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Updated: Jun 4, 2026
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Determination of Microbial Extracellular Enzyme Activity in Waters, Soils, and Sediments using High Throughput Microplate Assays
Published on: October 1, 2013
Screening for enzyme activity in turbid suspensions with scattered light
Robert Huber1, Helene Wulfhorst, Lukas Maksym
1AVT-Aachener Verfahrenstechnik, Biochemical Engineering, RWTH Aachen University, Worringerweg 1, D-52074 Aachen, Germany.
Biotechnology Progress
|February 9, 2011
Summary
A new BioLector method enables enzyme activity screening in turbid media by monitoring scattered light. This technique efficiently characterizes protease variants and their kinetics, aiding enzyme discovery for industrial applications.
Area of Science:
- Biotechnology
- Enzyme Engineering
- Bioprocess Monitoring
Background:
- Industries like detergent and food require efficient screening methods for enzyme variants in turbid media.
- Current enzyme activity measurement techniques face limitations with insoluble substrates and complex media.
Purpose of the Study:
- To present a novel method for semiquantitative determination of protease activity in various substrate suspensions.
- To enable online monitoring of enzyme kinetics using BioLector technology.
Main Methods:
- Utilized BioLector technology for online monitoring of scattered light and fluorescence signals.
- Measured the decrease in scattered light to track the hydrolysis of insoluble protein substrates.
- Determined kinetic parameters (V(max,app) and K(m,app)) from scattered light curves.
Main Results:
- Successfully monitored protease activity via scattered light reduction during substrate hydrolysis.
- Determined optimal pH for protease activity to be between 8 and 11.
- Compared activities of five subtilisin serine proteases, highlighting the method's utility for variant characterization.
Conclusions:
- The BioLector method provides an effective way to screen and characterize enzyme variants in turbid media.
- This technique facilitates the comparison of enzyme activities and determination of kinetic parameters.
- The method is applicable to various enzyme systems, including those involved in cellulose decomposition.
