CN102174445A - Acinetobacter sp.Bap30 capable of effectively degrading benzo(a)pyrene and application thereof - Google Patents
Acinetobacter sp.Bap30 capable of effectively degrading benzo(a)pyrene and application thereof Download PDFInfo
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Abstract
The invention discloses a bacterium for degrading benzo(a)pyrene and application thereof. The bacterium is acinetobacter sp.Bap30, CGMCC No.4586. 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 28.66%. 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 cane sugar and maltose. 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 48.87% 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
Technical field
The present invention relates to environment hardly degraded organic substance process field, particularly a kind of benzo [a] pyrene high-efficiency degradation bacterium series 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 to provides a kind of efficient benzo [a] pyrene degradation bacteria and application thereof at the inefficient difficult problem of benzo [a] pyrene biological degradation.
Acinetobacter calcoaceticus Acinetobacter sp.Bap30 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 № 4586.
Strains A cinetobacter sp.Bap30 takes a sample from the sewage wastewater of dependents' district, school district, China Agricultural University east, a strain gram negative bacterium that obtains through domestication, separation and purifying.
Morphological specificity: bacterial strain Bap30 behind the cultivation 16h, is grown to the circular bacterium colony of xanchromatic 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, size is 1.0～3.0 μ m, the catalase reaction result is positive, the oxydase reaction result is negative.
According to its morphological specificity and physiological and biochemical property and 16S rRNA gene order thereof, identify that this bacterial strain is an acinetobacter calcoaceticus, the 16S rRNA of this bacterial strain has as the nucleotide sequence shown in sequence is represented, and sequence length is 1390bp.
CGAGCGGAGAGAGGTAGCTTGCTACTGATCTTAGCGGCGGACGGGTGAGT
AATGCTTAGGAATCTGCCTATTAGTGGGGGACAACATTTCGAAAGGAATGC
TAATACCGCATACGTCCTACGGGAGAAAGCAGGGGATCTTCGGACCTTGCG
CTAATAGATGAGCCTAAGTCGGATTA---GCTAG-TTGGTGGGGTAAAGGCCT
ACCAAGGCGACGATCTGTAGCGGGTCTGAGAGGATGATCCGCCACACTGG
GACTGAGACACGGCCCAGACTCCTACGGGAGGCAGCAGTGGGGAATATTG
GACAATGGGCGCAAGCCTGATCCAGCCATGCCGCGTGTGTGAAGAAGGCC
TTATGGTTGTAAAGCACTTTAAGCGAGGAGGAGGCTACTTTAGTTAATACCT
AGAGATAGTGGACGTTACTCGCAGAATAAGCACCGG-CTAACTCTGTGCCA
GCAGCCGCGGTAATACAGAGGGTGCAAGCGTTAATCGGATTTACTGGGCGT
AAAGCGCGCGTAGGCGGCTAATTAAGTCAAATGTGAAATCCCCGAGCTTAA
CTTGGGAATTGCATTCGATACTGGTTAGCTAGAGTGTGGGAGAGGATGGTA
GAATTCCAGGTGTAGCGGTGAAATGCGTAGAGATCTGGAGGAATACCGATG
GCGAAGGCAGCCATCTGGCCTAACACTGACGCTGAGGTGCGAAAGCATGG
GGAGCAAACAGGATTAGATACCCTGGTAGTCCATGCCGTAAACGATGTCTA
CTAGCCGTTGGGGCCTTTGAGGCTTTAGTGGCGCAGCTAACGCGATAAGTA
GACCGCCTGGGGAGTACGGTCGCAAGACTAAAACTCAAATGAATTGACGG
GGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGATGCAACGCGAAG
AACCTTACCTGG-CCTTGACATAGTAAGAACTTTCCAGAGATGGATTGGTGC
CTTCGGGAACTTACATACAGGTGCTGCATGGCTGTCGTCAGCTCGTGTCGT
GAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTTTCCTTATTTGCC
AGCGAGTAATGTCGGGAACTTTAAGGATACTGCCAGTGACAAACTGGAGG
AAGGCGGGGACGACGTCAAGTCATCATGGCCCTTACGGCCAGGGCTACAC
ACGTGCTACAATGGTCGGTACAAAGGGTTGCTACCTAGCGATAGGATGCTA
ATCTCAAAAAGCCGATCGTAGTCCGGATTGGAGTCTGCAACTCGACTCCAT
GAAGTCGGAATCGCTAGTAATCGCGGATCAGAATGCCGCGGTGAATACGTT
CCCGGGCCTTGTACACACCGCCCGTCACACCATGGGAGTTTGTTGCACCAG
AAGTAGCTAGCCTAACTGCAAAGA
According to its morphological specificity and physiological and biochemical property and 16S rRNA gene order thereof, identify that this bacterial strain is acinetobacter calcoaceticus Acinetobacter sp..
This bacterial strain 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 28.66%.
This bacterial strain benzo [a] pyrene of all degrading in the wider pH value scope of neutrality or meta-acid or meta-alkalescence, the pH value of the suitableeest growth and degraded is 5.5-9.5, is preferably 5.5-7.0.
The optimum inoculation amount of this strains for degrading benzo [a] pyrene is 0.5%-10%, is preferably 2%-5%.
The best liquid amount of this strains for degrading benzo [a] pyrene is 5mL/50mL-30mL/50mL, is preferably 5mL/50mL-15mL/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 sucrose, 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 strains A cinetobacter sp.Bap30.
Acinetobacter calcoaceticus 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 28.66%, compares with existing bacterial strain, and degradation rate improves greatly.
(2) when benzo [a] pyrene and phenanthrene existed jointly, this bacterial strain reached 48.87% 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 strains A cinetobacter sp.Bap30 and the degradation curve of benzo [a] pyrene
Fig. 2 is the influences of different initial pH values to strains A cinetobacter sp.Bap30 degraded benzo [a] pyrene.
Fig. 3 is the influence of different vaccination amount to strains A cinetobacter sp.Bap30 degraded benzo [a] pyrene.
Fig. 4 is the influences of different liquid amounts to strains A cinetobacter sp.Bap30 degraded benzo [a] pyrene.
Fig. 5 is the degradation effect of strains A cinetobacter sp.Bap30 to benzo [a] pyrene and luxuriant and rich with fragrance mixed system.
Embodiment
The invention will be further described below in conjunction with specific embodiment, but the present invention is not limited to following examples.
Among the following embodiment, if no special instructions, be ordinary method.
Embodiment 1: the performance of the separation purification method of strains A cinetobacter sp.Bap30 and degraded pyrene
One, the separation purification method of efficient degradation benzo [a] pyrene bacterial strain Bap30
This method has following steps:
1,10mg benzo [a] pyrene is dissolved in the 50mL acetone soln, making benzo [a] pyrene final concentration is 200mg/L;
2, get an amount of absorbent cotton, be soaked in the acetone soln of above-mentioned benzo [a] pyrene, placement is spent the night;
3, the inferior daily string bag wraps up absorbent cotton, and ties down the string bag with blocky long rope, throws in the water drain of dependents' district, school district, China Agricultural University east;
4, place after 21 days, absorbent cotton is taken out, be placed in the 250mL triangular flask that 50mL domestication substratum is housed, the effective constituent of domestication substratum is (NH
4)
2SO
41g, K
2HPO
42g, MgSO
47H
2O 0.5g, NaCl0.1g, FeCl
30.5g, CaCl
20.5g, benzo [a] pyrene 0.04g, distilled water 1000mL, the pH value is 7.0;
5, above-mentioned triangular flask is placed 160r/min, 37 ℃ of shaking table shaking culture;
6, through after some generations tame repeatedly, get the 1mL nutrient solution and evenly coat on the inorganic salt flat board that contains 200mg/L benzo [a] pyrene, the main component of minimal medium is: (NH
4)
2SO
41g, K
2HPO
42g, MgSO
47H
2O 0.5g, NaCl 0.1g, FeCl
30.5g, CaCl
20.5g, agar 20g, distilled water 1000mL, the pH value is 7.0;
7,37 ℃, cultivated 3-5 days, 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.
Single bacterium colony behind the purifying is inoculated in benzo [a] the pyrene inorganic salt liquid substratum, and the lucifuge shaking culture is 20 days in 160r/min, 37 ℃ of shaking tables, chooses the highest bacterial strain Bap30 of degradation rate 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 № 4586.
Two, efficient degrading bacterial strain Bap30 is to the degradation property of benzo [a] pyrene
Embodiment 1: the performance of the separation purification method of strains A cinetobacter sp.Bap30 and degraded pyrene
Above-mentioned bacterial strains Bap30 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 was taken a sample respectively the 0th, 4,8,12,16,20 day same time, the residual quantity of nectar degree and benzo [a] pyrene in the mensuration nutrient solution, experimental result is seen Fig. 1.As can be 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, and during by the 20th day, the OD value of bacterial strain reaches 0.195, the degradation rate of benzo [a] pyrene reaches the highest, is 28.66%.
The degradation bacteria that present embodiment explanation separation domestication obtains 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 strains A cinetobacter sp.Bap30 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) pH value is respectively 5.5,6.0,7.0,8.0,9.0,9.5, benzo [a] pyrene starting point concentration is 40mg/L, inoculum size is 5% (V/V), 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, the results are shown in Figure 2.
As seen from Figure 2, be 6.0 o'clock at pH, degradation bacteria is the strongest to the Degradation of benzo [a] pyrene, degradation rate is 17.89% after 7 days, at pH is that 9.5 o'clock degradation rates are subjected to bigger inhibition, is 1.57% only, and bacterial strain easier utilization degraded benzo [a] pyrene in the slant acidity environment is described.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 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% inorganic salt nutrient solution 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 98.03%, 93.27%, 87.46%, 85.09% and 89.21% (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, 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 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 83.28%, 85.09%, 86.57%, 91.21%, 94.83%, 97.46% (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 and the influence of degraded benzo [a] pyrene
Bacterial strain Bap30 is seeded to the minimal medium 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, 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, measure nectar degree and benzo [a] pyrene residual quantity in the nutrient solution after 7 days, calculate degradation rate, the result is as shown in table 1.As shown in Table 1, degradation rate is than higher when adopting sucrose, maltose and glucose as carbon source, 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.Result in the table 1 shows that also sucrose is best common metabolism carbon source.
Table 1 adds the influence of carbon source to strains A cinetobacter sp.Bap30 growth and benzo [a] pyrene degradation rate
Inoculation to sucrose concentration is respectively 0mg/L, 10mg/L, 40mg/L, 100mg/L, 500mg/L, 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 sucrose of different concns to the influence of strain cell growth with the degraded of benzo [a] pyrene
As shown in Table 2, when sucrose concentration is 10mg/L, the degradation efficiency of benzo [a] pyrene with do not add control sucrose and be more or less the same; When concentration of sucrose was increased to 100mg/L, the degradation rate of benzo [a] pyrene increased to 40.87%; When concentration of sucrose was increased to 500mg/L, the degradation rate of benzo [a] pyrene sharply dropped to 13.42%.The sucrose that has added 10-100mg/L can promote thalli growth, improves the degradation rate of bacterial strain to pyrene, adds excessive sucrose, 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 is 20 days in 160r/min, 37 ℃ of shaking tables, measures nectar degree and benzo [a] pyrene residual quantity in the nutrient solution, calculates degradation rate.The influence that table 3 is degraded to strain cell growth and benzo [a] pyrene for the phenanthrene that adds different concns.
The influence that table 3 is degraded to strain cell growth and benzo [a] pyrene for the phenanthrene that adds different concns
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 Bap30 degradable 48.87% after 20 days had improved 20.2% 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.
Claims (8)
1. acinetobacter calcoaceticus Acinetobacter sp.Bap30 preserves and is numbered CGMCC № 4586.
2. the application of the described acinetobacter calcoaceticus Acinetobacter of claim 1 sp.Bap30 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 28.66%.
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 acinetobacter calcoaceticus Acinetobacter of claim 1 sp.Bap30.
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| CN104745515A (en) * | 2015-04-07 | 2015-07-01 | 华中农业大学 | Acinetobacter sp. for degrading polycyclic aromatic hydrocarbon and application of acinetobacter sp. |
| CN105695360A (en) * | 2016-03-18 | 2016-06-22 | 中国科学院广州地球化学研究所 | Phenanthrene-degrading strain Acinetobacter tandoii LJ-5 and application thereof |
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| CN105695360A (en) * | 2016-03-18 | 2016-06-22 | 中国科学院广州地球化学研究所 | Phenanthrene-degrading strain Acinetobacter tandoii LJ-5 and application thereof |
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