|
KMID : 1007520110200020561
|
|
Food Science and Biotechnology
2011 Volume.20 No. 2 p.561 ~ p.565
|
|
|
Expression, Purification, and Characterization of Human Intestinal Maltase Secreted from Pichia pastoris
|
|
Ryu Hwa-Ja
Seo Eun-Seong
Kang Hee-Kyoung
Kim Young-Min
Kim Do-Man
|
|
|
Abstract
|
|
|
|
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 (≥90%) up to 65oC. The kinetic parameters Km and Vmax were 3.3±0.25 mM maltose and 61.9±2 U/mg, respectively.
|
|
|
KEYWORD
|
|
|
α-glucosidase, maltase, diabetes, Pichia pastoris, expression in yeast
|
|
|
FullTexts / Linksout information
|
|
|
|
|
|
Listed journal information
|
|
 
|