Journal of Applied Phycology

, Volume 27, Issue 5, pp 2059–2074 | Cite as

Bioassay-guided fractionation approach for determination of protein precursors of proteolytic bioactive metabolites from macroalgae

  • Stéphanie Bondu
  • Claudie Bonnet
  • Julie Gaubert
  • Éric Deslandes
  • Sylvie L. Turgeon
  • Lucie BeaulieuEmail author
5th Congress of the International Society for Applied Phycology


In the last decades, an upsurge in the occurrence of chronic diseases caused by hypertension and oxidative stress has been observed. The objective of this research was to isolate and characterize groups of antioxidant and anti-hypertensive natural bioactive peptides originating from hydrolyzed proteins of the red seaweeds Solieria chordalis and Palmaria palmata, the green seaweed Ulva lactuca and the brown seaweed Saccharina longicruris. Enzymatic hydrolysis by trypsin and chymotrypsin was performed in order to release bioactive algal peptides. Three antioxidant assays, 2,2-diphenyl-1-picrylhydrazyl (DPPH), ferric ion reducing antioxidant power (FRAP) and oxygen radical absorbance capacity (ORAC), were used to determine the activity of seaweed peptides. Angiotensin-converting enzyme (ACE) inhibition assay was used to evaluate the anti-hypertensive potential. Additionally, liquid chromatography-tandem mass spectrometry (LC-MS/MS) analyses and database searches allowed for protein identification, indicating where these bioactive peptides originated from. Fractions of red seaweeds, especially S. chordalis, produced by chymotrypsin and trypsin hydrolysis, exhibited higher in vitro antioxidant and ACE inhibitory activities than the parent proteins. Size exclusion chromatography highlighted two groups of bioactive peptides: (i) peptides with molecular weight (MW) between 1,400 and 3,200 Da displaying antioxidant activities and (ii) smaller peptides (MW <1,000 Da) displaying both antioxidant and ACE inhibitory activities. LC-MS/MS analyses of the antioxidant subfractions of P. palmata and S. chordalis were performed, and several peptide sequences were elucidated, using tandem mass spectrometry. Of all identified peptides of S. chordalis, 61 and 43 % came from the hydrolysis of ribulose-1, 5-biphosphate carboxylase/oxygenase (RuBiSCo) enzymes. Phycoerythrin, elongation factors, photosystems and cytochrome oxidase clusters represented between 3 and 14 % of identified peptides in S. chordalis fractions. This is the first time, to the author’s knowledge, that distinction between several groups of bioactive peptides, within seaweed protein hydrolysates, has been reported. In addition, proteins of origin were also identified.


ACE inhibitory activity Antioxidant capacity Hydrolysates Peptide amino acid identification RuBisCo enzymes Seaweeds 



The authors wish to thank Organic Ocean Inc. (QC, Canada), Dr. Éric Tamigneaux (Merinov, Grande-Rivière, QC, Canada) for providing the seaweeds, Dr. Laurie-Ève Rioux (Laval University, QC, Canada) for her expertise in monosaccharide analyses, the Proteomics platform of the Eastern Quebec Genomics Center (Quebec, Canada) for the in-solution digest, mass spectrometry experiments and database searching, the NMR platform of University of Western Brittany (Brest, France) for NMR technical expertise, Mrs. Catherine Boisvert and Catherine Doucet for their involvement in statistical analysis and Mrs. Maria Pacheco-Oliver (National Research Council, Montreal) for her presubmission review of this paper. Authors would also like to thank the Institute of Nutrition and Functional Foods (INAF, QC, Canada), the Natural Sciences and Engineering Research Council (NSERC) of Canada, Ministère de l’Agriculture, des Pêcheries et de l’Alimentation du Québec (MAPAQ), and the University of Québec at Rimouski (Rimouski, QC, Canada) for their financial support.


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Copyright information

© Springer Science+Business Media Dordrecht 2014

Authors and Affiliations

  • Stéphanie Bondu
    • 1
    • 2
  • Claudie Bonnet
    • 1
    • 2
  • Julie Gaubert
    • 1
    • 2
  • Éric Deslandes
    • 3
  • Sylvie L. Turgeon
    • 1
  • Lucie Beaulieu
    • 1
    • 2
    Email author
  1. 1.Département des sciences des aliments et de nutrition, Institut sur la nutrition et les aliments fonctionnels (INAF)Université LavalQuébecCanada
  2. 2.Collectif de Recherche Appliquée aux Bioprocédés et à la chimie de l’Environnement (CRABE)Université du Québec à RimouskiRimouskiCanada
  3. 3.Laboratoire des Sciences de l’Environnement Marin (LEMAR), Institut Universitaire Européen de la MerUniversité de Bretagne Occidentale, Technopôle de Brest-IroisePlouzanéFrance

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