Diacetyl-chitobiose (GlcNAc)(2) was produced exclusively using a Vibrio furnissii chitinase (chi E, 89 kDa) hydrolysis of chitin treated with dimethylsulfoxide containing lithium chloride. This method represents a novel way for the preparation of colloidal chitin. Continuous diafiltration was used for the efficient recycling of enzyme and synthesis of (GlcNAc)(2) on a preparative scale. Chitooligosaccharides were also synthesized in a transglycosylation reaction employing lysozyme from hen-egg white with diacetyl-chitobiose produced by a Vibrio furnissii chitinase (chi E, 89 kDa) as a substrate in 50% 2-ethoxy ethyl ether co-solvent system. The co-solvent systems studied were triethyl phosphate, 2-ethoxy ethyl ether and acetone. (c) 2005 Elsevier Inc. All rights reserved.