CN102174446A - Bacillus pumilus strain.Bap9 capable of effectively degrading benzo(a)pyrene and application thereof - Google Patents

Bacillus pumilus strain.Bap9 capable of effectively degrading benzo(a)pyrene and application thereof Download PDF

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CN102174446A
CN102174446A CN 201110047402 CN201110047402A CN102174446A CN 102174446 A CN102174446 A CN 102174446A CN 201110047402 CN201110047402 CN 201110047402 CN 201110047402 A CN201110047402 A CN 201110047402A CN 102174446 A CN102174446 A CN 102174446A
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pyrene
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倪晋仁
朱婷婷
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Peking University
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Abstract

The invention discloses a bacterium for degrading benzo(a)pyrene and application thereof. The bacterium is bacillus pumilus strain.Bap9, CGMCC No.4585. The strain can grow with benzo(a)pyrene as the unique carbon source and energy, undergoes shake culture at the temperature of 37 DEG C for 20 days in the inorganic salt culture medium with benzo(a)pyrene concentration being 40mg/L and has degradation rate toward benzo(a)pyrene being 27.30%. The degradation rate of the strain toward benzo(a)pyrene can be effectively improved by adding defined amount of co-metabolic carbon sources, such as sodium acetate. When another polycyclic aromatic hydrocarbon phenanthrene is taken as a co-metabolic substrate and exists by being mixed with benzo(a)pyrene, the degradation rate of the strain toward benzo(a)pyrene can be improved to 53.70% and meanwhile, the strain can realize complete removal of phenanthrene. The strain provided by the invention can provide new microorganism resources for degradation of polycyclic aromatic hydrocarbons in the water environment or soil environment.

Description

But the bacillus pumilus and the application thereof of a kind of efficient degradation benzo [a] pyrene
Technical field
The invention belongs to the environmental pollutant processing technology field, be specifically related to a kind of benzo [a] pyrene efficient degrading bacteria and application thereof.
Background technology
Benzo [a] pyrene is a kind of have obvious teratogenesis, carcinogenic, mutagenic organic compound, and it is the polycyclic arene compound by a phenyl ring and a pyrene molecule be combined into.Benzo [a] pyrene mainly results from the incomplete combustion of hydrocarbon polymers such as oil, coal, timber, geseous fuel and paper and the thermolysis effect in reduction process thereof; Therefore, benzo [a] pyrene extensively is present in flue gas that burning such as coal tar, all kinds of carbon black and coal, oil produces, smoke from cigarette, the vehicle exhaust, and in the industrial sewages such as coking, oil refining, pitch, plastics.Benzo in the surface water [a] pyrene is except discharge of industrial wastes, mainly from scrubbing atmospheric rainwater, aqua storage tank and pipeline coatings leaching etc.People's long-term exposure can cause chronic poisoning in the environment that contains benzo [a] pyrene; Studies show that, the every increase by 1% of the benzo in the living environment [a] pyrene content, the mortality ratio of lung cancer promptly rises 5%.Benzo [a] pyrene (BaP) and aflatoxin and nitrosamine have been listed in the toxic organic pollutant Black List of preferential control by EPA at present, are the three big carcinogenic substances that the whole world is known as.
In recent years, along with a large amount of uses of oil and petroleum products, the benzo in the environment [a] pyrene has the trend of being on the increase.The hydrolysis and the photodissociation speed of benzo under natural environmental condition [a] pyrene are all very slow, must be caused bigger harm as not handling.With respect to chemistry and physical treatment method, characteristics such as microbial method is few owing to having a secondary pollution, and cost is low, and is simple to operate and being widely used; Its migration at pollutent transforms and even finally occupies critical role in the disappearance, is the main approach that multiring aromatic hydrocarbon substance is removed in the environment.Microorganism can finally be translated into nontoxic, harmless inorganic substance CO with the permineralization of benzo [a] pyrene 2And water, be considered to remove the best approach of benzo in the environment [a] pyrene.Yet, the bacterial species of finding with benzo [a] pyrene degradation capability is single at present, and degradation rate and metabolic activity are lower, degradation capability is not strong, therefore excavate benzo [a] the pyrene degradation bacteria that more has strong katabolism ability, and be applied to the removal process of environmental pollutant, will have great Significance for Environment.
Summary of the invention
The objective of the invention is provides a kind of benzo [a] pyrene efficient degrading bacteria and application thereof at the inefficient difficult problem of above-mentioned benzo [a] pyrene biological degradation.
Bacillus pumilus Bacillus pumilus strain.Bap9 provided by the invention is preserved in China Committee for Culture Collection of Microorganisms common micro-organisms center (being called for short CGMCC) on January 26th, 2011, and preserving number is CGMCC № 4585.
Bacterial strain Bacillus pumilus strain.Bap9 takes a sample from the soil of barbecue place, Peking University smooth spring Xin Yuan dormitory area, a strain gram positive bacterium that obtains through domestication, separation and purifying.
Morphological specificity: bacterial strain Bap9 behind the cultivation 16h, is grown to the circular bacterium colony of cheese look on the beef-protein medium under 37 ℃, colony edge is neat, and surface wettability is smooth; By behind the gramstaining at microscopically, thalline is tiny short pole shape, is close to spherically, size is 0.5~2.0um, the catalase reaction result is positive.
According to its morphological specificity and physiological and biochemical property and 16S rRNA gene order thereof, identify that this bacterial strain is a bacillus pumilus, its 16S rRNA has as the nucleotide sequence shown in sequence is represented, and sequence length is 1402bp.
TCGAGCGGACAGAAGGGAGCTTGCTCCCGGATGTTAGCGGCGGACGGGTGAGTAACA
CGTGGGTAACCTGCCTGTAAGACTGGGATAACTCCGGGAAACCGGAGCTAATACCGGA
TAGTTCCTTGAACCGCATGGTTCAAGGATGAAAGACGGTTTCGGCTGTCACTTACAGA
TGGACCCGCGGCGCATTAGCT--AGTTGGTGGGGTAATGGCTCACCAAGGCGACGATGC
GTAGCCGACCTGAGAGGGTGATCGGCCACACTGGGACTGAGACACGGCCCAGACTCC
TACG-GGAGGCAGCAGTAGGGAATCTTCCGCAATGGACGAAAGTCTGACGGAGCAAC
GCCGC-GTGAG-TGATGAAGGTTTTCGGATCGTAAAGCTCTGTTGTTAGGGAAGAACAA
GTGCGAGAGTAACTGCTCGCACCTTGACGGTACCTAACCAGAAAGCCACGGCTAACTA
CGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTTGTCCGG-AATTATTGGGCGT
AAAGGGCTCGCAGGCGGTTTCTTAAGTCTGATGTGAAAGCCCCCGGCTCAACC-GGGG
AGGGTCATTGGAAACTGGGAAACTTGAGTGCAGAAGAGGAGAGTGGAATTCCACGTG
TAGCGGTGAAATGCGTAGAGATGTGGAGGAACACCAGTGGCGAAGGCGACTCTCTGG
TCTGTAACTGACGCTGAGGAGCGAAAGCGTGGGGAGCGAACAGGATTAGATACCCTG
GTAGTCCACGCCGTAAACGATGAGTGCTAAGTGTTAGGGGGTTTCCGCCCCTTAGTGCT
GCAGCTAACGCATTAAGCACTCCGCCTGGGGAGTACGGTCGCAAGACTGAAACTCAA
AGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAACG
CGAAGAACCTTACCAGGTCTTGACATCCTCTGACAACCCTAGAGATAGGGCTTTCCCTT
CGGGGACAGAGTGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGG
GTTAAGTCCCGCAACGAGCGCAACCCTTGATCTTAGTTGCCAGCATTTAGTTGGGCACT
CTAAGGTGACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAATCATCAT
GCCCCTTATGACCTGGGCTACACACGTGCTACAATGGACAGAACAAAGGGCTGCAAGA
C-CGCAAGGTTTAGCCAATCCCATAAATCTGTTCTCAGTTCGGATCGCAGTCTGCAACT
CGACTGCGTGAAGCTGGAATCGCTAGTAATCGCGGATCAGCATGCCGCGGTGAATACG
TTCCCGGGCCTTGTACACACCGCCCGTCACACCACGAGAGTTTGCAACACCCGAAGTC
GGTGAGGTAACCTTTATGGA
This bacterium can be a growth and breeding in the environment of 0~200mg/L in benzo [a] pyrene concentration, under aerobic condition, in the minimal medium, this bacterium can be sole carbon source and energy growth and breeding with benzo [a] pyrene, and the degradation rate to benzo [a] pyrene (starting point concentration is 40mg/L) in 20 days reaches 27.3%.
This bacterial strain benzo [a] pyrene of all degrading in the wider pH value scope of neutrality or slant acidity, the pH value of the suitableeest growth and degraded is 5.5-9.5, is preferably 6.0-8.0.
The best liquid amount of this strains for degrading benzo [a] pyrene is 5mL/50mL-30mL/50mL, is preferably 5mL/50mL-15mL/50mL.
The optimum inoculation amount of strains for degrading is 5%, and best liquid amount is 5mL/50mL.
Add an amount of common metabolism substrate, can effectively promote the growth of this bacterial strain, improve the degradation rate of this bacterial strain benzo [a] pyrene.Described metabolism substrate altogether can be sucrose, maltose, glucose, sodium-acetate or Zulkovsky starch.
Above-mentioned metabolism substrate altogether is preferably sodium-acetate, and the optimal concentration scope is 40-100mg/L.
Add an amount of common metabolism substrate phenanthrene, also can promote of the degraded of this bacterial strain benzo [a] pyrene.Luxuriant and rich with fragrance optimal concentration scope of adding is 20-40mg/L.
Another purpose of the present invention is to provide a kind of microbiobacterial agent that is used for efficient degradation benzo [a] pyrene, and its activeconstituents is described bacterial strain Bacillus pumilus strain.Bap9.
Bacillus pumilus of the present invention and application thereof have following beneficial effect compared with prior art:
(1) bacterial strain provided by the invention is under the pure culture condition, and the clearance to benzo [a] pyrene in 20 days can reach 27.30%, compares with existing bacterial strain, and degradation rate improves greatly.
(2) when benzo [a] pyrene and luxuriant and rich with fragrance when existing jointly, this bacterial strain reached 53.70% to the clearance of benzo [a] pyrene in 20 days, and the clearance of phenanthrene is reached 100%.
Utilize these characteristics, bacterial strain of the present invention can be used for single high ring polycyclic aromatic hydrocarbon benzo [a] pyrene or the high ring polycyclic aromatic hydrocarbon mixture pollutent in the degradation water environment, perhaps is used for the biological restoration of edatope; This bacterial strain provides a kind of new germ plasm resource for high ring polycyclic aromatic hydrocarbon is total to metabolic mechanism research simultaneously.
Description of drawings
Fig. 1 is the growth of bacterial strain Bacilluspumilus strain.Bap9 and the degradation curve of benzo [a] pyrene
Fig. 2 is the influences of different initial pH values to bacterial strain Bacillus pumilus strain.Bap9 degraded benzo [a] pyrene.
Fig. 3 is the influence of different vaccination amount to bacterial strain Bacillus pumilus strain.Bap9 degraded benzo [a] pyrene.
Fig. 4 is the influences of different liquid amounts to bacterial strain Bacillus pumilus strain.Bap9 degraded benzo [a] pyrene.
Fig. 5 is the degradation effect of bacterial strain Bacillus pumilus strain.Bap9 to benzo [a] pyrene and luxuriant and rich with fragrance mixed system.
Embodiment
The invention will be further described below in conjunction with concrete enforcement.
Embodiment 1: the performance of the separation of bacterial strain Bacillus pumilus strain.Bap9 and degraded benzo [a] pyrene
One, the separation purification method of efficient degradation benzo [a] pyrene bacterial strain Bap9
This method has following steps:
1, take by weighing the 10g pedotheque, add in the triangular flask that 100mL domestication substratum is housed, the effective constituent of domestication substratum is: (NH 4) 2SO 41g, K 2HPO 42g, MgSO 47H 2O 0.5g, NaCl 0.1g, FeCl 30.5g, CaCl 20.5g, benzo [a] pyrene 0.04g, distilled water 1000mL, the pH value is 7.0;
2, above-mentioned triangular flask is placed 160r/min, 37 ℃ of 1 weeks of shaking table shaking culture, be transferred in the fresh domestication substratum, continue to cultivate 1 week, 3 times repeatedly by 10% inoculum size;
Draw the nutrient solution 1mL after taming repeatedly, picking feature difference, the vigorous and stable bacterium colony of growth from the flat board, line separates to obtain pure strain repeatedly on the beef extract-peptone solid plate, the pure strain of separation and purification gained is inoculated on benzo [a] the pyrene inorganic salt solid plate, switching is 3 times continuously, chooses the fastest bacterial strain Bap9 of the speed of growth as the research bacterial strain.And this bacterial strain is preserved in China Committee for Culture Collection of Microorganisms common micro-organisms center (be called for short CGMCC) on January 26th, 2011, preserving number is CGMCC № 4585.
Two, efficient degrading bacterial strain Bap9 is to the degradation property of benzo [a] pyrene
Above-mentioned bacterial strains Bap9 is seeded to the inorganic salt nutrient solution that contains benzo [a] pyrene 40mg/L., and (composition of this substratum is: (NH 4) 2SO 41g, K 2HPO 42g, MgSO 47H 2O 0.5g, NaCl 0.1g, FeCl 30.5g, CaCl 20.5g, benzo [a] pyrene 0.04g, distilled water 1000mL, the pH value is 7.0) in, lucifuge shaking culture in 160r/min, 37 ℃ of shaking tables, in the 0th, 4,8,12,16,20 day the same time residual quantity of nectar degree and benzo [a] pyrene in the sampling and measuring nutrient solution respectively, experimental result is seen Fig. 1.As seen from Figure 1, bacterial strain is 4-12d to the quick degradative phase of benzo [a] pyrene, to the later stage (12-20d) of degraded, bacterial strain tends towards stability to the degradation curve of benzo [a] pyrene, during by the 20th day, the OD value of bacterial strain reaches 0.223, and the degradation rate of benzo [a] pyrene reaches 27.30%.
The present embodiment explanation separates the resulting degradation bacteria of domestication and can utilize benzo [a] pyrene to carry out growth and breeding as the sole carbon source and the energy, and has the ability of efficient degradation benzo [a] pyrene.
Embodiment 2: different initial pH values are to the growth of bacterial strain Bacillus pumilus strain.Bap9 and the influence of degraded benzo [a] pyrene
(composition of this substratum is: (NH to regulate minimal medium 4) 2SO 41g, K 2HPO 42g, MgSO 47H 2O 0.5g, NaCl 0.1g, FeCl 30.5g, CaCl 20.5g, benzo [a] pyrene 0.04g, distilled water 1000mL) the pH value be respectively 5.5,6.0,7.0,8.0,9.0,9.5, observe the growing state of bacterial strain Bap9 and to the degradation property of benzo [a] pyrene.Benzo [a] pyrene starting point concentration is 40mg/L, and inoculum size is 5% (V/V), and shaking culture is 7 days in 160r/min, the 37 ℃ of shaking tables, measures the residual rate of bacterium OD value and benzo [a] pyrene, result such as Fig. 2.
As seen from Figure 2, be 8.0 o'clock at pH, degradation bacteria is the strongest to the Degradation of benzo [a] pyrene, and degradation rate is 10.92% after 7 days, is that 5.5 o'clock degradation rates are subjected to bigger inhibition at pH, only is 0.74%.The nectar kilsyth basalt of bacterial strain under different pH values is bright, and the pH value is consistent with degradation rate to the influence of microorganism growth.In weakly alkaline environment, bacterial strain can both be degraded to benzo [a] pyrene, for it provides assurance in different pH environmental applications in acidity.
Embodiment 3: the different vaccination amount is to the growth of bacterial strain Bacillus pumilus strain.Bap9 and the influence of degraded benzo [a] pyrene
(composition of this substratum is: (NH to be respectively 0.5%, 1.0%, 2.0%, 5.0% and 10.0% minimal medium in inoculum size 4) 2SO 41g, K 2HPO 42g, MgSO 47H 2O 0.5g, NaCl 0.1g, FeCl 30.5g, CaCl 20.5g, benzo [a] pyrene 0.04g, distilled water 1000mL, the pH value is 7.0) in, the residual rate of benzo [a] pyrene is respectively 98.31%, 94.26%, 91.75%, 89.94% and 92.86% (see figure 3) after 7 days.As shown in Figure 3, in the certain limit, inoculum size is big more, and the nectar degree is high more, and the degradation rate of bacterial strain is more and more higher in the certain limit, but inoculum size is not to be the bigger the better, and when inoculum size is 10.0%, is that 5.0% o'clock degradation rate is low than inoculum size.
Embodiment 4: different liquid amounts are to the growth of bacterial strain Bacillus pumilus strain.Bap9 and the influence of degraded benzo [a] pyrene
(composition of this substratum is: (NH to add the inorganic salt nutrient solution with different liquid amount (5ml/50ml, 10ml/50ml, 15ml/50ml, 20ml/50ml, 25ml/50ml, 30ml/50ml) in triangular flask 4) 2SO 41g, K 2HPO 42g, MgSO 47H 2O 0.5g, NaCl 0.1g, FeCl 30.5g, CaCl 20.5g, benzo [a] pyrene 0.04g, distilled water 1000mL, the pH value is 7.0), benzo [a] pyrene starting point concentration is 40mg/L, inoculum size is 5% (V/V), shaking culture is 7 days in 160r/min, 37 ℃ of shaking tables, and the residual rate of benzo [a] pyrene is respectively 87.63%, 89.94%, 91.24%, 94.06%, 97.95%, 99.14% (see figure 4), the height that dissolved oxygen concentration in the substratum is described directly influences microbial growth and metabolism, liquid amount is few more, and the nectar degree is high more, and dissolved oxygen amount is big more, help this bacterial strain and produce active substance, so degradation rate is high more.
Embodiment 5: add carbon source to the growth of bacterial strain Bacillus pumilus strain.Bap9 and the influence of degraded benzo [a] pyrene
(composition of this substratum is: (NH to the minimal medium that contains benzo [a] pyrene 40mg/L with inoculation 4) 2SO 41g, K 2HPO 42g, MgSO 47H 2O 0.5g, NaCl 0.1g, FeCl 30.5g, CaCl 20.5g, benzo [a] pyrene 0.04g, distilled water 1000mL, the pH value is 7.0) in, in nutrient solution, add sucrose, glucose, maltose, sodium-acetate and the Zulkovsky starch conduct that final concentration is 40mg/L respectively and be total to metabolism substrate, shaking culture is 7 days in 160r/min, 37 ℃ of shaking tables, measures nectar degree and benzo [a] pyrene residual quantity in the nutrient solution, calculates degradation rate.Table 1 is the influences of the different carbon sources of 40mg/L to strain cell growth and the degraded of benzo [a] pyrene for adding final concentration.As shown in Table 1, degradation rate is the highest when adopting sodium-acetate as carbon source, adding sucrose, glucose and maltose has restraining effect to strains for degrading benzo [a] pyrene, and the effect of adding Zulkovsky starch is least obvious, as seen selects suitable carbon source most important to the degradation rate that improves bacterial strain.
Table 1 adds the influence of carbon source to bacterial strain Bacillus pumilus strain.Bap9 growth and benzo [a] pyrene degradation rate
Inoculation is respectively 0mg/L, 10mg/L, 40mg/L, 100mg/L, 500mg/L to sodium-acetate concentration, and benzo [a] pyrene concentration is that (composition of this substratum is: (NH for the minimal medium of 40mg/L 4) 2SO 41g, K 2HPO 42g, MgSO 47H 2O 0.5g, NaCl 0.1g, FeCl 30.5g, CaCl 20.5g benzo [a] pyrene 0.04g, distilled water 1000mL, the pH value is 7.0) in, shaking culture was measured nectar degree and benzo [a] pyrene residual quantity in the nutrient solution after 20 days in 160r/min, 37 ℃ of shaking tables, calculated degradation rate, and the result is as shown in table 2.
Table 2 adds the sodium-acetate of different concns to the influence of strain cell growth with the degraded of benzo [a] pyrene
Figure BSA00000440933600062
Figure BSA00000440933600071
As shown in Table 2, when sodium-acetate concentration was 10mg/L, the degraded of benzo [a] pyrene was than the added-time has not improved 4.95%; When the concentration of sodium-acetate was 40mg/L, the degradation rate of benzo [a] pyrene had improved 8.77%; When the concentration of sodium-acetate was increased to 500mg/L, the degradation rate of benzo [a] pyrene sharply dropped to 13.12%.The sodium-acetate that has added 10-40mg/L can promote thalli growth, improves the degradation rate of bacterial strain to pyrene, adds excessive sodium-acetate, and bacterial strain utilizes quick-acting carbon sources in a large number, and its utilization to benzo [a] pyrene has been subjected to inhibition.
An amount of carbon source is added in the explanation of this example can promote the growth of bacterial strain, and improves its degradation rate to benzo [a] pyrene.
Embodiment 5: add being total to the influence of metabolism substrate phenanthrene to growth degraded benzo [a] pyrene of bacterial strain
Inoculation is respectively 0mg/L, 20mg/L, 40mg/L, 60mg/L, 100mg/L to luxuriant and rich with fragrance concentration, and benzo [a] pyrene concentration is that (composition of this substratum is: (NH for the minimal medium of 40mg/L 4) 2SO 41g, K 2HPO 42g, MgSO 47H 2O 0.5g, NaCl 0.1g, FeCl 30.5g, CaCl 20.5g benzo [a] pyrene 0.04g, distilled water 1000mL, the pH value is 7.0) in, shaking culture was measured nectar degree and benzo [a] pyrene residual quantity in the nutrient solution after 20 days in 160r/min, 37 ℃ of shaking tables, calculated degradation rate, and the result is as shown in table 3.
The influence that table 3 is degraded to strain cell growth and benzo [a] pyrene for the phenanthrene that adds different concns
Figure BSA00000440933600072
As shown in Table 3, add the phenanthrene (20-40mg/L) of low concentration, can improve benzo [a] pyrene degradation rate, supposition be because, the carbon source of easier utilization is provided for bacterial strain on the one hand, promoted strain growth, on the other hand, bacterial strain produces inducible enzyme in the process of metabolism phenanthrene, can promote the expression of strains for degrading benzo [a] pyrene gene, thereby improve degradation rate.When the concentration of phenanthrene surpassed 60mg/L, the nectar degree decreased, and its degraded to benzo [a] pyrene also is suppressed.
Is 40mg/L with inoculation to luxuriant and rich with fragrance concentration, and benzo [a] pyrene concentration is that (composition of this substratum is: (NH for the minimal medium of 40mg/L 4) 2SO 41g, K 2HPO 42g, MgSO 47H 2O 0.5g, NaCl 0.1g, FeCl 30.5g, CaCl 20.5g, luxuriant and rich with fragrance 0.04g, benzo [a] pyrene 0.04g, distilled water 1000mL, the pH value is 7.0) in, shaking culture in 160r/min, 37 ℃ of shaking tables, at the 0th, 4,8,12,16,20 day same time sampling, the residual quantity of nectar degree and benzo [a] pyrene was calculated degradation rate in the mensuration nutrient solution respectively.As shown in Figure 5, when altogether the concentration of metabolism substrate phenanthrene was 40mg/L, benzo [a] pyrene of bacterial strain Bap9 degradable 53.70% after 20 days had improved 26.4% when being single carbon source with benzo [a] pyrene, phenanthrene can be degraded fully about the 12nd day.
The growth that an amount of metabolism substrate altogether can promote bacterial strain is added in the explanation of this example, the expression of inducible strain degrading genes, improve its degradation rate to benzo [a] pyrene, simultaneously, the bacterial strain high ring polycyclic aromatic hydrocarbon mixture that can be used for degrading, perhaps be used for soil organisms and repair, and provide a kind of new germ plasm resource for high ring polycyclic aromatic hydrocarbon is total to metabolic mechanism research.
Figure ISA00000440933800011

Claims (8)

1. bacillus pumilus Bacillu spumilus strain.Bap9, CGMCC № 4585.
2. the application of the described bacillus pumilus Bacillus of claim 1 pumilus strain.Bap9 in the degraded of benzo [a] pyrene.
3. application according to claim 2 is characterized in that: can be sole carbon source and energy growth and breeding with benzo [a] pyrene, the degradation rate to benzo [a] pyrene (starting point concentration 40mg/L) in 20 days reaches 27.3%.
4. according to claim 2 or 3 described application, it is characterized in that: the optimum inoculation amount of this strains for degrading benzo [a] pyrene is 0.5%-10%, is preferably 2%-5%.
5. according to the arbitrary described application of claim 2-4, it is characterized in that: the best liquid amount of this strains for degrading benzo [a] pyrene is 5mL/50mL-30mL/50mL, is preferably 5mL/50mL-15mL/50mL.
6. application according to claim 2 is characterized in that: add an amount of common metabolism substrate, can effectively promote the growth of this bacterial strain, improve the degradation rate of this bacterial strain to benzo [a] pyrene.Described metabolism substrate altogether can be sucrose, maltose, glucose, sodium-acetate or Zulkovsky starch.
7. application according to claim 2 is characterized in that: add an amount of common metabolism substrate phenanthrene, also can promote the degraded of this bacterial strain to benzo [a] pyrene.Luxuriant and rich with fragrance optimal concentration scope of adding is 0-100mg/l, is preferably 20-40mg/L.
8. the microbiobacterial agent of benzo [a] pyrene that is used to degrade, its activeconstituents is the described bacillus pumilus Bacillus of claim 1 pumilus strain.Bap9.
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CN102491533A (en) * 2011-12-12 2012-06-13 沈阳化工大学 Method for co-degrading polycyclic aromatic hydrocarbon by using two strains
CN102491533B (en) * 2011-12-12 2013-04-03 沈阳化工大学 Method for co-degrading polycyclic aromatic hydrocarbon by using two strains
CN103667120A (en) * 2013-11-27 2014-03-26 云南省烟草农业科学研究院 Bacillus pumilus, method for acquiring strain and application of strain in orientated degradation of nitrosoamine specifically in tobacco
CN103667120B (en) * 2013-11-27 2015-07-22 云南省烟草农业科学研究院 Bacillus pumilus, method for acquiring strain and application of strain in orientated degradation of nitrosoamine specifically in tobacco
CN109182202A (en) * 2018-09-30 2019-01-11 中北大学 A kind of bacillus HT2 and its application
CN112620342A (en) * 2020-10-20 2021-04-09 广西博世科环保科技股份有限公司 Method for synergistic remediation of soil polluted by high-ring polycyclic aromatic hydrocarbon by using biosurfactant
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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