title: Biochemistry and physiology of the β class carbonic anhydrase (Cpb) from Clostridium perfringens strain 13.
abstract: The carbonic anhydrase (Cpb) from Clostridium perfringens strain 13, the only carbonic anhydrase encoded in the genome, was characterized both biochemically and physiologically. Heterologously produced and purified Cpb was shown to belong to the type I subclass of the β class, the first β class enzyme investigated from a strictly anaerobic species of the domain Bacteria. Kinetic analyses revealed a two-step, ping-pong, zinc-hydroxide mechanism of catalysis with Km and kcat/Km values of 3.1 mM CO₂ and 4.8 × 10⁶ s⁻¹ M⁻¹, respectively. Analyses of a cpb deletion mutant of C. perfringens strain HN13 showed that Cpb is strictly required for growth when cultured in semidefined medium and an atmosphere without CO₂. The growth of the mutant was the same as that of the parent wild-type strain when cultured in nutrient-rich media with or without CO₂ in the atmosphere, although elimination of glucose resulted in decreased production of acetate, propionate, and butyrate. The results suggest a role for Cpb in anaplerotic CO₂ fixation reactions by supplying bicarbonate to carboxylases. Potential roles in competitive fitness are discussed.
authors.affiliation: Department of Biochemistry and Molecular Biology, Pennsylvania State University, University Park, Pennsylvania, USA.
authors.forename: R Siva Sai
authors.initials: RS
authors.lastname: Kumar
authors.forename: William
authors.initials: W
authors.lastname: Hendrick
authors.forename: Jared B
authors.initials: JB
authors.lastname: Correll
authors.forename: Andrew D
authors.initials: AD
authors.lastname: Patterson
authors.forename: Stephen B
authors.initials: SB
authors.lastname: Melville
authors.forename: James G
authors.initials: JG
authors.lastname: Ferry
mesh_terms: D001426:Bacterial Proteins / Q000378:metabolism*; D002245:Carbon Dioxide; D002256:Carbonic Anhydrases / Q000145:classification / Q000235:genetics / Q000378:metabolism*; D003016:Clostridium perfringens / Q000201:enzymology*; D008962:Models, Theoretical; D010802:Phylogeny
chemical_list: D001426:Bacterial Proteins; D002245:Carbon Dioxide; D002256:Carbonic Anhydrases