Journal
FEBS OPEN BIO
Volume 6, Issue 5, Pages 484-493Publisher
WILEY
DOI: 10.1002/2211-5463.12061
Keywords
(D)-beta-acetylthioisobutyric acid; Escherichiacoli; esterase; inhibition; membrane protein; Pseudomonas aeruginosa
Categories
Funding
- European Union (FP6 Marie Curie EST project ANTIBIOTARGET) [MEST-CT-2005-020278]
- German Research Foundation (DFG) within the Collaborative Research Center 1208 'Identity and Dynamics of Membrane Systems'
Ask authors/readers for more resources
Pseudomonasaeruginosa strain 1001 produces an esterase (EstA) that can hydrolyse the racemic methyl ester of -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.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available