Expression, Isolation, Purification, and Biochemical Properties of Trehalose-6-phosphate Hydrolase from Thermoresistant Strain Bacillus sp. GP16

D. V. Karelov, V. V. Krasikov, M. A. Surjik, and L. M. Firsov^*

Konstantinov St. Petersburg Institute of Nuclear Physics, Russian Academy of Sciences, Gatchina 188350, Leningrad Region, Russia; E-mail: lfirsov@mail.wplus.net

^* To whom correspondence should be addressed.

Received July 24, 2002; Revision received October 29, 2002

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Here we describe cloning, expression, and purification of the enzyme trehalose-6-phosphate hydrolase from thermoresistant strain Bacillus sp. GP16. Principal biochemical properties of the enzyme at different pH and temperature values were determined. Entropy and enthalpy of activation of the enzyme for substrates trehalose-6-phosphate and p-nitrophenyl glucoside were calculated, and the dependence of the kinetic parameters from ionic strength was established.

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KEY WORDS: trehalose-6-phosphate hydrolase, cloning of a gene, gene expression, catalysis, temperature dependence, determination of Michaelis constants

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