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Characterization of cross-linked enzyme aggregates (CLEAs) of the fusion protein FUS-PepN_PepX and their application for milk protein hydrolysis

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Abstract

The use of general and specific exopeptidases is of great interest for the hydrolysis of food proteins. Protein hydrolysates with a high degree of hydrolysis and, therefore, a reduced bitterness and improved antioxidative capacity can be produced due to the synergistic specificities of the general aminopeptidase PepN and the proline-specific peptidase PepX. These two activities were previously combined in a fusion protein and the latter showed both specific activities. However, due to its solubility an application of the fusion protein in continuous processes will be complicated in the future. Therefore, the aim of this study was the production, characterization and use of cross-linked enzyme aggregates (CLEAs) from the fusion protein (FUS-PepN_PepX CLEAs). The FUS-PepN_PepX CLEAs produced had activity for both specific enzymes. The biochemical characteristics determined (e.g., pH and temperature optima, environmental conditions) showed that the CLEAs are suitable for application in a complex matrix, such as food protein hydrolysates. The relative degree of hydrolysis of a prehydrolyzed casein solution was increased by 100% and the hydrolysate obtained showed a strong antioxidative capacity (ABTS-IC50 value: 7.85 µg mL−1). The stability against NaCl and the possibility of using ethanol as a microbial hurdle as well as the size of the FUS-PepN_PepX CLEAs seem promising for an application in an enzyme membrane reactor in the future. In summary, using these CLEAs, casein hydrolysates with a high degree of hydrolysis, a potentially reduced bitterness and high antioxidative capacity can be produced.

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Acknowledgements

The authors thank Nina Pfahler and Wolfgang Claaßen (University of Hohenheim, Institute of Food Science and Biotechnology, Department of Biotechnology and Enzyme Science) for their help during the bioreactor cultivation. Additionally, the authors thank Alena Kussler and Lena Nesensohn (University of Hohenheim, Institute of Food Science and Biotechnology, Department of Biotechnology and Enzyme Science) for performing preliminary experiments. Finally the authors thank Prof. Dr. Lutz Fischer (University of Hohenheim, Institute of Food Science and Biotechnology, Department of Biotechnology and Enzyme Science) for giving the opportunity to perform this work in his department.

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  1. Carina Braun and Jacob Ewert have contributed equally to the paper and should both be considered as first authors.

Authors and Affiliations

  1. Department of Biotechnology and Enzyme Science, Institute of Food Science and Biotechnology, University of Hohenheim, Garbenstr. 25, 70599, Stuttgart, Germany

    Carina Braun, Jacob Ewert & Timo Stressler

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  1. Carina Braun
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Correspondence to Timo Stressler.

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Braun, C., Ewert, J. & Stressler, T. Characterization of cross-linked enzyme aggregates (CLEAs) of the fusion protein FUS-PepN_PepX and their application for milk protein hydrolysis. Eur Food Res Technol 243, 1815–1828 (2017). https://doi.org/10.1007/s00217-017-2885-3

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  • DOI: https://doi.org/10.1007/s00217-017-2885-3

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