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Purification, crystallization and preliminary X-ray analysis of rice BGlu1 β-glucosidase with and without 2-deoxy-2-fluoro-β-D-glucoside by Watchalee Chuenchor; Salila Pengthaisong; Jirundon Yuvaniyama; Rodjana Opassiri; Jisnuson Svasti; James R. Ketudat Cairns is a Biochemistry, Genetics and Molecular Biology article available to read on EtoBox.
What is Purification, crystallization and preliminary X-ray analysis of rice BGlu1 β-glucosidase with and without 2-deoxy-2-fluoro-β-D-glucoside about?
Rice (Oryza sativa) BGlu1 -glucosidase was expressed in Escherichia coli with N-terminal thioredoxin and hexahistidine tags and purified by immobilized metal-affinity chromatography (IMAC). After removal of the N-terminal tags, cation-exchange and S-200 gel-filtration chromatography yielded a 50 kDa BGlu1 with >95% purity. The free enzyme and a complex with 2,4-dinitrophenyl-2-deoxy-2-fluoro--d-glucopyranoside inhibitor were crystallized by microbatch and hanging-drop vapour diffusion. Small tetragonal crystals of BGlu1 with and without inhibitor grew in 18%(w/v) PEG 8000 with 0.1 M sodium cacodylate pH 6.5 and 0.2 M zinc acetate. Crystals of BGlu1 with inhibitor were streak-seeded into 23%(w/v) PEG MME 5000, 0.2 M ammonium sulfate, 0.1 M MES pH 6.7 to yield larger crystals. Crystals with and without inhibitor diffracted to 2.15 and 2.75 A ̊resolution, respectively, and had isomorphous orthorhombic unit cells belonging to space group P2 1 2 1 2 1 .
Who reads Purification, crystallization and preliminary X-ray analysis of rice BGlu1 β-glucosidase with and without 2-deoxy-2-fluoro-β-D-glucoside?
It is typically read by researchers, students, and practitioners in Biochemistry, Genetics and Molecular Biology.
- Author
- Watchalee Chuenchor; Salila Pengthaisong; Jirundon Yuvaniyama; Rodjana Opassiri; Jisnuson Svasti; James R. Ketudat Cairns
- Publisher
- John Wiley and Sons; International Union of Crystallography; John Wiley and Sons Ltd; International Union of Crystallography (IUCr) (ISSN 1744-3091)
- Published
- 2006
- Language
- EN
- Field
- Biochemistry, Genetics and Molecular Biology (Life Sciences)