Details zur Publikation

Kategorie Textpublikation
Referenztyp Zeitschriften
DOI 10.1128/JB.05120-11
Titel (primär) Identification and characterization of a Re-Citrate synthase in Dehalococcoides strain CBDB1
Autor Marco-Urrea, E.; Paul, S.; Khodaverdi, V.; Seifert, J.; von Bergen, M.; Kretzschmar, U.; Adrian, L.
Quelle Journal of Bacteriology
Erscheinungsjahr 2011
Band/Volume 193
Heft 19
Seite von 5171
Seite bis 5178
Sprache englisch

The genome annotations of all sequenced Dehalococcoides strains lack a citrate synthase although physiological experiments have indicated that such an activity should be encoded. We here report that a Re-face specific citrate synthase is synthesized by Dehalococcoides strain CBDB1 and that this function is encoded by the gene cbdbA1708 (NCBI accession number CAI83711), previously annotated as homocitrate synthase. Gene cbdbA1708 was heterologously expressed in E. coli and the recombinant enzyme was purified. The enzyme catalyzed the condensation of oxaloacetate and acetyl-CoA to citrate. The protein did not have homocitrate synthase activity, was inhibited by citrate, and Mn2+ was needed for full activity. The stereospecificity of the heterologously expressed citrate synthase was determined by ESI LC/MS. Citrate was synthesized from [2-13C]acetyl-CoA and oxaloacetate by the Dehalococcoides recombinant citrate synthase and then converted to acetate and malate by commercial citrate lyase plus malate dehydrogenase. The formation of unlabeled acetate and 13C-labeled malate proved the Re-face specific activity of the enzyme. Shotgun proteome analyses of cell extracts of strain CBDB1 demonstrated that cbdbA1708 is expressed in strain CBDB1.
dauerhafte UFZ-Verlinkung
Marco-Urrea, E., Paul, S., Khodaverdi, V., Seifert, J., von Bergen, M., Kretzschmar, U., Adrian, L. (2011):
Identification and characterization of a Re-Citrate synthase in Dehalococcoides strain CBDB1
J. Bacteriol. 193 (19), 5171 - 5178 10.1128/JB.05120-11