Abstract
Recombinant barley α-amylase 1 isozyme was constitutively secreted by Saccharomyces cerevisiae. The enzyme was purified to homogeneity by ultrafiltration and affinity chromatography. The protein had a correct N-terminal sequence of His-Gln-Val-Leu-Phe-Gln-Gly-Phe-Asn-Trp, indicating that the signal peptide was efficiently processed. The purified α-amylase had an enzyme activity of 1.9 mmol maltose/mg protein/min, equivalent to that observed for the native seed enzyme. The k cat/K m was 2.7 × 102 mM−1.s−1, consistent with those of α-amylases from plants and other sources.
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Wong, D.W.S., Batt, S.B. & Robertson, G.H. Characterization of Active Barley α-Amylase 1 Expressed and Secreted by Saccharomyces cerevisiae. J Protein Chem 20, 619–623 (2001). https://doi.org/10.1023/A:1013712101741
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DOI: https://doi.org/10.1023/A:1013712101741