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KMID : 0613820040140030435
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Journal of Life Science
2004 Volume.14 No. 3 p.435 ~ p.440
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Overexpression of the gct Gene Encoding 4-α-Glucanotransferase of a Hyperthermophilic Archaeon, Thermococcus litoralis
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Jeon Beong-Sam
Park Jeong-Won
Shin Gab-Gyun
Kim Beom-Kyu
Kim Hee-Kyu
Song Jae-Young
Cho Young-Su
Cha Jae-Young
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Abstract
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The gene encoding a extremely thermostable 4-α-glucanotransferase from a hyperthermophilic archaeon, Thermococcus litoralis, was cloned, sequenced and expressed in Escherichia coli. The amino acid sequence of the enzyme was distantly related to other functionally-related ones, such as D-enzymes. The enzyme is of industrial interest because of a novel activity of producing cycloamylose and is also important for fundamental studies of protein, sugar-metabolizing enzymes. In this paper, the overexpression of 4-α-glucanotransferase in E. coli was carried out expression vector system with lac and T7 promoters. The enzyme was successfully overexpressed, and purified by the heat treatment of a cell-free extract, successive Butyl-Toyopearl and Mono Q chromatographies. The purified recombinant enzyme showed the same specific activity and the same mobility in SDS-PAGE as natural enzyme
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KEYWORD
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Overexpression, 4-α-Glucanotransferase, Hyperthermophilic archaeon, Thermococcus litoralis, Thermostable enzyme
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