Identifying the negative cooperation between major inhibitors of cellulase activity and minimizing their inhibitory potential during hydrolysis of acid-pretreated corn stover.

Soluble compounds produced during the enzymatic hydrolysis of lignocelluloses hampers cellulose conversion. Cellobiose and vanillin most severely inhibited the effect of cellobiohydrolase I. A concentration-dependent negative cooperative effect was found between cellobiose and vanillin. The combined inhibitory effect was about 83.5% of the cellobiose and 88.1% of the vanillin when their concentration was 20 mg/ml. However, the negative synergy could be eliminated by excessive enzyme loading. Differences in their binding sites on the catalytic domain of cellobiohydrolase I lead to negative synergistic inhibition, which should be considered in devising strategies to alleviate this effect. Combined ß-glucosidase and PEG addition at an appropriate dose was feasible to balance cost and hydrolytic efficiency. To achieve efficient hydrolysis, especially at high solid concentrations, it is important to understand the synergistic inhibition between these inhibitors.