Alkaline protease의 생산능력이 강한 Bacillus sp. CW-1121을 토양으로부터 분리하였으며 이 균주에 의한 효소생산은 배지에 유기질소원으로 casein, 무기질소원으로 $(NH_4){_2}HPO_4$, 무기염류로는 $MgSO_4$ 그리고 탄소원으로서 fructose를 첨가하였을 때 최대활성을 나타내었다. 본 정제효소의 최적 작용 pH와 온도는 pH 10.0과 $40^{circ}C$였으며 pH 9.0~11.0의 범위와 $50^{circ}C$ 이하에서 안정하였다. 금속이온 중 $Mg^{2+}$, $K^2$ 등에 의해 활성이 증대되었으나 $Hg^{2+}$, $Fe^{2+}$ 등에 의해서는 강한 활성저해가 관찰되었다. 저해제 중 2,4-DNP, ACA, PCMB 등에 의해서는 활성저해가 관찰되지 않았으나 EDTA에 서는 강한 저해작용을 나타내어 본 효소는 활성단에 금속이온이 관여하는 metallo enzyme으로 추정되었다. 이 정제효소의 $K_m$치는 $1.25{ imes}10^{-4}M/1$, $V_{max}$ $7.14{mu}g/min$이었다.
An alkaline protease producting bacteria was isolated from soil and identified as Bacillus sp. CW-1121. It was found that the production of alkaline protease reached to maximum when casein, $(NH_4)_2HPO_4$, $MgSO_4$, fructose as organic and inorganic nitrogen, inorganic salt and carbon source added in culture medium. The optimum temperature and pH for the purified enzyme activity were $40^{circ}C$ and pH 10.0 respectively. The enzyme was relatively stable in the pH range of pH 9.0~11.0 and at the temperature below $50^{circ}C$. The activity of the purified enzyme was inhibited by $Hg^{2+}$ and $Fe^{2+}$, whereas $Mn^{2+}$ and $K^+$ gave rather activating effects on the enzyme activity. PCMB, 2,4-DNP, ${varepsilon}$-aminocaproic acid did not show inhibitory effect in the proteolytic activity of alkaline protease but ethylenediaminetetraacetic acid inhibited the enzyme activity. The reaction of this enzyme followed typical Michaels Menten kinetics with the $K_m$ value of $1.25{ imes}10^{-4}M/1$ with the $V_{max}$ of $7.14{mu}g/min$ for casein.
