We report the first example of the engineered expression of tetrameric β-galactosidase containing varying numbers of active monomers. Specifically, by combining wild-type and single-nucleotide-polymorphism plasmids, tetrameric β-galactosidase was expressed <i>in vitro</i> with 1~4 active monomers. Using a single-molecule array, the kinetics of individual enzyme molecules were studied and revealed four distinct populations, corresponding to the number of active monomers in the enzyme. Based on the correlation between enzyme configuration and activity, we investigated the rare event of mistranslation and measured an accurate <i>in vitro</i> mistranslation frequency.

Expression of multimeric enzymes with tunable activity and its application in the study of mistranslation

256th National Meeting (2018)

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