Aspergillus fumigatus이 생산하는 Alkaline protease의 특성과 작용양상

Characteristics and Action Pattern of Alkaline Protease produced from Aspergillus fumigatus

Alkaline protease 생성능이 강한 Aspergillus fumigatus 균주를 토양에서 분리하고, 생성효소를 정제하여 특성을 조사한 결과 최적 pH는 9.0, pH안정성은 $pH\;8.0{\sim}10.0$, 최적온도는 $50^{\circ}C$였으며, $50^{\circ}C$이하의 온도에서 안정하나 그 이상의 온도에서는 급격한 효소 불활성화를 보였고, 금속염 $Mn^{++},\;Cu^{++},\;Ba^{++},\;Mg^{++}$ 등에 의해서 활성이 다소 증대되나 $K^+,\;Fe^{+++},\;Ag^{++},\;Pb^{++},\;Na^+,\;Ca^{++},\;Hg^+,\;Zn^{++}$에 의해 저해를 받았다. 활성저해제인 EDTA, 2,4-DNP, ${\varepsilon}-amino$ caproic acid에는 큰 저해를 받지 않으나, PCMB에 많은 저해를 받는 것으로 미루어 활성 부위가 SH기인 cystein protease로 추정되었다. Km값은 $8.33{\times}10^{-4}mole/{\ell}$, Vmax는 $47.62{\mu}g/min$였으며, casein과 hemoglobin을 trypsin보다 더 잘 분해하고, casein을 hemoglobin보다 잘 분해하였다.

This experiment was conducted to investigate the characteristics of alkaline protease from Aspergillus fumigatus which was isolated from soil as a superior strain for the production of the alkaline protease. The optimum temperature for enzyme activity was $50^{\circ}C$ and optimum pH was 9.0. The enzyme was stable at pH 8.0 to 10.0 and thermal inactivation was shown $30^{\circ}C$. The activity of the enzyme was increased by the addition of $Mn^{++},\;Cu^{++},\;Ba^{++},\;Mg^{++},\;$wheras it was inhibitied by $K^+,\;Fe^{+++},\;Ag^+,\;Pb^{++},\;Na^+,\;Ca^{++},\;Hg^+,\;Zn^{++}$. EDTA. 2, 4-DNP, ${\varepsilon}-amino$ caproic acid did not show inhibitory effect on the proteolytic activity of alkaline protease but P-chloromercuribenzoic acid inhibited the enzyme activity, indicating that reactive sulfhydryl group is required for the enzymatic activity. The reaction of this enzyme followed typical Michael-Menten Kinetics with the Km value of $8.33{\times}10^{-4}mole/{\ell}$ with the Vmax of $47.62{\mu}g/min$. This enzyme had stronger proteolytic activity than trypsin on substrate such as casin and hemoglibin.