Enzymatic hydrolysis: bridging the gap between micro and nano scales
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Enzymatic hydrolysis: bridging the gap between micro and nano scales

Enzymatic hydrolysis: bridging the gap between micro and nano scales

In the current climate of climate change and depletion of fossil fuels, the search for sustainable energy solutions is imperative. Lignocellulosic biomass, made up of the non-food parts of plants and residues from the wood industry, is diverse in its origins and above all renewable: it appears to be a relevant alternative to fossil carbon. However, its chemical and structural complexity makes it costly to transform by biotechnological means into products of interest, which is why it has been described as a recalcitrant raw material. It is therefore vital to develop innovative strategies to deconstruct it efficiently. Although a great deal of research has identified recalcitrance factors at the nanometric scale, such as lignin content and cellulose crystallinity, the study of recalcitrance at the scale of plant cells and tissues remains little explored.

In this study, we overcame the experimental and mathematical challenges associated with segmenting and monitoring the deconstruction of plant cell walls by developing a 4D (space + time) imaging method. This is based on confocal fluorescence microscopy imaging combined with automated 4D image processing to identify, monitor and quantify the evolution of the morphology of individual cells. Thanks to this approach, which overcomes the limitations of conventional methods prone to bias and cell delimitation errors, we were able to acquire 3D images of poplar wood samples (chosen as the model species) during their transformation by enzymatic hydrolysis.

Beyond this significant technical development, our study has led to several major results and discoveries. Firstly, enzymatic deconstruction on a cellular scale mainly leads to a reduction in cell wall volume, rather than changes in surface area or accessible surface area. Furthermore, the 3D compactness of cell walls prior to hydrolysis correlates with volumetric deconstruction, with this correlation being modulated by enzymatic activity. Finally, we established a positive correlation between the volumetric deconstruction of cell walls and cellulose conversion, thus linking mechanisms at nano- and micro-metric scales for the first time.

Establishing this quantitative relationship between scales highlights the impact and importance of studying enzymatic deconstruction at the cellular and tissue scales, paving the way for future studies of plant cell wall deconstruction at these specific scales. Given the methodological advances and the implications for research into the mechanisms underlying the enzymatic deconstruction of lignocellulosic biomass, we believe that our work will generate widespread interest in fields such as plant biotechnology, enzymatic biochemistry and computational biotechnology.

 

2024-11-28 Article 4D.jpg

Read: Refahi Y, Zoghlami A, Viné T, Terryn C, Paës G (2024) Plant cell wall enzymatic deconstruction: Bridging the gap between micro and nano scales. Bioresource Technol. 414, 131551. https://doi.org/10.1016/j.biortech.2024.131551

Contacts: Dr Yassin Refahi, yassin.refahi@inrae.fr & Dr Gabriel Paës, gabriel.paes@inrae.fr

Modification date: 02 December 2024 | Publication date: 28 November 2024 | By: Y. Refahi / G. Paës