氯氰菊酯高效降解菌的筛选鉴定及其降解特性

摘要/Abstract

摘要： 从四川省雅安市茶园土壤中分离到1株能高效降解氯氰菊酯的细菌B-1。根据其形态、生理生化特征和16S rDNA序列分析(登录号HQ009796)，将该菌鉴定为Bacillus licheniformis。菌株B-1以共代谢方式降解氯氰菊酯，对20mg/L氯氰菊酯72h降解率达93.93%，对100mg/L氯氰菊酯240h降解率为70.35%，能检测到其降解中间产物3-苯氧基苯甲酸(3-PBA)；菌株B-1对20mg/L溴氰菊酯、氟氯氰菊酯和氰戊菊酯72h降解率分别为95.62%、79.73%和55.67%；菌株B-1对氯氰菊酯降解作用源于其所产生降解酯酶，为非诱导酶，存在于细胞内外，其主要分泌于胞外，同工酯酶电泳进一步表明其胞内外均可能有相同活性降解酯酶带；菌株B-1培养液对小白鼠的急性毒性实验证明其属于实际无毒。

关键词: 氯氰菊酯, 生物降解, 地衣芽孢杆菌, 降解特性

Abstract: A cypermethrin-degrading strain named B-1 was isolated from soil samples collected from a tea garden in Ya’an, Sichuan province. According to its morphological, physiological and biochemical characteristics and 16S rDNA sequence (GenBank Accession No. HQ009796 ), B-1 was identified as Bacillus licheniformis. B-1 degraded cypermethrin by a co-metabolic mechanism and the degradation rate was 93.93% for 20 mg/L cypermethrin in 72 h and 70.35% for 100 mg/L cypermethrin in 240 h. 3-PBA was detected as an intermediate product during the degradation process. The degradation rates for deltamethrin, cyfluthrin and fenvalerate at a concentration of 20 mg/L were 95.62%, 79.73% and 55.67% in 72 h, respectively. B-1 could produce a non-inducible esterase that degrades cypermethrin. The enzyme was found both inside and outside the cells mainly as an extracellular product. Isozyme electrophoresis analysis suggested that intracellular and extracellular esterase extracts from strain B-1 had the same esterase electrophoretic pattern. Acute toxicity tests showed that culture supernatant of strain B-1 had no observed toxicity for mice.

Key words: cypermethrin, biodegradation, Bacillus licheniformis, degradation characteristics

中图分类号:

Q939.1

赖?文，刘书亮*，赵?楠，袁怀瑜. 氯氰菊酯高效降解菌的筛选鉴定及其降解特性[J]. 食品科学, 2012, 33(21): 157-163.

参考文献

[1] Al-MAKKAWY H K , MADBOULY M D . Persistence and accumulation of some organic insecticides in Nile water and fish [J]. Resour. Conserv. Recy., 1999, 27(1):105-115.
[2] WOLANSKY M J , HARRILL J A . Neurobehavioral toxicology of pyrethroid insecticides in adult animals: A critical review [J]. Neurotoxicol. Teratol., 2008, 30:55-78.
[3] SINGH P B , SINGH V. Cypermethrin induced histological changes in gonadotrophic cells, liver, gonads, plasma levels of estradiol-1 and 11-ketotestosterone, and sperm motility in Heteropneustes fossilis (Bloch) [J]. Chemosphere, 2008, 72(3):422-431.
[4] Zhang Jun, Zhu Wei, Zheng Yifan, et al. The antiandrogenic activity of pyrethroid pesticides cyfluthrin and β-cyfluthrin [J]. Reprod. Toxicol., 2008, 25(4):491-496.
[5] Qu Jianhua, Hong Xia, Chen Jianfeng, et al. Fenvalerate inhibits progesterone production through cAMP-dependent signal pathway[J]. Toxicol. Lett., 2008, 176(1):31-39.
[6] KOLACZINSKI J H , CURTIS C F . Chronic illness as a result of low-level exposure to synthetic pyrethroid insecticides: a review of the debate [J]. Food Chem. Toxicol., 2004, 42(5):697-706.
[7] KOCAMAN A Y , TOPAKTA M . The in vitro genotoxic effects of a commercial formulation of alpha-cypermethrin in human peripheral blood lymphocytes [J]. Environ. Mol. Mutagen., 2009, 50(1):27.
[8] ARTHUR F H , CAMPPBELL J F. Distribution and efficacy of pyrethrin aerosol to control Tribolium confusum (Coleoptera: Tenebrionidae) in food storage facilities [J]. J. Stored. Prod. Res., 2008, 44(1):58-64.
[9] 周升扬. 出口蔬菜农药残留分析与安全性评价[D]. 山东: 山东农业大学, 2006.
[10] GONZALEZ-RODRIGUEZ R M , RIAL-OTERO R, CANCHO-GRANDE B, et al. Occurrence of fungicide and insecticide residues in trade samples of leafy vegetable [J]. Food Chem., 2008, 107:1342-1347.
[11] Zhang Zhiyong, Liu Xianjin, Hong Xiaoyue. Effects of home preparation on pesticide residues in cabbage [J]. Food Control, 2007, 39(8):887-891.
[12] BISHNU A, CHAKRABARTI K, CHAKRABORTY A, et al. Pesticide residue level in tea ecosystems of Hill and Dooars regions of West Bengal, India [J]. Environ. Monit. Assess., 2009, 149(1):457-464.
[13] FRIBERG-JENSEN U, WENDT-RASCH L, WOIN P, et al. Effects of the pyrethroid insecticide, cypermethrin, on a freshwater community studied under field conditions. I. Direct and indirect effects on abundance measures of organisms at different trophic levels [J]. Aquat Toxicol., 2003, 63(4):357-371.
[14] 《主要贸易国家和地区食品中农兽药残留限量标准》编委会. 主要贸易国家和地区食品中农兽药残留限量标准. 食品卷 (上下册). 北京: 中国标准出版社, 2006.
[15] 李顺鹏. 拟除虫菊酯类农药降解菌改良农作物品质的应用: 中国, 101057591. 2007-10-24.
[16] GRANT R J, DANIELL T J, Betts W B. Isolation and identification of synthetic pyrethroid-degrading bacteria [J]. J. Appl. Microbiol., 2002, 92(3):534-540.
[17] 刘君寒, 王兆守, 何健等. 一株氯氰菊酯降解菌的分离和鉴定[J]. 南京农业大学学报, 2007, 30(3):68-72.
[18] 丁海涛, 李顺鹏. 拟除虫菊酯类农药残留降解菌的筛选及其生理特性研究[J]. 土壤学报, 2003, 40(1):123-129.
[19] MALONEY S E . Degradation of insecticides and herbicides by fungi [J]. J. Chem. Technol. Biot., 1998, 71(4):360-361.
[20] 王兆守. 茶叶上拟除虫菊酯类农药降解菌的分离及其特性[J]. 生态学报, 2005, 25(7):1824-1827.
[21] 徐莲,张丽萍,刘怡辰,等. 功夫菊酯降解菌GF-3的筛选鉴定及其降解特性研究[J]. 农业环境科学学报. 2009, 28(7):1545-1551.
[22] Liang Weiqu, Wang Zhuoyi, Li He, et al. Purification and characterization of a novel pyrethroid hydrolase from Aspergillus niger ZD11[J]. J. Agric. Food Chem.,2005,53(19):7415-7420.
[23] MALONEY S E , MAULE A , SMITH A R . Purification and preliminary characterization of permethrinase from a pyrethroid-transforming strain of Bacillus cereus[J]. Appl. Environ. Microb., 1993,59(7):2007.
[24] Guo Peng, Wang Baozhao, Hang Baojian, et al. Pyrethroid-degrading Sphingobium sp. JZ-2 and the purification and characterization of a novel pyrethroid hydrolase[J]. Int. Biodeter. Biodegr.,2009,63(8):1107-1112.
[25] R E 布坎南, N E 吉本斯等.《伯杰细菌鉴定手册》第八版[M]. 中国科学院微生物研究所《伯杰细菌鉴定手册》翻译组, 译. 北京: 科学出版社,1984.729-732.
[26] 许育新, 孙纪全, 李晓慧, 等. 两株菌对氯氰菊酯及其降解产物3-PBA的协同代谢研究[J]. 微生物学报, 2007,47(5):834-837.
[27] 熊庆娥.《植物生理学实验教程》[M]. 四川: 科学技术出版社, 2003: 115-118.
[28] 林淦, 王兆守, 林海清,等. 氯氰菊酯的酶促降解[J]. 西北农业学报, 2006,15(001):64-67.
[29] 张艳辉. 拟除虫菊酯降解基因的克隆[D]. 北京:中国农业科学院, 2007.
[30] GB15193.3-2003,《食品安全性毒理学评价程序和方法》[S].