Expression crystallization and preliminary X-ray crystallographic analysis of Xoo2316 a predicted 6-­phosphogluconolactonase from Xanthomonas oryzae pv. oryzae

摘要

Xanthomonas oryzae pv. oryzae (Xoo) causes bacterial blight, which is one of the most devastating diseases of rice (Oryza sativa L.) in many rice-growing countries. The coding sequence of Xoo2316 (a predicted 6-­phosphogluconolactonase; 6PGL) from Xoo was cloned and expressed in Escherichia coli. 6PGL is an enzyme that is involved in the second step of the pentose phosphate pathway, which is essential for the synthesis of nucleotide sugars and NADPH, the main source of reducing power. The protein was purified and crystallized in order to elucidate the molecular basis for its enzymatic reaction. Native crystals diffracted to 2.4 Å resolution and belonged to the orthorhombic space group P212121, with unit-cell parameters a = 40.0, b = 65.1, c = 78.8 Å. A monomer exists in the asymmetric unit with a corresponding V M of 1.93 ų Da⁻¹ and a solvent content of 36.5%.