Potassium acetate is a good buffer used in high throughput screening for cellulose-degrading enzymes, can we replace it with sodium acetate or other buffer?
Dear Farah,
Yes, you can. For more details, please see attached publication (Figure 2):
Kračun et al. Biotechnology for Biofuels (2015) 8:70
DOI 10.1186/s13068-015-0250-y
A new generation of versatile chromogenic
substrates for high-throughput analysis of
biomass-degrading enzymes
Stjepan Krešimir Kračun1†
, Julia Schückel1†
, Bjørge Westereng1,2,3, Lisbeth Garbrecht Thygesen3
,
Rune Nygaard Monrad4
, Vincent G H Eijsink2 and William George Tycho Willats1*
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.
Keywords: Chromogenic substrates, Carbohydrate-active enzymes, High-throughput screening, Biomass degradation, Plant cell walls, Lytic polysaccharide monooxygenases
Hoping this will be helpful,
Rafik
Thank you very much Prof, It is really helpful.
Very Cordially!
Thank you very much Prof John H.T. Luong. I have checked that NA and K are used as a buffer in high throughput screening for cellulose-degrading enzymes.
Sincerely!
Yes you can use it. I work with cellulolytic yeast and i do use Na-acetate, but a friend that worked with bacteria used K-acetate. Either of the two can be used, but i read that its always good to continue for a particular research work with any of the two you decided to use.
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