TY  - JOUR
AU  - Kovacic, Filip
AU  - Bleffert, Florian
AU  - Caliskan, Muttalip
AU  - Wilhelm, Susanne
AU  - Granzin, Joachim
AU  - Batra-Safferling, Renu
AU  - Jaeger, Karl-Erich
TI  - A membrane-bound esterase PA2949 from Pseudomonas aeruginosa is expressed and purified from Escherichia coli
JO  - FEBS Open Bio
VL  - 6
IS  - 5
SN  - 2211-5463
CY  - Cambridge
PB  - Elsevier on behalf of the Federation of European Biochemical Societies
M1  - FZJ-2016-06319
SP  - 484 - 493
PY  - 2016
AB  - 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.
LB  - PUB:(DE-HGF)16
UR  - <Go to ISI:>//WOS:000375915100012
C6  - pmid:27419054
DO  - DOI:10.1002/2211-5463.12061
UR  - https://juser.fz-juelich.de/record/821075
ER  -