Catalytic activities of a cocaine hydrolase engineered from human butyrylcholinesterase against (+)- and (-)-cocaine

Liu Xue, Shurong Hou, Wenchao Yang, Lei Fang, Fang Zheng, Chang Guo Zhan

Research output: Contribution to journalArticlepeer-review

13 Scopus citations


It can be argued that an ideal anti-cocaine medication would be one that accelerates cocaine metabolism producing biologically inactive metabolites via a route similar to the primary cocaine-metabolizing pathway, i.e., hydrolysis catalyzed by butyrylcholinesterase (BChE) in plasma. However, wild-type BChE has a low catalytic efficiency against naturally occurring (-)-cocaine. Interestingly, wild-type BChE has a much higher catalytic activity against unnatural (+)-cocaine. According to available positron emission tomography (PET) imaging analysis using [11C](-)-cocaine and [11C](+)- cocaine tracers in human subjects, only [11C](-)-cocaine was observed in the brain, whereas no significant [11C](+)-cocaine signal was observed in the brain. The available PET data imply that an effective therapeutic enzyme for treatment of cocaine abuse could be an exogenous cocaine-metabolizing enzyme with a catalytic activity against (-)-cocaine comparable to that of wild-type BChE against (+)-cocaine. Our recently designed A199S/F227A/S287G/A328 W/Y332G mutant of human BChE has a considerably improved catalytic efficiency against (-)-cocaine and has been proven active in vivo. In the present study, we have characterized the catalytic activities of wild-type BChE and the A199S/F227A/S287G/A328 W/Y332G mutant against both (+)- and (-)-cocaine at the same time under the same experimental conditions. Based on the obtained kinetic data, the A199S/F227A/S287G/A328 W/Y332G mutant has a similarly high catalytic efficiency (kcat/KM) against (+)- and (-)-cocaine, and indeed has a catalytic efficiency (kcat/K M = 1.84 × 109 M-1 min-1) against (-)-cocaine comparable to that (kcat/KM = 1.37 × 109 M-1 min-1) of wild-type BChE against (+)-cocaine. Thus, the mutant may be used to effectively prevent (-)-cocaine from entering brain and producing physiological effects in the enzyme-based treatment of cocaine abuse.

Original languageEnglish
Pages (from-to)57-62
Number of pages6
JournalChemico-Biological Interactions
Issue number1
StatePublished - Mar 25 2013

Bibliographical note

Funding Information:
This work was supported in part by the NIH (Grants R01 DA032910, R01 DA013930, and R01 DA025100 to Zhan) and the NSF (Grant CHE-1111761 to Zhan). The authors also acknowledge the Computer Center at University of Kentucky for supercomputing time on a Dell X-series Cluster with 384 nodes or 4,768 processors.


  • Cholinesterase
  • Cocaine addiction
  • Drug overdose
  • Enzyme therapy
  • Hydrolase

ASJC Scopus subject areas

  • Toxicology


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