title: Kinetic behaviour of recombinant Fusarium solani lipases using monomolecular films: Effect of the heterologous expression.
abstract: Two lipases from Fusarium solani, FSL and FSL2, were efficiently expressed in Pichia pastoris. To check the influence of the expression on interfacial properties of FSL and to study kinetic properties of FSL2, interfacial parameters of FSL2, native FSL, untagged recombinant and tagged recombinant forms of FSL were compared using the monomolecular film technique. Kinetic study on the dependence of the stereoselectivity of these lipases on the surface pressure was performed using three dicaprin isomers spread in the form of monomolecular films at the air-water interface. The FSL2 seems to have an important penetration power with a preference for adjacent ester groups and the heterologous expression accompanied or not with the N-His-tag extension on the FSL were found to modify the pressure preference and increase the catalytic hydrolysis rate of three dicaprin isomers. The heterologous expression was found to preserve the FSL regioselectivity without affecting its stereospecificity at high and low surface pressure. The evaluation of the recombinant expression Effects on Catalysis (REC), the N-Tag Effects on Catalysis (TEC), and the N-Tag and Recombinant expression Effects on Catalysis (TREC) showed that the heterologous expression was more efficient than the presence of the N-terminal tag extension on the FSL.
authors.affiliation: Laboratoire de Biochimie et de Génie Enzymatique des Lipases, ENIS route de Soukra, BPW 3038 Sfax, Tunisie.
authors.forename: Raida
authors.initials: R
authors.lastname: Jallouli
authors.affiliation: Laboratoire de Biochimie et de Génie Enzymatique des Lipases, ENIS route de Soukra, BPW 3038 Sfax, Tunisie.
authors.forename: Madiha
authors.initials: M
authors.lastname: Bouali
authors.affiliation: Laboratoire de Biochimie et de Génie Enzymatique des Lipases, ENIS route de Soukra, BPW 3038 Sfax, Tunisie.
authors.forename: Youssef
authors.initials: Y
authors.lastname: Gargouri
authors.affiliation: Laboratoire de Biochimie et de Génie Enzymatique des Lipases, ENIS route de Soukra, BPW 3038 Sfax, Tunisie. Electronic address: sofiane_bezzine@yahoo.com.
authors.forename: Sofiane
authors.initials: S
authors.lastname: Bezzine
mesh_terms: D005227:Fatty Acids / Q000737:chemistry; D005656:Fungal Proteins / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics; D005670:Fusarium / Q000201:enzymology*; D007700:Kinetics; D008049:Lipase / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics; D008567:Membranes, Artificial; D010713:Phosphatidylcholines / Q000737:chemistry; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D013379:Substrate Specificity
chemical_list: D005227:Fatty Acids; D005656:Fungal Proteins; D008567:Membranes, Artificial; D010713:Phosphatidylcholines; D011994:Recombinant Proteins; D008049:Lipase
keywords: Fusarium solani lipases; Heterologous expression; Monolayer studies; Polyhistidine tag