The present study deals with the development of an efficient ABE fermentation process using mixed substrate strategy for butanol production wherein no acetone was produced. For this, glucose was supplemented in the medium containing glycerol as main substrate which leads to a higher butanol production of 17.75 g/L in 72 h by Clostridium acetobutylicum KF158795. Moreover, the high cell inoculum also resulted in an increased ABE productivity of 0.46 g/L/h. Further, industrial scalability of the process was also successfully validated in a 300 L fermenter. Furthermore, potential of the Polymeric (PolyRMem) and Zeolite (ZeoMem) membranes for separation of butanol from fermentation broth was also studied by testing the pervaporation performance through which the butanol was successfully recovered. (C) 2013 Elsevier Ltd. All rights reserved.