Abstract
Background: Enzymes that degrade or modify polysaccharides are widespread in pro- and eukaryotes and have
multiple biological roles and biotechnological applications. Recent advances in genome and secretome sequencing,
together with associated bioinformatic tools, have enabled large numbers of carbohydrate-acting enzymes to be
putatively identified. However, there is a paucity of methods for rapidly screening the biochemical activities of these
enzymes, and this is a serious bottleneck in the development of enzyme-reliant bio-refining processes.
Results: We have developed a new generation of multi-coloured chromogenic polysaccharide and protein substrates
that can be used in cheap, convenient and high-throughput multiplexed assays. In addition, we have produced substrates
of biomass materials in which the complexity of plant cell walls is partially maintained.
Conclusions: We show that these substrates can be used to screen the activities of glycosyl hydrolases, lytic
polysaccharide monooxygenases and proteases and provide insight into substrate availability within biomass. We
envisage that the assays we have developed will be used primarily for first-level screening of large numbers of putative
carbohydrate-acting enzymes, and the assays have the potential to be incorporated into fully or semi-automated robotic
enzyme screening systems
multiple biological roles and biotechnological applications. Recent advances in genome and secretome sequencing,
together with associated bioinformatic tools, have enabled large numbers of carbohydrate-acting enzymes to be
putatively identified. However, there is a paucity of methods for rapidly screening the biochemical activities of these
enzymes, and this is a serious bottleneck in the development of enzyme-reliant bio-refining processes.
Results: We have developed a new generation of multi-coloured chromogenic polysaccharide and protein substrates
that can be used in cheap, convenient and high-throughput multiplexed assays. In addition, we have produced substrates
of biomass materials in which the complexity of plant cell walls is partially maintained.
Conclusions: We show that these substrates can be used to screen the activities of glycosyl hydrolases, lytic
polysaccharide monooxygenases and proteases and provide insight into substrate availability within biomass. We
envisage that the assays we have developed will be used primarily for first-level screening of large numbers of putative
carbohydrate-acting enzymes, and the assays have the potential to be incorporated into fully or semi-automated robotic
enzyme screening systems
| Original language | English |
|---|---|
| Article number | 70 |
| Journal | Biotechnology for Biofuels |
| Volume | 8 |
| Number of pages | 16 |
| ISSN | 1754-6834 |
| DOIs | |
| Publication status | Published - 23 Apr 2015 |