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Catalysts 2018, 8(1), 24; https://doi.org/10.3390/catal8010024

Over-Expression of the Thermobifida fusca β-Glucosidase in a Yarrowia lipolytica Transformant to Degrade Soybean Isoflavones

1
Department of Applied Chemistry, Providence University, Taichung 43301, Taiwan
2
Department of Cosmetic Science, Providence University, Taichung 43301, Taiwan
3
Department of Biochemical Science and Technology, National Taiwan University, Taipei 10617, Taiwan
4
Xtremes Pure Company, Taipei 10652, Taiwan
*
Author to whom correspondence should be addressed.
Received: 4 January 2018 / Revised: 12 January 2018 / Accepted: 12 January 2018 / Published: 14 January 2018
(This article belongs to the Special Issue Novel Enzyme and Whole-Cell Biocatalysis)
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Abstract

A gene (bgl) encoding a β-glucosidase in thermophilic actinomycete Thermobifida fusca NTU 22 was cloned into a Yarrowia lipolytica expression system. Heterologous expression resulted in extracellular β-glucosidase production with activity as high as 630 U/mL in a Hinton flask culture filtrate. This recombinant β-glucosidase was purified 9.2-fold from crude culture filtrate by DEAE-Sepharose FF column chromatography as measured by its increase in specific activity. The overall yield of the purified enzyme was 47.5%. The molecular weight of the purified β-glucosidase estimated by SDS-PAGE was 45 kDa, which agreed with the predicted molecular weight based on the nucleotide sequence. About 15% enzyme activity loss was observed after the enzyme was heat-treated at 50 °C for 180 min. It was also found that the activity of the enzyme was inhibited by Hg2+, Cu2+, Ba2+, Ag+, p-chloromercuribenzene, and iodoacetate. The β-glucosidase from T. fusca had the most activity for daidzein-7-glucoside and genistein-7-glucoside among the tested flavonoid glycosides, but there was moderate or little activity for luteolin-7-glucoside, cyanidine-3-glucoside, and quercetin-3-glucoside. These properties are important for the soybean isoflavone applications of this β-glucosidase. View Full-Text
Keywords: Thermobifida fuscal; β-glucosidase; Yarrowia lipolytica; daidzin; genistin Thermobifida fuscal; β-glucosidase; Yarrowia lipolytica; daidzin; genistin
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Chen, W.-L.; Yang, Y.-M.; Guo, G.-W.; Chen, C.-Y.; Huang, Y.-C.; Liu, W.-H.; Huang, K.-F.; Yang, C.-H. Over-Expression of the Thermobifida fusca β-Glucosidase in a Yarrowia lipolytica Transformant to Degrade Soybean Isoflavones. Catalysts 2018, 8, 24.

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