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Bacterial botin protein ligase assays

Biotin protein ligase (BPL or BirA) is responsible for biotinylation of biotin-dependent proteins.  It is an

essential enzyme and an attractive target for anti-bacterial drug discovery.

The Bacterial Biotin Protein Ligase Assay is based on measurement of the pyrophosphate generated from

the BirA reaction using hydroxylamine in the place of protein as a biotin receptor.  The pyrophosphate is

converted to phosphate by pyrophosphatase and the phosphate is detected by light absorbance at 650 nm.

The assay reactions and detection can be performed by using 384-well or 96-well assay plates.

Alternatively, the assay reaction can be carried out in Eppendorf tubes and the signal is measured using a

cuvette. The high throughput assay can be used for screening inhibitors of Bacterial BirA in drug discovery

research.  It may also be used for characterization of Bacterial BirA.