Abstract
A new α-glucosidase from Shiraia sp. SUPER-H168 under solid-state fermentation was purified by alcohol precipitation and anion-exchange and by gel filtration chromatography. The optimum pH and temperature of the purified α-glucosidase were 4.5 and 60 °C, respectively, using p-nitrophenyl-α-glucopyranoside (α-pNPG) as a substrate. Ten millimoles of sodium dodecyl sulfate, Fe2+, Cu2+, and Ag+ reduced the enzyme activity to 0.7, 7.6, 26.0, and 6.2 %, respectively, of that of the untreated enzyme. The K m, V max, and k cat/K m of the α-glucosidase were 0.52 mM, 3.76 U mg−1, and 1.3 × 104 L s−1 mol−1, respectively. K m with maltose was 0.62 mM. Transglycosylation activities were observed with maltose and sucrose as substrates, while there was no transglycosylation with trehalose. DNA and its corresponding full-length cDNA were cloned and analyzed. The α-glucosidase coding region consisted of a 2997-bp open reading frame encoding a 998-amino acid protein with a 22-amino acid signal peptide; one 48-bp intron was located. The α-glucosidase was a monomeric protein with a predicted molecular mass of 108.2 kDa and a predicted isoelectric point of 5.08. A neighbor-joining phylogenetic tree demonstrated that Shiraia sp. SUPER-H168 α-glucosidase is an ascomycetes α-glucosidase. This is the first report of α-glucosidase from a filamentous fungus that had good glycoside hydrolysis with maltose and α-pNPG, transglycosylation and conversion activity of maltose into trehalose.
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Acknowledgements
This work was financially supported by the National High Technology and special funds for science and technology innovation from the Science and Technology Department of Jiangsu province (Grant No. BY2010117), the National Natural Science Foundation of China (Grant No. 21275066), and fundamental research of doctor of philosophy at 2014 (Grant No. 2050205), Project of graduate student scientific research and innovation of Jiangsu province at 2015 (Grant No. 205020502), Six talent peak program at 2016 (Grant No. SWYY-126).
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Gao, R., Deng, H., Guan, Z. et al. Purification, characterization and gene analysis of a new α-glucosidase from shiraia sp. SUPER-H168. Ann Microbiol 67, 65–77 (2017). https://doi.org/10.1007/s13213-016-1238-y
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DOI: https://doi.org/10.1007/s13213-016-1238-y