Characterization of a metalloprotease from an isolate Bacillus thuringiensis 29-126 in animal feces collected from a zoological garden in Japan

An extracellular metalloprotease, Btmp, was partially purified from the culture supernatant of Bacillus thuringiensis 29-126, isolated from animal feces collected in a zoological garden in Japan, by ultrafiltration, ammonium sulfate precipitation, and a set of chromatography on Sephadex G-75 and High-Q. The molecular mass of the protease was estimated to be 60 kDa by SDS-PAGE. The enzyme showed optimum activity at $50^{\circ}C$ and pH 6.0, and had a half-life of 14 min at $50^{\circ}C$. The enzyme activity was not influenced by $Na^+$, $K^+$, $As^+$, $Mg^{+2}$, $Ca^{2+}$, $Ba^{2+}$, and phenylmethylsulfonyl fluoride, but it was moderately inhibited by $Zn^{+2}$ at a concentration of 1.0 mM, while the activity was significantly inhibited to less than 50 % by $Cu^{2+}$, $Co^{2+}$, $Cd^{2+}$, and ethylenediaminetetraacetic acid. Interestingly, the enzyme was activated to 178 % by 1.0 mM of $Mn^{2+}$. From these results, it may be suggested that the protease is a novel extracellular manganeseactivated metalloprotease.