Crystallization and X-ray analysis of D-threonine aldolase from Chlamydomonas reinhardtii

摘要

D-Threonine aldolase from the green alga Chlamydomonas reinhardtii (CrDTA) catalyzes the interconversion of several beta-hydroxy-D-amino acids (e.g. D-threonine) and glycine plus the corresponding aldehydes. Recombinant CrDTA was overexpressed in Escherichia coli and purified to homogeneity; it was subsequently crystallized using the hanging-drop vapour-diffusion method at 295 K. Data were collected and processed at 1.85 angstrom resolution. Analysis of the diffraction pattern showed that the crystal belonged to space group P1, with unit-cell parameters a = 64.79, b = 74.10, c = 89.94 angstrom, alpha = 77.07, beta = 69.34, gamma = 71.93 degrees. The asymmetric unit contained four molecules of CrDTA. The Matthews coefficient was calculated to be 2.12 angstrom(3) Da(-1) and the solvent content was 41.9%.