Open AccessCrystallization of dihydrodipicolinate synthase from a clinical isolate of Streptococcus pneumoniae
Author(s) -
Sibarani Natalia E.,
Gorman Michael A.,
Dogovski Con,
Parker Michael W.,
Perugini Matthew A.
Publication year - 2010
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s174430910904771x
Subject(s) - streptococcus pneumoniae , crystallization , microbiology and biotechnology , biology , chemistry , organic chemistry , antibiotics
Dihydrodipicolinate synthase (DHDPS; EC 4.2.1.52) catalyzes the rate‐limiting step in the ( S )‐lysine biosynthesis pathway of bacteria and plants. Here, the cloning of the DHDPS gene from a clinical isolate of Streptococcus pneumoniae (OXC141 strain) and the strategy used to express, purify and crystallize the recombinant enzyme are described. Diffracting crystals were grown in high‐molecular‐weight PEG precipitants using the hanging‐drop vapour‐diffusion method. The best crystal, from which data were collected, diffracted to beyond 2.0 Å resolution. Initially, the crystals were thought to belong to space group P 4 2 2 1 2, with unit‐cell parameters a = 105.5, b = 105.5, c = 62.4 Å. However, the R factors remained high following initial processing of the data. It was subsequently shown that the data set was twinned and it was thus reprocessed in space group P 2, resulting in a significant reduction in the R factors. Determination of the structure will provide insight into the design of novel antimicrobial agents targeting this important enzyme from S. pneumoniae .