The renewable biofuels derived from lignocellulosic biomass (LCB) through hydrolysis is a promising alternative of fossil fuel and it creates carbon balance of the ecosystem by recycling the emitted CO2 into biomass production. LCB mainly comprises of cellulose, hemicellulose and lignin with a small percentage of pectin, protein, extractive and ash. Prior to hydrolysis of LCB, pretreatment can be performed by different methods namely physical (e.g. mechanical reduction, pyrolysis and extrusion), chemical (e.g. acid, alkaline, oxidative pretreatment and ozonolysis), physiochemical (e.g. steam pretreatment and ammonia fiber explosion (AFEX)) and biological pretreatment. The pretreatment process is one key step to remove lignin and hemicellulose attributing to an improvement in the LCB hydrolysis efficiency. Enzymatic hydrolysis is preferred over acid hydrolysis that produces inhibitory products (e.g. furfural, hydroxymethylfurfural (HMF), acetic acid, formic acid and levulinic acid) of subsequent fermentation process. Different detoxification methods (physical, chemical and biological) are employed to remove the inhibitors. However, enzymatic hydrolysis rate and yield depend on several factors such as concentration and quality of substrate, pretreatment methods, cellulase enzymes and reaction conditions (e.g. temperature, pH and mixing). Upcoming amalgamated techniques with both hydrolysis and fermentation, such as separate enzymatic hydrolysis and fermentation (SHF), simultaneous saccharification and fermentation (SSF), non isothermal simultaneous saccharification and fermentation (NSSF), simultaneous saccharification and co-fermentation (SSCF), simultaneous saccharification, filtration and fermentation (SSFF), two chamber bioreactor separated by a membrane filter, and consolidated bioprocessing are the key areas that require detailed analysis for further technology improvement prior to commercialization of hydrolysis process. © 2015 by Nova Science Publishers, Inc. All rights reserved.