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Improved Hydrolysis of Organophosphorus Compounds by Engineered Human Prolidases

[ Vol. 24 , Issue. 7 ]

Author(s):

Hyeongseok Yun, Sungrae Lee, Sumi Kim, Jiyeon Yu, Nari Lee, Jinhee Lee, Nam Doo Kim, Chiho Yu and Jaerang Rho*   Pages 617 - 625 ( 9 )

Abstract:


Background: Human prolidase has weak hydrolytic activity for toxic organophosphorus compounds including diisopropyl fluorophosphates (DFP), chemical warfare nerve agents and pesticides.

Objectives: In order to use human prolidase as a catalytic bioscavenger against toxic organophosphorus compound exposure, protein engineering is an important issue to improve the catalytic activity of human prolidase towards the hydrolysis of toxic organophosphorus compounds.

Method: We developed two human prolidase mutants, A252R and P365R, with a single amino acid substitution using in silico analysis based on the sequence, protein structure and stability to improve the catalytic activity of human prolidase towards DFP hydrolysis.

Results: Our results showed that the catalytic efficiencies of A252R and P365R towards DFP hydrolysis were 1.23- and 1.36-fold increases, respectively, than that of the wild type, while the prolidase activities of A252R and P365R towards Leu-Pro hydrolysis were 0.88- and 0.78-fold decreases that of the wild type, respectively, indicating that substitution mutations of A252R and P365R in human prolidase show improved hydrolytic activity for toxic organophosphorus compounds.

Conclusion: We report here that by introducing either the A252R or P365R substitution mutation, the structural changes affecting catalytic turnover rate and substrate binding affinity are valuable in improving the catalytic activity of human prolidase towards toxic organophosphorus compound hydrolysis.

Keywords:

Prolidase, bioscavenger, organophosphorus compound, diisopropyl fluorophosphate, in silico analysis, mutant.

Affiliation:

Department of Microbiology and Molecular Biology, Chungnam National University, Daejeon 34134, Department of Microbiology and Molecular Biology, Chungnam National University, Daejeon 34134, Department of Microbiology and Molecular Biology, Chungnam National University, Daejeon 34134, Department of Microbiology and Molecular Biology, Chungnam National University, Daejeon 34134, Department of Microbiology and Molecular Biology, Chungnam National University, Daejeon 34134, Department of Microbiology and Molecular Biology, Chungnam National University, Daejeon 34134, New Drug Development Center, DGMIF, 80, Cheombok-ro, Dong-gu, Daegu 41061, Agency for Defense Development, P.O. Box 35, Yuseong, Daejeon 34186, Department of Microbiology and Molecular Biology, Chungnam National University, 99 Daehak-ro, Yuseong-gu, Daejeon 34134

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