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Crystal structures of Ophiostoma piceae sterol esterase: Structural insights into activation mechanism and product release

DOI: 10.1016/j.jsb.2014.07.007 DOI Help
PMID: 25108239 PMID Help

Authors: Javier Gutierrez-fernandez (Departamento de Cristalografía y Biología Estructural, Instituto de Química-Física “Rocasolano”, CSIC) , María Eugenia Vaquero (Departamento de Biología Medioambiental, Centro de Investigaciones Biológicas, CSIC) , Alicia Prieto (Departamento de Biología Medioambiental, Centro de Investigaciones Biológicas, CSIC) , Jorge Barriuso (Departamento de Biología Medioambiental, Centro de Investigaciones Biológicas, CSIC) , María Jesús Martínez (Departamento de Biología Medioambiental, Centro de Investigaciones Biológicas, CSIC) , Juan A. Hermoso (Departamento de Biología Medioambiental, Centro de Investigaciones Biológicas, CSIC)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Journal Of Structural Biology , VOL 187 (3) , PAGES 215 - 222

State: Published (Approved)
Published: September 2014
Diamond Proposal Number(s): 5027

Abstract: Sterol esterases are able to efficiently hydrolyze both sterol esters and triglycerides and to carry out synthesis reactions in the presence of organic solvents. Their high versatility makes them excellent candidates for biotechnological purposes. Sterol esterase from fungus Ophiostoma piceae (OPE) belongs to the family abH03.01 of the Candida rugosa lipase-like proteins. Crystal structures of OPE were solved in this study for the closed and open conformations. Enzyme activation involves a large displacement of the conserved lid, structural rearrangements of loop alpha 16–alpha 17, and formation of a dimer with a large opening. Three PEG molecules are placed in the active site, mimicking chains of the triglyceride substrate, demonstrating the position of the oxyanion hole and the three pockets that accommodate the sn-1, sn-2 and sn-3 fatty acids chains. One of them is an internal tunnel, connecting the active center with the outer surface of the enzyme 30 Å far from the catalytic Ser220. Based on our structural and biochemical results we propose a mechanism by which a great variety of different substrates can be hydrolyzed in OPE paving the way for the construction of new variants to improve the catalytic properties of these enzymes and their biotechnological applications.

Journal Keywords: Fungal Lipase/Esterase; Activation Mechanism

Subject Areas: Biology and Bio-materials


Instruments: I03-Macromolecular Crystallography

Other Facilities: ESRF