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학술연구/단체지원/교육 등 연구자 활동을 지속하도록 DBpia가 지원하고 있어요.
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섬유소를 비롯한 단백질, 지질, 녹말을 분해할 수 있는 세균을 된장으로부터 분리하여 동정한 결과 Bacillus subtilis로 분류 되어, Bacillus subtilis CK-2로 명명하였다. B. subtilis CK-2는 40~45℃의 비교적 높은 온도 범위와 pH 6~9의 넓은 pH 범위, 그리고 NaCl 0~3%(w/v) 범위에서 잘 자랐으며, 높은 자가분해효소 활성을 갖는 것을 알 수 있다. B. subtilis CK-2가 분비하는 가수분해효소들은 세균의 생장과 거의 비례적으로 나타났으며 세포 외 활성을 나타내는 1차 대사산물로 확인하였다.
B. subtilis CK-2가 생산하는 cellulase의 반응 최적 온도는 55℃이며, 50℃이하의 온도에서 안정하였다. 최적 반응 pH는 9.0이었으며 pH 5.0~10.0의 범위에서 높은 효소활성을 보였다. 이 효소의 활성은 여러 가지 금속이온 중 Co2+에의해 촉진하는 것을 확인하였으며, 0.45%(w/v)농도에서 최대 효소활성 보였다.
amylase의 반응 최적 온도는 50℃이었으며 50℃이하의 온도에서 안정하였다. 최적 반응 pH는 5.0이었으며 pH 4.0~5.0의 범위에서 높은 효소활성을 보였다. amylase 또한 Co2+이온에 의해 활성이 촉진 하였으며, 0.2%(w/v)에서 최대 효소활성을 보였다.
protease의 반응 최적 온도는 50℃이었으며 50℃이하의 온도에서 안정하였다. 최적 pH는 8.0이었으며 pH 7.0~8.5의 범위에서 높은 효소활성을 보였다. protease의 효소활성은Mn2+이온에 의해 촉진 하였으며, Mn2+ 0.1%(w/v)에서 최대 효소활성 보였다.
전통 된장에서 분리한 B. subtilis CK-2는 다양한 자가분해 효소의 활성은 40~65℃의 비교적 높은 온도 범위와 pH 5~11의 넓은 pH 범위에서 높은 자가분해효소 활성을 보였다.
이상의 결과로부터 B. subtilis CK-2는 농수임산물 폐기물이나 음식물 폐기물의 퇴비화, 사료 생산 등에 유용하게 이용될 수 있을 것으로 생각 한다.
B. subtilis CK-2가 생산하는 cellulase의 반응 최적 온도는 55℃이며, 50℃이하의 온도에서 안정하였다. 최적 반응 pH는 9.0이었으며 pH 5.0~10.0의 범위에서 높은 효소활성을 보였다. 이 효소의 활성은 여러 가지 금속이온 중 Co2+에의해 촉진하는 것을 확인하였으며, 0.45%(w/v)농도에서 최대 효소활성 보였다.
amylase의 반응 최적 온도는 50℃이었으며 50℃이하의 온도에서 안정하였다. 최적 반응 pH는 5.0이었으며 pH 4.0~5.0의 범위에서 높은 효소활성을 보였다. amylase 또한 Co2+이온에 의해 활성이 촉진 하였으며, 0.2%(w/v)에서 최대 효소활성을 보였다.
protease의 반응 최적 온도는 50℃이었으며 50℃이하의 온도에서 안정하였다. 최적 pH는 8.0이었으며 pH 7.0~8.5의 범위에서 높은 효소활성을 보였다. protease의 효소활성은Mn2+이온에 의해 촉진 하였으며, Mn2+ 0.1%(w/v)에서 최대 효소활성 보였다.
전통 된장에서 분리한 B. subtilis CK-2는 다양한 자가분해 효소의 활성은 40~65℃의 비교적 높은 온도 범위와 pH 5~11의 넓은 pH 범위에서 높은 자가분해효소 활성을 보였다.
이상의 결과로부터 B. subtilis CK-2는 농수임산물 폐기물이나 음식물 폐기물의 퇴비화, 사료 생산 등에 유용하게 이용될 수 있을 것으로 생각 한다.
목차
List of Figures ························································ ⅳList of Tables ·························································· ⅵⅠ. 서론 ································································ 1Ⅱ. 실험재료 및 방법 ················································ 41. 실험재료 ··························································· 41) 공시균주 ························································· 42) 사용배지 ···················································· 43) 시약 및 기기 ···················································· 42. 실험방법 ··························································· 51) 세균의 분리 ······················································ 5(1) 균주 분리 및 선정 ············································ 5(2) 분리균주의 동정 ·············································· 62) 조효소의 특성 ··················································· 7(1) 조효소액 제조 ················································· 7(2) 효소의 활성 측정 ·········································· 7① Cellulase 활성 측정 ·········································· 7② Amylase 활성 측정 ·········································· 8③ Protease활성 측정 ··········································· 8(3) 효소 반응의 최적온도 ······································ 10(4) 효소 반응의 최적 pH ······································· 10(5) 효소의 열안정성 ············································· 10(6) 효소활성에 대한 금속이온의 영향 ························ 11Ⅲ. 결과 및 고찰 ···················································· 121. 분리균주 유기중합체 분해능 확인 ··························· 122. 분리 및 동정 ····················································· 143. 분리균주의 최적배양조건 ······································ 161) 최적 배양 온도 ··············································· 162) 최적 배양 pH ················································· 164. 분리세균의 생장에 따른 세포외 효소 활성 변화 ··········· 185. 조효소의 특성 ··················································· 201) Cellulase ························································ 20(1) 최적 온도 ····················································· 20(2) 열안정성 ······················································ 22(3) 금속이온에 따른 영향 ······································ 24(4) Co2+ 금속이온 농도에 따른 영향 ·························· 26(5) 최적 pH ······················································ 282) Amylase ························································ 30(1) 최적 온도 ····················································· 30(2) 열안정성 ····················································· 32(3) 금속이온에 따른 영향 ·········································34(4) Co2+ 금속이온 농도에 따른 영향 ·························· 36(5) 최적 pH ······················································ 383) Protease ························································· 40(1) 최적온도 ······················································ 40(2) 열안전성 ······················································ 42(3) 금속이온에 따른 영향 ·····································
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