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Given the need for new therapeutics, we intend to investigate enzymes of cell wall biosynthesis as novel drug targets. Alanine racemase, a ubiquitous enzyme among bacteria and absent in humans, provides the essential cell wall precursor, D-alanine, which forms part of the tetrapeptide crosslinking the peptidoglycan layer.<\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>Results<\/jats:title>\n            <jats:p>The alanine racemases gene from <jats:italic>S. pneumoniae<\/jats:italic> (<jats:italic>alr<\/jats:italic>\n              <jats:sub>\n                <jats:italic>SP<\/jats:italic>\n              <\/jats:sub>) was amplified by PCR and cloned and expressed in <jats:italic>Escherichia coli<\/jats:italic>. The 367 amino acid, 39854 Da dimeric enzyme was purified to electrophoretic homogeneity and preliminary crystals were obtained. Racemic activity was demonstrated through complementation of an <jats:italic>alr<\/jats:italic> auxotroph of <jats:italic>E. coli<\/jats:italic> growing on L-alanine. In an alanine racemases photometric assay, specific activities of 87.0 and 84.8 U mg<jats:sup>-1<\/jats:sup> were determined for the conversion of D- to L-alanine and L- to D-alanine, respectively.<\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>Conclusion<\/jats:title>\n            <jats:p>We have isolated and characterized the alanine racemase gene from the opportunistic human pathogen <jats:italic>S. pneumoniae<\/jats:italic>. The enzyme shows sufficient homology with other alanine racemases to allow its integration into our ongoing structure-based drug design project.<\/jats:p>\n          <\/jats:sec>","DOI":"10.1186\/1471-2180-7-40","type":"journal-article","created":{"date-parts":[[2007,5,18]],"date-time":"2007-05-18T20:19:55Z","timestamp":1179519595000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":20,"title":["Purification and preliminary crystallization of alanine racemase from Streptococcus pneumoniae"],"prefix":"10.1186","volume":"7","author":[{"given":"Ulrich","family":"Strych","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Milya","family":"Davlieva","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Joseph P","family":"Longtin","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Eileen L","family":"Murphy","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hookang","family":"Im","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Michael J","family":"Benedik","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kurt L","family":"Krause","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2007,5,17]]},"reference":[{"issue":"2","key":"357_CR1","doi-asserted-by":"publisher","first-page":"187","DOI":"10.1128\/MMBR.65.2.187-207.2001","volume":"65","author":"MJ Jedrzejas","year":"2001","unstructured":"Jedrzejas MJ: Pneumococcal virulence factors: structure and function. 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