Oxidation of colourless dye precursors with laccase enzyme provided simultaneous "in situ" generation and fixation of a pigment on amino groups pre-functionalized cotton fabric. Aromatic amine moieties of 2,5-diaminobenzenesulfonic acid introduced onto tosylated cotton were coupled and copolymerised with a phenolic compound catechol into coloured product covalently fixed on the fabric upon oxidation with laccase. The controlled amination of cellulose in a first step and subsequent colouration allowed for Lip to 95% pigment fixation on the fabric. Electrochemical studies were performed to elucidate the mechanism of the pigment formation. The pigment was further isolated from the acid hydrolysate of the dyed cellulose fabric to confirm the covalent fixation and to further elucidate the pigment structure by means of FTIR, MS, H-1 and C-13 NMR analysis. An oligomeric pigment has been identified composed by up to six phenolic units. (C) 2009 Elsevier Inc. All rights reserved.