title: 3D structure elucidation of thermostable L2 lipase from Thermophilic Bacillus sp. L2.
abstract: The crystallization of proteins makes it possible to determine their structure by X-ray crystallography, and is therefore important for the analysis of protein structure-function relationships. L2 lipase was crystallized by using the J-tube counter diffusion method. A crystallization consisting of 20% PEG 6000, 50 mM MES pH 6.5 and 50 mM NaCl was found to be the best condition to produce crystals with good shape and size (0.5 × 0.1 × 0.2 mm). The protein concentration used for the crystallization was 3 mg/mL. L2 lipase crystal has two crystal forms, Shape 1 and Shape 2. Shape 2 L2 lipase crystal was diffracted at 1.5 Å and the crystal belongs to the orthorhombic space group P2(1)2(1)2(1), with unit-cell parameters a = 72.0, b = 81.8, c = 83.4 Å, α = β = γ = 90°. There is one molecule per asymmetric unit and the solvent content of the crystals is 56.9%, with a Matthew's coefficient of 2.85 Å Da(-1). The 3D structure of L2 lipase revealed topological organization of α/β-hydrolase fold consisting of 11 β-strands and 13 α-helices. Ser-113, His-358 and Asp-317 were assigned as catalytic triad residues. One Ca(2+) and one Zn(2+) were found in the L2 lipase molecule.
authors.affiliation: Faculty of Biotechnology and Biomolecular Sciences, Enzyme and Microbial Technology Research, Universiti Putra Malaysia, UPM Serdang, Selangor 43400, Malaysia.|Institute of Bioscience, Enzyme and Microbial Technology Research, Universiti Putra Malaysia, UPM Serdang, Selangor 43400, Malaysia.
authors.forename: Raja Noor Zaliha
authors.initials: RNZ
authors.lastname: Raja Abd Rahman
authors.affiliation: Faculty of Biotechnology and Biomolecular Sciences, Enzyme and Microbial Technology Research, Universiti Putra Malaysia, UPM Serdang, Selangor 43400, Malaysia.
authors.forename: Fairolniza
authors.initials: F
authors.lastname: Mohd Shariff
authors.affiliation: Institute of Bioscience, Enzyme and Microbial Technology Research, Universiti Putra Malaysia, UPM Serdang, Selangor 43400, Malaysia.|Faculty of Science, Enzyme and Microbial Technology Research, Universiti Putra Malaysia, UPM Serdang, Selangor 43400, Malaysia.
authors.forename: Mahiran
authors.initials: M
authors.lastname: Basri
authors.affiliation: Faculty of Biotechnology and Biomolecular Sciences, Enzyme and Microbial Technology Research, Universiti Putra Malaysia, UPM Serdang, Selangor 43400, Malaysia.|Institute of Bioscience, Enzyme and Microbial Technology Research, Universiti Putra Malaysia, UPM Serdang, Selangor 43400, Malaysia.
authors.forename: Abu Bakar
authors.initials: AB
authors.lastname: Salleh
mesh_terms: D001407:Bacillus / Q000201:enzymology*; D001426:Bacterial Proteins / Q000737:chemistry*; D002118:Calcium / Q000737:chemistry*; D018360:Crystallography, X-Ray; D008049:Lipase / Q000737:chemistry*; D017434:Protein Structure, Tertiary; D015032:Zinc / Q000737:chemistry*
chemical_list: D001426:Bacterial Proteins; D008049:Lipase; D015032:Zinc; D002118:Calcium