Poster Presentation 29th Annual Lorne Proteomics Symposium 2024

Unveiling Bioactive Peptides in Caseins: An Integrated Approach Using Peptidomics and Bioinformatics (#147)

Yasuri Amarasekara 1 2 , Rachana Pathak 1 2 , Narsimha Reddy 1 3 , Helena Nevalainen 1 4 , Junior Te'o 1 4 , Anwar Sunna 1 2
  1. Australian Research Council Industrial Transformation Training Centre for Facilitate Advancement of Australia's Bioactives (FAAB), North Ryde, NSW 2109, Australia
  2. Macquarie University, School of Natural Sciences, North Ryde, NSW 2109, Australia
  3. Western Sydney University, School of Science, Parramatta, NSW 2150, Australia
  4. Change Foods , Palo Alto, CA 94306, USA

Bioactive peptides (BAPs) are specific protein fragments (2-20 amino acids long) with the ability to positively impact crucial biological functions and physiological responses in living organisms.1,2 Bovine caseins (CNs) found in milk are recognised as a rich reservoir of BAPs with potential applications as supplements in both food and pharmaceutical formulations.3 As these bioactive peptides are encrypted within the primary structure of CNs, they can be produced via in vitro simulated gastrointestinal digestion.4 However, the conventional method, along with subsequent bioactivity assays, is considered inefficient for large-scale industrial production due to the cost implications.

In addressing these constraints, we present the application of peptidomics and bioinformatics methodologies, representing analytical and computational techniques that can expedite the transition of BAPs from research to market. These tools have emerged as a strategic method for generating known and novel peptide sequences from proteins. In this study, bovine CNs were hydrolysed using simulated gastric (pepsin, pH 2-3) and intestinal (trypsin and chymotrypsin, pH 7.5) digestions in a sequential manner. Peptide sequences were identified using high-resolution tandem mass spectrometry. The sequences obtained were further analysed using in silico tools (PeptideRanker, BIOPEP, molecular modelling, and molecular docking) to predict their novelty and respective bioactivities. Computational models identified two new BAPs predicted to be non-toxic and with considerable anti-hypertensive activity, antioxidation, and anti-cancer functionality. In silico methods were utilised to pre-select the most promising peptides for chemical synthesis and further experimental evaluation.

This work demonstrates the efficiency of using peptidomics and bioinformatics as an unbiased screening strategy to identify and characterise a wide array of BAPs.

 

  1. (1) Sharma, S.; Singh, R.; Rana, S. Bioactive Peptides: A Review. Int. J. Bioautomation 2011, 15 (4), 223–250. https://doi.org/10.1093/fqs/fyx006.
  2. (2) Daliri, E. B. M.; Oh, D. H.; Lee, B. H. Bioactive Peptides. Foods 2017, 6 (5), 1–21. https://doi.org/10.3390/foods6050032.
  3. (3) Miquel, E.; Alegría, A.; Barberá, R.; Farré, R. Casein Phosphopeptides Released by Simulated Gastrointestinal Digestion of Infant Formulas and Their Potential Role in Mineral Binding. Int. Dairy J. 2006, 16 (9), 992–1000. https://doi.org/10.1016/j.idairyj.2005.10.010.
  4. (4) Lemes, A. C.; Sala, L.; Ores, J. D. C.; Braga, A. R. C.; Egea, M. B.; Fernandes, K. F. A Review of the Latest Advances in Encrypted Bioactive Peptides from Protein-Richwaste. Int. J. Mol. Sci. 2016, 17 (6). https://doi.org/10.3390/ijms17060950.