Optimization of laccase enzyme from Flammulina velutipes using response surface methodology box-behnken design

Laccase enzyme is a ligninolytic enzyme which has an ability to oxidize phenolic compounds of lignocellulosic biomass. The aim of the study was to identify, optimize a thermo-tolerant isolate and laccase enzyme production using Response Surface Methodology- Box Behnken design respectively. The strain S3 was isolated from the compost soil pit and the crude laccase enzyme was extracted from the strain. The parameters such as pH, incubation period and CuSO 4 concentration has been optimized using Box-Behnken design and the outcome shows that the enzyme with 4.71 U mL − 1 was stable at pH 6 and 8, the optimum CuSO 4 concentration was 100 mM and the incubation period was 7 days at the temperature of 46֯ C. The genomic DNA of the strain was amplified using ITS-1 and 4 primers which showed a product size of 700 bp. The amplicon was sequenced and the blast analysis showed a sequence similarity of 96.6% with the Flammulina velutipes . The protein was isolated and the SDS analysis showed a molecular weight of 120 kDa.The crude laccase enzyme produced from the strain S3 Flammulina velutipes was formulated and optimized at varying parameters to increase the yield of laccase enzyme. We conclude that our results will assist to a wide application in bio-fuel production.