Rational design of organophosphorus hydrolase for altered substrate specificities.
Organophosphorus hydrolase (OPH) is a bacterial enzyme that hydrolyzes a broad variety of OP neurotoxins, including chemical warfare agents and many widely used pesticides. OPH has extremely high hydrolytic efficiency with different phosphotriester and phophothiolester pesticides (k(cat) = 50-15,000 s(-1)) as well as phosphorofluorates such as DFP and the chemical warfare agents sarin and soman (k(cat) = 50-11,000 s(-1)). In contrast, the enzyme has much lower catalytic capabilities for phosphonothioate neurotoxins such as acephate or the chemical warfare agent VX [O-ethyl S-(2-diisopropyl aminoethyl) methylphosphonothioate] (k(cat) = 0.3-20 s(-1)). Different metal-associated forms of the enzyme have demonstrated varying hydrolytic capabilities for each of the OP neurotoxins, and the activity of OPH (Co2+) is consistently higher than that of OPH (Zn2+) by five- to 20-fold. Protein engineering strategies have exploited these metal-induced catalytic differences, and other slight modifications to the opd gene have resulted in significant enhancement of the rates of detoxification of the thioate pesticides and chemical warfare agents. In order to develop practical applications of OPH, other experiments have focused on improvement of enzyme production, localization, stability, and shelf-life, as well as efficient catalysis of substrates of interest.
Di Sioudi
B D
BD
Department of Biochemistry and Biophysics, Texas A&M University, College Station 77843-2128, USA.
Miller
C E
CE
Lai
K
K
Grimsley
J K
JK
Wild
J R
JR
D000818:Animals; D043303:Aryldialkylphosphatase; D015195:Drug Design; D004950:Esterases / Q000138:chemical synthesis* / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology; D006801:Humans; D006868:Hydrolysis; D015202:Protein Engineering / Q000379:methods; D013379:Substrate Specificity
D004950:Esterases; D043303:Aryldialkylphosphatase