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In the current study, extracellular lipase production from thermotolerant Bacillus subtilis TTP-06, which was isolated from Tattapani Hot Spring in Himachal Pradesh, India, was optimized using response surface approach. The interactions of six different parameters, including glucose and peptone concentrations, pH, temperature, incubation duration, and olive oil concentration, were examined using the Plackett-Burman Design (PBD). Central Composite Design (CCD) was used to establish the best values for the positive factors. The importance of the quadratic model with a determination coefficient (R2) value of 0.7846 was revealed by statistical analysis of variance (ANOVA) results with a p-value of <0.05. The optimal combination of glucose (2.75 % w/v) and peptone (1.6 %) in a pH 7.5 medium incubated at 55°C for 24 h with 1 % (v/v) inoculum produced the highest amount of lipase activity (8.889 U/ml). The lipase produced under optimized conditions was further used for the degradation of a bioplastic film.