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Suppression of protein inactivation during freezing by minimizing pH changes using ionic cryoprotectants
Ľ. Krausková, J. Procházková, M. Klašková, L. Filipová, R. Chaloupková, S. Malý, J. Damborský, D. Heger,
Language English Country Netherlands
Document type Journal Article
- MeSH
- Ions chemistry MeSH
- Hydrogen-Ion Concentration MeSH
- Cryopreservation methods MeSH
- Cryoprotective Agents chemistry MeSH
- Freeze Drying methods MeSH
- Proteins chemistry MeSH
- Buffers MeSH
- Solutions chemistry MeSH
- Freezing MeSH
- Publication type
- Journal Article MeSH
Freezing and lyophilization are often used for stabilization of biomolecules; however, this sometimes results in partial degradation and loss of biological function in these molecules. In this study we examined the effect of freezing-induced acidity changes on denaturation of the model enzyme haloalkane dehalogenase under various experimental conditions. The effective local pH of frozen solutions is shown to be the key causal factor in protein stability. To preserve the activity of frozen-thawed enzymes, acidity changes were prevented by the addition of an ionic cryoprotectant, a compound which counteracts pH changes during freezing due to selective incorporation of its ions into the ice. This approach resulted in complete recovery of enzyme activity after multiple freeze-thaw cycles. We propose the utilization of ionic cryoprotectants as a new and effective cryopreservation method in research laboratories as well as in industrial processes.
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- $a Freezing and lyophilization are often used for stabilization of biomolecules; however, this sometimes results in partial degradation and loss of biological function in these molecules. In this study we examined the effect of freezing-induced acidity changes on denaturation of the model enzyme haloalkane dehalogenase under various experimental conditions. The effective local pH of frozen solutions is shown to be the key causal factor in protein stability. To preserve the activity of frozen-thawed enzymes, acidity changes were prevented by the addition of an ionic cryoprotectant, a compound which counteracts pH changes during freezing due to selective incorporation of its ions into the ice. This approach resulted in complete recovery of enzyme activity after multiple freeze-thaw cycles. We propose the utilization of ionic cryoprotectants as a new and effective cryopreservation method in research laboratories as well as in industrial processes.
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- $a Chaloupková, Radka $u Loschmidt Laboratories, Department of Experimental Biology, Faculty of Science, Masaryk University, Kamenice 5/A13, 625 00 Brno, Czech Republic; Research Centre for Toxic Compounds in the Environment RECETOX, Faculty of Science, Masaryk University, Kamenice 5/A29, 625 00 Brno, Czech Republic.
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- $a Damborský, Jiří $u Loschmidt Laboratories, Department of Experimental Biology, Faculty of Science, Masaryk University, Kamenice 5/A13, 625 00 Brno, Czech Republic; Research Centre for Toxic Compounds in the Environment RECETOX, Faculty of Science, Masaryk University, Kamenice 5/A29, 625 00 Brno, Czech Republic.
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- $a Heger, Dominik $u Department of Chemistry, Faculty of Science, Masaryk University, Kamenice 5/A8, 625 00 Brno, Czech Republic; Research Centre for Toxic Compounds in the Environment RECETOX, Faculty of Science, Masaryk University, Kamenice 5/A29, 625 00 Brno, Czech Republic. Electronic address: hegerd@chemi.muni.cz.
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