Publication: Expression and characterization of a recombinant stevioside hydrolyzing β-glycosidase from Enterococcus casseliflavus
Issued Date
2019-11-01
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ISSN
10465928
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2-s2.0-85069729738
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Mahidol University
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SCOPUS
Bibliographic Citation
Protein Expression and Purification. Vol.163, (2019)
Suggested Citation
Bootsakorn Boonkaew, Somsiri Udompaisarn, Dumrongkiet Arthan, Jamorn Somana Expression and characterization of a recombinant stevioside hydrolyzing β-glycosidase from Enterococcus casseliflavus. Protein Expression and Purification. Vol.163, (2019). doi:10.1016/j.pep.2019.105449 Retrieved from: https://repository.li.mahidol.ac.th/handle/20.500.14594/50048
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Title
Expression and characterization of a recombinant stevioside hydrolyzing β-glycosidase from Enterococcus casseliflavus
Abstract
© 2019 Elsevier Inc. The demand for steviol glycosides, non-caloric sweet components of Stevia rebaudiana Bertoni (stevia) leaves, has increased considerably as a benefit to enhance human health. However, the supply has remained challenging due to limited production, with the lack of a specific steviol glycoside hydrolyzing enzyme. In this study, a novel β-glucosidase (EcBgl) from Enterococcus casseliflavus was cloned and expressed in Escherichia coli. An EcBgl consists of 721 amino acids corresponding to a molecular mass of 79.37 kDa. The EcBgl was purified to homogeneity, followed by enzyme characterization. The enzyme showed optimum pH and temperature at 6.0 and 37 °C, and exhibited the kinetic constants kcat/Km for pNPG and kcat/Km for stevioside of 8583 mM−1s−1 and 95.41 mM−1s−1, respectively. When compared to the stevioside hydrolyzing β-glycosidases previously reported, EcBgl was found to be the most efficient enzyme. EcBgl also rendered hydrolysis of the stevioside to produce rubusoside, a rare steviol glycoside with a pharmaceutical solubilizing property, by cleaving at the glucose moiety. In addition, the enzyme demonstrated substantial resistance against amygdalin, so it served as a potential enzyme in agricultural and pharmaceutical applications.