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Logo of biotbiofuelBioMed CentralBiomed Central Web Sitesearchsubmit a manuscriptregisterthis articleBiotechnology for Biofuels
 
Biotechnol Biofuels. 2012; 5: 51.
Published online Jul 24, 2012. doi:  10.1186/1754-6834-5-51
PMCID: PMC3432595
Use of substructure-specific carbohydrate binding modules to track changes in cellulose accessibility and surface morphology during the amorphogenesis step of enzymatic hydrolysis
Keith Gourlay,1 Valdeir Arantes,1 and Jack N Saddlercorresponding author1
1Forest Products Biotechnology/Bioenergy Group, Department of Wood Science, Faculty of Forestry, University of British Columbia, 2424 Main Mall, Vancouver, BC, V6T 1Z4, Canada
corresponding authorCorresponding author.
Keith Gourlay: kg08/at/interchange.ubc.ca; Valdeir Arantes: valdeir.arantes/at/ubc.ca; Jack N Saddler: jack.saddler/at/ubc.ca
Received May 16, 2012; Accepted July 2, 2012.
Abstract
Background
Cellulose amorphogenesis, described as the non-hydrolytic “opening up” or disruption of a cellulosic substrate, is becoming increasingly recognized as one of the key steps in the enzymatic deconstruction of cellulosic biomass when used as a feedstock for fuels and chemicals production. Although this process is thought to play a major role in facilitating hydrolysis, the lack of quantitative techniques capable of accurately describing the molecular-level changes occurring in the substrate during amorphogenesis has hindered our understanding of this process.
Results
In this work, techniques for measuring changes in cellulose accessibility are reviewed and a new quantitative assay method is described. Carbohydrate binding modules (CBMs) with specific affinities for crystalline (CBM2a) or amorphous (CBM44) cellulose were used to track specific changes in the surface morphology of cotton fibres during amorphogenesis. The extents of phosphoric acid-induced and Swollenin-induced changes to cellulose accessibility were successfully quantified using this technique.
Conclusions
The adsorption of substructure-specific CBMs can be used to accurately quantify the extent of changes to cellulose accessibility induced by non-hydrolytic disruptive proteins. The technique provided a quick, accurate and quantitative measure of the accessibility of cellulosic substrates. Expanding the range of CBMs used for adsorption studies to include those specific for such compounds as xylan or mannan should also allow for the accurate quantitative tracking of the accessibility of these and other polymers within the lignocellulosic biomass matrix.
Keywords: Amorphogenesis, Cellulose Accessibility, Swollenin, Cellulose Disruption, Carbohydrate Binding Modules, Enzymatic Hydrolysis, Biofuels
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