Insight into the role of antioxidant in microbial lignin degradation: Ascorbic acid as a fortifier of lignin-degrading enzymes

Background: Microbial-driven lignin depolymerization has emerged as a promising approach for lignin degradation. However, this process is hindered by the limited activity of lignin-degrading enzymes. Antioxidants are crucial for maintaining redox homeostasis in living cells, which can impact the efficiency of enzymes. Ascorbic acid (AA) is well-known for its antioxidant properties, while Trametes versicolor is a commonly used lignin-degrading fungus capable of secreting laccase (Lac) and manganese peroxidase (MnP). Thus, AA was selected as model antioxidant and added into the culture medium of T. versicolor to examine the effect of antioxidants on the activity of lignin-degrading enzymes in the fungus. Results: The presence of AA resulted in a 4.9-fold increase in the Lac activity and a 3.9-fold increase in the MnP activity, reaching 10736 U/L and 8659 U/L, respectively. This increase in enzyme activity contributed to a higher lignin degradation rate from 17.5% to 35.2%, consistent with observed morphological changes in the lignin structure. Furthermore, the addition of AA led to a reduction in the molecular weights of lignin and an increase in the content of degradation products with lower molecular weight, indicating more thorough degradation of lignin. Proteomics analysis suggested that the enhancement in enzyme activity was more likely to attributed to the reinforcement of AA on oxidative protein folding and transportation, rather than changes in enzyme expression. Conclusions: The addition of AA enhanced the performance of enzymes responsible for lignin degradation in terms of enzyme activity, degradation rate, lignin structural change, and product mapping. This study offers a feasible strategy for enhancing the activity of lignin-degrading enzymes in the fungus and provides insights into the role of antioxidant in microbial lignin degradation.