Isolasi dan Optimasi Protease Bakteri Termofilik dari Sumber Air Panas Tangkuban Perahu Bandung

Abstract

Protease is an enzyme widely used in food industry. Thermophilic bacteria produce protease that can withstand high temperature. The research was aimed to isolate thermophilic bacteria and characterize protease of isolates with highest proteolytic index. Isolation was carried out on 33 isolates from three hot springs. Isolation was done by using pour plate method. Protease activity was measured by Bergmeyer, and protease characterization was conducted at pH 6-10 and temperature of 50oC- 90oC. The results showed that two isolates from Sipaholon hot spring, eleven isolates from Tangkuban Perahu hot spring, and two isolates from Ciseeng hot spring established clear areas around their colonies. T8 isolate derived from Tangkuban Perahu hot spring showed the highest proteolytic index of 33.5 and was used for further testing. The highest proteolytic activity of T8 isolate was obtained at 21th hour amounted to 3.8375 x 10-3 U/mL. Protease activity increased in ammonium sulphate 60% fraction, which was 7.7257 x 10-3 U/mL. Statistical analysis showed that pH and temperature affected protease activity. The optimum protease activity was obtained at pH 8 Clark & Lubs buffer and temperature of 60oC.