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000821075 1001_ $$0P:(DE-Juel1)131480$$aKovacic, Filip$$b0$$eCorresponding author
000821075 245__ $$aA membrane-bound esterase PA2949 from Pseudomonas aeruginosa is expressed and purified from Escherichia coli
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000821075 520__ $$aPseudomonas aeruginosa strain 1001 produces an esterase (EstA) that can hydrolyse the racemic methyl ester of b-acetylthioisobutyrate to produce the (D)-enantiomer, which serves as a precursor of captopril, a drug used for treatment of hypertension. We show here that PA2949 from P. aeruginosa PA01, a homologue of EstA, can efficiently be expressed in an enzymatically active form in E. coli. The enzyme is membrane-associated as demonstrated by cell fractionation studies. PA2949 was purified to homogeneity after solubilisation with the nonionic detergent, Triton X-100, and was shown to possess a conserved esterase catalytic triad consisting of Ser137–His258–Asp286. Our results should allow the development of an expression and purification strategy to produce this biotechnologically relevant esterase in a pure form with a high yield.
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000821075 7001_ $$0P:(DE-HGF)0$$aCaliskan, Muttalip$$b2
000821075 7001_ $$0P:(DE-HGF)0$$aWilhelm, Susanne$$b3
000821075 7001_ $$0P:(DE-Juel1)131965$$aGranzin, Joachim$$b4
000821075 7001_ $$0P:(DE-Juel1)131950$$aBatra-Safferling, Renu$$b5
000821075 7001_ $$0P:(DE-Juel1)131457$$aJaeger, Karl-Erich$$b6
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