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

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 α16-α17, 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.

Evolutionary history of versatile-lipases from Agaricales through reconstruction of ancestral structures

“…Comparison of the structures of different lipases from the C. rugosa- like family indicates the importance of both the shape and amino acid composition of the internal tunnel, and their relationship with the kinetic properties of these proteins [3, 7, 8]. The analysis of putative intramolecular tunnels in the ancestral lipases showed that its size increased along evolution (Fig.…”

“…They are able not only to catalyze hydrolysis reactions under aqueous conditions, but also to carry out synthesis reactions, such as esterification and transesterification, in the presence of organic solvents [2]. All of them display the α/β-hydrolase fold, with their catalytic machinery formed by a catalytic triad (serine, histidine and glutamic acid) and the oxyanion hole [3]. These enzymes are present from microbes to plants and animals, and are widely applicable in industrial processes, such as biofuels, oleochemical, food, detergents, cosmetics, pharmaceutical, textile and paper industry, due to their versatility and stability to harsh conditions [4].…”

A Novel CalB-Type Lipase Discovered by Fungal Genomes Mining

“…The pPIC9:PlicBconstruct was transformed into P . pastoris KM71 and GS115 strainsas described before [ 13 ].Activity screening was performed using single colonies from the transformation plates grown in 20 mL YEPS medium in 100 mL flasks with 0.5% w/v of methanol to induce gene expression. The flasks were incubated at 28°C and 250rpm and 0.5% w/v methanol was added daily for maintaining the induction.…”

Classification of lipolytic enzymes and their biotechnological applications in the pulping industry

“…(Ghosh et al 1991), plants (Pringle and Dickstein 2004), and animals (Finer et al 2004), and may be applied in valuable biological processes. The efficient hydrolysis of both triglycerides and sterol esters using sterol esterase from Ophiostoma piceae has been successfully applied for pitch control in the pulp and paper industry (Coloma et al 2015;Gutiérrez-Fernández et al 2014;Calero-Rueda et al 2002). Steryl and cholesteryl esterases from Pseudomonas sp.…”