Crystals of Thermoactinomyces vulgaris alpha-amylase II, which is a new type of cu-amylase having hydrolysis activities for pullulan and cyclodextrins, have been obtained and diffraction data to 2.9 Angstrom resolution were collected. The crystal belongs to an orthorhombic system with cell dimensions of a = 119.5 Angstrom, b = 120.6 Angstrom, and c = 114.6 Angstrom and a space group of P2(1)2(1)2(1). Two or three protein molecules are expected to exist in an asymmetric unit. (C) 1995 Academic Press, Inc.