Journal article
Expression, purification, crystallization and preliminary X-ray diffraction analysis of dihydrodipicolinate synthase from Bacillus anthracis in the presence of pyruvate
JE Voss, SW Scally, NL Taylor, C Dogovski, MR Alderton, CA Hutton, JA Gerrard, MW Parker, RCJ Dobson, MA Perugini
Acta Crystallographica Section F Structural Biology and Crystallization Communications | INT UNION CRYSTALLOGRAPHY | Published : 2009
Abstract
Dihydrodipicolinate synthase (DHDPS) catalyses the first committed step in the lysine-biosynthesis pathway in bacteria, plants and some fungi. In this study, the expression of DHDPS from Bacillus anthracis (Ba-DHDPS) and the purification of the recombinant enzyme in the absence and presence of the substrate pyruvate are described. It is shown that DHDPS from B. anthracis purified in the presence of pyruvate yields greater amounts of recombinant enzyme with more than 20-fold greater specific activity compared with the enzyme purified in the absence of substrate. It was therefore sought to crystallize Ba-DHDPS in the presence of the substrate. Pyruvate was soaked into crystals of Ba-DHDPS prep..
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Awarded by Defense Threat Reduction Agency
Funding Acknowledgements
We would firstly like to acknowledge the support and assistance of the friendly staff at the Bio21 Collaborative Crystallographic Centre at CSIRO Molecular and Health Technologies, Parkville, Melbourne. We would also like to thank all members of the Perugini laboratory for helpful discussions during the preparation of this manuscript. Finally, we acknowledge the Defense Threat Reduction Agency ( DTRA; DTRA Project ID AB07CBT004) and the Australian Research Council for providing an Australian Postdoctoral Fellowship for MAP and a Federation Fellowship for MWP.