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 65°C. The kinetic parameters K m and V max were 3.3±0.25 mM maltose and 61.9±2 U/mg, respectively.
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Ryu, HJ., Seo, ES., Kang, HK. et al. Expression, purification, and characterization of human intestinal maltase secreted from Pichia pastoris . Food Sci Biotechnol 20, 561–565 (2011). https://doi.org/10.1007/s10068-011-0079-5
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DOI: https://doi.org/10.1007/s10068-011-0079-5