Flavourzyme, an Enzyme Preparation with Industrial
Relevance: Automated Nine-Step Purification and Partial Characterization
of Eight Enzymes
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Abstract
Flavourzyme
is sold as a peptidase preparation from Aspergillus
oryzae. The enzyme preparation is widely
and diversely used for protein hydrolysis in industrial and research
applications. However, detailed information about the composition
of this mixture is still missing due to the complexity. The present
study identified eight key enzymes by mass spectrometry and partially
by activity staining on native polyacrylamide gels or gel zymography.
The eight enzymes identified were two aminopeptidases, two dipeptidyl
peptidases, three endopeptidases, and one α-amylase from the <i>A. oryzae</i> strain ATCC 42149/RIB 40 (yellow koji mold). Various
specific marker substrates for these Flavourzyme enzymes were ascertained.
An automated, time-saving nine-step protocol for the purification
of all eight enzymes within 7 h was designed. Finally, the purified
Flavourzyme enzymes were biochemically characterized with regard to
pH and temperature profiles and molecular sizes