This video demonstrates an assay to screen enzymes effective in degrading complex polysaccharides in plant biomass using insoluble chromogenic biomass or ICB. The enzymes hydrolyze ICB to release soluble dye-linked oligosaccharides, and the enzyme activity is assessed by colorimetric analysis of the reaction product.
Protocol
1. Chromogenic Assay with ICB Substrates in a 96-well Format Enzyme reaction Add the 150 µl 100 mM sodium acetate buffer, pH 4.5 and 5 µl 31 U/ml endo-xylanase solution to each well of the assay kit plate containing red ICB-wheat straw (final enzyme concentration in the well: 1 U/ml). NOTE: The assay kit plates (96-well filter plates containing ICB substrates) are manufactured as described in literature (see List of Materials). The …
Declarações
The authors have nothing to disclose.
Materials
Assay kit plates
Glycospot
Customized assay kit plates
350 ml receiver plate spacer block for vacuum manifold
Insoluble Chromogenic Biomass-Based Enzyme Screening: An Assay to Evaluate the Degradation Activity of Plant Biomass-Degrading Enzymes. J. Vis. Exp. (Pending Publication), e21177, doi: (2023).