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Updated: Jan 9, 2026

Monitoring the Reductive and Oxidative Half-Reactions of a Flavin-Dependent Monooxygenase using Stopped-Flow Spectrophotometry
Published on: March 18, 2012
Comparative Characterization of Oxidative Enzymes for Arabinoxylan and Protein Cross-Linking via Ferulic Acid and
Katharina Hoefler1,2, Ulrich Sukop2, Elisabeth Reiter1
1Institute for Animal Nutrition and Feed, AGES - Austrian Agency for Health and Food Safety, Spargelfeldstraße 191, 1220 Vienna, Austria.
Abstract:
This study comparatively investigated various oxidoreductases (laccase, peroxidase, tyrosinase, and glucose oxidase) and their combinations for their conversion capability of available connection points, tyrosine in proteins and ferulic acid (FA) in arabinoxylans, as useful cross-linking tools. Therefore, substrate specificity and pH- and temperature-dependent activity were studied in different substrate ratios. Enzyme characteristics varied notably across standard assays, where even substrate-dependent shifts in pH optima occurred. Combining enzymes significantly reduced the Km for laccase with tyrosinase (0.504 to 0.238 mM) for tyrosine, whereas for FA, the Km increased from 0.057 to 0.107 mM but decreased for peroxidase with glucose oxidase from 0.250 to 0.045 mM. The substrate ratio was found to be crucial to target homo- or heterocross-linking and the most effective simultaneous substrate conversion was reached by combining peroxidase with glucose oxidase at a 1:5 (FA:tyrosine) ratio. These outcomes provide valuable insights into the cross-linking behavior of oxidoreductases, supporting a rational selection for food structure improvements.
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