Biotechnology for Biofuels

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Measurement of saccharifying cellulase

Douglas E Eveleigh*, Mary Mandels, Raymond Andreotti and Charles Roche

Author Affiliations

US Army Natick Development Center, Natick, Massachusetts 01760, USA

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Biotechnology for Biofuels 2009, 2:21 doi:10.1186/1754-6834-2-21

Published: 1 September 2009

Abstract

This article sets forth a simple cellulase assay procedure. Cellulose is variable in nature, insoluble and resistant to enzymatic attack. As a result there have been a bevy of bewildering cellulase assays published that yielded irrational results. Certain protocols focused on the rapidity of the assay while ignoring that only the most readily susceptible cellulose regions were being hydrolyzed. Other assays simplified the system by using modified soluble substrates and yielded results that bore no relationship to the real world hydrolysis of insoluble cellulose. In this study Mandels, Andreotti and Roche utilized a common substrate, Whatman filter paper. Hydrolysis of a 50 mg sample of the paper was followed to roughly 4% degradation, which circumvented the problems of attack of only the most susceptible zones. This common hydrolysis target range also resulted in some balance with regard to the interaction of the several cellulase components. The method was subsequently widely adopted.

Douglas E Eveleigh