Overexpression, crystallization and preliminary X-ray crystallographic analysis of pyridoxal biosynthesis lyase PdxS from Pyrococcus horikoshii.

摘要: Pyridoxal biosynthesis lyase (PdxS) is an important player in the of pyridoxal 5'-phosphate (PLP), biologically active form vitamin B(6). PLP cofactor involved metabolic pathway amine-containing natural products such as amino acids and sugars. PdxS catalyzes condensation ribulose 5-phosphate (Ru5P), glyceraldehyde 3-phosphate (G3P) ammonia, while glutamine amidotransferase (PdxT) production ammonia from glutamine. PdxT a complex, synthase, widely exist eubacteria, archaea, fungi plants. To facilitate further structural comparisons among proteins, analysis Pyrococcus horikoshii encoded by Ph1355 gene was initiated. P. overexpressed Escherichia coli crystallized at 296 K using 2-methyl-2,4-pentanediol precipitant. Crystals diffracted to 2.61 A resolution belonged monoclinic space group P2(1), with unit-cell parameters = 59.30, b 178.56, c 109.23 A, β 102.97°. The asymmetric unit contained six monomers, corresponding V(M) 2.54 A(3) Da(-1) solvent content 51.5% volume.