KMID : 1007520110200020561
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Food Science and Biotechnology 2011 Volume.20 No. 2 p.561 ~ p.565
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Expression, Purification, and Characterization of Human Intestinal Maltase Secreted from Pichia pastoris
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Ryu Hwa-Ja
Seo Eun-Seong Kang Hee-Kyoung Kim Young-Min Kim Do-Man
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Abstract
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A gene encoding human intestinal maltase (HMA) was successfully expressed in Pichia pastoris under the control of the methanol-induced alcohol oxidase (AOX1) promoter. The secreted recombinant HMA fused with a His6-tag was produced (150 U/L) and was easily purified from culture supernatants in a 3-step diafiltration, ultrafiltration, and affinity column chromatography protocol. The specific activity of the purified HMA was 16.8 U/mg. Endoglycosidase H digestion of the protein showed that the recombinant HMA was N-glycosylated. The purified HMA was maximally active at pH 6.5 and stable (¡Ã90%) up to 65oC. The kinetic parameters Km and Vmax were 3.3¡¾0.25 mM maltose and 61.9¡¾2 U/mg, respectively.
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KEYWORD
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¥á-glucosidase, maltase, diabetes, Pichia pastoris, expression in yeast
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