发明名称 |
High-activity mutants of butyrylcholinesterase for cocaine hydrolysis and method of generating the same |
摘要 |
A novel computational method and generation of mutant butyrylcholinesterase for cocaine hydrolysis is provided. The method includes molecular modeling a possible BChE mutant and conducting molecular dynamics simulations and hybrid quantum mechanical/molecular mechanical calculations thereby providing a screening method of possible BChE mutants by predicting which mutant will lead to a more stable transition state for a rate determining step. Site-directed mutagenesis, protein expression, and protein activity is conducted for mutants determined computationally as being good candidates for possible BChE mutants, i.e., ones predicted to have higher catalytic efficiency as compared with wild-type BChE. In addition, mutants A199S/A328W/Y332G, A199S/F227A/A328W/Y332G, A199S/S287G/A328W/Y332G, A199S/F227A/S287G/A328W/Y332G, and A199S/F227A/S287G/A328W/E441D all have enhanced catalytic efficiency for (−)-cocaine compared with wild-type BChE. |
申请公布号 |
US8835150(B1) |
申请公布日期 |
2014.09.16 |
申请号 |
US201313767418 |
申请日期 |
2013.02.14 |
申请人 |
University of Kentucky Research Foundation |
发明人 |
Zhan Chang-Guo;Cho Hoon;Tai Hsin-Hsiung |
分类号 |
C12N9/16;C07H21/04 |
主分类号 |
C12N9/16 |
代理机构 |
Stites & Harbison PLLC |
代理人 |
Stites & Harbison PLLC ;Decker Mandy Wilson |
主权项 |
1. A nucleic acid molecule comprising a nucleic acid sequence which encodes a butyrylcholinesterase variant peptide, said nucleic acid sequence consisting of SEQ ID NO: 25. |
地址 |
Lexington KY US |