재조합 균주 Bacillus subtilis가 생산하는 Strept-omyces albus KSM-35 Amylase의 정제 및 특성 = Purification and Characterization of Amylase from Bacillus subtilis Cells Haboring the Recombinant Plasmid PASA240

다국어 초록 (Multilingual Abstract)

Amylase encoded by the samy gene of Streptomyces albus KSM-35 was produced from Bacillus subtilis LKS88 carrying a recombinant plasmid PASA240. The amylase was purified 14.6-folds to homogeneity by salting out with ammonium sulfate, and by some chromatographic procedures using DEAE-Toyopearl, Sephadex G-75, and hydroxyapatite.
The molecular weight of purified enzyme was estimated to be 50.000 by SDS-polyacrylamide gel electrophoresis analysis. The optimum temperature and pH of the amylase was 500 and 6.0, repectively. The amylase activity was stimulated by the addition of Na+ ion, whereas it was inhibited by 10 mM Al3+, Cu2+ ions, and 1 mM EDTA. Enzyme inhibitors, such as the ß-mercaptoethanol, concanavalin A,
hydoroxylamine-HCl, and acetic anhydride, did not affect the amylase activity. Maltotetraose was detected as the major product during the soluble starch hydrolysis, and small amounts of maltopentaose.
maltotriose, and maltose were also found. The general properties like pH, temperature dependences, and starch hydrolysis patterns of Bacillus-expressed amylase maintained the original properties of the s albus KSM-35 amylase.

목차 (Table of Contents)

• I.서론
• II. 재료 및 방법
• 1. 사용 균주
• 2. 조효소액 제조
• 3. 효소 정제
• 4. 단백질 정량
• 5, Amylase 활성 측정
• 6. 분자량 측정
• 7. 온도의 영향
• 8. pH의 영향
• 9. 금속 이온 및 저해제의 영향
• 10. 전분 가수 분해물의 HPLC 분석
• III. 결과 및 고찰
• 1. 효소의 정제
• 2. B.subtilis LKS88 (pASA240] amylase 분자량 측정
• 3. 반응 최적 온도
• 4. 반응 최적 pH
• 5. 금속 이온 및 저해제의 영향
• 6. 전분 가수 분해물의 분석
• IV. 요약