Bacterial expression of the bifunctional alpha-amylase/subtilisin inhibitor from barley

The bifunctionai α-amylaBe/subtilisin inhibitor (BASI) is an endogenous inhibitor of the high pi cereal α-amylases encoded by the amyl genes. Evaluation of the potential role of this protein in malting and brewing would be greatly assisted by the availability of large quantities of the protein. We have produced the protein by expression of the barley gene in bacteria. The barley gene was cloned into a pMAL vector and expressed as a fusion protein. The purified fusion protein was successfully cleaved with a specific protease to release the native BASI protein. The BASI produced by bacterial expression will be a useful source of the protein for studies of interactions with barley α-amylases and studies of the influence of this protein on malting and brewing.