The characterization of a (nutritionally important) proline iminopeptidase from Eikenella corrodens.
Eikenella corrodens generates energy primarily through the oxidative deamination of specific amino acids, a process that is coupled to dissimilatory nitrate reduction to nitrite. Cell yields resulting from chemostat-growth of the organism in simple, chemically defined media containing varying amounts of proline confirm that this amino acid is a likely source of energy for E. corrodens in the oral environment. The importance of proline in ATP generation by the organism is reflected in molar growth yields, which showed that biomass production per mole of this amino acid was significantly higher than that for other amino acids. The organism was found to express, constitutively, the enzyme proline iminopeptidase, which releases proline from the N-terminus of small peptides. The enzyme was partially purified and characterized and found to exist in the cytoplasm as a 35 kDa monomer. Inhibition studies showed that the enzyme, although classified as a serine protease, also appears to require thiol groups for activity, a finding which is consistent with previous reports. The enzyme obeyed Michaelis-Menten kinetics and was found to have a Km value of 0.223 mM for the substrate proline-p-nitroanilide.
Gully
N J
NJ
Microbiology Laboratory, Dental School, Adelaide University, North Terrace, Adelaide, South Australia 5005.
Rogers
A H
AH
D000255:Adenosine Triphosphate / Q000096:biosynthesis; D000596:Amino Acids / Q000378:metabolism; D000626:Aminopeptidases / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000302:isolation & purification*; D000814:Aniline Compounds / Q000378:metabolism; D003470:Culture Media; D003593:Cytoplasm / Q000201:enzymology; D003641:Deamination; D016124:Eikenella corrodens / Q000201:enzymology* / Q000254:growth & development; D006801:Humans; D007700:Kinetics; D008970:Molecular Weight; D009566:Nitrates / Q000378:metabolism; D009573:Nitrites / Q000378:metabolism; D010084:Oxidation-Reduction; D011392:Proline / Q000378:metabolism; D013696:Temperature
D000596:Amino Acids; D000814:Aniline Compounds; D003470:Culture Media; D009566:Nitrates; D009573:Nitrites; C019498:4-nitroaniline; D000255:Adenosine Triphosphate; D011392:Proline; D000626:Aminopeptidases; C022450:prolyl aminopeptidase