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[1]易忠权,王 睿 周潮洋 陈雪婷 洪 青 何 健 闫 新** 李顺鹏.以菌株Pseudomonas putida KT2440为宿主构建多功能农药降解菌[J].应用与环境生物学报,2016,22(06):1145-1149.[doi:10.3724/SP.J.1145.2016.01006]
 YI Zhongquan,WANG Rui,et al.Construction of a multifunctional pesticide-degrading strain using Pseudomonas putida KT2440 as the host[J].Chinese Journal of Applied & Environmental Biology,2016,22(06):1145-1149.[doi:10.3724/SP.J.1145.2016.01006]
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以菌株Pseudomonas putida KT2440为宿主构建多功能农药降解菌()
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《应用与环境生物学报》[ISSN:1006-687X/CN:51-1482/Q]

卷:
22卷
期数:
2016年06期
页码:
1145-1149
栏目:
研究论文
出版日期:
2016-12-25

文章信息/Info

Title:
Construction of a multifunctional pesticide-degrading strain using Pseudomonas putida KT2440 as the host
作者:
易忠权1 2 王 睿1 周潮洋1 陈雪婷1 洪 青1 何 健1 闫 新1** 李顺鹏1
1南京农业大学生命科学学院 南京 210095 2东南大学医学院附属盐城医院中心实验室 盐城 224000
Author(s):
YI Zhongquan1 2 WANG Rui1 ZHOU Chaoyang1 CHEN Xueting1 HONG Qing1 HE Jian1 YAN Xin1** & LI Shunpeng1
1College of Life Science, Nanjing Agricultural University, Nanjing 210095, China 2Department of Central Laboratory, Affiliated Yancheng Hospital, School of Medicine, Southeast University, Yancheng 224000, China
关键词:
同源重组农药微生物降解Pseudomonas putida KT2440基因工程菌
Keywords:
Homologous recombination microbial degradation of pesticides contamination Pseudomonas putida strain KT2440 genetically engineered microorganism
分类号:
X172
DOI:
10.3724/SP.J.1145.2016.01006
摘要:
通过富集培养法分离到的农药降解菌株存在稳定性差、底物谱窄、安全性不明等问题. Pseudomonas putida KT2440分离自根际土壤,是国际上公认的环境安全型菌株. 利用同源重组方法将4种农药降解酶基因pytH、mpd、chd和amyA同时整合到P. putida KT2440的染色体上,获得了一株无抗性基因残留的工程菌P. putida KT-pmca. 工程菌KT-pmca能够同时降解拟除虫菊酯类杀虫剂、有机磷类杀虫剂、百菌清和氯代酰胺类4种农药,百菌清降解率大于95%,甲氰菊酯、毒死蜱和敌稗的降解率均达到80%以上. 此外,传代实验证明4种降解酶基因能够在遗传中稳定存在. 本研究表明工程菌KT-pmca可为微生物原位修复消除农药污染提供新的菌株资源. (图6 表2 参20)
Abstract:
Microbial in situ remediation is considered as a promising method to eliminate the pollution of pesticides. Generally, pesticide-degrading microorganisms are isolated from pesticide-contaminated sites and are then employed to remediate the pollution of pesticides. The shortcomings of these strains are narrow substrate spectrum, unstable biodegrading ability and unknown safety. Pseudomonas putida strain KT2440 is isolated from rhizosphere soil and is generally recognized as safe. This study aimed to construct a multifunctional pesticide-degrading microorganism using P. putida strain KT2440 as the host, with a hope of making a promising candidate for in situ bioremediation. Pesticide-degrading enzyme encoding genes pytH, mpd, chd and ampA were integrated into the chromosome of P. putida strain KT2440 via homologous recombination. The recombinant strain KT-pmca could degrade four pesticides including pyrethroids, organophosphorus, chlorothalonil and amide. Among them, the degradation rate of chlorothalonil was more than 95% and the degradation rate of chlorpyrifos, fenpropathrin and stam exceeded 80%. Furthermore, pesticide-degrading phenotype of P. putida strain KT-pmca was quite stable. This study showed that the recombinant strain KT-pmca could provide a promising strain for the in situ remediation of pesticides contamination.

参考文献/References:

1 李顺鹏. 环境生物学[M]. 北京: 中国农业出版社, 2002 [Li SP. Environmental Biology [M]. Beijing: China Agriculture Press, 2002] 2 Langlois BE, Collins JA, Sides KG. Some factors affecting degradation of organochlorine pesticides by bacteria [J]. J?Dairy?Sci, 1970, 53 (12): 1671-1675 3 Mulchandani P, Chen W, Mulchandani A, Wang J, Chen L. Amperometric microbial biosensor for direct determination of organophosphate pesticides using recombinant microorganism with surface expressed organophosphorus hydrolase [J]. Biosens Bioelectron, 2001, 16 (7): 433-437 4 Gerhardson B. Biological substitutes for pesticides [J]. Trends?Biotechnol, 2002, 20 (8): 338-343 5 Jiang J, Zhang R, Li R, Gu JD, Li S. Simultaneous biodegradation of methyl parathion and carbofuran by a genetically engineered microorganism constructed by mini-Tn5 transposon [J]. Biodegradation, 2007, 18 (4): 403-412 6 Liu Z, Hong Q, Xu JH, Wu J, Li SP. Construction of a genetically engineered microorganism for degrading organophosphate and carbamate pesticides [J]. Int Biodeter Biodegr,?2006,?58 (2): 65–69 7 武俊, 徐敬亮, 洪青, 李顺鹏. 遗传稳定型六六六、多菌灵降解基因工程菌构建[J]. 微生物学报, 2008, 48 (1): 45-50 [Wu J, Xu JL, Hong Q, Li SP. Construction of a genetically engineered and stable strain of degrading γ-hexachlorocyclohexane and carbendazim by transposon mini-Tn5 [J]. Acta Microbiol. Sin, 2008, 48 (1):45-50] 8 Timmis KN. Pseudomonas putida:a cosmopolitan opportunist par excellence [J]. Environ Microbiol, 2002, 4 (12): 779-781 9 Walker AW, Keasling JD. Metabolic engineering of Pseudomonas putida for the utilization of parathion as a carbon and energy source [J]. Biotechnol Bioeng, 2002, 78 (7): 715-721 10 顾立峰,何健,黄星,贾开志,李顺鹏. 多功能降解菌Pseudomonas putida KT2440-DOP的构建与降解特性研究[J]. 微生物学报, 2006, 46 (5): 763-766 [Gu LF, He J, Huang X, Jia KZ, Li SP. Construction of a versatile degrading bacteria Pseudomonas putida KT24 40 -DOP and its degrading characteristics [J]. Acta Microbiol. Sin, 2006, 46 (5): 763-766] 11 Zuo ZQ, Gong T, Che Y, Liu RH, Xu P, Jiang H, Qiao CL, Song CJ, Yang C. Engineering?Pseudomonas putida KT2440 for simultaneous degradation of organophosphates and pyrethroids and its application in bioremediation of soil [J]. Biodegradation, 2015, 26 (3): 223-233 12 Guo P, Wang BZ, Zheng JW, Hang BJ, Li L, He J, Li SP. Sphingobium faniae sp.nov., a pyrethroid-degrading bacterium isolated from activated sludge treating wastewater from pyrethroid manufacture [J]. Int J Syst Evol Microbiol, 2009, 60 (2): 408-412 13 Li XH, He J, Li SP. Isolation of a chlorpyrifos-degrading bacterium, Sphingomonas sp. strain Dsp-2, and cloning of the mpd gene [J]. Res Microbiol, 2007, 158 (2): 143-149 14 Wang GL, Li R, Li SP, Jiang JD. A novel hydrolytic dehalogenase for the chlorinated aromatic compound chlorothalonil [J]. J Bacteriol, 2010, 192 (11): 2737-2745 15 Zhang J, Yin JG, Hang BJ, Cai S, He J, Zhou SG, Li SP. Cloning of a novel arylamidase gene from Paracoccus sp. strain FLN-7 that hydrolyzes amide pesticides [J]. Appl Environ Microb, 2012, 78 (14): 4848-4855 16 Geize RVD, Hessels GI, Gerwen RV, Meijden PVD, Dijkhuizen L. Unmarked gene deletion mutagenesis of kstD, encoding 3-ketoasteroid Delta1-dehydrogenase, in Rhodococcus erythropolis SQ1 using sacB as counter-selectable marker [J]. FEMS Microbiol Lett, 2001, 205 (2): 197-202 17 Ho SN, Hunt HD, Horton RM, Pullen JK, Pease LR. Site-directed mutagenesis by overlap extension using the polymerase chain reaction [J]. Gene, 1989, 77 (1): 51-59 18 Horton RM, Hunt HD, Ho SN, Pullen JK, Pease LR. Engineering hybrid genes without the use of restriction enzymes: gene splicing by overlap extension [J]. Gene, 1989, 77 (1): 61-68 19 Horton RM, Cai Z, Ho SN, Pease LR. Gene splicing by overlap extension: tailor-made genes using the polymerase chain reaction [J]. Biotechniques, 1990, 8 (5): 528-535 20 Takeo M, Ohara A, Sakae S, Okamoto Y, Kitamura C, Kato D, Negoro S. Function of a Glutamine Synthetase-Like Protein in Bacterial Aniline Oxidation via γ-Glutamylanilide [J]. J Bacteriol, 2013, 195 (19): 4406-4414

更新日期/Last Update: 2016-12-30