Crystallization and preliminary X-ray diffraction analysis of a thermostable endo-1,5-alpha-L-arabinanase from Bacillus thermodenitrificans TS-3.
A thermostable endo-1,5-alpha-L-arabinanase ABN-TS from Bacillus thermodenitrificans TS-3 with a molecular weight of 35 kDa was crystallized by the hanging-drop vapour-diffusion method using sodium citrate as a precipitant. The crystals were loop-mounted in a cryoprotectant solution containing 28%(w/v) sucrose and 1 M sodium citrate pH 6.0 and flash-cooled. Sucrose was selected as the most suitable cryoprotectant. The crystal belonged to the orthorhombic space group P2(1)2(1)2(1), with unit-cell parameters a = 40.3, b = 77.8, c = 89.7 angstroms. The calculated VM based on one molecule per asymmetric unit was 2.0 angstroms3 Da(-1). A complete data set from a frozen crystal was collected to 1.9 angstroms resolution using synchrotron radiation at SPring-8. A molecular-replacement solution was obtained using the structure of alpha-arabinanase 43A from Cellvibrio japonicus.