Navigating Side Reactions for Robust Colorimetric Detection of Galactose Oxidase Activity.

ABSTRACT: Colorimetric assays are a rapid, scalable technique well suited to enzyme activity screening. However, side reactions or chromogenic reagent instability can result in false positives or false negatives that compromise the accuracy of such assays. Here, we identify three classes of compounds incompatible with the 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) colorimetric assay for galactose oxidase activity. Dark green ABTS^·+ cationic radicals indicating enzyme activity can get quenched to yield colorless solutions or couple with substrates to form differently colored adducts, thus preventing accurate colorimetric measurements. These side reactions limit the utility of the ABTS assay and introduce uncertainty in the substrate scope to which it is applicable. We ha

SUBMITTER: Koo YS 

PROVIDER: S-EPMC11912124 | biostudies-literature | 2025 Mar

REPOSITORIES: biostudies-literature