Immobilization on macroporous resin makes E. coli RutB a robust catalyst for production of (-) Vince lactam.

摘要: A novel (+) γ-lactamase gene (rutB) was cloned from Escherichia coli JM109 and expressed in E. BL21 (DE3), the recombinant protein characterized. The optimal conditions for enzyme were pH 7.0 temperature 30 °C, which indicated that it a mesophilic protein. free purified deactivated when incubated at 50 °C min. However, kcat value of RutB its about 2.5 times archaeal Sulfolobus sofataricus (85 °C). After immobilization on macroporous resin using glutaraldehyde cross-linkage, thermostability crude greatly enhanced deactivating raised to 70 °C. immobilization, minimal substrate inhibition concentration also improved 0.75 1.5 M. concentrations immobilized determined be 250 mg/ml M, initial reaction velocity response variable batch transformations. This resins provides another feasible approach preparation optically active Vince lactam. As member isochorismatase superfamily, demonstrated typical showed catalytic promiscuity.