Flavourzyme, an enzyme preparation with industrial relevance – Automated nine-step purification and partial characterization of eight enzymes

Publication Type

Journal contribution (peer reviewed)

Authors

Merz, M.; Eisele, T.; Berends, P.; Appel, D.; Rabe, S.; Blank, I.; Stressler, T.; Fischer, L.

Year of publication

2015

Published in

Journal of Agricultural and Food Chemistry

DOI

10.1021/acs.jafc.5b01665

Flavourzyme is sold as 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. In the present study, we 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 alpha-amylase from the Aspergillus oryzae strain ATCC 42149/RIB 40 (Yellow koji mold). We ascertained various specific marker substrates for these Flavourzyme enzymes. An automated, time-saving nine-step protocol for the purification of all eight enzymes within seven hours was designed. Finally, the purified Flavourzyme enzymes were biochemically characterized regarding pH and temperature profiles and molecular sizes.