Replication Data for: Hydrolysis activity measurements do not depend on the aeration conditions

Hydrolysis, often the rate-limiting step in waste valorization processes (e.g., anaerobic digestion), is a complex process, influenced by various factors. One of these factors is the presence of oxygen. While the effect of oxygen on hydrolysis itself is well-documented, its impact on hydrolysis activity monitoring has received less attention. This study focuses on the influence of aeration conditons on fluorescence-based, 96-well plate reader assays. More specifically the oxygen depletion rate of each well and the resulting hydrolysis activity is measured. The oxygen depletion rate was significantly higher when flushing the plate reader device with nitrogen. However, the hydrolysis activity (protease and cellulase) of both aerobic and anaerobic sludge samples, and a pure enzyme was not influenced by the aeration conditions. This result suggests that oxygen does not affect the functionality of the enzymes themselves, but may influence microbial contributors to enzyme production. These findings confirm that hydrolysis activity monitoring tools yield consistent results regardless of aeration conditions. Consequently, both aerobic and anaerobic samples can be reliably assessed under standard aerobic conditions, typical for plate reader devices, without risking overestimation of anaerobic sample activities.