Y. Fuke et H. Matsuoka, THE PURIFICATION AND CHARACTERIZATION OF PROLYL AMINOPEPTIDASE FROM PENICILLIUM-CAMEMBERTI, Journal of dairy science, 76(9), 1993, pp. 2478-2484
Prolyl aminopeptidase [EC 3.4.11.5] from the cell-free extract of Peni
cillium camemberti was purified about 2800-fold by chromatographic tec
hniques. The purity of the enzyme was confirmed by electrophoretic ana
lysis. The molecular mass of the enzyme was estimated to be 270,000 Da
by gel filtration. The enzyme had a maximum activity at pH 7.0 and 45
-degrees-C, and Pro-p-naphthylamide was the substrate; the enzyme was
stable up to 50-degrees-C. The enzyme cleaved Pro-amino acid bond when
the Pro residue was at the amino-terminal. The enzyme was completely
inactivated by p-chloromercuribenzoic acid and reduced to approximatel
y 50% activity by diisopropyl fluorophosphate. The Michaelis-Menten co
nstant was estimated to be .25 mM and the maximum velocity to be .56 m
M/min per ml.