Isolation, identification, and mode of action of antibacterial peptides derived from egg yolk hydrolysate
ABSTRACT: Egg yolk is a coproduct of egg white processing. The protein hydrolysis of egg yolks to exhibit antimicrobial activity is a strategy for its valorization. The objective of this study is to fractionate antibacterial peptides from pepsin-hydrolyzed egg yolks using flash chromatography. In ad...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2023-07-01
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| Series: | Poultry Science |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S0032579123002171 |
| _version_ | 1797802303219040256 |
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| author | Thippawan Pimchan Fu Tian Kanjana Thumanu Sureelak Rodtong Jirawat Yongsawatdigul |
| author_facet | Thippawan Pimchan Fu Tian Kanjana Thumanu Sureelak Rodtong Jirawat Yongsawatdigul |
| author_sort | Thippawan Pimchan |
| collection | DOAJ |
| description | ABSTRACT: Egg yolk is a coproduct of egg white processing. The protein hydrolysis of egg yolks to exhibit antimicrobial activity is a strategy for its valorization. The objective of this study is to fractionate antibacterial peptides from pepsin-hydrolyzed egg yolks using flash chromatography. In addition, the mode of actions of the fractionated peptides were elucidated and plausible antibacterial peptides were reported. The fraction 6 (F6) obtained from a C18-flash column exhibited antibacterial activity against Staphylococcus aureus ATCC 29213 and Salmonella typhimurium TISTR 292 at minimal inhibitory concentration (MIC) values of 0.5 to 1 mmol/L (Leucine equivalent). The fractionated peptides induced DNA leakage as monitored by 260 nm. Propidium iodide and SYTO9 staining observed under a confocal microscope suggested the disintegration of cell membranes. Synchrotron-based Fourier-transform infrared spectroscopy analysis revealed that the egg yolk peptides at 1 × MIC induced an alteration of phospholipids at cell membranes and modified conformation of intracellular proteins and nucleic acids. Scanning electron microscopy revealed obvious cell ruptures when S. aureus was treated at 1 × MIC for 4 h, whereas damage of cell membranes and leakage of intracellular components were also observed for the transmission electron microscopy. Egg yolk peptides showed no hemolytic activity in human erythrocytes at concentrations up to 4 mmol/L. Peptide identification by LC-MS/MS revealed 3 cationic and 10 anionic peptides with 100% sequence similarity to apolipoprotein-B of Gallus gallus with hydrophobicity ranging from 27 to 75%. The identified peptide KGGDLGLFEPTL exhibited the highest antibacterial activity toward S. aureus at MIC of 2 mmol/L. Peptides derived from egg yolk hydrolysate present significant potential as antistaphylococcal agents for food and/or pharmaceutical application. |
| first_indexed | 2024-03-13T05:03:39Z |
| format | Article |
| id | doaj.art-4b47d45e5c8e4b8988478d2d11fc3053 |
| institution | Directory Open Access Journal |
| issn | 0032-5791 |
| language | English |
| last_indexed | 2024-03-13T05:03:39Z |
| publishDate | 2023-07-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Poultry Science |
| spelling | doaj.art-4b47d45e5c8e4b8988478d2d11fc30532023-06-17T05:17:17ZengElsevierPoultry Science0032-57912023-07-011027102695Isolation, identification, and mode of action of antibacterial peptides derived from egg yolk hydrolysateThippawan Pimchan0Fu Tian1Kanjana Thumanu2Sureelak Rodtong3Jirawat Yongsawatdigul4School of Food Technology, Institute of Agricultural Technology, Suranaree University of Technology, Nakhon Ratchasima, 30000, ThailandSchool of Food Technology, Institute of Agricultural Technology, Suranaree University of Technology, Nakhon Ratchasima, 30000, ThailandSynchrotron Light Research Institute (Public Organization), Nakhon Ratchasima, 30000, ThailandSchool of Preclinical Sciences, Institute of Science, Suranaree University of Technology, Nakhon Ratchasima, 30000, ThailandSchool of Food Technology, Institute of Agricultural Technology, Suranaree University of Technology, Nakhon Ratchasima, 30000, Thailand; Corresponding author:ABSTRACT: Egg yolk is a coproduct of egg white processing. The protein hydrolysis of egg yolks to exhibit antimicrobial activity is a strategy for its valorization. The objective of this study is to fractionate antibacterial peptides from pepsin-hydrolyzed egg yolks using flash chromatography. In addition, the mode of actions of the fractionated peptides were elucidated and plausible antibacterial peptides were reported. The fraction 6 (F6) obtained from a C18-flash column exhibited antibacterial activity against Staphylococcus aureus ATCC 29213 and Salmonella typhimurium TISTR 292 at minimal inhibitory concentration (MIC) values of 0.5 to 1 mmol/L (Leucine equivalent). The fractionated peptides induced DNA leakage as monitored by 260 nm. Propidium iodide and SYTO9 staining observed under a confocal microscope suggested the disintegration of cell membranes. Synchrotron-based Fourier-transform infrared spectroscopy analysis revealed that the egg yolk peptides at 1 × MIC induced an alteration of phospholipids at cell membranes and modified conformation of intracellular proteins and nucleic acids. Scanning electron microscopy revealed obvious cell ruptures when S. aureus was treated at 1 × MIC for 4 h, whereas damage of cell membranes and leakage of intracellular components were also observed for the transmission electron microscopy. Egg yolk peptides showed no hemolytic activity in human erythrocytes at concentrations up to 4 mmol/L. Peptide identification by LC-MS/MS revealed 3 cationic and 10 anionic peptides with 100% sequence similarity to apolipoprotein-B of Gallus gallus with hydrophobicity ranging from 27 to 75%. The identified peptide KGGDLGLFEPTL exhibited the highest antibacterial activity toward S. aureus at MIC of 2 mmol/L. Peptides derived from egg yolk hydrolysate present significant potential as antistaphylococcal agents for food and/or pharmaceutical application.http://www.sciencedirect.com/science/article/pii/S0032579123002171egg yolkprotein hydrolysateantibacterial peptideflash chromatographymechanism |
| spellingShingle | Thippawan Pimchan Fu Tian Kanjana Thumanu Sureelak Rodtong Jirawat Yongsawatdigul Isolation, identification, and mode of action of antibacterial peptides derived from egg yolk hydrolysate Poultry Science egg yolk protein hydrolysate antibacterial peptide flash chromatography mechanism |
| title | Isolation, identification, and mode of action of antibacterial peptides derived from egg yolk hydrolysate |
| title_full | Isolation, identification, and mode of action of antibacterial peptides derived from egg yolk hydrolysate |
| title_fullStr | Isolation, identification, and mode of action of antibacterial peptides derived from egg yolk hydrolysate |
| title_full_unstemmed | Isolation, identification, and mode of action of antibacterial peptides derived from egg yolk hydrolysate |
| title_short | Isolation, identification, and mode of action of antibacterial peptides derived from egg yolk hydrolysate |
| title_sort | isolation identification and mode of action of antibacterial peptides derived from egg yolk hydrolysate |
| topic | egg yolk protein hydrolysate antibacterial peptide flash chromatography mechanism |
| url | http://www.sciencedirect.com/science/article/pii/S0032579123002171 |
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