Cloning expression crystallization and preliminary X-ray crystallographic analysis of β-ketoacyl-ACP synthase III (FabH) from Xanthomonas oryzae pv. oryzae

摘要

The bacterial β-ketoacyl-ACP synthase III (KASIII) encoded by the gene fabH (Xoo4209) from Xanthomonas oryzae pv. oryzae, a plant pathogen, is an important enzyme in the elongation steps of fatty-acid biosynthesis. It is expected to be one of the enzymes responsible for bacterial blight (BB), a serious disease that results in huge production losses of rice. As it represents an important target for the development of new antibacterial drugs against BB, determination of the crystal structure of the KAS III enzyme is essential in order to understand its reaction mechanism. In order to analyze the structure and function of KAS III, the fabH (Xoo4209) gene was cloned and the enzyme was expressed and purified. The KASIII crystal diffracted to 2.05 Å resolution and belonged to the orthorhombic space group P21212, with unit-cell parameters a = 69.8, b = 79.5, c = 62.3 Å. The unit-cell volume of the crystal is compatible with the presence of a single monomer in the asymmetric unit, with a corresponding Matthews coefficient V M of 2.27 ų Da⁻¹ and a solvent content of 45.8%.