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G. Yu. Lomakina, Yu. A. Modestova, N. N. Ugarova

The thermostability enhancement of firefly luciferase Luciola mingrelica by site-directed mutagenesis of non-concervative residies Cys62 and Cys146

Abstract

Mutant forms of L.mingrelica firefly luciferase with point mutations Cys62Ser and Cys146Ser were prepared by site-directed mutagenesis. Mutations were not influenced the catalytic and spectral properties of the enzyme. Fast (k1) and slow (k2) inactivation rate constants at 37°C for wild type enzyme and its mutants with or without DTT were determined. Several times increase of mutant forms thermostability on both stages of the inactivation was shown. For wild type enzyme three–fold decrease of k2 value in the presence of DTT was shown while k1 and k2 values for mutant forms were not changed. Thus it was concluded the significance of Cys62 and Cys146 residues in the L.mingrelica firefly luciferase inactivation process and their replacements to Ser resulted in the enzyme stabilization.
Moscow University Chemistry Bulletin.
2008, Vol. 49, No. 2, P. 81
   

Copyright (C) Chemistry Dept., Moscow State University, 2002
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