Polyhistidine-containing organophosphorus hydrolase with outstanding properties |
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Authors: | Elena Efremenko Ilya Lyagin Yulia Votchitseva Maria Sirotkina Sergey Varfolomeyev |
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Affiliation: | 1. Department of Chemical Enzymology, Faculty of Chemistry, The M. V. Lomonosov Moscow State University, Lenin's Hills 1/11, Moscow, 119992, Russiaefremenko@enzyme.chem.msu.ru;3. Department of Chemical Enzymology, Faculty of Chemistry, The M. V. Lomonosov Moscow State University, Lenin's Hills 1/11, Moscow, 119992, Russia |
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Abstract: | The catalytic and physical–chemical properties of organophosphorus hydrolase (OPH) modified by the addition of an N-terminal dodecahistidine tag (His12-OPH) have been investigated. Introduction of the His12-tag caused a 30- and 74-fold increase in catalytic efficiency of the enzyme with parathion and methyl parathion, respectively, compared to OPH. Concurrently, the His12-OPH had a more alkaline pH-optimum and extended temperature range than OPH and OPH modified with a hexahistidine tag. A study of His12-OPH thermostability showed that the enzyme had a tendency to oligomerise. This resulted in a decrease in the enzymatic activity of His12-OPH at temperatures <50°C, but provided the enzyme with much higher thermostability at temperatures >50°C, compared to OPH. |
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Keywords: | Organophosphorus hydrolase polyhistidine sequence oligomerisation thermostability |
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