Cloning, expression, purification and crystallization of dihydroxybutanone phosphate synthase from Magnaporthe grisea.
Dihydroxybutanone phosphate synthase (DS) catalyzes a commitment step in riboflavin biosynthesis where ribulose 5-phosphate is converted to dihydroxybutanone phosphate and formate. DS was cloned from the pathogenic fungus Magnaporthe grisea (using functional complementation of an Escherichia coli DS knockout mutant) and expressed in E. coli. The purified protein crystallized in space group P2(1)2(1)2. Diffraction data extending to 1.5, 1.0 and 1.8 A resolution were collected from crystals that were divalent cation free, soaked in Zn(2+) or soaked in Mg(2+), respectively.
Liao
D I
DI
DuPont Central Research and Development, Experimental Station, Wilmington, Delaware 19880, USA.
Viitanen
P V
PV
Jordan
D B
DB
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D003460:Crystallization; D018360:Crystallography, X-Ray; D017931:DNA Primers; D019751:Intramolecular Transferases / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D020082:Magnaporthe / Q000201:enzymology*; D008969:Molecular Sequence Data
D017931:DNA Primers; D019751:Intramolecular Transferases; C066493:L-3,4-dihydroxy-2-butanone-4-phosphate synthase