Description: A membrane-bound esterase PA2949 from Pseudomonas aeruginosa is expressed and purified from Escherichia coli

This title appears in the Scientific Report : 2016

A membrane-bound esterase PA2949 from Pseudomonas aeruginosa is expressed and purified from Escherichia coli

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,...

Personal Name(s):	Kovacic, Filip (Corresponding author)
	Bleffert, Florian / Caliskan, Muttalip / Wilhelm, Susanne / Granzin, Joachim / Batra-Safferling, Renu / Jaeger, Karl-Erich
Contributing Institute:	Institut für Molekulare Enzymtechnologie (HHUD); IMET Strukturbiochemie; ICS-6
Published in:	FEBS Open Bio, 6 (2016) 5, S. 484 - 493
Imprint:	Cambridge Elsevier on behalf of the Federation of European Biochemical Societies 2016
PubMed ID:	27419054
DOI:	10.1002/2211-5463.12061
Document Type:	Journal Article
Research Program:	Biotechnology Physical Basis of Diseases
Link:	OpenAccess OpenAccess
	Publikationsportal JuSER

Please use the identifier: http://dx.doi.org/10.1002/2211-5463.12061 in citations.
Please use the identifier: http://hdl.handle.net/2128/12856 in citations.

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.