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Production, Crystallization and Structure Determination of C. difficile PPEP-1 via Microseeding and Zinc-SAD
Published on: December 30, 2016
Protein preparation, crystallization and preliminary X-ray analysis of imidazolonepropionase from Bacillus subtilis
Yamei Yu1, Lanfen Li, Xiaofeng Zheng
1The National Laboratory of Protein Engineering and Plant Genetic Engineering, College of Life Sciences, Peking University, Beijing 100871, PR China.
Abstract:
Imidazolonepropionase (EC 3.5.2.7) is the third enzyme of the histidine degradation pathway that has been conserved from bacteria to eukaryotes. The enzyme is the only one with unknown three-dimensional structure in this pathway. In this work, Bacillus subtilis imidazolonepropionase (HutI) was expressed in E. coli and purified to homogeneity. After thrombin digestion, high quality crystals were obtained by hanging-drop vapor diffusion method. The best crystal diffracted to 2.0 A and belonged to the space group P2(1) with unit-cell parameters a = 57.73 A, b = 106.34 A, c = 66.47 A, beta = 89.93 degrees .

