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Updated: Aug 12, 2026
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Matrix-assisted Laser Desorption/Ionization Time of Flight (MALDI-TOF) Mass Spectrometric Analysis of Intact Proteins Larger than 100 kDa
Published on: September 9, 2013
Profiling Hydrolysis Rates of αS1-Casein to αS1-I-Casein During Cheese Ripening by Intact-Protein Liquid
Sebastiaan Dolman1, Marjolein van der Eijk1, Loes Bevers1
1dsm-firmenich, Delft, the Netherlands.
Rapid Communications in Mass Spectrometry : RCM
|August 11, 2026
Summary
Bovine chymosins hydrolyze αS1-casein to αS1-I-casein faster than camel chymosins during mozzarella ripening. Intact-protein LC-HRMS accurately tracks these differences, revealing enzyme-specific impacts on cheese quality and texture development.
Area of Science:
- Food Science
- Proteomics
- Enzymology
Background:
- Chymosin activity influences cheese texture, flavor, and quality.
- Specific hydrolysis of αS1-casein to αS1-I-casein is key for early cheese texture.
- Chymosin variant differences impact this hydrolysis rate.
Purpose of the Study:
- To monitor αS1-casein to αS1-I-casein hydrolysis during mozzarella ripening.
- To compare the proteolytic activity of bovine and camel chymosins.
- To assess the impact of chymosin variants on cheese maturation.
Main Methods:
- Intact-protein liquid chromatography-high-resolution mass spectrometry (LC-HRMS) was utilized.
- Mozzarella cheese samples from different ripening stages were analyzed.
- Hydrolysis kinetics and phosphorylation variants of αS1-casein were quantified.
Main Results:
- Bovine chymosins showed significantly higher αS1-casein hydrolysis rates than camel chymosins.
- Distinct phosphorylation variants were identified and quantified.
- Consistent differences in proteolytic activity between bovine and camel chymosins were observed.
Conclusions:
- LC-HRMS effectively tracks αS1-casein hydrolysis kinetics during cheese ripening.
- Bovine chymosins exhibit faster proteolytic activity, leading to quicker αS1-I-casein formation.
- Enzyme-specific effects on cheese texture and quality are highlighted, underscoring the value of advanced protein analysis.
