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Expression, purification, and characterization of human intestinal maltase secreted from Pichia pastoris

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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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Authors and Affiliations

  1. The Research Institute for Catalysis and School of Biological Sciences and Technology, Chonnam National University, Gwangju, 500-757, Korea

    Hwa-Ja Ryu, Eun-Seong Seo, Hee-Kyoung Kang & Doman Kim

  2. Eco-Friendly Biomaterial Research Center and AI Control Material Research Center, Korea Research Institute of Bioscience and Biotechnology, Jeongeup, Jeonbuk, 580-185, Korea

    Young-Min Kim

Authors
  1. Hwa-Ja Ryu
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  2. Eun-Seong Seo
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  3. Hee-Kyoung Kang
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  4. Young-Min Kim
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  5. Doman Kim
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Correspondence to Doman Kim.

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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

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