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Computational Design and Crystal Structure of a Highly Efficient Benzoylecgonine Hydrolase
Author(s) -
Chen Xiabin,
Deng Xingyu,
Zhang Yun,
Wu Yanan,
Yang Kang,
Li Qiang,
Wang Jiye,
Yao Weixuan,
Tong Junsen,
Xie Tian,
Hou Shurong,
Yao Jianzhuang
Publication year - 2021
Publication title -
angewandte chemie
Language(s) - English
Resource type - Journals
eISSN - 1521-3757
pISSN - 0044-8249
DOI - 10.1002/ange.202108559
Subject(s) - benzoylecgonine , chemistry , metabolite , hydrolase , enzyme , biochemistry , stereochemistry
Abstract Benzoylecgonine (BZE) is the major toxic metabolite of cocaine and is responsible for the long‐term cocaine‐induced toxicity owing to its long residence time in humans. BZE is also the main contaminant following cocaine consumption. Here, we identified the bacterial cocaine esterase (CocE) as a BZE‐metabolizing enzyme (BZEase), which can degrade BZE into biological inactive metabolites (ecgonine and benzoic acid). CocE was redesigned by a reactant‐state‐based enzyme design theory. An encouraging mutant denoted as BZEase2, presented a >400‐fold improved catalytic efficiency against BZE compared with wild‐type (WT) CocE. In vivo, a single dose of BZEase2 (1 mg kg −1 , IV) could eliminate nearly all BZE within only two minutes, suggesting the enzyme has the potential for cocaine overdose treatment and BZE elimination in the environment by accelerating BZE clearance. The crystal structure of a designed BZEase was also determined.