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M. I. Koksharov, N. N. Ugarova
The thermostability enhancement of Luciola mingrelicafirefly luciferase by random mutagenesis
Abstract
Firefly luciferase has a wide range of applications in a
number of areas of biotechnology and molecular biology, but its application is
often limited by the rapid inactivation at elevated temperatures. After four
consecutive rounds of random mutagenesis we obtained a Luciola mingrelica
firefly luciferase mutant with significantly increased thermostability. Amino
acid substitutions obtained also resulted in an increase of specific activity
and lowered a Michaelis constant for ATP 8-fold, which shows higher catalytic
efficiency of the mutant. Random mutagenesis was shown to be a highly efficient
approach for increasing luciferase stability.
Copyright (C) Chemistry Dept., Moscow State University, 2002
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