Computational Design and Crystal Structure of a Highly Efficient Benzoylecgonine Hydrolase

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‐...

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Published in:Angewandte Chemie International Edition 2021-09, Vol.60 (40), p.21959-21965
Main Authors: Chen, Xiabin, Deng, Xingyu, Zhang, Yun, Wu, Yanan, Yang, Kang, Li, Qiang, Wang, Jiye, Yao, Weixuan, Tong, Junsen, Xie, Tian, Hou, Shurong, Yao, Jianzhuang
Format: Article
Language:English
Subjects:
Benzoic acid
benzoylecgonine
Biocompatibility
CHARMM rotamer force field
Cocaine
cocaine detoxification
Computer applications
Contaminants
Crystal structure
enzyme therapy
Enzymes
Esterase
Hydrolase
Metabolites
Overdose
protein design
Toxicity
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Summary: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. Benzoylecgonine (BZE) is the major toxic metabolite of cocaine and is responsible for cocaine‐induced long‐term toxicity owing to its long residence time in humans. We computationally identified, re‐designed, and crystallized a novel BZEase. The obtained BZEase mutant shows a 427‐fold improved catalytic efficiency toward BZE compared to WT BZEase. In vivo tests prove that the BZEase mutant is a promising treatment for BZE detoxification.
ISSN:1433-7851
1521-3773
DOI:10.1002/anie.202108559