Rational design of organophosphorus hydrolase with high catalytic efficiency for detoxifying a V-type nerve agent.
V-type nerve agents, known as VX, are organophosphate (OP) compounds, and show extremely toxic effects on human and animals by causing cholinergic overstimulation of synapses. The bacterial organophosphorus hydrolase (OPH) has attracted much attention for detoxifying V-type agents through hydrolysis of the P-S bond. However, low catalytic efficiency of OPH has limited the practical use of the enzyme. Here we present rational design of OPH with high catalytic efficiency for a V-type nerve agent. Based on the model structure of the enzyme and substrate docking simulation, we predicted the key residues that appear to enhance the access of the substrate to the active site of the enzyme, and constructed numerous OPH mutants. Of them, double mutant, L271/Y309A, was shown to exhibit a 150-fold higher catalytic efficiency for VX than the wild-type.
Department of Biological Sciences, Korea Advanced Institute of Science and Technology (KAIST), 291 Daehak-ro, Yuseong-gu, Daejeon 305-701, Republic of Korea.
Young-Su
YS
Jeong
Department of Biological Sciences, Korea Advanced Institute of Science and Technology (KAIST), 291 Daehak-ro, Yuseong-gu, Daejeon 305-701, Republic of Korea.
Jung Min
JM
Choi
Department of Biological Sciences, Korea Advanced Institute of Science and Technology (KAIST), 291 Daehak-ro, Yuseong-gu, Daejeon 305-701, Republic of Korea.
Hyun-Ho
HH
Kyeong
Genofocus, Inc., Daejeon 305-500, Republic of Korea.
Jae-Youl
JY
Choi
Genofocus, Inc., Daejeon 305-500, Republic of Korea.
Eui-Joong
EJ
Kim
Department of Biological Sciences, Korea Advanced Institute of Science and Technology (KAIST), 291 Daehak-ro, Yuseong-gu, Daejeon 305-701, Republic of Korea. Electronic address: hskim76@kaist.ac.kr.
Hak-Sung
HS
Kim
D000818:Animals; D043303:Aryldialkylphosphatase / Q000737:chemistry* / Q000235:genetics; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics; D002384:Catalysis; D020134:Catalytic Domain; D002619:Chemical Warfare Agents / Q000378:metabolism*; D015195:Drug Design; D005417:Flavobacterium / Q000201:enzymology*; D006801:Humans; D008956:Models, Chemical; D062105:Molecular Docking Simulation; D009154:Mutation; D009946:Organothiophosphorus Compounds / Q000378:metabolism*; D011487:Protein Conformation; D013379:Substrate Specificity
D043303:Aryldialkylphosphatase; D001426:Bacterial Proteins; D002619:Chemical Warfare Agents; D009946:Organothiophosphorus Compounds; C009680:VX-agent
Organophosphates; Organophosporus hydrolase; Rational design; V-type agent