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An extracellular agarase was purified from Bacillus sp. BI-3, a thermophilic agar-degradingbacterium isolated from a hot spring in Indonesia. The purified agarase revealed a single bandon sodium dodecyl sulfate-polyacrylamide gel electrophoresis, with an apparent molecularmass of 58 kDa. The optimum pH and temperature of the agarase were 6.4 and 70oC,respectively. The activity of the agarase was stable at high temperatures, and more than 50%activity was retained at 80oC for 15 min. Furthermore, the enzyme was stable in the pH rangeof 5.8–8.0, and more than 60% of the residual activity was retained. Significant activation ofthe agarase was observed in the presence of K+, Na+, Ca2+, Mg2+, and Sr2+; on the other hand,Ba2+, Zn2+, Cu2+, Mn2+, Co2+, Fe2+, and EDTA inhibited or inactivated the enzyme activity. Thecomponents of the hydrolytic product analyzed by thin-layer chromatography showed thatthe agarase mainly produced neoagarobiose. This study is the first to present evidence ofagarolytic activity in aerobic thermophilic bacteria.

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