A new strategy for cellulases application in high temperature industrial scenarios with syringic acid assisting

• Published in: Global change biology. Bioenergy 2023-06, Vol.15 (6), p.821-833
• Main Authors: Ran, Qiuping, Huang, Mengtian, Li, Huanan, Chen, Bixin, He, Lili, Cui, Xinxin, Liu, Jiashu, Jiang, Zhengbing
• Format: Article
• Language: English
• Subjects: Acids; activity; Biofuels; Biomass; Cellulase; Cellulose; Enzymes; High temperature; Hydrolysis; Lignin; Lignocellulose; molecular dynamic simulation; Molecular dynamics; Plasmids; Protein expression; Proteins; secondary structure; syringic acid; Temperature requirements; Thermal stability; thermostability
• ISSN: 1757-1693; 1757-1707
• DOI: 10.1111/gcbb.13052

In the process of bioethanol production, more stable and active cellulase in high temperature condition is required. In this study, syringic acid was applied in cellulase hydrolysis system. At 70°C, TvEG3 activity increased 201.36%, CtBglA activity decreased 72.79% by syringic acid. With syringic acid assisting, TvEG3 thermostability was improved, CtBglA thermostability was reduced. Syringic acid scarcely affected CtCBH. In hydrolysis system with the cellulases containing TvEG3, CtCBH, and CtBglA, the reducing sugar yield improved by 28.37% with syringic acid assisting. With the molecular dynamic simulation in syringic acid system, the backbone root‐mean‐square deviation (RMSD) and the residue root‐mean‐square fluctuation (RMSF) of TvEG3, CtCBH reduced, while the RMSD and RMSF of CtBglA increased. The reduction in the number of secondary structures, especially α‐helix, caused the structure of CtBglA in the presence of syringic acid to collapse at high temperature. More secondary structures in TvEG3 and more α‐helix in CtCBH in the presence of syringic acid make them more stable at high temperatures. These means syringic acid can stabilize TvEG3 and CtCBH structure, destabilize CtBglA structure at high temperature. In summary, this study not only provides insight into cellulase hydrolysis at high temperature with syringic acid assisting but also demonstrates the promoting mechanism of syringic acid. Syringic acid was found to boost cellulase hydrolysis at high temperature (70°C). The increased a‐helix content and secondary structure number revealed the improvement of thermostability of TvEG3 and CtCBH. The reduction in the number of secondary structures, especially a‐helix, caused the structure of CtBglA in the presence of syringic acid to collapse at high temperature. Syringic acid has the potential to stabilize the high temperature activity of cellulase by slowing down thermal denaturation.