The chemical modification of cellulose (Cs)-chitosan (Ch)-electrospun non-woven (ESNW) composite fabrics is described. Since the as-spun Cs/Ch-composite ESNW (Cs : Ch ¼ 4 : 6) deforms in water, an insolublilization procedure using an alkaline-ethanol solution was developed to extend the applications in an aqueous environment. The fine fiber surface was modified with bifunctional isocyanate in order to introduce the -NCO groups on the ESNW surfaces. The -NCO groups were condensed with N,N-(diethyl)ethylene diamine to generate a cationic diethylaminoethyl (DEAE)-ESNW. An enzyme, aminoacylase-I, was immobilized onto the cationic matrix under mild conditions. The immobilized enzyme was subjected to the stereo-specific recognition of N -acetyl-methionines. The results suggest that the chemically modified Cs/Ch-ESNW is a useful support matrix for biological catalysts.Electrospinning is a unique technique for the fabrication of polymer solutions into fine fibers, which are referred to as electrospun non-woven fabrics (ESNWs). The electrospun fibers often have average diameters ranging from submicron to nano-scale. 1,2 Our research group has reported the preparation of chitosan (Ch) 3,4 and cellulose (Cs) 5 nanofibers via a conventional electrospinning method for the first time. We then continued a series of applied research studies using the composite ESNWs which were prepared from polysaccharides-synthetic phosphorylated poly(amino acid)s for crystallization of the calcium phosphate in an aqueous environment. 6,7 More recently, we have reported the immobilization of an enzyme on the chemically modified hydroxypropyl cellulose ESNWs. 8 Cellulose and chitosan (Scheme 1) are two widely available biomaterials, which have undergone more research studies. 9 Our recent report on the direct electrospinning of cellulose-chitosan composite ESNWs is promising for the production of composite fibers with average nano-scale diameters. 10 The as-spun Cs/Ch-ESNW in water tends to swell or become distorted into a thin film due to the presence of trifluoroacetic acid (TFA) salts, which were derived from the spinning solvent. An appropriate post-spun treatment