Immobilization of Trametes versicolor laccase on chitosan/halloysite as a biocatalyst in the Remazol Red RR dye

Abstract

In this study, the laccase obtained from Trametes versicolor was immobilized onto the chitosan(CTS)/halloysite (HNT) beads. In the immobilization step, the effects of chitosan (1–3% w/v), halloysite (0–2% w/v), glutaraldehyde (0.5–1.5% v/v) and enzyme concentrations (1–3%) on loading and immobilization efficiency were investigated. SEM, FT-IR, XRD, TGA and XPS analyses were performed to examine the structure of beads. In addition, the effects of parameters such as pH (4–10), temperature (25–55 °C), storage life on the activity of free and immobilized laccase were also investigated. The activities of free and immobilized laccase preserved 23% and 56% of its initial activity at the end of 59 days of storage. The effects of mediators such as 2.2-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS), 1-Hydroxybenzotriazole hydrate (HBT), 2,2,6,6-Tetramethyl-1-piperidinyloxy (TEMPO) and violuric acid (VLA) on the dye removal efficiency were investigated. Reusability of the CTS/HNT/Lac in the presence of HBT and VLA mediators, which enable the highest dye removal, was tested. After 15 cycles, 42% and 54% dye removal were achieved with the CTS/HNT/Lac in the medium containing HBT and VLA, and 42% and 49% of the activity is preserved, respectively. This study showed that CTS/HNT/Lac can be used repeatedly for Remazol Red RR dye removal.