Bacillus subtilis BK-17을 이용한 혈전용해효소의 생산 = Production of fibriloytic enzyme from Bacillus subtilis BK-17

다국어 초록 (Multilingual Abstract)

The production of fibrinolytic enzyme from Bacillus subtilis BK-17 was studied in flask and bioreactor.
In the flask culture, soybean flour was the best nitrogen source and the concentration of 1.5% gave the best enzyme production. For carbon source and inorganic salt, 0.5% D-glucose and 0.05% Na_(2)HPO_(4)gave the best enzyme production. Optimal temperature and initial pH were 37℃ and 9, respectively. Enzyme production was greatly enhanced with increasing agitation speed in the range of 100∼300rpm or decreasing flack working volume in the range of 25∼100ml. the maximum enzyme production under tne optimal flask culture condition was 1200 unit/ml with urokinase as the standard.
In the bioreactor culture, cell growth, enzyme activity, glucose concentration, pH and DO were monitored. With proper preculture no lag phase was observed in cell growth. During exponential phase, DO and pH rapidly decreased and then slowly slowed down. The enzyme production started at last exponential phase and continued for 5∼6hr. During this production period rapid glucose consumption was observed. When the medium concentration was increased by two- or three-fold the enzyme production was increased. With high glucose concentration, agitation speed and air flow fate, enzyme production was enhanced. The maximum enzyme production under the optimal bioreactor culture condition was 1050 unit/ml and some further investigation was needed to enhance the enzyme production.

목차 (Table of Contents)

• 목차
• Ⅰ. 서론
• 1. 연구배경과 목적 = 1
• 2. 본 연구의 범위 = 6
• 3. 문헌 연구 = 8
• 3-1. 혈전용해효소 = 8
• 3-2. fibrinolytic enzyme의 연구동향 = 11
• 3-3. Protease = 14
• Ⅱ. 실험방법
• 1. 균주 및 배양조건 = 17
• 1-1. 균주 및 배지 = 17
• 1-2. 배양조건 = 17
• 2. 분석 방법 = 20
• 2-1. 효소 활성 측정 = 20
• 2-2. 생균수의 측정 = 23
• 2-3. Glucose 농도의 측정 = 23
• Ⅲ. 결과 및 토의
• 1. 플라스크 배양 = 26
• 1-1. 영양 조건의 영향 = 26
• 1-1-1. L-broth에서의 성장 = 26
• 1-1-2. 유기 질소원의 영향 = 28
• 1-1-3. 무기 질소원의 영향 = 33
• 1-1-4. 탄소원의 영향 = 33
• 1-1-5. 무기염류의 영향 = 38
• 1-1-6. Inducer의 영향 = 41
• 1-2. 환경 조건의 영향 = 44
• 1-2-1. 초기 pH의 영향 = 44
• 1-2-2. 온도의 영향 = 46
• 1-2-3. Working volume의 영향 = 48
• 1-2-4. 교반 속도의 영향 = 48
• 1-3. 기타 발효 조건의 영향 = 51
• 1-3-1. 접종량의 영향 = 51
• 1-3-2. 소포제의 영향 = 51
• 2. Bioreactor 배양 = 54
• 2-1. 배지농도의 영향 = 54
• 2-2. Glucose 농도의 영향 = 61
• 2-3. 교반 속도와 air 유량의 영향 = 61
• Ⅳ. 결론 = 65
• Ⅴ. 참고문헌 = 67
• ABSTRACT = 73