Two Heat Resistant Endopeptidases from Pseudomonas Species with Destabilizing Potential during Milk Storage
- Veronika VolkVeronika VolkInstitute of Food Science and Biotechnology, Department of Biotechnology and Enzyme Science, University of Hohenheim, Garbenstrasse 25, 70599 Stuttgart, GermanyMore by Veronika Volk
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- Claudia GlückClaudia GlückInstitute of Food Science and Biotechnology, Department of Biotechnology and Enzyme Science, University of Hohenheim, Garbenstrasse 25, 70599 Stuttgart, GermanyMore by Claudia Glück
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- Sebastian LeptihnSebastian LeptihnInstitute of Microbiology, Department of Microbiology, University of Hohenheim, Garbenstrasse 30, 70599 Stuttgart, GermanyMore by Sebastian Leptihn
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- Jacob EwertJacob EwertInstitute of Food Science and Biotechnology, Department of Biotechnology and Enzyme Science, University of Hohenheim, Garbenstrasse 25, 70599 Stuttgart, GermanyMore by Jacob Ewert
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- Timo Stressler*Timo Stressler*E-mail: [email protected]. Tel.: +49 711 459 22576.Institute of Food Science and Biotechnology, Department of Biotechnology and Enzyme Science, University of Hohenheim, Garbenstrasse 25, 70599 Stuttgart, GermanyMore by Timo Stressler
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- Lutz FischerLutz FischerInstitute of Food Science and Biotechnology, Department of Biotechnology and Enzyme Science, University of Hohenheim, Garbenstrasse 25, 70599 Stuttgart, GermanyMore by Lutz Fischer
Abstract
In the current study, the extracellular endopeptidases from Pseudomonas lundensis and Pseudomonas proteolytica were investigated. The amino acid sequence identity between both endopeptidases is 68%. Both endopeptidases were purified to homogeneity and partially characterized. They were classified as metallopeptidases with a maximum activity at pH 10.0 (P. lundensis) or 8.5 (P. proteolytica) at 35 °C. Both remained active in skim milk with 39.7 ± 2.4% and 24.5 ± 3.3%, respectively, of the initial enzyme activity after UHT processing (138 °C for 20 s), indicating the relevance for milk destabilization. The transition points in buffer were determined at 50 °C (P. lundensis) and 43 °C (P. proteolytica) using circular dichroism spectroscopy. The loss of the secondary structure at different temperatures was correlated with residual peptidase activities after heat treatment. The ability to destabilize UHT milk was proven by supplementation of skim milk with endopeptidase and storage for 4 weeks.
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This article is cited by 4 publications.
- Yu Wang, Yingwang Deng, Jialei Sun, Wenjing Cai, Xue Han. The effect of extracellular protease secreted by Pseudomonas fluorescens W3 on the quality of UHT milk. Journal of Food Processing and Preservation 2022, 46 (7) https://doi.org/10.1111/jfpp.16660
- Laura Quintieri, Leonardo Caputo, Milena Brasca, Francesca Fanelli. Recent Advances in the Mechanisms and Regulation of QS in Dairy Spoilage by Pseudomonas spp.. Foods 2021, 10 (12) , 3088. https://doi.org/10.3390/foods10123088
- Veronika Volk, Nicole Graw, Timo Stressler, Lutz Fischer. An indirect ELISA system for the detection of heat-stable Pseudomonas endopeptidases (AprX) in milk. Journal of Dairy Science 2021, 104 (5) , 5185-5196. https://doi.org/10.3168/jds.2020-19790
- Christopher Maier, Christopher Huptas, Mario von Neubeck, Siegfried Scherer, Mareike Wenning, Genia Lücking. Genetic Organization of the aprX-lipA2 Operon Affects the Proteolytic Potential of Pseudomonas Species in Milk. Frontiers in Microbiology 2020, 11 https://doi.org/10.3389/fmicb.2020.01190