Bacterial infections have become more difficult to treat as communication between bacteria allows for growth and resistance. Bacteria use autoinducer molecules to take a census count of neighboring bacteria in the colony. The autoinducer molecules are synthesized inside the cell by a dedicated set of enzymes and are released outside to aid in intercellular communication. This mechanism of communication, known as quorum sensing, allows bacteria to coordinate their activities that include secretion of virulence factors and formation of biofilms. EsaI is an AHL Synthase enzyme in the plant pathogen Pantoea stewartii, which uses S-adenosyl-L-methionine and 3-oxohexanoyl-ACP to make 3-oxohexanoyl-HSL autoinducer molecules inside the cell. We expressed EsaI AHL Synthase as a his-tagged protein and purified it using Ni-NTA chromatography. The purity of EsaI was confirmed by SDS-PAGE gel electrophoresis. EsaI enzyme will be subjected to mechanistic studies in the future.

Expression and purification of EsaI AHL synthase enzyme

243rd National Meeting (2012)

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