Background: The increasing interest in replacing petroleum-based products by more sustainable materials in the packaging sector gives relevance to cellulose as a biodegradable natural resource. Moreover, its properties can be modified physically, chemically or biotechnologically in order to obtain new bioproducts. Refined cotton linters with high cellulose content were treated with hydrolytic (cellulases) and oxidative (LPMO and Laccase_Tempo) enzymes to evaluate their effect on fibre properties and in improving mechanical fibrillation. Results: Cellulases released cellooligosaccharides, reducing fibre length and partially degrading cellulose. They also improved mechanical fibrillation yielding up to 18% of nanofibrillated cellulose (NFC). LPMO introduced a slight amount of COOH groups in cellulose fibres, releasing cellobionic acid to the effluents. The action of cellulases was improved after LPMO treatment; however, the COOH groups created disappeared from fibres. After mechanical fibrillation of LPMO–cellulase-treated cotton linters a 23% yield of NFC was obtained. Laccase_Tempo treatment also introduced COOH groups in cellulose fibres from cotton, yielding 10% of NFC. Degree of polymerization was reduced by Laccase_Tempo, while LPMO treatment did not significantly affect it but produced a higher reduction in fibre length. The combined treatment with LPMO and cellulase provided films with higher transparency (86%), crystallinity (92%), smoothness and improved barrier properties to air and water than films casted from non-treated linters and from commercial NFC. Conclusions: The combined enzymatic treatment with LPMO and cellulases boosted mechanical fibrillation of cotton linters, improving the NFC production and providing bioproducts with high transparency and high barrier properties
