) in solution add 6) in the cone mouth bottle in, each 6ml.Cultivate respectively 8day; 15day; 30day.Condition: 30 ℃, 200r/min;
12) extracted organic phase and dewatering uses high performance liquid phase to measure luxuriant and rich with fragrance content.
The organophosphorus pesticide tolerance:The bacterial strain that screens take 20 μ g/L organophosphorus pesticides in the inorganic salt of sole carbon source, fermented 7 days, bacterial strain still can be bred, and has good tolerance.
This shows, the bacterial strain that filters out can be grown in the inorganic salt solution of high density phenanthrene well, and has the ability of degrading polycyclic aromatic hydrocarbons; The bacterial strain that filters out has good tolerance in the inorganic salt solution of high density organophosphorus pesticide.
The above be only the specific embodiment of the present invention, but protection scope of the present invention is not limited to this, and any variation or replacement of expecting without creative work is within all should being encompassed in protection scope of the present invention.Therefore, protection scope of the present invention should be as the criterion with the protection domain that claims were limited.
Claims (3)
1. a genus bacillus, is characterized in that preserving number is CGMCC 6900, is preserved in China Committee for Culture Collection of Microorganisms common micro-organisms culture presevation administrative center.
2. the effect of genus bacillus claimed in claim 1 in degrading polycyclic aromatic hydrocarbons, preferred, described polycyclic aromatic hydrocarbons contains phenanthrene.
3. application according to claim 2, described genus bacillus can contain organophosphorus pesticide OP-DDT, PP-DDE, PP-DDD, PP-DDT, at least a in parathion-methyl and thiophos, two kinds or three kinds or whole situations are carried out degrading polycyclic aromatic hydrocarbons.
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| CN201310000489.8A CN103087948B (en) | 2013-01-04 | 2013-01-04 | Bacillus megatherium capable of degrading polycyclic aromatic hydrocarbon and application thereof |
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| CN103087948B CN103087948B (en) | 2014-12-03 |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105018458A (en) * | 2015-07-31 | 2015-11-04 | 湖南大学 | Compound microbial agent as well as preparation method and application thereof |
| CN106698678A (en) * | 2017-01-13 | 2017-05-24 | 江苏大学 | Application of Bacillus in degrading aromatic compounds |
| CN107937321A (en) * | 2018-01-09 | 2018-04-20 | 南京工业大学 | A kind of bacillus subtilis and its application in the luxuriant and rich with fragrance degraded of polycyclic aromatic hydrocarbon pollutant |
| CN113336334A (en) * | 2021-05-31 | 2021-09-03 | 辽宁大学 | Bacillus firmus and application thereof in degrading naphthalene, anthracene, phenanthrene and fluorene |
-
2013
- 2013-01-04 CN CN201310000489.8A patent/CN103087948B/en not_active Expired - Fee Related
Non-Patent Citations (4)
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| 李丽等: "枯草芽孢杆菌对多环芳烃的降解能力研究", 《河北农业科学》 * |
| 李晶等: "PhoR / Pho P双因子调控系统在枯草芽孢杆菌NCD-2菌株解磷能力中的功能分析", 《中国农业科技导报》 * |
| 王海峰等: "一株枯草芽孢杆菌分离鉴定及其降解稠油特性", 《深圳大学学报理工版》 * |
| 郑传进等: "巨大芽孢杆菌解磷能力的研究", 《江西农业大学学报（自然科学版）》 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105018458A (en) * | 2015-07-31 | 2015-11-04 | 湖南大学 | Compound microbial agent as well as preparation method and application thereof |
| CN106698678A (en) * | 2017-01-13 | 2017-05-24 | 江苏大学 | Application of Bacillus in degrading aromatic compounds |
| CN107937321A (en) * | 2018-01-09 | 2018-04-20 | 南京工业大学 | A kind of bacillus subtilis and its application in the luxuriant and rich with fragrance degraded of polycyclic aromatic hydrocarbon pollutant |
| CN107937321B (en) * | 2018-01-09 | 2020-04-03 | 南京工业大学 | Bacillus subtilis and application thereof in degradation of polycyclic aromatic hydrocarbon pollutant phenanthrene |
| CN113336334A (en) * | 2021-05-31 | 2021-09-03 | 辽宁大学 | Bacillus firmus and application thereof in degrading naphthalene, anthracene, phenanthrene and fluorene |
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| Publication number | Publication date |
|---|---|
| CN103087948B (en) | 2014-12-03 |
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