A New Versatile Microarray-based Method for High Throughput Screening of Carbohydrate-active Enzymes

Abstract

Background: There is a growing discrepancy between the putative identification and the empirical characterization of carbohydrate-active enzymes (CAZymes). Results: We have developed a new versatile and high throughput microarray-based method for screening CAZymes. Conclusion:The method is a powerful addition to the enzyme screening toolbox. Significance: The technique enables the rapid screening of CAZymes and facilitates our biological understanding and industrial utilization.

Branched Pectic Galactan in Phloem-Sieve-Element Cell Walls: Implications for Cell Mechanics

“…AIR extracts from different plant materials, with more defined oligosaccharides from partial degradation/extraction, or with chemically synthesized glycans. Such arrays provide a platform for studying plant cell wall polysaccharides with molecular probes such as MAbs and for identifying enzymatic activity (Vidal-Melgosa et al, 2015). Welldefined, single-compound oligosaccharides provide the highest resolution possible when characterizing protein-carbohydrate interactions, provided that the binding interactions are sufficiently strong to be detected.…”

A Synthetic Glycan Microarray Enables Epitope Mapping of Plant Cell Wall Glycan-Directed Antibodies

“…Although exact substrate specificities of glycosyl hydrolases acting on cell wall glycans can only be determined by structural characterization of the reaction products after incubation of oligo-or polysaccharides, these data demonstrate that the synthetic plant glycan microarray platform provides a useful tool, in combination with well-characterized mAbs, to collect valuable information on the substrate specificities of glycosyl hydrolases in a high-throughput fashion. As our platform provides specific information on the enzyme's substrate specificities, it represents a powerful extension of previous polysaccharide-based array platforms for the identification of new glycosyl hydrolases (Vidal-Melgosa et al 2015;Walker et al 2017). …”

White biotechnology: State of the art strategies for the development of biocatalysts for biorefining

“…This illustrates an essential point that must be addressed in order to truly access enzyme diversity. In this respect, it is noteworthy that some progress in this direction has been made, especially in the area of high throughput cloning and expression (Groisillier et al , 2010, Vincentelli et al , 2011, but also with regard to high throughput characterization (Kracun et al , 2015, Vidal-Melgosa et al , 2015. Finally, it is noteworthy that another problem that plagues sequence-based metagenomics is the incorrect annotation of sequences in public databases.…”

“…Therefore, there is an increasing tendency to develop screens that deploy intact or pretreated biomass as the substrate in high throughput screens (Chundawat et al , 2008, Navarro et al , 2010, Song et al, 2010. In this respect further developments might arise from the field of nanotechnology, as illustrated by a study that focused on the enzyme degradation of xyloglucan-cellulose nanofilms (Cerclier et al , 2013) or by the development of glycan chips (Vidal-Melgosa et al, 2015). It is conceivable that with further development, such approaches might find applications in the field of high-throughput enzyme screening.…”