NoteA rapid procedure for the in situ assay of periplasmic, PQQ-dependent methanol dehydrogenase in intact single bacterial colonies

- Development of an in situ colony assay for PQQ-linked methanol dehydrogenase using nitroblue tetrazolium (NBTZ).
- The assay is suitable for robotic screening techniques.
- The NBTZ in situ assay can also detect heterogeneity in populations of bacterial colonies containing methanol dehydrogenase.

Mechanistic details of methanol oxidation catalyzed by the periplasmically-located pyrroloquinoline quinone-dependent methanol dehydrogenase of methylotrophs can be elucidated using site-directed mutants. Here, we present an in situ colony assay of methanol dehydrogenase, which allows robotic screening of large populations of intact small colonies, and regrowth of colonies for subsequent analysis.

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