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A Sensitive and Robust Method for Direct Determination of Lipolytic Activity in Natural Milk Environment

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Abstract

Microbial lipases may be produced during milk storage and processing. This can lead to organoleptic changes in the corresponding dairy products. Thus, monitoring of lipase activity in milk is desirable. Turbidity of milk prevents a direct photometric measurement of lipase activity using chromophore- or fluorophore-based assays. Laborious pretreatments or alternative analytical methods normally have to be used. With the method for the determination of lipolytic activity (MeDeLi) proposed here, it is possible to measure lipase activity directly in the natural milk utilizing tailored fluorometric substrates. Only a defatting step is carried out initially for the MeDeLi. Then, the conversion of added lipase substrate is carried out in the unmodified milk without addition of any solutions or any enzyme extraction procedure which may influence the enzyme activity. Thereafter, the milk sample is treated with two solutions to remove the turbidity of milk by dissolution. A valid and sensitive fluorometric measurement is then possible. The applicability of the MeDeLi was demonstrated in comparison with tests published previously: The limit of detection for lipolytic activity measured by MeDeLi was the lowest, with 41 pkat/mL. Raw milk, milk products, and spoiled milk samples were also investigated with the MeDeLi.

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Funding

This research project was supported by the Federal Ministry for Economic Affairs and Energy (via AiF) and the FEI (Forschungskreis für Ernährungsindustrie e.V., Bonn): Project AiF 16588 N.

Conflict of Interest

Manuel Krewinkel declares that he has no conflict of interest. Claudia Baur declares that she has no conflict of interest. Bertolt Kranz declares that he has no conflict of interest. Mario von Neubeck declares that he has no conflict of interest. Mareike Wenning declares that she has no conflict of interest. Siegfried Scherer declares that he has no conflict of interest . Marina Stoeckel declares that she has no conflict of interest. Jörg Hinrichs declares that he has no conflict of interest. Lutz Fischer declares that he has no conflict of interest. This article does not contain any studies with human participants or animals performed by any of the authors.

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Authors and Affiliations

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

    Manuel Krewinkel, Claudia Baur, Bertolt Kranz & Lutz Fischer

  2. Lehrstuhl für Mikrobielle Ökologie, Zentralinstitut für Ernährungs- und Lebensmittelforschung (ZIEL), Wissenschaftszentrum Weihenstephan, Technische Universität München, Weihenstephaner Berg 3, 85354, Freising, Germany

    Mario von Neubeck, Mareike Wenning & Siegfried Scherer

  3. Department of Soft Matter Science and Dairy Technology, Institute of Food Science and Biotechnology, University of Hohenheim, Garbenstr. 21, 70599, Stuttgart, Germany

    Marina Stoeckel & Jörg Hinrichs

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  1. Manuel Krewinkel
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  2. Claudia Baur
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  3. Bertolt Kranz
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  4. Mario von Neubeck
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  5. Mareike Wenning
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  6. Siegfried Scherer
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  7. Marina Stoeckel
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  8. Jörg Hinrichs
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  9. Lutz Fischer
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Correspondence to Lutz Fischer.

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Krewinkel, M., Baur, C., Kranz, B. et al. A Sensitive and Robust Method for Direct Determination of Lipolytic Activity in Natural Milk Environment. Food Anal. Methods 9, 646–655 (2016). https://doi.org/10.1007/s12161-015-0233-4

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  • DOI: https://doi.org/10.1007/s12161-015-0233-4

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