Abstract
In this paper, the cgt gene encoding cyclodextrin glycosyltransferase (CGTase) from Bacillus stearothermophilus was cloned into pWB980 plasmid for extracellular expression in Bacillus subtilis SCK6. Through adding a six-histidine affinity tag fused to the C-terminus, the recombinant CGTase could be purified by nickel ion affinity chromatography, and its molecular weight was approximately 76 kDa on SDS-PAGE. Then, the enzymatic properties were determined, and results were as follows: the optimum temperature and pH were identified as 40 ℃ and pH 5.0, respectively. CGTase had good tolerance to metal ions of Mn2+, Ca2+, and Mg2+. The enzyme activity was activated by Na+, Al3+, Fe3+, and Ni+, and it was remarkably inhibited by Cu2+ and Zn2+. To improve the aqueous solubility of rutin, CGTase was used to catalyze the transglycosylation reaction, and the conversion rate could reach as high as 80.13% under optimal conditions. Furthermore, the reaction mixture was treated with glucoamylase and microporous adsorbent resin. The yield of glycosyl-rutin was 56.1%, and its purity was 74.3%, which further improved the value of the product.
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Acknowledgements
This research was financially supported by the National Natural Science Foundation of China (31600639), Science and Technology Project Founded by the Education Department of Jiangxi Province (GJJ202305).
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Conceptualization: JZ and XL; data curation: WS and MZ; funding acquisition: JZ and XL; investigation: WS, MZ and YZ; supervision: JZ and XL; writing original draft: WS and MZ.
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Song, W., Zhang, M., Li, X. et al. Heterologous expression of cyclodextrin glycosyltransferase from Bacillus stearothermophilus in Bacillus subtilis and its application in glycosyl rutin production. 3 Biotech 13, 84 (2023). https://doi.org/10.1007/s13205-023-03510-5
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DOI: https://doi.org/10.1007/s13205-023-03510-5