Purification and enzymatic characterization of a novel β-1,6-glucosidase from Aspergillus oryzae

Journal of Bioscience and Bioengineering
Akira WatanabeK Gomi

Abstract

In this study, among the 10 genes that encode putative β-glucosidases in the glycoside hydrolase family 3 (GH3) with a signal peptide in the Aspergillus oryzae genome, we found a novel gene (AO090038000425) encoding β-1,6-glucosidase with a substrate specificity for gentiobiose. The transformant harboring AO090038000425, which we named bglH, was overexpressed under the control of the improved glaA gene promoter to form a small clear zone around the colony in a plate assay using 4-methylumbelliferyl β-d-glucopyranoside as the fluorogenic substrate for β-glucosidase. We purified BglH to homogeneity and enzymatically characterize this enzyme. The thermal and pH stabilities of BglH were higher than those of other previously studied A. oryzae β-glucosidases, and BglH was stable over a wide temperature range (4°C-60°C). BglH was inhibited by Hg(2+), Zn(2+), glucono-δ-lactone, glucose, dimethyl sulfoxide, and ethanol, but not by ethylenediaminetetraacetic acid. Interestingly, BglH preferentially hydrolyzed gentiobiose rather than other oligosaccharides and aryl β-glucosides, thereby demonstrating that this enzyme is a β-1,6-glucosidase. To the best of our knowledge, this is the first report of the purification and characterization of ...Continue Reading

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Citations

Mar 28, 2018·The Biochemical Journal·Hideyuki TakahashiTakumi Takeda
Nov 30, 2019·Applied Microbiology and Biotechnology·Tomohiko MatsuzawaKatsuro Yaoi
Jan 13, 2016·The Journal of Biological Chemistry·Tomohiko MatsuzawaKatsuro Yaoi
Apr 26, 2020·Scientific Reports·Lummy Maria Oliveira MonteiroMaria de Lourdes Teixeira de Moraes Polizeli
Jan 29, 2021·Journal of Food Science and Technology·Min SuChunhong Piao

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