Production and purification of laccase by Bacillus sp. using millet husks and its pesticide degradation application.

Abstract

Lignocellulosic agricultural bi-products, pearl millet (PM) and finger millet (FM) husks, were used for the production of laccase using Bacillus sp. PS under solid-state fermentation (SSF). Abiotic variables such as substrate (PM, FM) concentration (1-5%), incubation time (24-96h) and pH (5-10) were optimized using Response surface methodology (RSM) to maximize the laccase production. The predicted model showed maximum laccase activity of 402 U/mL appearing after 96h of incubation with PM 2.0g/L and FM 1.5g/L at pH 7.0. Single protein band on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) confirmed homogeneity of the laccase with a molecular weight of 63-75kDa. The partially purified laccase effectively degraded the pesticides (Tricel, 71.8 ± 3.5 and Phoskill 77.3 ± 3.4%) within 5days of incubation (40°C) in pH 7.0. The pesticide degradation was further confirmed by high-performance liquid chromatography (HPLC) and the chromatograms showed the single dominant peaks at retention time 2.482 (tricel) and 2.608 (phoskill) min, respectively. Pesticide-degrading laccase was produced by Bacillus sp. PS under SSF reveals the utilization of low-cost bi-substrates for enhanced laccase production.