Production of chitosanase from termophylic bacteria isolated from Bora Hotspring

The production of chitosanase enzyme was carried out by utilizing isolate of thermophilic bacteria from Bora hot spring in order to determine the optimum condition of its production process. B1211 bacterial isolates with the highest Chitinolytic Index values of 1.71 were used in crude chitosanase production. The crude chitosanase production was performed using a Completely Randomized Design (RAL) consisting of four independent variables, in the form of chitosan substrate concentration (0.25%, 0.50%, 0.75%, 1.00%), agitation speed (60, 90, 120, and 150 rpm), fermentation time (1, 3, 5, 7, and 9 days), and fermentation temperature (30, 40, 50, and 60°C). The activity of chitosanase as dependent variable was determined by Schales method, which referred to standard curve of D-glucosamine. The results showed that 1% chitosan substrate concentration yielded crude chitosanase with the highest activity of 0.408 U/mL and dissolved protein content of 9.887 mg/mL. Furthermore, the best agitation speed (90 rpm) resulted in 0.385 U/mL chitosanase activity and dissolved protein content of 23.74 mg/mL. In the fermentation time treatment, the 5-day fermentation time resulted in the highest chitosanase activity of 0.384 U/mL with dissolved protein content of 36.52 mg/mL. The fermentation temperature determination showed that the temperature of 60 °C as the preferred treatment with chitosanase activity of 0.569 U/mL and dissolved protein content of 5.752 mg/mL.