DSpace About DSpace Software
 

DSpace Biblioteca Universidad de Talca (v1.5.2) >
Dirección de Programas de Investigación y Asistencia Técnica (DIAT) >
Artículos en publicaciones ISI - Universidad de Talca >

Please use this identifier to cite or link to this item: http://dspace.utalca.cl/handle/1950/8911

Title: Structural characterization and substrate specificity of VpAAT1 protein related to ester biosynthesis in mountain papaya fruit
Authors: Morales-Quintana, L.
Fuentes, L.
Gaete-Eastman, C.
Herrera, R.
Moya-Leon, M.A.
Keywords: Alcohol acyltransferase
Docking simulation
Ester biosynthesis
Molecular modelling
Vasconcellea pubescens
Issue Date: Feb-2011
Publisher: ELSEVIER SCIENCE INC
Citation: JOURNAL OF MOLECULAR GRAPHICS & MODELLING Volume: 29 Issue: 5 Pages: 635-642 DOI: 10.1016/j.jmgm.2010.11.011
Abstract: The aroma in fruits is an important attribute of quality that influences consumer's acceptance. This attribute is a complex character determined by a set of low molecular weight volatile compounds. In mountain papaya fruit (Vasconcellea pubescens) the aroma is determined mainly by esters, which are produced through an esterification reaction catalyzed by the enzyme alcohol acyltransferase (AAT) that utilizes alcohols and acyl-CoAs as substrates. In order to understand the molecular mechanism involved in the production of esters in this fruit, an AAT gene which has been previously cloned and characterized from mountain papaya (VpAAT1) was expressed in yeasts, and the highest enzyme activity of the recombinant protein was obtained when the enzyme was tested for its ability to produce benzyl acetate. On the other hand, to gain insight the mechanism of action at the molecular level, a structural model for VpAAT1 protein was built by comparative modelling methodology, which was validated and refined by molecular dynamics simulation. The VpAAT1 structure consists of two domains connected by a large crossover loop, with a solvent channel in the center of the structure formed between the two domains. Residues H166 and D170, important for catalytic action, displayed their side chains towards the central cavity of the channel allowing their interaction with the substrates. The conformational interaction between the protein and several ligands was explored by molecular docking simulations, and the predictions obtained were tested through kinetic analysis. Kinetic results showed that the lowest K(M) values were obtained for acetyl-CoA and benzyl alcohol. In addition, the most favorable predicted substrate orientation was observed for benzyl alcohol and acetyl CoA, showing a perfect coincidence between kinetic studies and molecular docking analysis. (C) 2010 Elsevier Inc. All rights reserved.
Description: Morales-Quintana, L (Morales-Quintana, Luis)1; Fuentes, L (Fuentes, Lida)1; Gaete-Eastman, C (Gaete-Eastman, Carlos)1; Herrera, R (Herrera, Raul)1; Moya-Leon, MA (Alejandra Moya-Leon, Maria)1Univ Talca, Inst Biol Vegetal & Biotecnol, Lab Fisiol Vegetal & Genet Mol, Talca, Chile
URI: http://dspace.utalca.cl/handle/1950/8911
ISSN: 1093-3263
Appears in Collections:Artículos en publicaciones ISI - Universidad de Talca

Files in This Item:

File Description SizeFormat
TEXTO COMPLETO.htm3.68 kBHTMLView/Open

Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.

 

Valid XHTML 1.0! DSpace Software Copyright © 2002-2009  The DSpace Foundation - Feedback