Abstract
Bioactive peptides derived from chia (Salvia hispanica) seed with antioxidant, antihypertensive, and anti-inflammatory activities have been well documented; however, few studies describe the antimicrobial properties of these peptides, which is of great interest not only in the prevention of food-borne diseases but also food spoilage. The aim of this study was to generate chia seed peptides using microwave-assisted hydrolysis with sequential (alcalase + flavourzyme) enzymes (AF-MW), fractionate them into 3–10 and < 3 kDa fractions, and evaluate their potential antimicrobial activity towards Escherichia coli, Salmonella enterica, and Listeria monocytogenes. Overall, the peptide fraction < 3 kDa showed higher antimicrobial activity than both chia seed hydrolysate and peptide fraction 3–10 kDa. Furthermore, the < 3 kDa fraction showed remarkable increase in membrane permeability of E. coli (71.49% crystal violet uptake) and L. monocytogenes (80.10% crystal violet uptake). These peptides caused a significant extension in the lag phase, decreases in the maximum growth, and growth rate in the bacteria and promoted multiple indentations (transmembrane tunnels), membrane wrinkling, and pronounced deformations in the integrity of the bacterial cell membranes. Finally, a select group of peptides in the AF-MW < 3 kDa fraction contained 16 sequences with cationic and hydrophobic character, with seven of them sharing the exact same sequence (GDVIAIR) and eight of them having the amino acid K as either N- or C-terminal or both. In conclusion, our results indicate that bioactive peptides obtained from chia seed proteins by microwave and enzymatic hydrolysis could be employed as antimicrobial agents in foods and therapeutic applications.
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Acknowledgments
The authors would like to thank for the technical support from Christopher J. Gilpin, Laurie Mueller, Robert Seiler at the Life Science Microscopy Facility at Purdue University, and from Emma Doud at Proteomics Core Facility at the Indiana University School of Medicine. In addition, the authors express their appreciation to Uriel C. Urbizo Reyes (Protein Chemistry and Bioactive Peptides Laboratory, Department of Food Sciences) and Hansel Mina Cordoba (Food Safety Laboratory, Department of Food Sciences) of Purdue University, for their valuable technical assistance.
Funding
The present work was supported by the USDA-NIFA, Hatch Act formula funds (project 1019794) in the College of Agriculture, Purdue University.
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A. M. Liceaga contributed to the study conception and design. A. Deering assisted with the bacterial strains used in the study. Material preparation, data collection, and analysis were performed by J.E. Aguilar-Toalá, A. Deering, and A. M. Liceaga. The first draft of the manuscript was written by J.E. Aguilar-Toalá, and all the authors edited and commented on previous versions of the manuscript. All the authors read and approved the final manuscript.
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Aguilar-Toalá, J.E., Deering, A.J. & Liceaga, A.M. New Insights into the Antimicrobial Properties of Hydrolysates and Peptide Fractions Derived from Chia Seed (Salvia hispanica L.). Probiotics & Antimicro. Prot. 12, 1571–1581 (2020). https://doi.org/10.1007/s12602-020-09653-8
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DOI: https://doi.org/10.1007/s12602-020-09653-8