Abstract
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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Acknowledgments
This research was supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology (2010-0025345), and this work was also financially supported by the Ministry of Knowledge Economy (MKE) and Korea Institute for Advancement in Technology (KIAT) through the Workforce Development Program in Strategic Technology.
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Song, H., Park, E.J., Shin, YH. et al. Effect of polyhistidine-tagging site on the stability of recombinant alginate lyase from Streptomyces sp. ALG-5. Journal of Pharmaceutical Investigation 42, 15–19 (2012). https://doi.org/10.1007/s40005-012-0007-6
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DOI: https://doi.org/10.1007/s40005-012-0007-6