Evaluating the effect of glycation on lipase activity using boronate affinity chromatography and mass spectrometry.

Autor: van Schaick G; Leiden University Medical Center, Center for Proteomics and Metabolomics, Leiden, the Netherlands. Electronic address: g.van_schaick@lumc.nl., Pot S; Leiden University Medical Center, Center for Proteomics and Metabolomics, Leiden, the Netherlands., Schouten O; DSM Science & Innovation, Biodata & Translation, Center for Analytical Innovation, Delft, the Netherlands., den Hartog J; DSM Science & Innovation, Biodata & Translation, Center for Analytical Innovation, Delft, the Netherlands., Akeroyd M; DSM Science & Innovation, Biodata & Translation, Center for Analytical Innovation, Delft, the Netherlands., van der Hoeven R; DSM Science & Innovation, Biodata & Translation, Center for Analytical Innovation, Delft, the Netherlands., Bijleveld W; DSM Science & Innovation, Biodata & Translation, Center for Analytical Innovation, Delft, the Netherlands., Abello N; DSM Science & Innovation, Biodata & Translation, Center for Analytical Innovation, Delft, the Netherlands., Wuhrer M; Leiden University Medical Center, Center for Proteomics and Metabolomics, Leiden, the Netherlands., Olsthoorn M; DSM Science & Innovation, Biodata & Translation, Center for Analytical Innovation, Delft, the Netherlands., Dominguez-Vega E; Leiden University Medical Center, Center for Proteomics and Metabolomics, Leiden, the Netherlands.
Jazyk: angličtina
Zdroj: Food chemistry [Food Chem] 2023 Sep 30; Vol. 421, pp. 136147. Date of Electronic Publication: 2023 Apr 17.
DOI: 10.1016/j.foodchem.2023.136147
Abstrakt: Protein glycation may occur naturally when reducing sugars and proteins coexist, which is often the case for industrial enzymes. The impact of post-translational modifications on enzyme performance (e.g., stability or function) is often not predictable, highlighting the importance of having appropriate analytical methodologies to monitor the influence of glycation on performance. Here, a boronate affinity chromatography method was developed to enrich glycated species followed by mass spectrometry for structural characterization and activity assays for functional assessment. This approach was applied to a (temperature-stressed) lipase used for food applications revealing that storage at -20 °C and 4 °C resulted in minor glycation (below 9%), whereas storage at 25 °C led to a higher glycation level with up to four sugars per lipase molecule. Remarkably, activity measurements revealed that glycation did not reduce lipase activity or stability. Altogether, this novel strategy is a helpful extension to the current analytical toolbox supporting development of enzyme products.
Competing Interests: Declaration of Competing Interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Co-authors Olaf Schouten, Joost den Hartog, Michiel Akeroyd, Rob van der Hoeven, Wim Bijleveld, Nicolas Abello, and Maurien Olsthoorn are affiliated with Royal DSM, a global company active in health, nutrition, and bioscience.
(Copyright © 2023 The Author(s). Published by Elsevier Ltd.. All rights reserved.)
Databáze: MEDLINE
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