title: Isolation and characterization of aminopeptidase from Capnocytophaga granulosa ATCC 51502.
abstract: There is evidence that enzymes from the genus Capnocytophaga play a role in dental calculus formation. Although most of the species in the genus produce aminopeptidases, there is a paucity of data on the purification and characterization of the enzyme, except in the case of Capnocytophaga gingivalis. The aim of this study was to purify aminopeptidase from culture supernatant of Capnocytophaga granulosa ATCC 51502, a new species of the genus. Purification was performed using ammonium sulfate fractionation and two chromatographic steps. The aminopeptidase was purified 158,433-fold with a yield of 12.0%. The enzyme appeared to be a trimer with a molecular mass of 270 kDa. The optimal pH of the aminopeptidase was 6.5 and its activity was completely inhibited by incubation at 50 degrees C for 10 min. The enzyme showed maximum specificity for basic amino acids (Arg and Lys) and also hydrolyzed noncharged amino acids (Met, Leu and Ala). Ca(2+), Zn(2+) and Fe(3+) activated the enzyme, while EDTA, Ag(+), Hg(+) and Cu(2+) inhibited it. These results suggest that aminopeptidase of C. granulosa is different from that of C. gingivalis but similar to aminopeptidase B.
authors.affiliation: Department of Oral Health, Okayama University Graduate School of Medicine and Dentistry, Okayama, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Ohishi
authors.forename: T
authors.initials: T
authors.lastname: Yamamoto
authors.forename: T
authors.initials: T
authors.lastname: Tomofuji
authors.forename: N
authors.initials: N
authors.lastname: Tamaki
authors.forename: T
authors.initials: T
authors.lastname: Watanabe
mesh_terms: D000596:Amino Acids; D000626:Aminopeptidases / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000302:isolation & purification*; D002206:Capnocytophaga / Q000201:enzymology*; D005591:Chemical Fractionation; D002852:Chromatography, Ion Exchange; D003728:Dental Calculus / Q000382:microbiology; D004591:Electrophoresis, Polyacrylamide Gel; D007526:Isoelectric Point; D008970:Molecular Weight; D011480:Protease Inhibitors / Q000378:metabolism; D013379:Substrate Specificity
chemical_list: D000596:Amino Acids; D011480:Protease Inhibitors; D000626:Aminopeptidases