title: A new generation of versatile chromogenic substrates for high-throughput analysis of biomass-degrading enzymes.
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.
authors.affiliation: Department of Plant and Environmental Sciences, Thorvaldsensvej 40, Frederiksberg, C 1871 Denmark.
authors.forename: Stjepan Krešimir
authors.initials: SK
authors.lastname: Kračun
authors.affiliation: Department of Plant and Environmental Sciences, Thorvaldsensvej 40, Frederiksberg, C 1871 Denmark.
authors.forename: Julia
authors.initials: J
authors.lastname: Schückel
authors.affiliation: Department of Plant and Environmental Sciences, Thorvaldsensvej 40, Frederiksberg, C 1871 Denmark.|Department of Chemistry, Biotechnology and Food Science, Norwegian University of Life Sciences, Chr. M. Falsens vei 1., Aas, 1432 Norway.|University of Copenhagen, Faculty of Science, Rolighedsvej 23, Frederiksberg, C 1958 Denmark.
authors.forename: Bjørge
authors.initials: B
authors.lastname: Westereng
authors.affiliation: University of Copenhagen, Faculty of Science, Rolighedsvej 23, Frederiksberg, C 1958 Denmark.
authors.forename: Lisbeth Garbrecht
authors.initials: LG
authors.lastname: Thygesen
authors.affiliation: Novozymes A/S, Krogshoejvej 36, Bagsværd, 2880 Denmark.
authors.forename: Rune Nygaard
authors.initials: RN
authors.lastname: Monrad
authors.affiliation: Department of Chemistry, Biotechnology and Food Science, Norwegian University of Life Sciences, Chr. M. Falsens vei 1., Aas, 1432 Norway.
authors.forename: Vincent G H
authors.initials: VGH
authors.lastname: Eijsink
authors.affiliation: Department of Plant and Environmental Sciences, Thorvaldsensvej 40, Frederiksberg, C 1871 Denmark.
authors.forename: William George Tycho
authors.initials: WGT
authors.lastname: Willats
keywords: Biomass degradation; Carbohydrate-active enzymes; Chromogenic substrates; High-throughput screening; Lytic polysaccharide monooxygenases; Plant cell walls