Acid aspartyl proteinase of Penicillium roqueforti
was purified from
culture filtrates using FPLC ion-exchange chromatography on Mono Q and
size
exclusion chromatography on Superdex 75. The purity obtained allowed us
to
produce, using rabbits, a specific antiserum which was then used to develop
a
sandwich ELISA. The test was sensitive (detection limit,
0·25 ng/ml), reproducible
(CV, 3·1–6·9%) and closely correlated with
enzyme activity measurement (r=0·995,
P<0·001). This ELISA should be a valuable tool
for monitoring acid aspartyl
proteinase level in culture filtrates and for determining the
enzyme activity[ratio ]enzyme
protein ratio of acid aspartyl proteinase obtained after genetic recombinations
or
site-directed mutagenesis.