Journal of Applied Phycology

, Volume 28, Issue 5, pp 3101–3115 | Cite as

Evaluation of the in vitro biological activity of protein hydrolysates of the edible red alga, Palmaria palmata (dulse) harvested from the Gaspe coast and cultivated in tanks

  • Lucie BeaulieuEmail author
  • Maude Sirois
  • Éric Tamigneaux


The in vitro antioxidant activity and angiotensin-converting enzyme (ACE) inhibition potential of protein extracts of Palmaria palmata (dulse) red seaweed from the Gaspé coast (QC, Canada) were investigated. The effects of the algae geographical origin and cultivation conditions were also studied. The highest activity values were displayed by <10 kDa protein fraction hydrolyzed by chymotrypsin (HF) at concentrations as low as 0.1 to 5 mg mL−1. Algae harvested in Pabos (QC, Canada) were more biologically active than algae from Newport (QC, Canada), demonstrating, respectively, 67.74 ± 3.9 % and 49.10 ± 10.9 % ACE inhibition at a concentration of 5 mg mL−1. Algae collected in Newport, cultivated in tanks with or without nutrients (NC+, NC−), scored higher biological activity than algae grown in Newport (N) waters. The <10 kDa HF obtained from the algae batch NC+ demonstrated, through an oxygen radical absorbance capacity assay, the highest antioxidant value: 440.73 ± 43.21 μmol trolox equivalents g−1 at 0.4 mg mL−1. Using liquid chromatography-tandem mass spectrometry (LC-MS/MS) with database searches, protein precursors such as protein ribulose-1,5-biphosphate carboxylase/oxygenase (RuBiSCo) enzyme, allophycocyanins and phycocyanins, were identified in <10 kDa HF (P, N, NC−, NC+). Results show that protein hydrolysates from P. palmata demonstrate a high potential as healthy and functional ingredients and identified proteins could be targeted for crop improvement.


Rhodophyta Palmaria palmata Environmental growth conditions Hydrolysates ACE inhibitory activity Antioxidant capacity Peptide amino acid identification 



The authors wish to thank the Proteomics platform of the Eastern Quebec Genomics Centre (Quebec, Canada) for the mass spectrometry experiments and database searching, Mrs. Roxanne Brion-Roby, Mataëlle Onapin and Jacinthe Thibodeau for their technical expertise, Mrs. Catherine Boisvert and Catherine Doucet for their involvement in statistical analysis, Emilie Gouhier and the technical staff from Merinov for assistance with seaweed sampling and cultivation, Mrs. Maria Pacheco-Oliver (National Research Council, Montreal, Canada) and Claudie Bonnet (UQAR, Rimouski, Canada) for their involvement in the presubmission review of this paper. Authors would also like to thank the Institute of Nutrition and Functional Foods (INAF, QC, Canada), the Ministère de l’Agriculture, des Pêcheries et de l’Alimentation du Québec (MAPAQ), the University of Québec at Rimouski (Rimouski, QC, Canada), Fonds de recherche du Québec-Nature et technologies (FRQNT), Ministère de l’Éducation, de l’Enseignement supérieur et de la Recherche (MEESR) and the NSERC Industrial Research Chair Grants for their financial support.


  1. Abd El Baky H, El Baroty GS, Ibrahem EA (2015) Functional characters evaluation of biscuits sublimated with pure phycocyanin isolated from Spirulina and Spirulina biomass. Nutr Hosp 32:231–241PubMedGoogle Scholar
  2. Beaulieu L, Bondu S, Doiron K, Turgeon SL (2015) Isolation and characterization of antibacterial peptides from protein hydrolysates of the macroalgae Saccharina longicruris. J Funct Foods 17:685–697CrossRefGoogle Scholar
  3. Bixler HJ, Porse H (2011) A decade of change in the seaweed hydrocolloids industry. J Appl Phycol 23:321–335CrossRefGoogle Scholar
  4. Bligh EG, Dyer WJ (1959) A rapid method of total lipid extraction and purification. Can J Biochem Physiol 37:911–917Google Scholar
  5. Boisvert C, Beaulieu L, Bonnet C, Pelletier E (2015) Assessment of the antioxidant and antibacterial activities of three species of edible seaweeds. J Food Biochem 39:377–387CrossRefGoogle Scholar
  6. Bondu S, Bonnet C, Gaubert J, Deslandes E, Turgeon SL, Beaulieu L (2014) Bioassay-guided fractionation approach for determination of protein precursors of proteolytic bioactive metabolites from macroalgae. J Appl Phycol 27:2059–2074CrossRefGoogle Scholar
  7. Brand-Williams W, Cuvelier ME, Berset C (1995) Use of a free radical method to evaluate antioxidant activity. Lebensm Wiss Technol 28:25–30Google Scholar
  8. Cao G, Prior RL (1998) Comparison of different analytical methods for assessing total antioxidant capacity of human serum. Clin Chem 44:1309–1315Google Scholar
  9. Cao G, Alessio HM, Cutler RG (1993) Oxygen-radical absorbance capacity assay for antioxidants. Free Rad Biol Med 14:303–11Google Scholar
  10. Chopin T (2015) Marine aquaculture in Canada: well-established monocultures of finfish and shellfish and an emerging Integrated Multi-Trophic Aquaculture (IMTA) approach including seaweeds, other invertebrates, and microbial communities. Fisheries 40:28–31CrossRefGoogle Scholar
  11. Cian RE, Martinez-Augustin O, Drago SR (2012) Bioactive properties of peptides obtained by enzymatic hydrolysis from protein byproducts of Porphyra columbina. Food Res Int 49:364–372CrossRefGoogle Scholar
  12. Cian RE, Alaiz M, Vioque J, Drago SR (2013) Enzyme proteolysis enhanced extraction of ACE inhibitory and antioxidant compounds (peptides and polyphenols) from Porphyra columbina residual cake. J Appl Phycol 25:1197–1206CrossRefGoogle Scholar
  13. Cian RE, Garz AG, Betancur Ancona D, Chel Guerrero L, Drago SR (2015) Hydrolyzates from Pyropia columbina seaweed have antiplatelet aggregation, antioxidant and ACE I inhibitory peptides which maintain bioactivity after simulated gastrointestinal digestion. LWT - Food Sci Technol 64:881–888CrossRefGoogle Scholar
  14. Corey P, Kim JK, Duston J, Garbary DJ, Prithiviraj (2013) Bioremediation of Palmaria palmata and Chon ‐ drus crispus (Basin Head): effect of nitrate and ammonium ratio as nitrogen source on nutrient removal. J Appl Phycol 25:1349–1358Google Scholar
  15. Dattolo E, Ruocco M, Brunet C, Lorenti M, Lauritano C, D’Esposito D, De Luca P, Sanges R, Mazzuca S, Procaccini G (2014) Response of the seagrass Posidonia oceanica to different light environments: Insights from a combined molecular and photo-physiological study. Mar Env Res 101:225–236CrossRefGoogle Scholar
  16. Dudonné S, Vitrac X, Coutière P, Woillez M, Mérillon JM (2009) Comparative study of antioxidant properties and total phenolic content of 30 plant extracts of industrial interest using DPPH, ABTS, FRAP, SOD, and ORAC assays. J Agric Food Chem 57:1768–1774CrossRefPubMedGoogle Scholar
  17. Edwards MD, Dring MJ (2011) Open-sea cultivation trial of the red alga. Palmaria palmata from seeded tetraspores in Strangford Lough, Northern Ireland. Aquaculture 317:203–209CrossRefGoogle Scholar
  18. Faes VA, Viejo RM (2003) Structure and dynamics of a Palmaria palmata (Rhodophyta) population in northern Spain. J Phycol 39:1038–1049CrossRefGoogle Scholar
  19. FAO (2014) The state of world fisheries and aquaculture. FAO Fisheries and Aquaculture Department, Rome, p 243Google Scholar
  20. Fitzgerald C, Gallagher E, Tasdemir D, Hayes M (2011) Heart health peptides from macroalgae and their potential use in functional foods. J Agr Food Chem 59:6829–6836CrossRefGoogle Scholar
  21. Fitzgerald C, Mora-Soler L, Gallagher E, O’Connor P, Prieto J, Soler-Vila A, Hayes M (2012) Isolation and characterization of bioactive pro-peptides with in vitro renin inhibitory activities from the macroalga Palmaria palmata. J Agric Food Chem 60:7421–7427CrossRefPubMedGoogle Scholar
  22. Fitzgerald C, Gallagher E, O’Connor P, Prieto J, Mora-Soler L, Grealy M, Hayes M (2013) Development of a seaweed derived platelet activating factor acetylhydrolase (PAF-AH) inhibitory hydrolysate, synthesis of inhibitory peptides and assessment of their toxicity using the zebrafish larvae assay. Peptides 50:119–124CrossRefPubMedGoogle Scholar
  23. Fitzgerald C, Aluko RE, Hossain M, Rai DK, Hayes M (2014a) Potential of a renin inhibitory peptide from the red seaweed Palmaria palmata as a functional food ingredient following confirmation and characterization of a hypotensive effect in spontaneously hypertensive rats. J Agric Food Chem 62:8352–8356CrossRefPubMedGoogle Scholar
  24. Fitzgerald C, Gallagher E, Doran L, Auty M, Prieto J, Hayes M (2014b) Increasing the health benefits of bread: assessment of the physical and sensory qualities of bread formulated using a renin inhibitory Palmaria palmata protein hydrolysate. LWT - Food Sci Technol 56:398–405CrossRefGoogle Scholar
  25. Fleurence J (2004) Seaweed proteins. In: Yada RY (ed) Proteins in food processing. Woodhead Publishing, Cambridge, UK, pp 197–213CrossRefGoogle Scholar
  26. Galland-Irmouli AV, Fleurence J, Lamghari R, Luçon L, Rouxel C, Barbaroux O, Bronowicki JP, Villaume C, Guéant JL (1999) Nutritional value of proteins from edible seaweed Palmaria palmata (Dulse). J Nutr Biochem 10:353–359CrossRefPubMedGoogle Scholar
  27. Garbary DJ, Beveridge LF, Flynn AD, White KL (2012) Population ecology of Palmaria palmata (Palmariales, Rhodophyta) from harvested and non-harvested shores on Digby Neck, Nova Scotia, Canada. Algae 27:33–42CrossRefGoogle Scholar
  28. Harnedy PA, FitzGerald RJ (2011) Bioactive proteins, peptides, and amino acids from macroalgae. J Phycol 47:218–232CrossRefPubMedGoogle Scholar
  29. Harnedy PA, FitzGerald RJ (2013) In vitro assessment of the cardioprotective, anti-diabetic and antioxidant potential of Palmaria palmata protein hydrolysates. J Appl Phycol 25:1793–1803CrossRefGoogle Scholar
  30. Harnedy PA, Soler-Vila A, Edwards MD, FitzGerald RJ (2014) The effect of time and origin of harvest on the in vitro biological activity of Palmaria palmata protein hydrolysates. Food Res Int 62:746–752CrossRefGoogle Scholar
  31. Harnedy PA, O’Keeffe MB, FitzGerald RJ (2015) Purification and identification of dipeptidyl peptidase (DPP) IV inhibitory peptides from the macroalga Palmaria palmata. Food Chem 172:400–406CrossRefPubMedGoogle Scholar
  32. Hayakari M, Kondo Y, Izumi H (1978) A rapid and simple spectrophotometric assay of Angiotensinconverting enzyme. Anal Biochem 84:361–369Google Scholar
  33. Holdt SL, Kraan S (2011) Bioactive compounds in seaweed: functional food applications and legislation. J Appl Phycol 23:543–597CrossRefGoogle Scholar
  34. Jueterbock A, Tyberghein L, Verbruggen H, Coyer JA, Olsen JL, Hoarau G (2013) Climate change impact on seaweed meadow distribution in the North Atlantic rocky intertidal. Ecol Evol 3:1356–1373CrossRefPubMedPubMedCentralGoogle Scholar
  35. Keller A, Nesvizhskii AI, Kolker E (2002) Empirical statistical model to estimate the accuracy of peptide identifications made by MS/MS and database search. Anal Chem 74:5383–5392CrossRefPubMedGoogle Scholar
  36. Kelman D, Kromkowski Posner E, McDermid KJ, Tabandera NK, Wright PR, Wright AD (2012) Antioxidant activity of Hawaiian marine algae. Mar Drugs 10:403–416Google Scholar
  37. Kim KT, Rioux LE, Turgeon SL (2014) Alpha-amylase and alpha-glucosidase inhibition is differentially modulated by fucoidan obtained from Fucus vesiculosus and Ascophyllum nodosum. Phytochemistry 98:27–33CrossRefPubMedGoogle Scholar
  38. Licois A, Ngom MC, Hersant G, Corcuff R, Couture F, Tamigneaux E (2014) Projet PRÉFAB – Étude de la filière intégrée de laminaire au Québec. Merinov Rapport de Recherche-Développement no 14–06. 38 pGoogle Scholar
  39. Litzer S (2012) L’algoculture et ses contraintes géographiques en Chine. EchoGéo. doi: 10.4000/echogeo.12913 Google Scholar
  40. Lourenço SO, Barbarino E, De-Paula JC, da S Pereira LO, Lanfer Marquez UM (2002) Amino acid composition, protein content and calculation of nitrogen-to-protein conversion factors for 19 tropical seaweeds. Phycol Res 50:233–241Google Scholar
  41. Lu Y, Li Y, Yang Q, Zhang Z, Chen Y, Zhang S, Peng XX (2014) Suppression of glycolate oxidase causes glyoxylate accumulation that inhibits photosynthesis through deactivating Rubisco in rice. Physiol Plant 150:463–476CrossRefPubMedGoogle Scholar
  42. Marcus Y, Gurevitz M (2000) Activation of cyanobacterial RuBP-carboxylase/oxygenase is facilitated by inorganic phosphate via two independent mechanisms. Eur J Biochem 267:5995–6003CrossRefPubMedGoogle Scholar
  43. Marinho GS, Holdt SL, Angelidaki I (2015) Seasonal variations in the amino acid profile and protein nutritional value of Saccharina latissima cultivated in a commercial IMTA system. J Appl Phycol 25:1991–2000CrossRefGoogle Scholar
  44. Martinez B, Rico JM (2002) Seasonal variation of P content and major N pools in Palmaria palmata (Rhodophyta). J Phycol 38:1082–1089CrossRefGoogle Scholar
  45. Mishra K, Ojha H, Chaudhury NK (2012) Estimation of antiradical properties of antioxidants using DPPH•assay: a critical review and results. Food Chem 130:1036–1043Google Scholar
  46. Morgan KC, Wright JLC, Simpson FJ (1980) Review of chemical constituents of the red alga Palmaria palmata (Dulse). Econ Bot 34:27–50CrossRefGoogle Scholar
  47. Murthy MS, Radia P (1978) Eco-biochemical studies on some economical important intertidal algae from Port Okha (India). Bot Mar 24:417–422Google Scholar
  48. Nesvizhskii AI, Keller A, Kolker E, Aebersold R (2003) A statistical model for identifying proteins by tandem mass spectrometry. Anal Chem 75:4646–4658CrossRefPubMedGoogle Scholar
  49. Ngo DH, Vo TS, Ngo DN, Wijesekara I, Kim SK (2012) Biological activities and potential health benefits of bioactive peptides derived from marine organisms. Int J Biol Macromol 51:378–383CrossRefPubMedGoogle Scholar
  50. Pang S, Lüning K (2004) Tank cultivation of the red alga Palmaria palmata: effects of intermittent light on growth rate, yield and growth kinetics. J Appl Phycol 16:93–99CrossRefGoogle Scholar
  51. Pangestuti R, Kim SK (2011) Biological activities and health benefit effects of natural pigments derived from marine algae. J Funct Foods 3:255–266CrossRefGoogle Scholar
  52. Parjikolaei BR, Kloster L, Bruhn A, Rasmussen MB, Fretté XC, Christensen KV (2013) Effect of light quality and nitrogen availability on the biomass production and pigment content of Palmaria palmata (Rhodophyta). Chem Eng Trans 32:967–972Google Scholar
  53. Parry MAJ, Andralojc PJ, Scales JC, Salvucci ME, Carmo-Silva AE, Alonso H, Whitney SM (2013) Rubisco activity and regulation as targets for crop improvement. J Exp Bot 64:717–730CrossRefPubMedGoogle Scholar
  54. Perfeto PNM (1998) Relation between chemical composition of Grateloupia doryphore (Montagne) Howe, Gymnogongrus griffithsiae (Turner) Martius, and abiotic parameters. Acta Bot Bras 12:77–88Google Scholar
  55. Peteiro C, Freire Ó (2013) Biomass yield and morphological features of the seaweed Saccharina latissima cultivated at two different sites in a coastal bay in the Atlantic coast of Spain. J Appl Phycol 25:205–213CrossRefGoogle Scholar
  56. Rioux LE, Turgeon SL, Beaulieu M (2009) Effect of season on the composition of bioactive polysaccharides from the brown seaweed Saccharina longicruris. Phytochemistry 70:1069–1075CrossRefPubMedGoogle Scholar
  57. Shilov IV, Seymour SL, Patel AA, Loboda A, Tang WH, Keating SP, Hunter CL, Nuwaysir LM, Schaeffer DA (2007) The Paragon algorithm, a next generation search engine that uses sequence temperature values and feature probabilities to identify peptides from tandem mass spectra. Mol Cell Proteomics 6:1638–1655CrossRefPubMedGoogle Scholar
  58. Simpson BK, Haard NF (1985) The use of proteolytic enzymes to extract carotenoproteins from shrimp wastes (Vol. 7). Academic Press, San DiegoGoogle Scholar
  59. Stengel DB, Connan S, Popper ZA (2011) Algal chemodiversity and bioactivity: sources of natural variability and implications for commercial application. Biotech Adv 29:483–501CrossRefGoogle Scholar
  60. Suetsuna K (1998a) Purification and identification of angiotensin I converting enzyme inhibitors from the red alga Porphyra yezoensis. J Mar Biotech 6:163–167Google Scholar
  61. Suetsuna K (1998b) Separation and identification of angiotensin I converting enzyme inhibitory peptides from peptic digest of Hizikia fusiformis protein. Nippon Suisan Gakkaishi 64:862–866CrossRefGoogle Scholar
  62. Suetsuna K, Nakano T (2000) Identification of an antihypertensive peptide from peptic digest of wakame (Undaria pinnatifida). J Nutr Biochem 11:450–454CrossRefPubMedGoogle Scholar
  63. Suetsuna K, Maekawa K, Chen JR (2004) Antihypertensive effects of Undaria pinnatifida (wakame) peptide on blood pressure in spontaneously hypertensive rats. J Nutr Biochem 15:267–272CrossRefPubMedGoogle Scholar
  64. Tamigneaux E, Pedneault E, Gendron L (2014) Comparaison des rendements de l’algue brune Saccharina longicruris cultivée en milieu ouvert en Gaspésie et en lagune aux Îles-de-la-Madeleine. Merinov Rapport de Recherche-Développement no 14–04. 35 pGoogle Scholar
  65. TASTE project 2012.
  66. Udenigwe CC, Gong M, Wu S (2013) In silico analysis of the large and small subunits of cereal RuBisCO as precursors of cryptic bioactive peptides. Process Biochem 48:1794–1799CrossRefGoogle Scholar
  67. Van der Meer JP, Todd ER (1980) The life history of Palmaria palmata in culture. A new type for the Rhodophyta. Can J Bot 58:1250–1256CrossRefGoogle Scholar
  68. Wang T, Jónsdóttir R, Kristinsson HG, Hreggvidsson GO, Jónsson JO, Thorkelsson G, Ólafsdóttir G (2010) Enzyme-enhanced extraction of antioxidant ingredients from red algae Palmaria palmata. LWT - Food Sci Technol 43:1387–1393CrossRefGoogle Scholar
  69. Winberg P (2011) Scaling up for new opportunities in the practical use of algae (Applied Phycology), RIRDC Publication No. 11/174 RIRDC Project No. PRJ-007249. RIRDC, Canberra, AustraliaGoogle Scholar
  70. Yang Y, Marczak ED, Yokoo M, Usui H, Yoshikawa M (2003) Isolation and antihypertensive effect of angiotensin I-converting enzyme (ACE) inhibitory peptides from spinach Rubisco. J Agric Food Chem 51:4897–4902CrossRefPubMedGoogle Scholar
  71. Yuan YV, Walsh NA (2006) Antioxidant and antiproliferative activities of extracts from a variety of edible seaweeds. Food Chem Toxicol 44:1144–1150CrossRefPubMedGoogle Scholar
  72. Yuan YV, Bone DE, Carrington MF (2005) Antioxidant activity of dulse (Palmaria palmata) extract evaluated in vitro. Food Chem 91:485–494CrossRefGoogle Scholar
  73. Yuan YV, Westcott ND, Hu C, Kitts DD (2009) Mycosporine-like amino acid composition of the edible red alga, Palmaria palmata (dulse) harvested from the west and east coasts of Grand Manan Island, New Brunswick. Food Chem 112:321–328CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2016

Authors and Affiliations

  • Lucie Beaulieu
    • 1
    • 2
    Email author
  • Maude Sirois
    • 1
    • 2
  • Éric Tamigneaux
    • 3
  1. 1.Institut sur la nutrition et les aliments fonctionnels (INAF), Département des sciences des alimentsUniversité Laval, 2425 rue de l’AgricultureQuébecCanada
  2. 2.Collectif de Recherche Appliquée aux Bioprocédés et à la chimie de l’Environnement CRABEUniversité du Québec à RimouskiRimouskiCanada
  3. 3.École des pêches et de l’aquaculture du Québec, Cégep de la Gaspésie et des IlesQuebec Fisheries and Aquaculture Innovation CentreGrande-RivièreCanada

Personalised recommendations