糖基转移酶
糖基化
生物化学
酶
苷元
糖苷
化学
生物合成
立体化学
蛋白质工程
底物特异性
转移酶
基质(水族馆)
生物
生态学
作者
Kai-Zhi Jia,Liwen Zhu,Xudong Qu,Shengying Li,Yuemao Shen,Qingsheng Qi,Youming Zhang,Yuezhong Li,Ya‐Jie Tang
标识
DOI:10.1021/acssynbio.9b00318
摘要
The 4-O-β-d-glucopyranoside of DMEP ((−)-4′-desmethylepipodophyllotoxin) (GDMEP), a natural product from Podophyllum hexandrum, is the direct precursor to the topoisomerase inhibitor etoposide, used in dozens of chemotherapy regimens for various malignancies. The biosynthesis pathway for DMEP has been completed, while the enzyme for biosynthesizing GDMEP is still unclear. Here, we report the enzymatic O-glycosylation of DMEP with 53% conversion by exploring the substrate promiscuity and entrances of glycosyltransferases. Notably, we found 6 essential amino acid residues surrounding the putative substrate entrances exposed to the protein surface in UGT78D2, CsUGT78D2, and CsUGT78D2-like, and these residues may determine substrate specificity and high O-glycosylation activity toward DMEP. Our results provide an effective route for one-step synthesis of GDMEP. Identification of the key residues and entrances of glycosyltransferases will promote precise identification of glycosyltransferase biocatalysts for novel substrates and provide a rational basis for glycosyltransferase engineering.
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