%0 Journal Article
%A Kovacic, Filip
%A Bleffert, Florian
%A Caliskan, Muttalip
%A Wilhelm, Susanne
%A Granzin, Joachim
%A Batra-Safferling, Renu
%A Jaeger, Karl-Erich
%T A membrane-bound esterase PA2949 from Pseudomonas aeruginosa is expressed and purified from Escherichia coli
%J FEBS Open Bio
%V 6
%N 5
%@ 2211-5463
%C Cambridge
%I Elsevier on behalf of the Federation of European Biochemical Societies
%M FZJ-2016-06319
%P 484 - 493
%D 2016
%X Pseudomonas 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.
%F PUB:(DE-HGF)16
%9 Journal Article
%U <Go to ISI:>//WOS:000375915100012
%$ pmid:27419054
%R 10.1002/2211-5463.12061
%U https://juser.fz-juelich.de/record/821075