Characterization of Thermophilic Protease from Bacteria Isolated from Tangkuban Perahu Crater
Maggy T. SUHARTONO, Budiasih WAHYUNTARI, Gunawan, A. SUWANTO, and Young Kok YANG
IUC Biotechnology IPB, Bogor Agricultural University Indonesia and Yonsei University, Korea
The aim of this study was to collect culturable thermophillic microorganisms potential for biotechnological products. Samples collected from Tangkuban Perahu crater in West Java, Indonesia, were grown in media suitable for Thermus sp. (Yeast extract 0.4%, polypepton 0.8%, NaC1 0.2%, and CaC12 0,1%). From 118 isolates capable of growing at 60℃, 6 isolates were found to be proteolytic. One isolate (TPS2) was particularly stable, showing proteolytic index higher than the known protease producer of Bacillus type. The bacteria was able to grow nicely up to 75℃. The stationary phase was reached after 6-10 hours of incubation, and the extracellular protease was produced optimally during this phase. The extracellular protease enzyme showed optimum pH at 6-8, optimum temperature at 75℃, and inhibited by typical serine protease inhibitor Phenyl Methyl Sulfo Fluoride at 5 mM, and the metallo protease inhibitor EDTA at 2 mM. The enzyme was stable, 75 % of the original activity was still maintained after 24 hours incubation at 70℃.
