However, the industrial conversion of lignocellulosic biomasses to such monomers faces two major bottlenecks: (i) the plant cell wall is highly recalcitrant to enzymatic degradation due to the presence of lignin which reduces enzyme accessibility to cellulose and hemicelluloses and (ii) a lack of high-throughput methods to rapidly screen for the enzymatic activities associated to these degradation capabilities in plant-degrading strains.įungi are the predominant source of enzymes currently being used on an industrial scale for the conversion of lignocellulosic biomass to platform molecules (Archer, 2000 Sims et al., 2010 Gusakov, 2011). These enzymes (Cell Wall-Degrading Enzymes, CWDE) are of great interest in the industrial conversion of lignocellulosic substrates into component sugars, which then serve as substrates for the synthesis of biofuels and other highly valuable platform molecules. Also, the results showed that the enzymatic activities measured were the same when doing the assay manually or using the automated method.Įnzymes secreted by filamentous fungi have a key role in the degradation of the most abundant biopolymers found in nature, that is cellulose and hemicelluloses. As highlighted in this study, it was shown that the CWDE activities gathered from the microbioreactor cultivations were similar or higher to those obtained from shake flasks cultures, with a lower standard deviation on the measured values, making this new method much faster than the traditional one and suitable for HT CWDE production thanks to its pipetting platform compatibility. The range of CWDE activities produced from the cultures was assayed using AZO-died and pNP-linked substrates in an SBS plate format using a Biomek FXp pipetting platform. Both approaches were compared in order to validate our new methodology. Fungi were grown in a microbioreactor in a high-throughput (HT) way to replace the fastidious shaking flask cultivations. In this study, wheat straw was selected as model substrate as it represents an important carbon source but yet poorly valorised to yield high added value products. This study aimed at developing a complete miniaturized high-throughput screening workflow for the evaluation of the Cell Wall-Degrading Enzyme (CWDE) activities produced by any fungal strain directly cultivated on raw feedstock in a submerged manner.
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