Cloning, expression, purification and crystallization of dihydroxybutanonephosphate synthase from Magnaporthe grisea

Dihydroxybutanone phosphate synthase (DS) catalyzes a commitment step in ri boflavin biosynthesis where ribulose 5-phosphate is converted to dihydroxyb utanone phosphate and formate. DS was cloned from the pathogenic fungus Mag naporthe grisea (using functional complementation of an Escherichia coli DS knockout mutant) and expressed in E. coli. The purified protein crystalliz ed in space group P2(1)2(1)2. Diffraction data extending to 1.5, 1.0 and 1. 8 Angstrom resolution were collected from crystals that were divalent catio n free, soaked in Zn2+ or soaked in Mg2+, respectively.