Isolation, characterization, and heterologous expression of a carboxylesterase of Pseudomonas aeruginosa PAO1. (Articolo in rivista)

Type

• Articolo in rivista (Classe)
• Prodotto della ricerca (Classe)

Label

• Isolation, characterization, and heterologous expression of a carboxylesterase of Pseudomonas aeruginosa PAO1. (Articolo in rivista) (literal)

Anno

• 2005-01-01T00:00:00+01:00 (literal)

Alternative label

• Pesaresi A., Devescovi G., Lamba D., Venturi V., Degrassi G. (2005)
  Isolation, characterization, and heterologous expression of a carboxylesterase of Pseudomonas aeruginosa PAO1.
  in Current microbiology (Print)
  (literal)

Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#autori

• Pesaresi A., Devescovi G., Lamba D., Venturi V., Degrassi G. (literal)

Pagina inizio

• 102 (literal)

Pagina fine

• 109 (literal)

Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#numeroVolume

• 50 (literal)

Rivista

• Current microbiology (Rivista)

Note

• ISI Web of Science (WOS) (literal)

Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#affiliazioni

• Pesaresi A. -SISSA,Trieste Devescovi G. -ICGEB, Trieste Lamba D. -IC-CNR, Trieste Venturi V. -ICGEB, Trieste Degrassi G. -ICGEB, Trieste (literal)

Titolo

• Isolation, characterization, and heterologous expression of a carboxylesterase of Pseudomonas aeruginosa PAO1. (literal)

Abstract

• We purified to homogeneity an intracellular esterase from the opportunistic pathogen Pseudomonas aeruginosa PAO1. The enzyme hydrolyzes p-nitrophenyl acetate and other acetylated substrates. The N-terminal amino acid sequence was analyzed and 11 residues, SEPLILDAPNA, were determined. The corresponding gene PA3859 was identified in the P. aeruginosa PAO1 genome as the only gene encoding for a protein with this N-terminus. The encoding gene was cloned in Escherichia coli, and the recombinant protein expressed and purified to homogeneity. According to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis and analytical gel filtration chromatography, the esterase was found to be a monomer of approximately 24 kDa. The experimentally determined isoelectric point was 5.2 and the optimal enzyme activity was at 55 degrees C and at pH 9.0. The esterase preferentially hydrolyzed short-chain fatty acids. It is inhibited by phenylmethylsulfonyl fluoride (PMSF) but not by ethylendiaminotetraacetic acid (EDTA). Native enzyme preparations typically showed a Michaelis constant (K(m)) and V(max) of 0.43 mM and 12,500 U mg(-1), respectively, using p-nitrophenyl acetate as substrate. Homology-based database searches clearly revealed the presence of the consensus GXSXG signature motif that is present in the serine-dependent acylhydrolase protein family. (literal)

Prodotto di

• Institute of Crystallography (IC) (Istituto)
• Studi strutturali e funzionali di proteine di interesse biomedico e biotecnologico (PM.P01.023.001) (Modulo)

Autore CNR

• DORIANO LAMBA (Unità di personale interno)
• Alessandro Pesaresi (Unità di personale esterno)

Incoming links:

Prodotto

• Institute of Crystallography (IC) (Istituto)
• Studi strutturali e funzionali di proteine di interesse biomedico e biotecnologico (PM.P01.023.001) (Modulo)

Autore CNR di

• DORIANO LAMBA (Unità di personale interno)
• Alessandro Pesaresi (Unità di personale esterno)

Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#rivistaDi

• Current microbiology (Rivista)