Effects of freezing rate on structural changes in l-lactate dehydrogenase during the freezing process

Abstract Freezing is a common method for improving enzyme storage stability. During the freezing process, the freezing rate is an important parameter that can affect protein stability. However, there is limited information on the denaturation mechanisms and protein conformational changes associated...

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Autores principales:	Haena Park, Jun-Young Park, Kyung-Min Park, Pahn-Shick Chang
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/3f1c0e626bfc41b5ac7e67f35cd10d69
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spelling	oai:doaj.org-article:3f1c0e626bfc41b5ac7e67f35cd10d692021-12-02T16:10:35ZEffects of freezing rate on structural changes in l-lactate dehydrogenase during the freezing process10.1038/s41598-021-93127-62045-2322https://doaj.org/article/3f1c0e626bfc41b5ac7e67f35cd10d692021-07-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-93127-6https://doaj.org/toc/2045-2322Abstract Freezing is a common method for improving enzyme storage stability. During the freezing process, the freezing rate is an important parameter that can affect protein stability. However, there is limited information on the denaturation mechanisms and protein conformational changes associated with the freezing rate. In this study, the effects of freezing rate on activity loss and conformational changes in a model enzyme, l-lactate dehydrogenase, were evaluated. Enzyme solutions were frozen at various rates, from 0.2 to 70.6 °C/min, and ice seeding was conducted to reduce supercooling. The results demonstrated that fast freezing results in activity loss, structural changes, and aggregation. The residual activities at freezing rates of 0.2, 12.8, and 70.6 °C/min were 77.6 ± 0.9%, 64.1 ± 0.4%, and 44.8 ± 2.0%, respectively. As the freezing rate increased, the degree of dissociation and unfolding increased significantly, as determined using blue native-polyacrylamide gel electrophoresis and fluorescence spectroscopy. Moreover, a large number of amyloid aggregates were detected in samples frozen at a fast freezing rate (70.6 °C/min). The enzyme inactivation mechanism induced by fast freezing was proposed in terms of increased dehydration at the enzyme surface and an ice/unfroze solution interface, which could be helpful to establish a common understanding of enzyme inactivation during the freezing process.Haena ParkJun-Young ParkKyung-Min ParkPahn-Shick ChangNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-10 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Haena Park Jun-Young Park Kyung-Min Park Pahn-Shick Chang Effects of freezing rate on structural changes in l-lactate dehydrogenase during the freezing process
description	Abstract Freezing is a common method for improving enzyme storage stability. During the freezing process, the freezing rate is an important parameter that can affect protein stability. However, there is limited information on the denaturation mechanisms and protein conformational changes associated with the freezing rate. In this study, the effects of freezing rate on activity loss and conformational changes in a model enzyme, l-lactate dehydrogenase, were evaluated. Enzyme solutions were frozen at various rates, from 0.2 to 70.6 °C/min, and ice seeding was conducted to reduce supercooling. The results demonstrated that fast freezing results in activity loss, structural changes, and aggregation. The residual activities at freezing rates of 0.2, 12.8, and 70.6 °C/min were 77.6 ± 0.9%, 64.1 ± 0.4%, and 44.8 ± 2.0%, respectively. As the freezing rate increased, the degree of dissociation and unfolding increased significantly, as determined using blue native-polyacrylamide gel electrophoresis and fluorescence spectroscopy. Moreover, a large number of amyloid aggregates were detected in samples frozen at a fast freezing rate (70.6 °C/min). The enzyme inactivation mechanism induced by fast freezing was proposed in terms of increased dehydration at the enzyme surface and an ice/unfroze solution interface, which could be helpful to establish a common understanding of enzyme inactivation during the freezing process.
format	article
author	Haena Park Jun-Young Park Kyung-Min Park Pahn-Shick Chang
author_facet	Haena Park Jun-Young Park Kyung-Min Park Pahn-Shick Chang
author_sort	Haena Park
title	Effects of freezing rate on structural changes in l-lactate dehydrogenase during the freezing process
title_short	Effects of freezing rate on structural changes in l-lactate dehydrogenase during the freezing process
title_full	Effects of freezing rate on structural changes in l-lactate dehydrogenase during the freezing process
title_fullStr	Effects of freezing rate on structural changes in l-lactate dehydrogenase during the freezing process
title_full_unstemmed	Effects of freezing rate on structural changes in l-lactate dehydrogenase during the freezing process
title_sort	effects of freezing rate on structural changes in l-lactate dehydrogenase during the freezing process
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/3f1c0e626bfc41b5ac7e67f35cd10d69
work_keys_str_mv	AT haenapark effectsoffreezingrateonstructuralchangesinllactatedehydrogenaseduringthefreezingprocess AT junyoungpark effectsoffreezingrateonstructuralchangesinllactatedehydrogenaseduringthefreezingprocess AT kyungminpark effectsoffreezingrateonstructuralchangesinllactatedehydrogenaseduringthefreezingprocess AT pahnshickchang effectsoffreezingrateonstructuralchangesinllactatedehydrogenaseduringthefreezingprocess
_version_	1718384405536309248

Effects of freezing rate on structural changes in l-lactate dehydrogenase during the freezing process

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