Removal of pentachlorophenol and phenanthrene from lignocellulosic biorefinery wastewater by a biocatalytic/biosurfactant system comprising cross-linked laccase aggregates and rhamnolipid

Abstract

Synthesis and characterization of highly active cross-linked laccase aggregates (CLLAs) were performed and evaluated for removal of pentachlorophenol and phenanthrene from lignocellulosic biorefinery wastewater. Laccase from Tramates versicolor MTCC 138 was insolubilized as CLLAs via precipitation with 70% ammonium sulphate and simultaneous cross-linking with 5 mM glutaraldehyde to obtain activity recovery of 89.1%. Compared to the free laccase, the pH and thermal stability of the prepared CLLAs were significantly higher. At a high temperature of 60 °C, free laccase had a half-life of 0.25 h, while CLLAs had a half-life of 6.2 h. In biorefinery wastewater (pH 7.0), the free and CLLAs were stored for 3 day at a temperature of 30 °C. Free laccase completely lost their initial activity after 60 h; however, the CLLAs retained 39% activity till 72 h. Due to its excellent stability, free laccase and CLLAs were assessed for removing pentachlorophenol and phenanthrene in wastewater. CLLAs could remove 51-58% of pentachlorophenol (PCP) and phenanthrene (PHE) in 24 h. Biosurfactants, including surfactin, sophorolipid, and rhamnolipid, were assessed for their aptitude to improve the removal of organic contaminants in wastewater. Biorefinery wastewater incubated with all surfactants enhanced PCP and PHE removal compared to the no-surfactant controls. Further, 1 μM rhamnolipid significantly amplified pentachlorophenol and phenanthrene removal to 81-93% for free laccase and CLLAs, respectively.

Keywords: Bioremediation; Cross-linked enzyme aggregates; Laccase; Pentachlorophenol; Rhamnolipid.