Direct selective hydroxylation, one of the most convenient and economical routes employed in C-H oxyfunctionalization has been widely utilized in various of fields, including medicine, chemical industry, materials science, and so on.
Despite remarkable progress in C-H hydroxylation, it is a great challenge to optionally access diverse hydroxylation products from a given substrate bearing multiple reaction sites of sp3 and sp2 C-H bonds.
Recently, a research team led by Prof. CONG Zhiqi from the Qingdao Institute of Bioenergy and Bioprocess Technology (QIBEBT) of the Chinese Academy of Sciences (CAS) has developed a new “ synergistic using protein engineering and exogenous molecules” approach to achieve multi-site highly regio- and enantioselective hydroxylation of alkylbenzenes.
The study was published in Angewandte Chemie International Edition.
In a series of multi-year experiments, the team obtained various combinations of P450BM3 mutants and DFSM, and uesd these specific combinations to access more than half of the possible hydroxylated products from each substrate with excellent regioselectivity (up to >99%), enantioselectivity (up to >99% ee), and high total turnover numbers (up to 80,963), which is far better than any natural or engineered P450 monooxygenases previously reported. The synergistic catalysis induced by the bound DFSMs was key to generating peroxygenase activity, product diversity, and selectivity.
Additionally, the results of crystal structure analysis, molecular dynamic simulations and theoretical calculations provided direct evidence of the synergistic effects between the engineered P450 variants and DFSMs during alkylbenzene hydroxylation, supporting the occurrence of accurately controlled regio- and enantioselectivity at various sites.
"The study findings not only demonstrate the power of synergistic use of engineered P450 enzymes and DFSMs to achieve regiodivergent and enantioselective hydroxylation of alkylbenzenes, but also highlight the potential of exogenous molecules in modulating enzymatic catalysis for biochemical transformations. " said corresponding author Prof. CONG Zhiqi from QIBEBT.
The study was supported by National Natural Science Foundation of China, Qingdao Innovative Leading Talent Project, National Key Research and Development Program of China and Program for Postdoctoral Innovative Talents of Shandong Province, China etc.
Divergent hydroxylation of alkylbenzenes catalyzed by synergistic use of P450BM3 variants and DFSMs. (Image by CONG Zhiqi and CHEN Jie)
(Text by CHEN Jie)
Contact:
KONG Fengru
Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of Sciences
Tel: 86-532-58261072
E-mail: kongfr@qibebt.ac.cn
