CRYSTALLIZATION AND PRELIMINARY CRYSTALLOGRAPHIC ANALYSIS OF AN AMYLOPULLULANASE FROM THE HYPERTHERMOPHILIC ARCHAEON PYROCOCCUS-WOESEI

The thermostable amylopullulanase from Pyrococcus woesei was crystalli zed. Crystals, suitable for a crystallographic analysis up to a size o f 0.6 mm in their longest dimension, have been obtained by the vapor d iffusion method in a solution containing polyethyleneglycol 4000 (PEG 4000), isopropanol, and Tris/Cl- buffer pH 7.5. Crystals grown under t hese conditions form hexagonal rods and diffract to a maximum resoluti on of 3 Angstrom. The crystals belong to the trigonal lattice type wit h the spacegroup P3(1)21 or P3(2)21, respectively, have the cell dimen sions a = b = 96.8% Angstrom, c = 196.2 Angstrom, alpha = beta = 90 de grees, gamma = 120 degrees. The crystals have a theoretical packing de nsity of 2.7 Angstrom(3)/Da, assuming one molecule with a molecular we ight of 88.8 kDa in the asymmetric unit. Furthermore the self-rotation analysis of the dataset revealed only crystallographic symmetries. Th e merged native data of two crystals resulted in a 88% complete datase t. (C) 1995 Wiley-Liss, Inc.