The thermostable GH3 family β-glucosidase DthBgl3 and thermostable GH78 family α-l-rhamnosidase DthRha from Dictyoglomus thermophilum DSM3960 were successfully immobilized by industrial amino resin 1000NH. The optimal reaction temperature and pH of 1000NH-DthBgl3 was 85 °C and pH 5.0. Over 65% residual enzyme activity maintained for 1000NH-DthBgl3 after a 3-h incubation under 85 °C, while about 20% residual enzyme activity maintained for free DthBgl3. The optimal reaction temperature and pH of 1000NH-DthRha was 95 °C and pH 6.5. Over 90% residual enzyme activity maintained for 1000NH-DthRha after a 3-h incubation under 90 °C, while about 22% residual enzyme activity maintained for free DthRha. Meanwhile, immobilized 1000NH-DthBgl3 and 1000NH-DthRha were successfully and could completely transform all major ingredient of 10 g/L total flavonoids extract from Epimedium (TFEE) into icaritin. After 15 cycles (45 h) of repeated use at 85 °C, 3 U 1000NH-DthBgl3 showed a molar conversion rate of 73.12%, initial activity of 30.09% and productivity of 124 mg/L/h, while 15 U 1000NH-DthRha showed a molar conversion rate of 88.50%, initial activity of 85.31% and productivity of 75 mg/L/h after ten cycles (60 h) of repeated use at 85 °C. The cooperation of two immobilized enzymes showed a molar conversion rate of 87.21% and productivity of 141 mg/L/h after 15 cycles of repeated use at 85 °C. This is the first report on immobilization of two thermostable glycosidases from thermophilic bacteria and their cooperated biocatalysis of all major ingredients of TFEE into icaritin with high productivity under a high substrate concentration.
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This work was supported by the National Natural Science Foundation of China (21978135), the National Key R & D Program of China (2016YFD0600805), and the Jiangsu “333” project of cultivation of high-level talents (BRA2015317).
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Dong, Y., Zhang, S., Lu, C. et al. Immobilization of Thermostable β-Glucosidase and α-l-Rhamnosidase from Dictyoglomus thermophilum DSM3960 and Their Cooperated Biotransformation of Total Flavonoids Extract from Epimedium into Icaritin. Catal Lett (2021). https://doi.org/10.1007/s10562-020-03522-3
- Amino resin
- Thermostable glycosidase