Butanol production from cellulosic material by anaerobic co-culture of white-rot fungus Phlebia and bacterium Clostridium in consolidated bioprocessing

•A fungal–bacterial co-culture could directly produce butanol from cellulose.•Inhibition of fungal ethanol fermentation increased the yield of butanol.•A synergistic effect of fungus and bacterium enhanced saccharification. Butanol production from lignocelluloses is desirable. Unfortunately, the known wild-types of butanol fermenting Clostridium bacteria are not capable of delignification and saccharification. Here we analyzed butanol production from cellulosic material using anaerobic co-culture of C. saccharoperbutylacetonicum with the white-rot fungus Phlebia sp. MG-60-P2. In consolidated bioprocessing, the co-culture synergistically produced butanol and enhanced saccharification. Knockout of the pyruvate decarboxylase gene from MG-60-P2 to produce transformant line KO77 led to inhibition of ethanol fermentation and high accumulation of saccharified cellobiose and glucose from cellulose. In co-culture of KO77 with C. saccharoperbutylacetonicum, enhanced butanol production was observed (3.2 g/L, compared with 2.5 g/L in co-culture of MG-60-P2 and C. saccharoperbutylacetonicum). We believe this is the first application of co-culture between white-rot fungus and Clostridium to produce butanol from cellulose; butanol production from lignocellulose by co-culture of C. saccharoperbutylacetonicum with Phlebia sp. MG-60-P2 and its transformant should be pursued.