Directed evolution of stereoselective enzymes meets click reactions: Asymmetric synthesis of chiral triazoles using a Cu(I)-compatible halohydrin dehalogenaseOA
We report for the first time the combination of directed evolution focused on enhancing and reversing the stereoselectivity of an enzyme with Cu(I)-mediated click chemistry(CuAAC),providing an asymmetric click approach for versatile chiral triazoles products.In this study,the halohydrin dehalogenase HheG was used as the enzyme which was evolved to induce a stereoselective ring-opening reaction of cyclic epoxides in the presence of NaN3 with the formation of chiral azido products.Two mutants of opposite stereopreference were generated,which convert cyclohexene oxide as well as cycloheptene oxide to(1S,2S)-2-azidocyclohexanol,(1R,2R)-2-azidocyclohexanol,(1S,2S)-2-azidocycloheptanol and(1R,2R)-2-azidocycloheptanol with essentially high stereoselectivity.The chiral products were then subjected to CuAAC in reactions with structurally different alkynes.Since HheG was found to be compatible with Cu(I),the process was also performed successfully in a unique 2-step one-pot process leading to various chiral triazoles.In order to understand the enhancement and reversal of the evolved enantioselectivity,QM and MD computations were performed.This approach harnesses the modifiability and high stereoselectivity of the evolved biocatalysts in combination with click chemistry.It holds great potential for diverse fields,particularly in the area of pharmaceuticals.
Shaixiao Tian;Xinying Ge;Qipeng Yan;Min Li;Qun Huang;Xinhua Zhang;Ming Ma;Bo Chen;Jian-bo Wang
Key Laboratory of Chemical Biology and Traditional Chinese Medicine Research(Ministry of Education)and Key Department Laboratory of Phytochemical R&D of Hunan Province,College of Chemistry and Chemical Engineering,Hunan Normal University,Changsha 410081,ChinaKey Laboratory of Chemical Biology and Traditional Chinese Medicine Research(Ministry of Education)and Key Department Laboratory of Phytochemical R&D of Hunan Province,College of Chemistry and Chemical Engineering,Hunan Normal University,Changsha 410081,ChinaKey Laboratory of Chemical Biology and Traditional Chinese Medicine Research(Ministry of Education)and Key Department Laboratory of Phytochemical R&D of Hunan Province,College of Chemistry and Chemical Engineering,Hunan Normal University,Changsha 410081,ChinaKey Laboratory of Chemical Biology and Traditional Chinese Medicine Research(Ministry of Education)and Key Department Laboratory of Phytochemical R&D of Hunan Province,College of Chemistry and Chemical Engineering,Hunan Normal University,Changsha 410081,ChinaSchool of Pharmacy and Bioengineering,Chongqing University of Technology,Chongqing 400054,ChinaKey Laboratory of Chemical Biology and Traditional Chinese Medicine Research(Ministry of Education)and Key Department Laboratory of Phytochemical R&D of Hunan Province,College of Chemistry and Chemical Engineering,Hunan Normal University,Changsha 410081,ChinaKey Laboratory of Chemical Biology and Traditional Chinese Medicine Research(Ministry of Education)and Key Department Laboratory of Phytochemical R&D of Hunan Province,College of Chemistry and Chemical Engineering,Hunan Normal University,Changsha 410081,ChinaKey Laboratory of Chemical Biology and Traditional Chinese Medicine Research(Ministry of Education)and Key Department Laboratory of Phytochemical R&D of Hunan Province,College of Chemistry and Chemical Engineering,Hunan Normal University,Changsha 410081,ChinaKey Laboratory of Chemical Biology and Traditional Chinese Medicine Research(Ministry of Education)and Key Department Laboratory of Phytochemical R&D of Hunan Province,College of Chemistry and Chemical Engineering,Hunan Normal University,Changsha 410081,China Key Laboratory of Multiple Organ Failure(Zhejiang University)Ministry of Education,Department of General Intensive Care Unit of the Second Affiliated Hospital and Institute of Pharmaceutical Biotechnology,Zhejiang University School of Medicine,Hangzhou 310058,China Zhejiang Provincial Key Laboratory for Microbial Biochemistry and Metabolic Engineering,Hangzhou 310058,China
化学化工
Directed evolutionChemo-enzymatic synthesisHalohydrin dehalogenasesChiral triazolesAsymmetric click reaction
《Green Synthesis and Catalysis》 2026 (1)
P.75-82,8
support from the National Key Research and Development Program of China(No.2023YFA0914100/2023YFA0914102)the National Natural Science Foundation of China(Nos.22077029 and 22034002)the Science Fund for Distinguished Young Scholars of Hunan Province(No.2021JJ10034)support from the National Natural Science Foundation of China(No.22276049)M.M.acknowledge the support from the National Natural Science Foundation of China(No.22276050).
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