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

Food Chemistry, vol. 421, pp. 136147

Abstract

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.

Authors 10

1. Guusje van Schaick corresponding Aachen Center for Proteomics and Metabolomics

   Leiden University Medical Center

   Affiliation as printed

   Leiden University Medical Center, Center for Proteomics and Metabolomics, Leiden, the Netherlands. Electronic address: g.van_schaick@lumc.nl

   Leiden University Medical Center, Center for Proteomics and Metabolomics, Leiden, the Netherlands

2. Sanne Pot Aachen Center for Proteomics and Metabolomics

   Leiden University Medical Center

   Affiliation as printed

   Leiden University Medical Center, Center for Proteomics and Metabolomics, Leiden, the Netherlands

3. Joost den Hartog
   Affiliation as printed

   DSM Science & Innovation, Biodata & Translation, Center for Analytical Innovation, Delft, the Netherlands

4. Michiel Akeroyd
   Affiliation as printed

   DSM Science & Innovation, Biodata & Translation, Center for Analytical Innovation, Delft, the Netherlands

5. Rob van der Hoeven
   Affiliation as printed

   DSM Science & Innovation, Biodata & Translation, Center for Analytical Innovation, Delft, the Netherlands

6. Wim Bijleveld
   Affiliation as printed

   DSM Science & Innovation, Biodata & Translation, Center for Analytical Innovation, Delft, the Netherlands

7. Nicolas Abello
   Affiliation as printed

   DSM Science & Innovation, Biodata & Translation, Center for Analytical Innovation, Delft, the Netherlands

8. Manfred Wuhrer Aachen Center for Proteomics and Metabolomics

   Leiden University Medical Center

   Affiliation as printed

   Leiden University Medical Center, Center for Proteomics and Metabolomics, Leiden, the Netherlands

9. Maurien M. A. Olsthoorn
   Affiliation as printed

   DSM Science & Innovation, Biodata & Translation, Center for Analytical Innovation, Delft, the Netherlands

10. Elena Domínguez‐Vega Aachen Center for Proteomics and Metabolomics

    Leiden University Medical Center

    Affiliation as printed

    Leiden University Medical Center, Center for Proteomics and Metabolomics, Leiden, the Netherlands