Over-Expression of the Thermobifida fusca β-Glucosidase in a Yarrowia lipolytica Transformant to Degrade Soybean Isoflavones
AbstractA 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
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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.
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(1):24.Chicago/Turabian Style
Chen, Wei-Lin; Yang, Yo-Ming; Guo, Gui-Wen; Chen, Cheng-Yu; Huang, Yu-Chun; Liu, Wen-Hsiung; Huang, Keh-Feng; Yang, Chao-Hsun. 2018. "Over-Expression of the Thermobifida fusca β-Glucosidase in a Yarrowia lipolytica Transformant to Degrade Soybean Isoflavones." Catalysts 8, no. 1: 24.
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