Abstract
Second generation biofuels are derived from inedible lignocellulosic biomass of food and non-food crops. Lignocellulosic biomass is mainly composed of cell walls that contain a large proportion of cellulosic and hemicellulosic polysaccharides. An interesting route to generate biofuels and bio-based materials is via enzymatic hydrolysis of cell wall polysaccharides into fermentable sugars, a process called saccharification. The released sugars can then be fermented to fuels, e.g., by use of yeast. To test the saccharification efficiency of lignocellulosic biomass on a lab-scale, a manual saccharification protocol was established that uses only small amounts of biomass and a low concentration of enzyme. This protocol can be used for different plant species like Arabidopsis thaliana, tobacco, maize and poplar. The low enzyme concentrations make it possible to detect subtle improvements in saccharification yield and to analyze the speed of hydrolysis. Although a specific acid and alkali pretreatment were included, the saccharification step can be preceded by any other pretreatment. Because no advanced equipment is necessary, this protocol can be carried out in many laboratories to analyze saccharification yield. The protocol was initially described in Van Acker et al. (2013).
Keywords: Saccharification, Lignocellulose, Biofuels
Materials and Reagents
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Acknowledgments
This work was supported by grants from the Multidisciplinary Research Partnership “Biotechnology for a sustainable economy” of Ghent University, the European Commission through the Directorate General Research within the 7th Framework Program RENEWALL (KBBE-2007-3-1-01) and MultiBioPro (grant agreement N° 311804) and the Agency for Innovation by Science and Technology (IWT). RV is indebted to the Research Foundation-Flanders (FWO) for a postdoctoral fellowship.
References
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