Stabilization of the restriction enzyme EcoRI dried with trehalose and other selected glass-forming solutes
The stabilization of the restriction enzyme EcoRI by its incorporation into aqueous glass-forming carbohydrate or polymer solutions, followed by vacuum-drying to low moisture, has been studied. Glass-forming solutes included trehalose, sucrose, lactose, maltose, raffinose, maltodextrin DE 10, and po...
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| Publicado: |
1997
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| Acceso en línea: | https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_87567938_v13_n5_p609_Rossi http://hdl.handle.net/20.500.12110/paper_87567938_v13_n5_p609_Rossi |
| Aporte de: |
| id |
paper:paper_87567938_v13_n5_p609_Rossi |
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| record_format |
dspace |
| institution |
Universidad de Buenos Aires |
| institution_str |
I-28 |
| repository_str |
R-134 |
| collection |
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) |
| topic |
Aromatic polymers Atmospheric humidity Catalyst activity Crystallization Glass transition Polysaccharides Sugar (sucrose) Vacuum applications Viscosity of liquids Enzyme stabilization Lactose Maltodextrin Maltose Polyvinylpyrrolidone Raffinose Trehalose Vacuum desiccation Enzymes glass glycerol raffinose sucrose trehalose type II site specific deoxyribonuclease article chemistry desiccation enzyme stability methodology temperature Deoxyribonuclease EcoRI Desiccation Enzyme Stability Glass Glycerol Raffinose Sucrose Temperature Trehalose |
| spellingShingle |
Aromatic polymers Atmospheric humidity Catalyst activity Crystallization Glass transition Polysaccharides Sugar (sucrose) Vacuum applications Viscosity of liquids Enzyme stabilization Lactose Maltodextrin Maltose Polyvinylpyrrolidone Raffinose Trehalose Vacuum desiccation Enzymes glass glycerol raffinose sucrose trehalose type II site specific deoxyribonuclease article chemistry desiccation enzyme stability methodology temperature Deoxyribonuclease EcoRI Desiccation Enzyme Stability Glass Glycerol Raffinose Sucrose Temperature Trehalose Rossi, Silvia Graciela Buera, María del Pilar Moreno de Colonna, Silvia Stabilization of the restriction enzyme EcoRI dried with trehalose and other selected glass-forming solutes |
| topic_facet |
Aromatic polymers Atmospheric humidity Catalyst activity Crystallization Glass transition Polysaccharides Sugar (sucrose) Vacuum applications Viscosity of liquids Enzyme stabilization Lactose Maltodextrin Maltose Polyvinylpyrrolidone Raffinose Trehalose Vacuum desiccation Enzymes glass glycerol raffinose sucrose trehalose type II site specific deoxyribonuclease article chemistry desiccation enzyme stability methodology temperature Deoxyribonuclease EcoRI Desiccation Enzyme Stability Glass Glycerol Raffinose Sucrose Temperature Trehalose |
| description |
The stabilization of the restriction enzyme EcoRI by its incorporation into aqueous glass-forming carbohydrate or polymer solutions, followed by vacuum-drying to low moisture, has been studied. Glass-forming solutes included trehalose, sucrose, lactose, maltose, raffinose, maltodextrin DE 10, and poly(vinylpyrrolidone) (molecular weight 40 000, PVP). Among the solutes examined, trehalose and sucrose protected the enzyme most effectively during storage at 37 and 45 °C. The restriction enzyme dried with trehalose or sucrose maintained its activity without detectable loss for at least 20 days at 37 °C and 12 days at 45 °C. In contrast, the activity of the enzyme dried with maltodextrin or PVP was reduced during vacuum desiccation and also it decreased remarkably during storage at the same temperatures. Stored (37/45 °C) vacuum-dried trehalose and sucrose systems were either a dense paste or a very viscous syrup, and this indicated that they were not glassy. Moreover, no relationship was found between the glass transition temperatures (T(g)) of the pure added solute and enzyme protection during storage, since, e.g., sucrose which has significantly lower T(g) values protected the enzyme much better than either maltose, lactose, maltodextrin, or PVP. The trisaccharide raffinose offered good protection of enzyme activity, and its role as a novel excipient matrix for labile enzyme stabilization deserves further investigation. The stability of enzyme EcoRI was rapidly lost when the vacuum-dried trehalose and sucrose systems were humidified to 58% relative humidity and stored at 45 °C, and this was attributed to disaccharide crystallization. The stabilization of the restriction enzyme EcoRI by its incorporation into aqueous glass-forming carbohydrate or polymer solutions, followed by vacuum-drying to low moisture, has been studied. Glass-forming solutes included trehalose, sucrose, lactose, maltose, raffinose, maltodextrin DE 10, and poly(vinylpyrrolidone) (molecular weight 40 000, PVP). Among the solutes examined, trehalose and sucrose protected the enzyme most effectively during storage at 37 and 45 °C. The restriction enzyme dried with trehalose or sucrose maintained its activity without detectable loss for at least 20 days at 37 °C and 12 days at 45 °C. In contrast, the activity of the enzyme dried with maltodextrin or PVP was reduced during vacuum desiccation and also it decreased remarkably during storage at the same temperatures. Stored (37/45 °C) vacuum-dried trehalose and sucrose systems were either a dense paste or a very viscous syrup, and this indicated that they were not glassy. Moreover, no relationship was found between the glass transition temperatures (Tg) of the pure added solute and enzyme protection during storage, since, e.g., sucrose which has significantly lower Tg values protected the enzyme much better than either maltose, lactose, maltodextrin, or PVP. The trisaccharide raffinose offered good protection of enzyme activity, and its role as a novel excipient matrix for labile enzyme stabilization deserves further investigation. The stability of enzyme EcoRI was rapidly lost when the vacuum-dried trehalose and sucrose systems were humidified to 58% relative humidity and stored at 45 °C, and this was attributed to disaccharide crystallization. |
| author |
Rossi, Silvia Graciela Buera, María del Pilar Moreno de Colonna, Silvia |
| author_facet |
Rossi, Silvia Graciela Buera, María del Pilar Moreno de Colonna, Silvia |
| author_sort |
Rossi, Silvia Graciela |
| title |
Stabilization of the restriction enzyme EcoRI dried with trehalose and other selected glass-forming solutes |
| title_short |
Stabilization of the restriction enzyme EcoRI dried with trehalose and other selected glass-forming solutes |
| title_full |
Stabilization of the restriction enzyme EcoRI dried with trehalose and other selected glass-forming solutes |
| title_fullStr |
Stabilization of the restriction enzyme EcoRI dried with trehalose and other selected glass-forming solutes |
| title_full_unstemmed |
Stabilization of the restriction enzyme EcoRI dried with trehalose and other selected glass-forming solutes |
| title_sort |
stabilization of the restriction enzyme ecori dried with trehalose and other selected glass-forming solutes |
| publishDate |
1997 |
| url |
https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_87567938_v13_n5_p609_Rossi http://hdl.handle.net/20.500.12110/paper_87567938_v13_n5_p609_Rossi |
| work_keys_str_mv |
AT rossisilviagraciela stabilizationoftherestrictionenzymeecoridriedwithtrehaloseandotherselectedglassformingsolutes AT bueramariadelpilar stabilizationoftherestrictionenzymeecoridriedwithtrehaloseandotherselectedglassformingsolutes AT morenodecolonnasilvia stabilizationoftherestrictionenzymeecoridriedwithtrehaloseandotherselectedglassformingsolutes |
| _version_ |
1768545902268514304 |
| spelling |
paper:paper_87567938_v13_n5_p609_Rossi2023-06-08T16:36:53Z Stabilization of the restriction enzyme EcoRI dried with trehalose and other selected glass-forming solutes Rossi, Silvia Graciela Buera, María del Pilar Moreno de Colonna, Silvia Aromatic polymers Atmospheric humidity Catalyst activity Crystallization Glass transition Polysaccharides Sugar (sucrose) Vacuum applications Viscosity of liquids Enzyme stabilization Lactose Maltodextrin Maltose Polyvinylpyrrolidone Raffinose Trehalose Vacuum desiccation Enzymes glass glycerol raffinose sucrose trehalose type II site specific deoxyribonuclease article chemistry desiccation enzyme stability methodology temperature Deoxyribonuclease EcoRI Desiccation Enzyme Stability Glass Glycerol Raffinose Sucrose Temperature Trehalose The stabilization of the restriction enzyme EcoRI by its incorporation into aqueous glass-forming carbohydrate or polymer solutions, followed by vacuum-drying to low moisture, has been studied. Glass-forming solutes included trehalose, sucrose, lactose, maltose, raffinose, maltodextrin DE 10, and poly(vinylpyrrolidone) (molecular weight 40 000, PVP). Among the solutes examined, trehalose and sucrose protected the enzyme most effectively during storage at 37 and 45 °C. The restriction enzyme dried with trehalose or sucrose maintained its activity without detectable loss for at least 20 days at 37 °C and 12 days at 45 °C. In contrast, the activity of the enzyme dried with maltodextrin or PVP was reduced during vacuum desiccation and also it decreased remarkably during storage at the same temperatures. Stored (37/45 °C) vacuum-dried trehalose and sucrose systems were either a dense paste or a very viscous syrup, and this indicated that they were not glassy. Moreover, no relationship was found between the glass transition temperatures (T(g)) of the pure added solute and enzyme protection during storage, since, e.g., sucrose which has significantly lower T(g) values protected the enzyme much better than either maltose, lactose, maltodextrin, or PVP. The trisaccharide raffinose offered good protection of enzyme activity, and its role as a novel excipient matrix for labile enzyme stabilization deserves further investigation. The stability of enzyme EcoRI was rapidly lost when the vacuum-dried trehalose and sucrose systems were humidified to 58% relative humidity and stored at 45 °C, and this was attributed to disaccharide crystallization. The stabilization of the restriction enzyme EcoRI by its incorporation into aqueous glass-forming carbohydrate or polymer solutions, followed by vacuum-drying to low moisture, has been studied. Glass-forming solutes included trehalose, sucrose, lactose, maltose, raffinose, maltodextrin DE 10, and poly(vinylpyrrolidone) (molecular weight 40 000, PVP). Among the solutes examined, trehalose and sucrose protected the enzyme most effectively during storage at 37 and 45 °C. The restriction enzyme dried with trehalose or sucrose maintained its activity without detectable loss for at least 20 days at 37 °C and 12 days at 45 °C. In contrast, the activity of the enzyme dried with maltodextrin or PVP was reduced during vacuum desiccation and also it decreased remarkably during storage at the same temperatures. Stored (37/45 °C) vacuum-dried trehalose and sucrose systems were either a dense paste or a very viscous syrup, and this indicated that they were not glassy. Moreover, no relationship was found between the glass transition temperatures (Tg) of the pure added solute and enzyme protection during storage, since, e.g., sucrose which has significantly lower Tg values protected the enzyme much better than either maltose, lactose, maltodextrin, or PVP. The trisaccharide raffinose offered good protection of enzyme activity, and its role as a novel excipient matrix for labile enzyme stabilization deserves further investigation. The stability of enzyme EcoRI was rapidly lost when the vacuum-dried trehalose and sucrose systems were humidified to 58% relative humidity and stored at 45 °C, and this was attributed to disaccharide crystallization. Fil:Rossi, S. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Buera, M.P. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Moreno, S. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. 1997 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_87567938_v13_n5_p609_Rossi http://hdl.handle.net/20.500.12110/paper_87567938_v13_n5_p609_Rossi |