氟氯氰菊酯降解菌的筛选与降解特性的研究

doi:10.13560/j.cnki.biotech.bull.1985.2016.09.016

摘要/Abstract

摘要： 从生产氟氯氰菊酯的农药厂排污口周围的泥土中,筛选分离出一株能高效降解氟氯氰菊酯的细菌YC-WZ5。采用富集培养法筛选菌株,利用其生理生化特征及16S rDNA基因序列分析鉴定菌株种类,利用气相色谱法测定培养液中氟氯氰菊酯浓度,研究菌株在不同条件下的降解能力,并用响应面法优化降解条件。菌株YC-WZ5经鉴定属于中间苍白杆菌（Ochrobactrum intermedium）。菌株YC-WZ5能在5 d内将50 mg/L氟氯氰菊酯完全降解。采用响应面法对菌株降解氟氯氰菊酯的条件进行了优化,得出最佳条件为30.9℃,pH为7.16和接菌量10.9%。菌株YC-WZ5在5 d内对初始浓度为100、200、300和400 mg/L的氟氯氰菊酯的降解率分别为84.5%、54.7%、40%和30.5%;对50 mg/L的氯氰菊酯、功夫菊酯,溴氰菊酯和联苯菊酯的降解率分别为99.83%、91.16%、84.98%和55.76%。菌株YC-WZ5具有高效降解氟氯氰菊酯的能力及良好的环境适应性。

关键词: 氟氯氰菊酯, 生物修复, 中间苍白杆菌, 高效降解

Abstract: Cyfluthrin-degrading strain YC-WZ5 was isolated from the long-term cyfluthrin-contaminated sludge near a pesticide factory by enriched culture. We used physiological properties and 16S rDNA sequence analysis to identify the isolated strain from enriched culture,and used GC to detect the cyfluthrin concentration of the culture in different circumstances. The degrading capacities of the strain under different conditions were studied,and response surface method was used to optimize the conditions for cyfluthrin degradation. The strain YC-WZ5 was identified to be Ochrobactrum intermedium. The 50 mg/L cyfluthrin was fully degraded by YC-WZ5 in 5 days. The conditions for cyfluthrin degradation was optimized as 30.9℃,pH7.16 and 10.9% inoculum size. The degradation rate with different initial concentration of cyfluthrin（100 mg/L,200 mg/L,300mg/L,and 400 mg/L）was 84.5%,54.7%,40% and 30.5% respectively. The degradation rate of cypermethrin,cyhalothrin,deltamethrin and bifenthrin by YC-WZ5 reached 99.83%,91.16%,84.98% and 55.76% respectively at the concentration of 50 mg/L. Conclusively,the strain YC-WZ5 has highly-efficient degradation of cyfluthrin and highly environmental adaptability.

Key words: cyfluthrin, bioremediation, Ochrobactrum intermedium, highly-efficient degradation

卫正, 冯为民, 史延华, 任磊, 闫艳春. 氟氯氰菊酯降解菌的筛选与降解特性的研究[J]. 生物技术通报, 2016, 32(9): 114-122.

WEI Zheng¹, FENG Wei-min¹, SHI Yan-hua², REN Lei¹, YAN Yan-chun¹. Isolation,Identification and Degradative Properties of Cyfluthrin-degrading Bacterial Strain[J]. Biotechnology Bulletin, 2016, 32(9): 114-122.

参考文献

[1] Hu GP, Zhao Y, Song FQ, et al. Isolation, identification and cyfluth-rin-degrading potential of a novel Lysinibacillus sphaericus strain, FLQ-11-1[J]. Research in Microbiology, 2014, 165：110-118.
[2] McCoy MR, Yang Z, Fu X, et al. Monitoring of total type II pyrethroid pesticides in citrus oils and water by converting to a common product 3-phenoxybenzoic acid[J]. Journal of Agricultural and Food Chemistry, 2012, 60（20）：50-65.
[3] Guvenc D, Aksoy A, Gacar A, et al. Evaluation of changes in monoamine levels and apoptosis induced by cyfluthrin in rats[J]. Toxicology Research, 2015, 3（5）：331-340.
[4] Yilmaz M, Rencuzogullari E, Canli M. The effects of cyfluthrin on some biomarkers in the liver and kidney of Wistar rats[J]. Envir-onmental Science and Pollution Research, 2015, 22：4747-4752.
[5] Soni I, Syed F, Bhatnagar P, Mathur R. Perinatal toxicity of cyfluthrin in mice：developmental and behavioral effects[J]. Human and Experimental Toxicology, 2011, 30：1096-1105.
[6] Chen SH, Dong YH, Chang CQ et al. Characterization of a novel cyfluthrin-degrading bacterial strain Brevibacterium aureum and its biochemical degradation pathway[J]. Bioresource Technology, 2013, 132：16-23.
[7] Zhai Y, Li K, Song JL, et al. Molecular cloning, purification and biochemical characterization of a novel pyrethroid-hydrolyzing carboxylesterase gene from Ochrobactrum anthropi YZ-1[J]. Journal of Hazardous Materials, 2012, 221-222：206-212.
[8] 胡桂琴, 宋凤琴, 尤民生, 等.氟氯氰菊酯降解菌FLQ5的筛选鉴定及降解特性[J].应用与环境生物学报, 2013, 19（6）：851-856.
[9] 张建云, 崔树军, 武秀琴, 等. 1株氟氯氰菊酯降解菌GZ-3的分离和鉴定[J].安徽农业科学, 2010, 38（13）：6635-6636, 6640.
[10] Saikia N, Gopal M. Biodegradation of β-cyfluthrin by fungi[J]. Journal of Agricultural and Food Chemistry, 2004, 52（2）：1220-1223.
[11] Liang WQ, Wang ZY, Li H, et al. Purification and characterization of a novel pyrethroid hydrolase from Aspergillus niger ZD11[J]. Journal of Agricultural and Food Chemistry, 2005, 53（19）：7415-7420.
[12] Fan XJ, Liu XL, Huang R, et al. Identification and characterization of a novel thermostable pyrethroid-hydrolyzing enzyme isolated through metagenomic approach[J]. Microbial Cell Factories, 2012, 11：33.
[13] 许育新, 戴青华, 李晓慧, 等.氯氰菊酯降解菌株CDT3的分离鉴定及生理特性研究[J].农业环境科学学报, 2004, 23（5）：958-963.
[14] 秦坤, 朱鲁生, 王金花.氯氰菊酯降解真菌的筛选及其降解特性研究[J].环境工程学报, 2010, 4（4）：950-954.
[15] George MG, Don JB, Noel RK, et al. Bergey’s manual of systematic bacteriology[M]. 2nd edition. New York：Springer Publishing Press, 2001.
[16] Sambrook J, Fritsch EF. Molecular cloning：A laboratory manual[M]. 3rd edition. New York：Cold Spring Harbor Laboratory Press, 1989.
[17] Wang JH, Ren L, Jia Y, et al. Degradation characteristics and metabolic pathway of 4-nitrophenol by a halotolerant bacterium Arthrobacter sp. CN2[J]. Toxicological and Environmental Chemistry, 2016, 98（2）：226-240.
[18] 曲杰.氯氰菊酯降解菌的筛选鉴定及降解机理[D].北京：中国农业科学院, 2011.
[19] 刘艳, 范丽薇, 王晓萍.氯嘧磺隆降解菌的分离鉴定及其降解特性[J].微生物学通报, 2010, 37（8）：1164-1168.
[20] 陈少华, 罗建军, 肖盈, 等. 一株拟除虫菊酯农药降解菌的分离鉴定及其降解特性与途径[J].环境科学学报, 2011, 31（8）：1616-1625.
[21] 李文, 彭香, 闫艳春, 等.硫丹降解菌的筛选及降解性能研究[J].山东农业科学, 2009, 1：67-70.
[22] Jilani S, Khan MA. Isolation, characterization and growth response of pesticides degrading bacteria[J]. Journal of Biology Science, 2004, 4（1）：15-20.
[23] 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（9）：71-74.
[24] 闫法军, 田相利, 王军, 等.刺参养殖池塘降解有机污染物常、低温芽孢杆菌的分离筛选[J].中国海洋大学学报, 2013, 43（6）：17-24.
[25] 唐鸿志.恶臭假单胞菌代谢尼古丁分子生物学研究[D].济南：山东大学, 2008.
[26] 李晶, 饶婷, 李巍, 等.恶臭假单胞菌Pseudomonas putida LY1共代谢降解苯酚和4-氯苯酚系统的降解动力学研究[J].环境科学学报, 2011, 31（10）：2109-2116.
[27] 沈小娟, 聂麦茜, 葛碧洲, 等.铜绿假单胞菌降解菲的产物鉴定及代谢过程[J].安全与环境学报, 2012, 11（2）：62-65.
[28] 史延华, 任磊, 闫艳春, 等.施氏假单胞菌YC-YH1对甲基对硫磷的降解及其代谢产物检测[J]. 农业环境科学学报, 2015, 34（11）：2097-2104.
[29] Yuan YJ, Lu ZX, Wu N. Isolation and preliminary characterization of a novel nicotine-degrading bacterium, Ochrobactrum intermedium DN2[J]. International Biodeterioration & Biodegradation, 2005, 56：45-50.
[30] 张静.草甘膦降解菌苍白杆菌G-1的分离鉴定及草甘膦氧化还原酶基因gox的克隆[D].南京：南京农业大学, 2009.
[31] 柏文琴, 李梅, 邱星辉, 等.苍白杆菌B2对甲基对硫磷降解途径研究[J].农药学学报, 2004, 6（4）：48-54.