title: Flavourzyme, an Enzyme Preparation with Industrial Relevance: Automated Nine-Step Purification and Partial Characterization of Eight Enzymes.
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 A. oryzae 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.
authors.affiliation: †Institute of Food Science and Biotechnology, Department of Biotechnology and Enzyme Science, University of Hohenheim, Garbenstrasse 25, D-70599 Stuttgart, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Merz
authors.affiliation: †Institute of Food Science and Biotechnology, Department of Biotechnology and Enzyme Science, University of Hohenheim, Garbenstrasse 25, D-70599 Stuttgart, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Eisele
authors.affiliation: §Nestlé Product Technology Centre, Lange Strasse 21, D-78224 Singen, Germany.
authors.forename: Pieter
authors.initials: P
authors.lastname: Berends
authors.affiliation: §Nestlé Product Technology Centre, Lange Strasse 21, D-78224 Singen, Germany.
authors.forename: Daniel
authors.initials: D
authors.lastname: Appel
authors.affiliation: §Nestlé Product Technology Centre, Lange Strasse 21, D-78224 Singen, Germany.
authors.forename: Swen
authors.initials: S
authors.lastname: Rabe
authors.affiliation: §Nestlé Product Technology Centre, Lange Strasse 21, D-78224 Singen, Germany.
authors.forename: Imre
authors.initials: I
authors.lastname: Blank
authors.affiliation: †Institute of Food Science and Biotechnology, Department of Biotechnology and Enzyme Science, University of Hohenheim, Garbenstrasse 25, D-70599 Stuttgart, Germany.
authors.forename: Timo
authors.initials: T
authors.lastname: Stressler
authors.affiliation: †Institute of Food Science and Biotechnology, Department of Biotechnology and Enzyme Science, University of Hohenheim, Garbenstrasse 25, D-70599 Stuttgart, Germany.
authors.forename: Lutz
authors.initials: L
authors.lastname: Fischer
mesh_terms: D001236:Aspergillus oryzae / Q000737:chemistry / Q000201:enzymology*; D010450:Endopeptidases / Q000737:chemistry* / Q000302:isolation & purification*; D004795:Enzyme Stability; D005656:Fungal Proteins / Q000737:chemistry* / Q000302:isolation & purification*; D006863:Hydrogen-Ion Concentration; D008970:Molecular Weight; D013696:Temperature
chemical_list: D005656:Fungal Proteins; D010450:Endopeptidases; C412470:flavourzyme
keywords: Aspergillus oryzae amylase; Aspergillus oryzae peptidases; Flavourzyme; automated purification; characterization; key enzymes; protein hydrolysis