title: Purification, biochemical and kinetic properties of recombinant Staphylococcus aureus lipase.
abstract: We have compared the purification procedures as well as the biochemical and kinetic properties of wild type (wt-SAL3), untagged recombinant (rec(-His)SAL3), and tagged recombinant (rec(+His)SAL3) purified forms of Staphylococcus aureus lipase (SAL3). We used the pH-stat method (with emulsified tributyrin and olive oil as substrates) and the monomolecular film technique (with the three dicaprin isomers spread in the form of monomolecular films at the air-water interface). The data obtained showed that the recombinant expression process as well as the presence of a his-tag at the N-terminus of recombinant SAL3 affects significantly many biochemical and catalytic properties. The effects of the heterologous expression process on the catalytic properties of the staphylococcal lipases are three times more deleterious than the presence of an N-terminal tag extension.
authors.affiliation: Laboratoire de Biochimie et de Génie Enzymatique des Lipases, Ecole Nationale d'Ingénieurs de Sfax, Sfax, Tunisia.
authors.forename: Habib
authors.initials: H
authors.lastname: Horchani
authors.forename: Ahmed
authors.initials: A
authors.lastname: Fendri
authors.forename: Hanen
authors.initials: H
authors.lastname: Louati
authors.forename: Adel
authors.initials: A
authors.lastname: Sayari
authors.forename: Youssef
authors.initials: Y
authors.lastname: Gargouri
authors.forename: Robert
authors.initials: R
authors.lastname: Verger
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D003001:Cloning, Molecular; D004075:Diglycerides / Q000378:metabolism; D004655:Emulsions; D004926:Escherichia coli; D006639:Histidine / Q000737:chemistry; D006868:Hydrolysis; D007218:Industrial Microbiology; D007700:Kinetics; D008049:Lipase / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D008958:Models, Molecular; D009842:Oligopeptides / Q000737:chemistry; D000069463:Olive Oil; D010938:Plant Oils / Q000737:chemistry; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D013211:Staphylococcus aureus / Q000737:chemistry / Q000201:enzymology*; D013329:Structure-Activity Relationship; D013379:Substrate Specificity; D013499:Surface Properties; D014280:Triglycerides / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D004075:Diglycerides; D004655:Emulsions; C471213:His-His-His-His-His-His; D009842:Oligopeptides; D000069463:Olive Oil; D010938:Plant Oils; D011993:Recombinant Fusion Proteins; D014280:Triglycerides; D006639:Histidine; D008049:Lipase; C005830:tributyrin