Production and purification of laccase by Bacillus sp. using millet husks and its pesticide degradation application
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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 (24–96 h) 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 96 h of incubation with PM 2.0 g/L and FM 1.5 g/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 63–75 kDa. The partially purified laccase effectively degraded the pesticides (Tricel, 71.8 ± 3.5 and Phoskill 77.3 ± 3.4%) within 5 days 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.
KeywordsBacillus sp. Finger millet Pearl millet Laccase Phoskill Tricel
This work is partially supported by the Department of Biotechnology, Mahendra Arts and Science College (Autonomous), and the Department of Science and Technology, Government of India (DST-FIST sponsored—Ref. No. SR/FST/College-232/2014).
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Conflict of interest
The authors declare that they do not have any conflict of interest.
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