Agritrop
Home

Identification and biochemical characterization of a GDSL-motif carboxylester hydrolase from Carica papaya latex

Abdelkafi Slim, Ogata Hiroshi Y., Barouh Nathalie, Fouquet Benjamin, Lebrun Régine, Pina Michel, Scheirlinckx Frantz, Villeneuve Pierre, Carrière Frédéric. 2009. Identification and biochemical characterization of a GDSL-motif carboxylester hydrolase from Carica papaya latex. Biochimica and Biophysica Acta. Molecular and Cell Biology of Lipids, 1791 (11) : pp. 1048-1056.

Journal article ; Article de revue à facteur d'impact
[img] Published version - Anglais
Access restricted to CIRAD agents
Use under authorization by the author or CIRAD.
document_551148.pdf

Télécharger (1MB)

Quartile : Q1, Sujet : BIOPHYSICS / Quartile : Q1, Sujet : BIOCHEMISTRY & MOLECULAR BIOLOGY / Quartile : Q2, Sujet : CELL BIOLOGY

Abstract : An esterase (CpEst) showing high specific activities on tributyrin and short chain vinyl esters was obtained from Carica papaya latex after an extraction step with zwitterionic detergent and sonication, followed by gel filtration chromatography. Although the protein could not be purified to complete homogeneity due to its presence in high molecular mass aggregates, a major protein band with an apparent molecular mass of 41 kDa was obtained by SDS-PAGE. This material was digested with trypsin and the amino acid sequences of the tryptic peptides were determined by LC/ESI/MS/MS. These sequences were used to identify a partial cDNA (679 bp) from expressed sequence tags (ESTs) of C. papaya. Based upon EST sequences, a full-length gene was identified in the genome of C. papaya, with an open reading frame of 1029 bp encoding a protein of 343 amino acid residues, with a theoretical molecular mass of 38 kDa. From sequence analysis, CpEst was identified as a GDSL-motif carboxylester hydrolase belonging to the SGNH protein family and four potential N-glycosylation sites were identified. The putative catalytic triad was localised (Ser35-Asp307-His310) with the nucleophile serine being part of the GDSL-motif. A 3D-model of CpEst was built from known X-ray structures and sequence alignments and the catalytic triad was found to be exposed at the surface of the molecule, thus confirming the results of CpEst inhibition by tetrahydrolipstatin suggesting a direct accessibility of the inhibitor to the active site. (Résumé d'auteur)

Mots-clés Agrovoc : Carica papaya, Latex, Estérase

Mots-clés géographiques Agrovoc : Congo

Classification Agris : Q01 - Food science and technology
Q04 - Food composition

Champ stratégique Cirad : Axe 3 (2005-2013) - Alimentation accessible et de qualité

Auteurs et affiliations

  • Abdelkafi Slim, CNRS (FRA)
  • Ogata Hiroshi Y., CNRS (FRA)
  • Barouh Nathalie, CIRAD-PERSYST-UMR IATE (FRA)
  • Fouquet Benjamin, Biohainaut (BEL)
  • Lebrun Régine, Institut de Microbiologie de la Méditerranée (FRA)
  • Pina Michel, CIRAD-PERSYST-UMR IATE (FRA)
  • Scheirlinckx Frantz, Biohainaut (BEL)
  • Villeneuve Pierre, CIRAD-PERSYST-UMR IATE (FRA) ORCID: 0000-0003-1685-1494
  • Carrière Frédéric, CNRS (FRA)

Autres liens de la publication

Source : Cirad - Agritrop (https://agritrop.cirad.fr/551148/)

View Item (staff only) View Item (staff only)

[ Page générée et mise en cache le 2020-10-27 ]