Accessory enzymes influence cellulase hydrolysis of the model substrate and the realistic lignocellulosic biomass

• Accessory enzymes (AE) deviate cellulase hydrolysis to the lignocellulose from FPA.
• AE can boost the cellulase activity on different substrates to varying degrees.
• The boost is realized apparently by the synergy/cohydrolysis of AE with cellulase.
• New insights are provided into AE, which is helpful to customize cellulase cocktails.

The potential of cellulase enzymes in the developing and ongoing “biorefinery” industry has provided a great motivation to develop an efficient cellulase mixture. Recent work has shown how important the role that the so-called accessory enzymes can play in an effective enzymatic hydrolysis. In this study, three newest Novozymes Cellic CTec cellulase preparations (CTec 1/2/3) were compared to hydrolyze steam pretreated lignocellulosic substrates and model substances at an identical FPA loading. These cellulase preparations were found to display significantly different hydrolytic performances irrelevant with the FPA. And this difference was even observed on the filter paper itself when the FPA based assay was revisited. The analysis of specific enzyme activity in cellulase preparations demonstrated that different accessory enzymes were mainly responsible for the discrepancy of enzymatic hydrolysis between diversified substrates and various cellulases. Such the active role of accessory enzymes present in cellulase preparations was finally verified by supplementation with β-glucosidase, xylanase and lytic polysaccharide monooxygenases AA9. This paper provides new insights into the role of accessory enzymes, which can further provide a useful reference for the rational customization of cellulase cocktails in order to realize an efficient conversion of natural lignocellulosic substrates.