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首页> 外文期刊>Journal of the Korean Society for Applied Biological Chemistry >Biodegradation of Tolclofos-methyl by Extracellular Secreted Organophosphorus Hydrolase in Recombinant Escherichia coli
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Biodegradation of Tolclofos-methyl by Extracellular Secreted Organophosphorus Hydrolase in Recombinant Escherichia coli

机译：胞外分泌有机磷水解酶在重组大肠杆菌中对甲苯酚磷的生物降解

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摘要

Fungicide tolclofos-methyl [(O,O-dimethyl O-(2,6-dichloro-4-methylphenyl) phosphorothioate)], belonging to the group of organophosphorus compounds, has been usually applied for the effective control of soil born diseases by Rhizoctonia solani. However, its excessive usages have leaded to the accumulation of this compound on soil and ginseng itself. Therefore, we practiced the actual degradation of tolclofos-methyl using organophosphorus hydrolase (OPH). A gene encoding OPH from Flavobacterium sp. strain ATCC 27551 was cloned and expressed in Escherichia coli with or without signal peptide-encoding sequences under control of a T7 promoter. High-level expression of recombinant OPH was verified by specific OPH activity assay. In addition, although the extracellular secretion of OPH in E. coli has never been reported till now, secretion of the recombinant OPH was observed when a signal peptide of the gene was truncated. Recombinant E. colt strain removed a maximum of 80% of the organophosphorus compound tolclofos-methyl, as determined by an in vitro assay The present study reports for the first time on the secretion of recombinant OPH not affected by signal peptide sequence in E. coli and biodegradation of tolclofos-methyl by extracellular secreted OPH from recombinant E. coli.
机译：属于有机磷化合物类的杀菌剂甲苯酚磷甲基[[O，O-二甲基O-（2,6-二氯-4-甲基苯基）硫代磷酸酯]]通常已被用于有效控制根瘤菌防治土壤致病性疾病索拉尼。然而，其过量使用导致该化合物在土壤和人参本身上的积累。因此，我们使用有机磷水解酶（OPH）进行了甲苯酚磷的实际降解。 Flavobacterium sp。编码OPH的基因。在T7启动子的控制下，克隆ATCC 27551菌株ATCC 27551并在大肠杆菌中表达，无论是否带有信号肽编码序列。通过特异性OPH活性测定法验证了重组OPH的高表达。另外，尽管迄今为止尚未报道大肠杆菌中OPH的细胞外分泌，但是当基因的信号肽被截短时，观察到重组OPH的分泌。通过体外试验确定，重组柯尔特氏菌菌株最多可去除80％的有机磷化合物甲苯酚磷。本研究首次报道了不受信号肽序列影响的重组OPH的分泌。重组大肠杆菌胞外分泌的OPH对甲苯酚磷的生物降解作用

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