title: The esterase from Alicyclobacillus acidocaldarius as a reporter enzyme and affinity tag for protein biosynthesis.
abstract: Esterase from thermophilic bacteria Alicyclobacillus acidocaldarius can be produced up to 200 microg/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.
authors.affiliation: Laboratorium für Biochemie, Universität Bayreuth, 95440 Bayreuth, Germany.
authors.forename: Dmitry E
authors.initials: DE
authors.lastname: Agafonov
authors.forename: Kersten S
authors.initials: KS
authors.lastname: Rabe
authors.forename: Michael
authors.initials: M
authors.lastname: Grote
authors.forename: Yiwei
authors.initials: Y
authors.lastname: Huang
authors.forename: Mathias
authors.initials: M
authors.lastname: Sprinzl
mesh_terms: D000345:Affinity Labels; D001483:Base Sequence; D017931:DNA Primers; D004591:Electrophoresis, Polyacrylamide Gel; D004950:Esterases / Q000378:metabolism*; D016937:Gram-Positive Endospore-Forming Rods / Q000201:enzymology*; D015202:Protein Engineering
chemical_list: D000345:Affinity Labels; D017931:DNA Primers; D004950:Esterases