본 연구는 효소의 활성을 저해하는 주위 환경, 특히 열로부터 효소의 활성을 장기간 유지할 수 있는 효소 안정화 시스템에 대한 것으로, 이 시스템은 poly(${\epsilon}-caprolactone$) (PCL) 마이크로 캡슐로, 파파인 효소를 모델 효소로 하여, poly(propylene glycol) (PPG) 층이 코어 효소층을 둘러싸고 있는 형태로 설계되어 있다. 공촛점 현미경 및 장기 열 안정도 결과를 분석해본 결과, 파파인 효소가 소수성 PPG로 둘러쌓여 있고, 배타적 볼륨 효과(exclusive volume effect)에 의해 안정화되어 있음을 밝힐 수 있었다. 이와 같이 향상된 효소의 열 안정도는 소수성 사슬이 긴 PPG를 사용할수록 증가됨을 알 수 있었으며, 이것은 효소와 PPG 계면 사이에서 PPG 층이 파파인 효소를 효과적인 형태 고정(conformational anchoring)을 통해 안정화한 것임을 알 수 있었다.
본 연구는 효소의 활성을 저해하는 주위 환경, 특히 열로부터 효소의 활성을 장기간 유지할 수 있는 효소 안정화 시스템에 대한 것으로, 이 시스템은 poly(${\epsilon}-caprolactone$) (PCL) 마이크로 캡슐로, 파파인 효소를 모델 효소로 하여, poly(propylene glycol) (PPG) 층이 코어 효소층을 둘러싸고 있는 형태로 설계되어 있다. 공촛점 현미경 및 장기 열 안정도 결과를 분석해본 결과, 파파인 효소가 소수성 PPG로 둘러쌓여 있고, 배타적 볼륨 효과(exclusive volume effect)에 의해 안정화되어 있음을 밝힐 수 있었다. 이와 같이 향상된 효소의 열 안정도는 소수성 사슬이 긴 PPG를 사용할수록 증가됨을 알 수 있었으며, 이것은 효소와 PPG 계면 사이에서 PPG 층이 파파인 효소를 효과적인 형태 고정(conformational anchoring)을 통해 안정화한 것임을 알 수 있었다.
This article describes an enzyme stabilization method that allows the use of enzymes irrespective of environmental factors, especially heat, while maintaining their activity for a long time. We have designed enzyme microcapsules that consist of papain enzyme cores, poly(propylene glycol) interlayers...
This article describes an enzyme stabilization method that allows the use of enzymes irrespective of environmental factors, especially heat, while maintaining their activity for a long time. We have designed enzyme microcapsules that consist of papain enzyme cores, poly(propylene glycol) interlayers, and poly(${\epsilon}-caprolactone$) walls. By confocal laser scanning microscopy measurements and the thermal stability of papain-loaded microcapsules, it is demonstrated that the papain is surrounded by a hydrophobic polyol layer and stabilized by the exclusive volume effect. In our study, improved thermal stability can be obtained by using more hydrophobic long-chained polyols, which is understood to be attributed to the effective formation of a hydrophobic polyol layer between the papain and the polymer wall by means of conformational anchoring in the interface.
This article describes an enzyme stabilization method that allows the use of enzymes irrespective of environmental factors, especially heat, while maintaining their activity for a long time. We have designed enzyme microcapsules that consist of papain enzyme cores, poly(propylene glycol) interlayers, and poly(${\epsilon}-caprolactone$) walls. By confocal laser scanning microscopy measurements and the thermal stability of papain-loaded microcapsules, it is demonstrated that the papain is surrounded by a hydrophobic polyol layer and stabilized by the exclusive volume effect. In our study, improved thermal stability can be obtained by using more hydrophobic long-chained polyols, which is understood to be attributed to the effective formation of a hydrophobic polyol layer between the papain and the polymer wall by means of conformational anchoring in the interface.
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대상 데이터
N, N-dimethylated Casein (Sigma), sodium phosphate dibasic anhydrous (Sigma), Trichloroacetic acid (TCA, ACS grade, Sigma-aidrich), Citiric acid (Monohydrate, Sigma), L~tyrosine (minimum 98% (TLC), Sigma), BCA protein assay kit were used for papain activity measurement. Fluorescein isothiocyanate (FITC) and rhodamine B isothiocyanate (RBITC) were purchased from Fluka.
Baker) and polyethylene glycol (PEG, 400 g/moL, Aldrich) were used for papain microcapsules preparation. Poly(vinyl alcohol) (PVA, Mw = 8.8×104 9.2×104 g/moL, degree of saponification 87~89%, Kuraray) was used as a microcapsule stabilizer. N, N-dimethylated Casein (Sigma), sodium phosphate dibasic anhydrous (Sigma), Trichloroacetic acid (TCA, ACS grade, Sigma-aidrich), Citiric acid (Monohydrate, Sigma), L~tyrosine (minimum 98% (TLC), Sigma), BCA protein assay kit were used for papain activity measurement.
후속연구
Their understandings give us a hypothesis that if the solvophobic effect is maximized, the enzymes would be much more stable. Therefore, in this study, we try to reveal the importance of the hydrophobic polyols in stabilizing enzymes and build up a microcapsule system that can enhance the thermal stability of the enzymes therein. This study is expected to provide another useful approach to enzyme stabilization.
Therefore, in this study, we try to reveal the importance of the hydrophobic polyols in stabilizing enzymes and build up a microcapsule system that can enhance the thermal stability of the enzymes therein. This study is expected to provide another useful approach to enzyme stabilization.
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