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Automated Filter Paper Assay for Determination of Cellulase Activity
- Source :
- Applied Biochemistry and Biotechnology. 107:689-704
- Publication Year :
- 2003
- Publisher :
- Springer Science and Business Media LLC, 2003.
-
Abstract
- Recent developments in molecular breeding and directed evolution have promised great developments in industrial enzymes as demonstrated by exponential improvements in beta-lactamase and green fluorescent protein (GFP). Detection of and screening for improved enzymes are relatively easy if the target enzyme is expressible in a suitable high-throughput screening host and a clearly defined and usable screen or selection is available, as with GFP and beta-lactamase. Fungal cellulases, however, are difficult to measure and have limited expressibility in heterologous hosts. Furthermore, traditional cellulase assays are tedious and time-consuming. Multiple enzyme components, an insoluble substrate, and generally slow reaction rates have plagued cellulase researchers interested in creating cellulase mixtures with increased activities and/or enhanced biochemical properties. Although the International Union of Pure and Applied Chemists standard measure of cellulase activity, the filter paper assay (FPA), can be reproduced in most laboratories with some effort, this method has long been recognized for its complexity and susceptibility to operator error. Our current automated FPA method is based on a Cyberlabs C400 robotics deck equipped with customized incubation, reagent storage, and plate-reading capabilities that allow rapid evaluation of cellulases acting on cellulose and has a maximum throughput of 84 enzyme samples per day when performing the automated FPA.
- Subjects :
- Paper
Bioengineering
Cellulase
Standard measure
Applied Microbiology and Biotechnology
Biochemistry
Substrate Specificity
Automation
chemistry.chemical_compound
Cellulose
Molecular Biology
Trichoderma reesei
biology
Filter paper
Substrate (chemistry)
Equipment Design
General Medicine
Directed evolution
biology.organism_classification
Industrial enzymes
Kinetics
chemistry
biology.protein
Biochemical engineering
Biotechnology
Subjects
Details
- ISSN :
- 02732289
- Volume :
- 107
- Database :
- OpenAIRE
- Journal :
- Applied Biochemistry and Biotechnology
- Accession number :
- edsair.doi.dedup.....c3c55e1941b28721713733e23a1a7299