2.4. Ligninolytic enzymes assay

During the degradation of RBBR to the two colors, three ligninolytic enzymes, laccase, manganese‐dependent peroxidase (MnP), and LiP were assessed in the medium of the respective color using a UV‐Vis spectrophotometer (n = 3). After the filtration of the mycelium via a syringe filter (0.45 μm), the enzymatic activity of the crude supernatant was measured. For laccase, 0.1 M sodium acetate (pH 4.5) and 1.5 mM 2,2‐azinobis‐3‐ethylbenzothiazoline‐6‐sulfonic acid (ABTS) were used ¹⁵. And 150 μL supernatant was added and mixed into buffer for 1 min. For MnP, 0.5 M sodium malonate (pH 4.5), 5 mM MnSO₄, 1 mM 2,6‐dimethoxylphenol, and 1 mM H₂O₂ were used ¹⁶. For LiP, 0.25 M sodium tartrate (pH 2.5), 10 mM veratryl alcohol and 5 mM H₂O₂ were used ¹⁷. One unit of the ligninolytic enzyme activity produced 1 μmol of reaction product per minute at room temperature; activity was expressed in U/mL ¹⁸, ¹⁹.