The Impact of Low-Temperature Inactivation of Protease AprX from <i>Pseudomonas</i> on Its Proteolytic Capacity and Specificity: A Peptidomic Study
The destabilization of UHT milk during its shelf life can be promoted by the residual proteolytic activity attributed to the protease AprX from <i>Pseudomonas</i>. To better understand the hydrolysis patterns of AprX, and to evaluate the feasibility of using low-temperature inactivation...
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MDPI AG
2023-02-01
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| Series: | Dairy |
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| Online Access: | https://www.mdpi.com/2624-862X/4/1/11 |
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| author | Chunyue Zhang Sjef Boeren Liming Zhao Etske Bijl Kasper Hettinga |
| author_facet | Chunyue Zhang Sjef Boeren Liming Zhao Etske Bijl Kasper Hettinga |
| author_sort | Chunyue Zhang |
| collection | DOAJ |
| description | The destabilization of UHT milk during its shelf life can be promoted by the residual proteolytic activity attributed to the protease AprX from <i>Pseudomonas</i>. To better understand the hydrolysis patterns of AprX, and to evaluate the feasibility of using low-temperature inactivation (LTI) for AprX, the release of peptides through AprX activity on milk proteins was examined using an LC-MS/MS-based peptidomic analysis. Milk samples were either directly incubated to be hydrolyzed by AprX, or preheated under LTI conditions (60 °C for 15 min) and then incubated. Peptides and parent proteins (the proteins from which the peptides originated) were identified and quantified. The peptides were mapped and the cleavage frequency of amino acids in the P1/P1′ positions was analyzed, after which the influence of LTI and the potential bitterness of the formed peptides were determined. Our results showed that a total of 2488 peptides were identified from 48 parent proteins, with the most abundant peptides originating from κ-casein and β-casein. AprX may also non-specifically hydrolyze other proteins in milk. Except for decreasing the bitterness potential in skim UHT milk, LTI did not significantly reduce the AprX-induced hydrolysis of milk proteins. Therefore, the inactivation of AprX by LTI may not be feasible in UHT milk production. |
| first_indexed | 2024-03-11T06:41:36Z |
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| institution | Directory Open Access Journal |
| issn | 2624-862X |
| language | English |
| last_indexed | 2024-03-11T06:41:36Z |
| publishDate | 2023-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Dairy |
| spelling | doaj.art-33c3c4f0fee5481fa944d606f64fd4a42023-11-17T10:31:48ZengMDPI AGDairy2624-862X2023-02-014115016610.3390/dairy4010011The Impact of Low-Temperature Inactivation of Protease AprX from <i>Pseudomonas</i> on Its Proteolytic Capacity and Specificity: A Peptidomic StudyChunyue Zhang0Sjef Boeren1Liming Zhao2Etske Bijl3Kasper Hettinga4Dairy Science and Technology, Food Quality and Design Group, Wageningen University and Research, P.O. Box 17, 6700 AA Wageningen, The NetherlandsLaboratory of Biochemistry, Wageningen University and Research Center, Stippeneng 4, 6708 WE Wageningen, The NetherlandsState Key Laboratory of Bioreactor Engineering, School of Biotechnology, East China University of Science and Technology, Shanghai 200237, ChinaDairy Science and Technology, Food Quality and Design Group, Wageningen University and Research, P.O. Box 17, 6700 AA Wageningen, The NetherlandsDairy Science and Technology, Food Quality and Design Group, Wageningen University and Research, P.O. Box 17, 6700 AA Wageningen, The NetherlandsThe destabilization of UHT milk during its shelf life can be promoted by the residual proteolytic activity attributed to the protease AprX from <i>Pseudomonas</i>. To better understand the hydrolysis patterns of AprX, and to evaluate the feasibility of using low-temperature inactivation (LTI) for AprX, the release of peptides through AprX activity on milk proteins was examined using an LC-MS/MS-based peptidomic analysis. Milk samples were either directly incubated to be hydrolyzed by AprX, or preheated under LTI conditions (60 °C for 15 min) and then incubated. Peptides and parent proteins (the proteins from which the peptides originated) were identified and quantified. The peptides were mapped and the cleavage frequency of amino acids in the P1/P1′ positions was analyzed, after which the influence of LTI and the potential bitterness of the formed peptides were determined. Our results showed that a total of 2488 peptides were identified from 48 parent proteins, with the most abundant peptides originating from κ-casein and β-casein. AprX may also non-specifically hydrolyze other proteins in milk. Except for decreasing the bitterness potential in skim UHT milk, LTI did not significantly reduce the AprX-induced hydrolysis of milk proteins. Therefore, the inactivation of AprX by LTI may not be feasible in UHT milk production.https://www.mdpi.com/2624-862X/4/1/11UHT milkshelf life<i>Pseudomonas</i>AprX proteaseenzyme inactivationpeptidomics |
| spellingShingle | Chunyue Zhang Sjef Boeren Liming Zhao Etske Bijl Kasper Hettinga The Impact of Low-Temperature Inactivation of Protease AprX from <i>Pseudomonas</i> on Its Proteolytic Capacity and Specificity: A Peptidomic Study Dairy UHT milk shelf life <i>Pseudomonas</i> AprX protease enzyme inactivation peptidomics |
| title | The Impact of Low-Temperature Inactivation of Protease AprX from <i>Pseudomonas</i> on Its Proteolytic Capacity and Specificity: A Peptidomic Study |
| title_full | The Impact of Low-Temperature Inactivation of Protease AprX from <i>Pseudomonas</i> on Its Proteolytic Capacity and Specificity: A Peptidomic Study |
| title_fullStr | The Impact of Low-Temperature Inactivation of Protease AprX from <i>Pseudomonas</i> on Its Proteolytic Capacity and Specificity: A Peptidomic Study |
| title_full_unstemmed | The Impact of Low-Temperature Inactivation of Protease AprX from <i>Pseudomonas</i> on Its Proteolytic Capacity and Specificity: A Peptidomic Study |
| title_short | The Impact of Low-Temperature Inactivation of Protease AprX from <i>Pseudomonas</i> on Its Proteolytic Capacity and Specificity: A Peptidomic Study |
| title_sort | impact of low temperature inactivation of protease aprx from i pseudomonas i on its proteolytic capacity and specificity a peptidomic study |
| topic | UHT milk shelf life <i>Pseudomonas</i> AprX protease enzyme inactivation peptidomics |
| url | https://www.mdpi.com/2624-862X/4/1/11 |
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