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Published on: February 21, 2025
Characterization of active Lentinula edodes glucoamylase expressed and secreted by Saccharomyces cerevisiae
Dominic W S Wong1, Sarah B Batt, Charles C Lee
1Western Regional Research Center, USDA-ARS, 800 Buchanan Street, Albany, California 94710, USA. dwsw@pw.usda.gov
Abstract:
The gene encoding Lentinula edodes glucoamylase (GLA) was cloned into Saccharomyces cerevisiae, expressed constitutively and secreted in an active form. The enzyme was purified to homogeneity by (NH4)2SO4 fractionation, anion exchange and affinity chromatography. The protein had a correct N-terminal sequence of WAQSSVIDAYVAS, indicating that the signal peptide was efficiently cleaved. The recombinant enzyme was glycosylated with a 2.4% carbohydrate content. It had a pH optimum of 4.6 and a pH 3.4-6.4 stability range. The temperature optimum was 50 degrees C with stability
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