苷元
化学
蛋白质工程
类黄酮
糖基化
定向进化
酶
活动站点
依托泊苷
糖苷
立体化学
生物化学
生物
突变体
基因
化疗
抗氧化剂
遗传学
作者
Fei-Fan Tan,Rui Zhu,Bin Xiong,Guimin Zhang,Wei Zhao,Kai-Zhi Jia
标识
DOI:10.1021/acssynbio.2c00032
摘要
Enzyme entrances, which function as the first molecular filters, influence substrate selectivity and enzymatic activity. Because of low binding affinities, engineering enzyme entrances that recognize non-natural substrates is a major challenge for artificial biocatalyst design. Here, the entrance of flavonoid glycosyltransferase UGT78D2 was engineered to promote the recognition of the aglycone of etoposide, a chemotherapeutic agent. We found that Q258, S446, R444, and R450, the key residues surrounding the substrate entrance, specifically guide the flux of etoposide aglycone, which has a high steric hindrance, into the active site; this activity was inferred to be determined by the entrance size and hydrophobic and electrostatic interactions. Engineering the coordination of Q258 and S446 to increase the entrance size and hydrophobic interaction between UGT78D2 and etoposide aglycone increased the affinity by 10.10-fold and the conversion by 10%. The entrance-engineering strategy applied in this study can improve the design of artificial biocatalysts.
科研通智能强力驱动
Strongly Powered by AbleSci AI