Computational redesign of penicillin acylase for cephradine synthesis with high kinetic selectivity
Computational redesign of penicillin acylase for cephradine synthesis with high kinetic selectivity
复制标题
用于高动力学选择性头孢拉定合成的青霉素酰化酶的计算重新设计
DOI：
10.1039/c8gc03420f
复制
发表时间：
2018-12-21
期刊：
影响因子：
9.8
通讯作者：
Zhu, Yushan
中科院分区：
文献类型：
--
作者：
He, Jinwen;Huang, Xiaoqiang;Zhu, Yushan
Computational redesign of native enzyme active sites for non-natural substrates offers a general approach for creating biocatalysts in green chemistry. Herein, a computational strategy was developed to discover selective mutants of penicillin acylase to catalyze the condensation reaction between D-dihydrophenylglycine methyl ester (DHME) and 7-aminodesacetoxy cephalosporanic acid (7-ADCA), producing cephradine in fully aqueous medium. The key feature of this strategy is that mutants favoring the binding of the near-attack conformation of cephradine in the catalytic orientation were computationally selected using a scoring function based on discounted folding energy relative to binding energy. Using this strategy, we obtained a penicillin acylase mutant (M142αF/F24βA/S67βA) with high kinetic selectivity that increased the synthesis/hydrolysis ratio (S/H) by more than 10-fold compared with the wild-type. In its immobilized form, the redesigned triple mutant attained up to 99% yield under industrial conditions. This study represents a breakthrough in enzymatic synthesis of cephradine and suggests that computational design strategies can adapt an enzyme to catalyze non-natural chemical transformations for green process development with industrial significance.
