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Staphylococcus aureus lipase: purification, kinetic characterization, crystallization and crystallographic study
Author(s) -
Tanaka Mutsumi,
Kamitani Shigeki,
Kitadokoro Kengo
Publication year - 2018
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 37
ISSN - 2053-230X
DOI - 10.1107/s2053230x18010506
Subject(s) - staphylococcus aureus , lipase , chemistry , escherichia coli , chromatography , affinity chromatography , enzyme , biochemistry , biology , bacteria , gene , genetics
Staphylococcus aureus lipase (SAL), a triacylglycerol esterase, is an important virulence factor in S. aureus and may be a therapeutic target for infectious diseases caused by S. aureus . For the purposes of anti‐SAL drug development using structure‐based drug design, X‐ray crystallographic analysis of SAL overexpressed in Escherichia coli was performed. The recombinant protein was purified using a three‐step protocol involving immobilized metal‐affinity chromatography, cation‐exchange chromatography and anion‐exchange chromatography flowthrough methods, yielding 40 mg of protein per litre of bacterial culture. Crystals were obtained using the sitting‐drop vapor‐diffusion technique. Diffraction data to 3.0 Å resolution were collected on the BL44XU beamline at SPring‐8 at the zinc peak of 1.2842 Å for SAD phasing. The crystals belonged to space group P 4 1 22 or P 4 3 22, with unit‐cell parameters a = 131.0, b = 131.0, c = 250.6 Å, and are likely to contain four SAL molecules (408 residues) per asymmetric unit.