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    Article

    Cloning, Prokaryotic Expression, and Enzyme Activity of a UDP‑Glucose Flavonoid 3‑O‑Glycosyltransferase from Mulberry (Morus alba L.) Leaves

    Xiaofeng Yu1, Jia Liu1, Jingqiong Wan1, Li Zhao1, Yaran Liu1, Yuan Wei1, Zhen Ouyang1★★ Corresponding author

    1. 1Department of Pharmaceutical Engineering, School of Pharmacy, Jiangsu University, Zhen Jiang, People’s Republic of, China.

    CORRESPONDENCE

    Zhen Ouyang

    Department of Pharmaceutical Engineering, School of Pharmacy, Jiangsu University, Zhen Jiang, People’s Republic of, China.

    zhenouyang@ujs.edu.cn

    Received: 07-09-2019; Revised: 31-10-2019; Accepted: 17-02-2020.

    Volume 16, Issue 69 · pp. 441–447 · PUBLISHED 15 June 2020 · DOI: 10.4103/pm.pm_396_19

    View on Pharmacogn. Mag. original site ↗

    ABSTRACT

    Objectives: In this article, MaUFGT, a cDNA encoding the UFGT from mulberry leaves, was cloned, codon optimized, and expressed in Escherichia coli to study its effect in vitro. Materials and Methods: Mulberry UDP- Glucose flavonoid‑3‑O‑glucanotransferase (MaUFGT) gene was reverse transcription‑polymerase chain reaction amplified using the cDNA obtained from young leaves of mulberry, and the full‑length MaUFGT gene was synthesized by codon‑optimized whole‑gene synthetic method. Then, the plasmid pCzn1/MaUFGT was constructed and heterologously expressed in E. coli. After denatured, renatured, and purified, the recombinant protein was used to evaluate its function in vitro by determining the final product by high‑performance liquid chromatography. Results: The target protein was in the range of 45–66 KD and mainly present in the form of inclusion bodies. The obtained protein was found to transfer UDP‑glucose glycosyl moieties to the 3‑hydroxyl group of quercetin or kaempferol to form the corresponding products in vitro. Conclusion: The MaUFGT was preliminarily proved to be involved in flavonoid 3‑O‑glucoside biosynthesis.

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      Yu, X., Liu, J., Wan, J., Zhao, L., Liu, Y., Wei, Y., & Ouyang, Z. (2020). Cloning, Prokaryotic Expression, and Enzyme Activity of a UDP‑Glucose Flavonoid 3‑O‑Glycosyltransferase from Mulberry (Morus alba L.) Leaves. Pharmacognosy Magazine, 16(69), 441–447. https://doi.org/10.4103/pm.pm_396_19