Wet oxidation pretreatment, enzymatic hydrolysis and simultaneous saccharification and fermentation of clover-ryegrass mixtures.

The potential of clover (Trifolium repens) and ryegrass (Lolium perenne) mixtures as raw materials for ethanol production was investigated. Wet oxidation, at 175, 185 or 195 degrees C during 10min at two different oxygen pressures and with either addition or no addition of sodium carbonate, was evaluated as pretreatment method for clover-ryegrass mixtures. The enzymatic hydrolysis of cellulose was significantly improved after pretreatment. The highest conversion efficiency, 93.6%, was achieved for the sample pretreated at 195 degrees C, 10min, 1.2MPa and no addition of Na(2)CO(3). For that sample, the overall glucose yield after pretreatment and hydrolysis was 75.5%. No inhibition of cellulose enzymatic conversion by the filtrates was observed. The simultaneous saccharification and fermentation of the pretreated material yielded cellulose conversions of 87.5 and 86.6%, respectively, with Saccharomyces cerevisiae and the filamentous fungus Mucor indicus, and revealed that no addition of nutrients is needed for the fermentation of clover-ryegrass hydrolysates.