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A High Throughput Screen for Biomining Cellulase Activity from Metagenomic Libraries
Published on: February 1, 2011
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Screening for Cellulolytic Plant Enzymes Using Colorimetric and Fluorescence Methods.
1Department of Arctic and Marine Biology, Faculty of Biosciences, Fisheries and Economics, UiT The Arctic University of Norway, Tromsø, Norway. kirsten.krause@uit.no.
Methods in Molecular Biology (Clifton, N.J.)
|July 4, 2020
Summary
This study presents a rapid and reliable method for measuring cellulolytic activity using carboxymethylcellulose (CMC). The technique offers accurate enzyme quantification in diverse samples, including plants, unaffected by glucose levels.
Area of Science:
- Biochemistry
- Enzymology
- Plant Science
Background:
- Cellulolytic activity is crucial in various biological processes.
- Existing methods for measuring cellulolytic activity can be costly, slow, or material-dependent.
Purpose of the Study:
- To develop an inexpensive, rapid, and reliable method for quantifying cellulolytic activity.
- To establish a versatile assay applicable to diverse sample types, including plant materials.
Main Methods:
- Utilized carboxymethylcellulose (CMC) as a substrate for cellulase digestion.
- Employed a "negative staining principle" with colorimetric or fluorescent stains to detect undigested CMC.
- Quantified cellulolytic activity by measuring the reduction in staining intensity, which is proportional to enzyme activity.
Main Results:
- The method provides a direct and sensitive detection of CMC digestion.
- Enzyme activity measurements are independent of endogenous glucose levels in the sample.
- The assay demonstrated applicability to a wide range of samples, including plant extracts.
Conclusions:
- The developed CMC-based assay is a cost-effective, fast, and dependable tool for measuring cellulolytic activity.
- This method offers a significant advantage for analyzing samples with varying glucose concentrations, such as plant tissues.
