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Biochemical basis for hydrolysis of organophosphorus by a marine bacterial prolidase
Xiao, YZ; Yang, J; Tian, XP; Wang, XX; Li, J; Zhang, S; Long, LJ;;
AbstractExtensive application of synthesized organophosphorus compounds (OPs) leads to pollutant accumulation and enhanced eco-toxicity. Hydrolysis of phosphotriester bonds catalyzed by evolved microbial enzymes is a key step for detoxification of OPs. Here, a new marine bacterial prolidase OPAA4301 exhibiting promiscuous phosphotriesterase activity was isolated and systematically characterized. The homo-tetrameric enzyme OPAA4301 can catalyze the hydrolysis of both amido bond and phosphotriester bond. Manganese ions were observed to be essential for its catalytic integrity, and in vitro substitution of manganese ions by different metal cofactors led to decreased activity. We also revealed cooperation pattern of metal ligands and substrate-binding residues on OP hydrolysis by mutational analysis. Metal-binding sites together with Arg418 in the large-binding pocket of the enzyme were found to be indispensable for catalytic ability. Substitution mutation of small- and large-binding pocket residues caused significant variation in phosphotriesterase activity, and leaving group sites appeared to be involved in the catalytic process as substrate affinity regulators. Our study gave an overall biochemical understanding on the organophosphorus hydrolysis pattern of the newly identified marine bacterial prolidase and provided ideas for protein engineering to expand its application in the bioremediation field. (C) 2016 Elsevier Ltd. All rights reserved.
Department[Xiao, Yunzhu; Yang, Jian; Tian, Xinpeng; Wang, Xiaoxue; Li, Jie; Zhang, Si; Long, Lijuan] Chinese Acad Sci, South China Sea Inst Oceanol, CAS Key Lab Trop Marine Bioresources & Ecol, RNAM Ctr Marine Microbiol,Guangdong Key Lab Marin, Guangzhou 510301, Guangdong, Peoples R China; [Xiao, Yunzhu; Yang, Jian; Wang, Xiaoxue; Zhang, Si; Long, Lijuan] Univ Chinese Acad Sci, Beijing 100049, Peoples R China
KeywordEnzyme Promiscuity Organophosphorus Bioremediation Organophosphorus Acid Anhydrolase Prolidase
Funding ProjectLMB
Document Type期刊论文
Recommended Citation
GB/T 7714
Xiao, YZ,Yang, J,Tian, XP,et al. Biochemical basis for hydrolysis of organophosphorus by a marine bacterial prolidase[J]. PROCESS BIOCHEMISTRY,2017,52:141-148.
APA Xiao, YZ.,Yang, J.,Tian, XP.,Wang, XX.,Li, J.,...& basis for hydrolysis of organophosphorus by a marine bacterial prolidase.PROCESS BIOCHEMISTRY,52,141-148.
MLA Xiao, YZ,et al."Biochemical basis for hydrolysis of organophosphorus by a marine bacterial prolidase".PROCESS BIOCHEMISTRY 52(2017):141-148.
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