A high-throughput assay to quantify protein hydrolysis in aerobic and anaerobic wastewater treatment processes
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ENVIRONMENTAL BIOTECHNOLOGY
A high-throughput assay to quantify protein hydrolysis in aerobic and anaerobic wastewater treatment processes Pieter Van Gaelen 1
&
Dirk Springael 2
&
Ilse Smets 1
Received: 27 April 2020 / Revised: 9 June 2020 / Accepted: 16 June 2020 # Springer-Verlag GmbH Germany, part of Springer Nature 2020
Abstract Proteins, an important fraction of the organic matter in wastewater, typically enter a treatment facility as high molecular weight components. These components are degraded by extracellular protein hydrolytic enzymes, denoted as proteases. Adequate protein hydrolysis monitoring is crucial, since protein hydrolysis is often a rate-limiting step in wastewater treatment. However, current monitoring tools lack a high sample throughput and reliable quantification. Here, we present an improved assay for high-throughput protein hydrolysis rate measurements in wastewater treatment applications. A BODIPY FL casein model substrate was implemented in a microplate format for continuous fluorescent quantification. Case studies on a conventional and a high-rate aerobic municipal wastewater treatment plant and a lab-scale, two-stage, anaerobic reactor provided proofof-concept. The assay presented in this study can help to obtain monitoring-based process insights, which will in turn allow improving biological performance of wastewater treatment installations in the future. Key points • Protein hydrolysis is a crucial step in biological wastewater treatment. • Quantification of the protein hydrolysis rate enables in-depth process knowledge. • BODIPY FL casein is a suitable model substrate for a protein hydrolysis assay. • High sample throughput was obtained with fluorescent hydrolysis quantification. Keywords Protein . Hydrolysis . Enzyme activity . Fluorescence . Wastewater treatment
Introduction Biological conversion of organic matter is the cornerstone of all aerobic and anaerobic wastewater treatment facilities. This
Electronic supplementary material The online version of this article (https://doi.org/10.1007/s00253-020-10751-4) contains supplementary material, which is available to authorized users. * Ilse Smets [email protected] 1
Bio- and Chemical Systems Technology, Reactor Engineering and Safety (CREaS), Department of Chemical Engineering, KU Leuven, Celestijnenlaan 200F, PO box 2424, B-3001 Leuven, Belgium
2
Division of Soil and Water Management, Department of Earth and Environmental sciences, KU Leuven, Kasteelpark Arenberg 20, B-3001 Leuven, Belgium
conversion process relies on the coordinated action of a mixed microbial community to metabolize the protein, carbohydrate, and lipid fractions in the organic matter. This organic matter typically enters the treatment systems as high molecular weight (MW) components (MW > 10,000 Da) or particulates in the micro- and millimeter range, while only molecules in the nanometer range (MW < 1000) can be taken up directly through the microbial cell membrane (Cadoret et al. 2002; Hashimoto et al. 2005). Therefore, the microbial community
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