一株土著B[a]P降解菌的筛选及降解特性研究
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摘要
通过在液体培养基中添加苯并[a]芘(B[a]P)作为唯一碳源反复驯化培养,从长期受石油烃和多环芳烃(PAHs)污染的土壤中分离出8株能够降解B[a]P的细菌,其中一株细菌(命名为BB-1)具有最大降解效果。采用16s RNA基因测序结果表明,BB-1与巨大芽孢杆菌(Bacillus megaterium)同源性为100%。为了研究BB-1的降解特性,在pH7.0、30℃条件下培养8 d,菌株B[a]P的降解率高达66.36%。为了考察培养基初始pH和外加蔗糖浓度对BB-1降解B[a]P的影响,30℃下振荡培养8 d,当设置培养基初始pH为4.0、6.0、8.0和10.0时,BB-1对B[a]P的降解率分别为12.14%、39.61%、55.21%和30.03%,可见pH为8.0时其降解效果最优;当添加0.1%蔗糖和0.5%蔗糖为外加碳源时,菌株BB-1对B[a]P的降解率分别为70.56%和74.89%,表明0.5%蔗糖的降解效果最优。
Eight bacteria strains were isolated from the soil contaminated with petroleum hydrocarbons and polycyclic aromatic hydrocarbons(PAHs)using repeated subcultures in liquid medium with added benzo[a]pyrene(B[a]P)as the sole carbon source. One of the bacterial strains named BB-1 had an optimal degradation effect(100% homologous to Bacillus megaterium)through the 16 s RNA gene sequencing method. In order to explore the degradation character of BB-1,the strain was cultured under pH =7.0,30 ° C for 8 days, after which the B[a]P degradation rate reached a maximum of 66.36%. In order to investigate the degradation rate of BB-1 under varying conditions of initial pH and additional sucrose concentration in the culture medium, the incubation was carried out at 30 °C for 8 days, with various values of initial pH including 4.0, 6.0, 8.0, and 10.0. The results showed that the degradation rates of B [a]P were 12.14%, 39.61%, 55.21%,and 30.03%, respectively. In other words, the highest degradation efficiency was achieved for pH 8.0. With the addition of 0.1% and 0.5%sucrose as an additional carbon source, the degradation rate of B [a]P reached 70.56% and 74.89%, respectively. It was thus clear that the highest degradation efficiency occurred in the media with the addition of 0.5% sucrose.
Eight bacteria strains were isolated from the soil contaminated with petroleum hydrocarbons and polycyclic aromatic hydrocarbons(PAHs)using repeated subcultures in liquid medium with added benzo[a]pyrene(B[a]P)as the sole carbon source. One of the bacterial strains named BB-1 had an optimal degradation effect(100% homologous to Bacillus megaterium)through the 16 s RNA gene sequencing method. In order to explore the degradation character of BB-1,the strain was cultured under pH =7.0,30 ° C for 8 days, after which the B[a]P degradation rate reached a maximum of 66.36%. In order to investigate the degradation rate of BB-1 under varying conditions of initial pH and additional sucrose concentration in the culture medium, the incubation was carried out at 30 °C for 8 days, with various values of initial pH including 4.0, 6.0, 8.0, and 10.0. The results showed that the degradation rates of B [a]P were 12.14%, 39.61%, 55.21%,and 30.03%, respectively. In other words, the highest degradation efficiency was achieved for pH 8.0. With the addition of 0.1% and 0.5%sucrose as an additional carbon source, the degradation rate of B [a]P reached 70.56% and 74.89%, respectively. It was thus clear that the highest degradation efficiency occurred in the media with the addition of 0.5% sucrose.
引文
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[2]Kanaly R A,Harayama S.Biodegradation of high-molecular-weight polycylic aromatic hydrocarbons by bacteria[J].Journal of Bacteriology,2000,182(8):2059-2067.
[3]Kipopoulou A M,Manoli E,Samara C.Bioconcentration of polycyclic aromatic hydrocarbon in vegetables grown in an industrial area[J].Environmental Pollution,1999,106(3):369-380.
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[5]苏慧,赵波,张素坤,等.广州市小学生多环芳烃内暴露水平[J].环境科学,2015,36(12):4567-4573.SU Hui,ZHAO Bo,ZHANG Su-kun,et al.Internal exposure levels of PAHs of primary school students in Guangzhou[J].Environmental Science,2015,36(12):4567-4573.
[6]周启星.复合污染生态学[M].北京:中国环境科学出版社,1995.ZHOU Qi-xing.Ecology of combined pollution[M].Beijing:China Environmental Science Press,1995.
[7]张雪娜,贾海滨,王伟,等.不同载体材料固定化Fusarium.sp修复老化污染土壤中HMW-PAHs的研究[J].环境科学学报,2017,37(3):1107-1114.ZHANG Xue-na,JIA Hai-bin,WANG Wei,et al.Impact of different immobilization materials for Fusarium.sp on the remediation of aged soils polluted with HMW-PAHs[J].Acta Scientiae Circumstantiae,2017,37(3):1107-1114.
[8]Boonchan S,Britz M L,Stanley G A.Degradation and mineralization of high-molecular-weight polycylic aromatic hydrocarbons by defined fungal-bacterial cocultures[J].Applied and Environmental Microbiology,2000,66(3):1007-1019.
[9]Rafin C,Veignie E,Fayeulle A,et al.Benzo[a]pyrene degradation using simultaneously combined chemical oxidation,biotreatment with Fusarium solani and cyclodextrins[J].Bioresource Technology,2009,100(12):3157-3160.
[10]包贞,潘志彦,杨晔,等.环境中多环芳烃的分布及降解[J].浙江工业大学学报,2003,31(5):528-544.BAO Zhen,PAN Zhi-yan,YANG Ye,et al.The distribution and decomposition of PAHs in the environment[J].Journal of Zhejiang University of Technology,2003,31(5):528-544.
[11]Jules G E,Pratap S,Ramesh A,et al.In utero exposure to benzo[a]pyrene predisposes offspring to cardiovascular dysfunction in later-life[J].Toxicology,2012,295(1/2/3):56-67.
[12]王红旗,王帅,宁少尉,等.土壤铅-苯并[a]芘复合污染对小麦种子生长的影响研究[J].环境科学,2011,32(3):886-894.WANG Hong-qi,WANG Shuai,NING Shao-wei,et al.Effects of combined pollution of lead and benzo[a]pyrene on seed growth of wheat in soils[J].Environmental Science,2011,32(3):886-894.
[13]Keith L H,Telliard W A.Priority pollutantsⅠ:A perspective review[J].Environ Sci Technol,1979,13(4):416-423.
[14]Gan S,Lau E V,Ng H K,Remediation of soils contaminated with polycyclic aromatic hydrocarbons(PAHs)[J].Journal of Hazardous Materials,2009,172:532-549.
[15]巩宗强,李培军,王新,等.真菌对土壤中苯并[a]芘的共代谢降解[J].环境科学研究,2001,14(6):36-39.GONG Zong-qiang,LI Pei-jun,WANG Xin,et al.Cometabolic degradation of benzo[a]pyrene in the soil by the introduced fungi[J].Research of Environmental Sciences,2001,14(6):36-39.
[16]顾平,张倩茹,周启星,等.一株苯并[a]芘高效降解真菌的筛选与降解特性[J].环境科学学报,2010,30(1):1-6.GU Ping,ZHANG Qian-ru,ZHOU Qi-xing,et al.Isolation of a fungal strain capable of degrading benzo[a]pyrene and its degradation characteristics[J].Acta Scientiae Circumstantiae,2010,30(1):1-6.
[17]Hadibarata T,Kristanti R A.Fate and cometabolic degradation of benzo[a]pyrene by white-rot fungus Armillaria sp.F022[J].Bioresource Technology,2012,107:314-318.
[18]孙铁珩,宋玉芳,许华夏,等.植物法生物修复PAHs和矿物油污染土壤的调控研究[J].应用生态学报,1999,10(2):225-229.SUN Tie-heng,SONG Yu-fang,XU Hua-xia,et al.Research of bioremediation of contaminated soil by PAHs and mineral oil with plants act[J].Chinese Journal of Applied Ecology,1999,10(2):225-229.
[19]盛下放,何琳燕,胡凌飞,等.苯并[a]芘降解菌的分离筛选及其降解条件的研究[J].环境科学学报,2005,25(6):791-795.SHENG Xia-fang,HE Lin-yan,HU Ling-fei,et al.Isolation of a benzo[a]pyrene-degrading strain and its degradation conditions[J].Acta Scientiae Circumstantiae,2005,25(6):791-795.
[20]吴蔓莉,陈凯丽,叶茜琼,等.堆肥-生物强化对重度石油污染土壤的修复作用[J].环境科学,2017,38(10):4412-4419.WU Man-li,CHEN Kai-li,YE Xi-qiong,et al.Remediation of petroleumcontaminated soil using a bioaugmented compost technique[J].Environmental Science,2017,38(10):4412-4419.
[21]侯树宇,张清敏,余海晨,等.多环芳烃芘高效降解菌的筛选及其降解性能的研究[J].南开大学学报:自然科学版,2006,39(2):71-74.HOU Shu-yu,ZHANG Qing-min,YU Hai-chen,et al.Optimized cultivation of high-efficient degradation bacterial strains and their degradation ability towards pyrene[J].Acta Scientiarum Naturalium Universitatis Nankaiensis,2006,39(2):71-74.
[22]周启星,魏树和,张倩茹,等.生态修复[M].北京:中国环境科学出版社,2006.ZHOU Qi-xing,WEI Shu-he,ZHANG Qian-ru,et al.Ecological remediation[M].Beijing:China Environmental Science Press,2006.
[23]胡凤钗,苏振成,孙健,等.高效芘降解菌N12的分离鉴定与降解特性[J].应用生态学报,2011,22(6):1566-1572.HU Feng-chai,SU Zhen-cheng,SUN Jian,et al.Isolation and identification of a highly efficient pyrene-degrading Mycobacterium sp.strain N12[J].Chinese Journal of Applied Ecology,2011,22(6):1566-1572.
[24]刘海滨,王翠苹,张志远,等.苯并[a]芘高效降解菌筛选及其降解特性研究[J].环境科学,2011,32(9):2696-2701.LIU Hai-bin,WANG Cui-ping,ZHANG Zhi-yuan,et al.Isolation of highly-effective benzo[a]pyrene degrading strain and its degradation capacity[J].Environmental Science,2011,32(9):2696-2701.
[25]陈思尹,吕卫光,张娟琴,等.萘降解菌的分离及其联合修复作用的研究进展[J].环境科学与技术,2017,40(7):152-159.CHEN Si-yin,L譈Wei-guang,ZHANG Juan-qin,et al.Review on isolation of naphthalene degradation bacterium in different habitats and the combined remediation[J].Environmental Science and Technology,2017,40(7):152-159.
[26]黄兴如,张彩文,张瑞杰,等.多环芳烃降解菌的筛选、鉴定及降解特性[J].微生物学通报,2016,43(5):965-973HUANG Xing-ru,ZHANG Cai-wen,ZHANG Rui-jie,et al.Isolation,identification and degrading properties of PAHs-degrading bacteria[J].Microbiology China,2016,43(5):965-973.
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