Enzymatic delignification using the high-redox potential thermostable laccase from the fungus Pycnoporus cinnabarinus and a chemical mediator (1-hydroxybenzotriazole) was investigated to improve totally chlorine-free (TCF) bleaching of Eucalyptus globulus kraft pulps. Different points of incorporation of the enzyme treatment into an industrial-type bleaching sequence (consisting of double oxygen, chelation and peroxide stages) were investigated in pressurized laboratory reactors. The best final pulp properties were obtained using an O-O-L-Q-PoP sequence, where a laccase-mediator stage (L) was incorporated between double oxygen and chelation. The worse results, when the enzymatic and chelation treatments were combined in a unique stage, seemed related to partial inhibition of laccase-mediator activity by the chelator. The new TCF sequence including the laccase stage permitted to improve eucalypt pulp delignification to values around kappa 5 (hexenuronic acid contribution over 50%) compared to kappa 7 using only TCF chemical reagents. In a similar way, the final brightness obtained, over 91% ISO, was 3-4 points higher than that obtained in the chemical sequences. Although technical and economic issues are to be solved, the results obtained show the feasibility of integrating a laccase-mediator treatment into a TCF sequence for bleaching eucalypt kraft pulp. (c) 2006 Society of Chemical Industry.