Open AccessCloning, expression, purification, crystallization and preliminary X‐ray studies of a pyridoxine 5′‐phosphate oxidase from Mycobacterium smegmatis
Author(s) -
Jackson Colin J.,
Taylor Matthew C.,
Tattersall David B.,
French Nigel G.,
Carr Paul D.,
Ollis David L.,
Russell Robyn J.,
Oakeshott John G.
Publication year - 2008
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309108011512
Subject(s) - mycobacterium smegmatis , flavin mononucleotide , escherichia coli , cofactor , biochemistry , pyridoxal , enzyme , chemistry , mycobacterium tuberculosis , biology , gene , medicine , tuberculosis , pathology
Pyridoxine 5′‐phosphate oxidases (PNPOxs) are known to catalyse the terminal step in pyridoxal 5′‐phosphate biosynthesis in a flavin mononucleotide‐dependent manner in humans and Escherichia coli . Recent reports of a putative PNPOx from Mycobacterium tuberculosis , Rv1155, suggest that the cofactor or catalytic mechanism may differ in Mycobacterium species. To investigate this, a putative PNPOx from M. smegmatis , Msmeg_3380, has been cloned. This enzyme has been recombinantly expressed in E. coli and purified to homogeneity. Good‐quality crystals of selenomethionine‐substituted Msmeg_3380 were obtained by the hanging‐drop vapour‐diffusion technique and diffracted to 1.2 Å using synchrotron radiation.