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Analytical and Bioanalytical Chemistry

, Volume 410, Issue 15, pp 3573–3586 | Cite as

Peptidomic strategy for purification and identification of potential ACE-inhibitory and antioxidant peptides in Tetradesmus obliquus microalgae

  • Carmela Maria Montone
  • Anna Laura Capriotti
  • Chiara Cavaliere
  • Giorgia La Barbera
  • Susy Piovesana
  • Riccardo Zenezini Chiozzi
  • Aldo Laganà
Research Paper
Part of the following topical collections:
  1. Discovery of Bioactive Compounds

Abstract

Microalgae are unicellular marine organisms that have promoted complex biochemical pathways to survive in greatly competitive marine environments. They could contain significant amounts of high-quality proteins which, because of their structural diversity, contain a range of yet undiscovered novel bioactive peptides. In this work, a peptidomic platform was developed for the separation and identification of bioactive peptides in protein hydrolysates. In this work, a peptidomic platform was developed for the extraction, separation, and identification of bioactive peptides in protein hydrolysates. Indeed, extraction of proteins from recalcitrant tissues is still a challenge due to their strong cell walls and high levels of non-protein interfering compounds. Therefore, seven different protein extraction protocols, based on mechanical and chemical methods, were tested in order to produce high-quality protein extracts. Proteins obtained by means of the best protocol, consisting of milling the recalcitrant tissue with glass beads, were subjected to enzymatic digestion with Alcalase® and subsequently the hydrolysate was purified by two-dimensional semi-preparative reversed phase liquid chromatography. Fractions were assayed for antioxidant and antihypertensive activities and only the most active ones were finally analyzed by RP nanoHPLC-MS/MS. Around 500 peptide sequences were identified in these fractions. The identified peptides were subjected to an in silico analysis by PeptideRanker algorithm in order to assign a score of bioactivity probability. Twenty-five sequenced peptides were found with potential antioxidant and angiotensin-converting-enzyme-inhibitory activities. Four of these peptides, WPRGYFL, GPDRPKFLGPF, WYGPDRPKFL, SDWDRF, were selected for synthesis and in vitro tested for specific bioactivity, exhibiting good values of antioxidant and ACE-inhibitory activity.

Graphical abstract

Workflow showing the entire peptidomic approach developed for identification of bioactive peptides in microalgae

Keywords

Protein extraction methods Peptidomics Microalgae Antioxidant peptides ACE-inhibitory peptides Off-line two-dimensional chromatography High resolution mass spectrometry 

Notes

Acknowledgements

This work has been carried out within the framework of the Research Project “Microalgae as a source of bioactive compounds: chromatographic fractionation of peptides and lipids and their mass spectrometric characterization,” supported by Sapienza, no. RM11715C82118E74.

Moreover, the authors wish to thank Prof. Francesca Pagnanelli for providing the microalgae samples.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest

Supplementary material

216_2018_925_MOESM1_ESM.xlsx (61 kb)
ESM 1 (XLSX 61 kb)
216_2018_925_MOESM2_ESM.pdf (832 kb)
ESM 2 (PDF 831 kb)

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Carmela Maria Montone
    • 1
  • Anna Laura Capriotti
    • 1
  • Chiara Cavaliere
    • 1
  • Giorgia La Barbera
    • 1
  • Susy Piovesana
    • 1
  • Riccardo Zenezini Chiozzi
    • 1
  • Aldo Laganà
    • 1
  1. 1.Department of ChemistrySapienza Università di RomaRomeItaly

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