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    Heterologous expression, kinetic characterization and molecular modeling of a new sn-1,3-regioselective triacylglycerol lipase from Serratia sp. W3

    Year: 2021

    Journal: Process Biochem., Volume 103, APR, page 87–97

    Authors: Eddehech, Ahlem; Rahier, Renaud; Smichi, Nabil; Arhab, Yani; Noiriel, Alexandre; Abousalham, Abdelkarim; Sayari, Adel; Zarai, Zied

    Organizations: Tunisian Ministry of Higher Education and Scientific Research [18PJEC08-04]; PHC-Maghreb program of the French Ministry of Foreign Affairs; Ministry of Higher Education, Research and Innovation [43791 T M, PHC: 01MAG20]

    Keywords: Serratia lipase expression; Catalytic activity; Interfacial properties; Regio-selectivity; Molecular modeling

    A recombinant Serratia sp. W3 lipase (rSmL) was expressed in E. coli and purified to homogeneity. The kinetic properties, regio-selectivity, and interfacial performances of rSmL were compared with those of the native lipase (nSmL). rSmL has a molecular mass of 67 kDa and a specific activity of 3530 U mg(-1) which is around 12-fold higher than that of nSmL (300 U mg(-1)) when using olive oil as the substrate. Both rSmL and nSmL were able to hydrolyze the ester bond at the sn-1 and sn-3 positions but exhibited a clear regio-preference towards the sn-3 position of the surface-coated triglycerides (TG), which were esterified with alpha-eleostearic acid at the sn-1/3 position or dicaprin isomers spread as monomolecular films. Molecular modeling and docking of TG into the active site of rSmL indicated that this regio-preference may be due to steric hindrance, created by the residues Ile(308) and Trp(311), with distances between the sn-1 reactive carbon and the catalytic Serine residue ranging from 3.15 to 5.47 angstrom. In contrast, the sn-3 positioning within the enzyme catalytic pocket resulted in shorter distances, ranging from 2.79 to 4.15 angstrom, therefore facilitating the hydrolysis at the sn-3 position.
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