Ozonolysis, as a lignocellulosic biomass pretreatment, goes back to 80s; however, in the last years it is becoming widespread again owing to its efficiency and mild operation conditions. Ozone reacts preferably with lignin than carbohydrates, promoting biomass destructuration and delignification, and so the sugar release by enzymatic hydrolysis. The hydrolysate from pretreated biomass has being used as sugars source for second-generation fuels production, mainly ethanol, methane and hydrogen. Short-chain carboxylic acids are the