title: Manipulation of the conformation and enzymatic properties of T1 lipase by site-directed mutagenesis of the protein core.
abstract: In silico and experimental investigations were conducted to explore the effects of substituting hydrophobic residues, Val, Met, Leu, Ile, Trp, and Phe into Gln 114 of T1 lipase. The in silico investigations accurately predicted the enzymatic characteristics of the mutants in the experimental studies and provided rationalization for some of the experimental observations. Substitution with Leu successfully improved the conformational stability and enzymatic characteristics of T1 lipase. However, replacement of Gln114 with Trp negatively affected T1 lipase and resulted in the largest disruption of protein stability, diminished lipase activity and inferior enzymatic characteristics. These results suggested that the substitution of a larger residue in a densely packed area of the protein core can have considerable effects on the structure and function of an enzyme. This is especially true when the residue is next to the catalytic serine as demonstrated with the Phe and Trp mutation.
authors.affiliation: Faculty of Science, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia.
authors.forename: Roswanira Abdul
authors.initials: RA
authors.lastname: Wahab
authors.forename: Mahiran
authors.initials: M
authors.lastname: Basri
authors.forename: Raja Noor Zaliha Raja Abdul
authors.initials: RN
authors.lastname: Rahman
authors.forename: Abu Bakar
authors.initials: AB
authors.lastname: Salleh
authors.forename: Mohd Basyaruddin Abdul
authors.initials: MB
authors.lastname: Rahman
authors.forename: Leow Thean
authors.initials: LT
authors.lastname: Chor
mesh_terms: D019295:Computational Biology; D004795:Enzyme Stability; D056506:Geobacillus / Q000201:enzymology; D008049:Lipase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D016297:Mutagenesis, Site-Directed / Q000379:methods*; D009154:Mutation; D009930:Organic Chemicals / Q000737:chemistry; D010938:Plant Oils / Q000378:metabolism; D011487:Protein Conformation; D012997:Solvents / Q000737:chemistry; D013501:Surface-Active Agents / Q000737:chemistry; D013696:Temperature
chemical_list: D009930:Organic Chemicals; D010938:Plant Oils; D012997:Solvents; D013501:Surface-Active Agents; D008049:Lipase