title: Expression, purification, crystallization and preliminary X-ray crystallographic analysis of tomato β-galactosidase 4.
abstract: Plant β-galactosidases play important roles in carbohydrate-reserve mobilization, cell-wall expansion and degradation, and turnover of signalling molecules during ripening. Tomato β-galactosidase 4 (TBG4) not only has β-galactosidase activity but also has exo-β-(1,4)-galactanase activity, and prefers β-(1,4)-galactans longer than pentamers as its substrates; most other β-galactosidases only have the former activity. Recombinant TBG4 protein expressed in the yeast Pichia pastoris was crystallized by the sitting-drop vapour-diffusion method using PEG 10,000 as a precipitant. The crystals belonged to the orthorhombic space group P212121, with unit-parameters a = 92.82, b = 96.30, c = 159.26 Å, and diffracted to 1.65 Å resolution. Calculation of the Matthews coefficient suggested the presence of two monomers per asymmetric unit (VM = 2.2 Å(3) Da(-1)), with a solvent content of 45%.
authors: Graduate School of Science, Osaka Prefecture University, Sakai, Osaka 599-8531, Japan.
authors: Masahiro
authors: M
authors: Eda
authors: Faculty of Biology-Oriented Science and Technology, Kinki University, Kinokawa, Wakayama 631-8505, Japan.
authors: Megumi
authors: M
authors: Ishimaru
authors: Graduate School of Science, Osaka Prefecture University, Sakai, Osaka 599-8531, Japan.
authors: Toshiji
authors: T
authors: Tada
mesh_terms: D000595:Amino Acid Sequence; D003460:Crystallization; D018360:Crystallography, X-Ray; D004591:Electrophoresis, Polyacrylamide Gel; D018551:Solanum lycopersicum / Q000201:enzymology*; D008969:Molecular Sequence Data; D001616:beta-Galactosidase / Q000737:chemistry* / Q000302:isolation & purification*
chemical_list: D001616:beta-Galactosidase
keywords: Solanum lycopersicum; TBG4; plant cell wall; β-galactosidase