Study On Asymmetric Oxidation Resolution Of Catalyzing 2-substituted Tetrahydroquinolines Derivatives By Whole Cell Of Pseudomonas Monteilii

Objective: Screening and obtaining high activity and high selectivity of monoamine oxidase producing strain for catalyzing the asymmetric oxidative resolution of 2-substituted tetrahydroquinoline derivatives,further understanding the enzymatic properties of monoamine oxidase in the strain and constructing the monoamine oxidase recombinant strain.Methods: 2-methyl-1,2,3,4-tetrahydroquinoline was used as template substrate.Suspension-whole cell catalysis was used to screen highly active biocatalysts.The reaction conditions were optimized,and 2-substituted tetrahydroquinoline and the synthesis of the 2-substituted tetrahydroquinoline derivatives with high optical activity;and the production of monoamine oxidase recombinant strain derived from Pseudomonas monteilii ZMU-T01 strain.Results: 1.The results showed that the whole cell viability of Pseudomonas monteilii ZMU-T01 strain had good catalytic activity on the template substrate 2-methyl-1,2,3,4-tetrahydroquinoline.At the substrate concentration of 4 m M,the cell concentration was 50 g cdw/L,p H 8.0,5 m L reaction system,the substrate conversion rate of 47%,the ee value of 90%.On this basis,eight similar structures were expanded to obtain chiral 2-substituted tetrahydroquinones with an ee of 89-99%.2.The optimum conditions for the reaction were as follows: substrate concentration was 2 m M,cell concentration was 50 g cdw/L,buffer solution was PBS(p H = 8.0,50 m M),and the optimal concentration of the reaction was determined by the control variable method.The reaction volume was 5 m L,the reaction temperature was 30 oC,the rotation speed was 250 rpm,and the reaction time was 24 h.3.In the optimum reaction system,the properties of 2-substituted tetrahydroquinoline substrates were investigated.When the substituents on the benzene ring were electron-with drawing groups,Pseudomonas monteilii ZMU-T01 cell was not catalyzed,When the 2-position substituents were electron-donating groups,the wholecells had good catalytic activity,and 8 high-optically active compounds were synthesized.All the structures were analyzed by NMR,chiral HPLC,optical rotation and HRMS Confirmation and evidence.4.Based on the sequence analysis of Pseudomonas monteilii ZMU-T01 whole cell gene as a template,two genes were designed by molecular biology technique,and a bio-catalyzed bacteria containing monoamine oxidase were successfully obtained.Conclusion: 1.Screening of the Pseudomonas monteilii ZMU-T01 strain can efficiently catalyze the asymmetric oxidative resolution of 2-substituted tetrahydroquinoline derivatives and synthesize 2-substituted tetrahydroquinoline derivatives with high optical activity and Which has a good substrate adaptation range.It provides an effective method for the green synthesis of chiral 2-substituted tetrahydroquinoline derivatives.2.The genome of Pseudomonas monteilii ZMU-T01 was analyzed and two gene sequences were designed.A genetically engineered strain with monoamine oxidase activity was cloned,which established a basis for the follow-up study.