Abstract
Improving the specificity of α-cyclodextrin glucanotransferase is a significant issue in the field of α-cyclodextrin production. In this study, a constructed Y167H mutant α-cyclodextrin glucanotransferase with enhanced α-cyclodextrin specificity was successfully expressed and purified. Single crystals were grown using PEG 4000 as a precipitating agent by the hanging-drop vapour-diffusion method at 293 K. The crystals exhibited two kinds of morphology in different crystallization conditions. The crystals diffracted to at least 2.2 Å resolution (space group P2₁2₁2₁), with unit-cell parameters a=65.69, b=78.70, c=137.00 Å. Assuming the asymmetric cell to be occupied by a monomer of 75 kDa, the unit cell contains 43.77% solvent with a crystal volume per protein mass, VM, of 2.19 Å3 Da(-1).