%D 2006 %0 ARTICLE %T Purification , crystallization and preliminary X-ray analysis of rice BGlu1 beta-glucosidase with and without 2-deoxy-2-fluoro-beta-D-glucoside . %J Acta Crystallograph . Sect . F Struct . Biol . Cryst Commun . %V 62 ( Pt 8 ) %P 798-801 %A Chuenchor W %A Pengthaisong S %A Yuvaniyama J %A Opassiri R %A Svasti J %A Ketudat Cairns %M pub16880561 %X Rice ( Oryza sativa ) BGlu1 beta-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-beta-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 angstroms resolution , respectively , and had isomorphous orthorhombic unit cells belonging to space group P2 ( 1 ) 2 ( 1 ) 2 ( 1 ) .