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KMID : 0369820120420010015
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Jorunal of Korean Pharmaceutical Sciences
2012 Volume.42 No. 1 p.15 ~ p.19
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Effect of polyhistidine-tagging site on the stability of recombinant alginate lyase from Streptomyces sp. ALG-5
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Song Hee-Sub
Park Eun-Ji
Shin Young-Hee
Kim Hee-Sook
Na Dong-Hee
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Abstract
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The purpose of this study was to investigate the effect of polyhistidine (His)-tagging site on the stability of alginate lyase from a marine bacterium Streptomyces species ALG-5 by the combined use of microchip electrophoresis and enzymatic depolymerizing activity assay. In microchip electrophoresis, C-terminally His-tagged alginate lyase (C-His-AL) was more stable than N-terminally His-tagged alginate lyase (N-His-AL) after the incubation in 50 mM potassium phosphate buffer (pH 7.0) at 37°C for 14 days. When the enzymatic depolymerizing activity of the same samples was measured, the activity of C-His-AL was not significantly changed for 14 days, whereas N-His-AL showed substantially declined activity after incubation. Consequently, this study demonstrated that the C-terminally His-tagging is more efficient than N-terminally His-tagging for preparing stable ALG-5 alginate lyase.
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KEYWORD
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Alginate lyase, Stability, Enzymatic depolymerizing activity, Microchip electrophoresis
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