A current barrier to the large-scale production of lignocellulosic biofuels is the cost associated with the energy and chemical inputs required for feedstock pretreatment and hydrolysis. The use of controlled partial biological degradation to replace elements of the current pretreatment technologies would offer tangible energy and cost benefits to the whole biofuel process. It has been known for some time from studies of wood decay that, in the early stages of growth in wood, brown rot fungi utilise a mechanism that causes rapid and extensive depolymerisation of the carbohydrate polymers of the wood cell wall. The brown rot hyphae act as delivery vectors to the plant cell wall for what is thought to be a combination of a localised acid pretreatment and a hydroxyl radical based depolymerisation of the cell wall carbohydrate polymers. It is this quality that we have exploited in the present work to enhance the saccharification potential of softwood forest residues for biofuel production. Here we show that after restricted exposure of pine sapwood to brown rot fungi, glucose yields following enzymatic saccharification are significantly increased. Our results demonstrate the potential of using brown rot fungi as a biological pretreatment for biofuel production. (C) 2010 Elsevier Ltd. All rights reserved.