The esterase from Alicyclobacillus acidocaldarius as a reporter enzyme and affinity tag for protein biosynthesis

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Abstract
Esterase from thermophilic bacteria Alicyclobacillus acidocaldarius can be produced up to 200 μg/ml by coupled in vitro transcription/translation system derived from Escherichia coli. The synthesized thermostable enzyme can be determined by photometrical and fluorescent assays at least up to 10−8 M concentration or by activity staining in the polyacrylamide gels. Enhanced green fluorescence protein‐esterase fusion protein was bound to a matrix with immobilized esterase inhibitor and purified by affinity chromatography. Thus, the esterase is suited as a reporter enzyme to monitor the expression of polypeptides coupled to its N‐terminus and simultaneously, as a cleavable tag for polypeptide purification.