Thermostability is among of the vital enzyme characteristics for industrial application. Brevibacillus thermoruber LII was obtained as a potential isolate from the previous research which screened the potential thermostable protease producing bacteria from Indonesian hotspring. The newly thermostable protease produced by thermophilic Brevibacillus thermoruber LII had been purified and characterized. It was predicted that the pure enzyme obtained from Brevibacillus thermoruber LII was homo hexameric, having molecular weight of 36 kDa unit protein and its native was 215 kDa. In addition, it was also a neutral metalo serine protease according to biochemical tests that it was totaly inhibited by PMSF (Phenylmethanesulfonyl fluoride) and EDTA (Ethylenediaminetetraacetic acid). It showed optimum activity at pH of 8 and active in acidic buffer (up to pH of 4). All of metal ion in the form of chloride salt (2.5 mM) which were tested on the enzyme enhanced the enzyme activity but Li Keywords: thermostable protease, Brevibacillus thermoruber, hotspring ABSTRAK Ketahanan terhadap panas merupakan salah satu sifat enzim yang penting untuk aplikasi industri. Brevibacillus themoruber LII diperoleh dari penelitian sebelumnya pada penapisan isolat potensial penghasil protease tahan panas dari sumber air panas Indonesia. Protease baru tahan panas yang dihasilkan oleh Brevibacillus thermoruber LII sudah dimurnikan dan dikarakterisasi. Diprediksi bahwa enzim murni yang dihasilkan oleh mikroorganisma tahan panas, Brevibacillus thermoruber LII merupakan heksamer dengan berat molekul unit yang sama yaitu 36 kDa dan nativenya 215 kDa.Lebih lanjut, protease ini merupakan protease metaloserin netral berdasarkan uji-uji biokimianya yang dihambat sepenuhnya oleh PMSF (Phenylmethanesulfonyl fluoride) and EDTA (Ethylenediaminetetraacetic acid). Enzim menunjukkan aktifitas optimum pada pH 8 dan aktif lebih baik pada pH bufer bersifat asam (sampai pH 4). . Enzim juga menunjukkan kestabilan terhadap pelarut organik. Nilai K M danV max enzim murni masing-masing adalah 27.2 mg/ml atau 0.362 -0.272 M dengan substrat HammersteinCasein (MM 75-100 kDa) dan 261.1 µg/menit/ ml. Kata Kunci: protease tahan panas, Brevibacillus thermoruber, sumber air panas
Transglutaminases (EC 2.3.2.13) have attracted a wide interest from both scientific and appliedpoints of view due to their capacity to cross-link protein substrates. Obtaining transglutaminasesderived from animals are extremely high cost process, which has hampered its wider applicationuntil the discovery of transglutaminase produced by microorganisms. In the early 1990, sincemicrobial transglutaminase have been found, many transglutaminase-producing microbial strainshave been isolated and the enzyme production processes have been optimized. This resulted in the increased uses of transglutaminases in the food industries. In the fisheries industry, MTGasehas successfully been used to improve the mechanical properties of surimi from various fishes.
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