Plant Foods for Human Nutrition

, Volume 73, Issue 2, pp 101–107 | Cite as

Peptides from Chia Present Antibacterial Activity and Inhibit Cholesterol Synthesis

  • Michele Silveira Coelho
  • Rosana Aparecida Manólio Soares-Freitas
  • José Alfredo Gomes Arêas
  • Eliezer Avila Gandra
  • Myriam de las Mercedes Salas-Mellado
Original Paper


In previous studies, it has not been reported that protein isolated from chia interferes favorably with antibacterial activity, and reduces cholesterol synthesis. The objective of this study was to determine whether commonly used commercial microbial proteases can be utilized to generate chia protein-based antibacterial and hypocholesterolemic hydrolysates/peptides, considering the effects of protein extraction method. Alcalase, Flavourzyme and sequential Alcalase-Flavourzyme were used to produce hydrolysates from chia protein (CF), protein-rich fraction (PRF) and chia protein concentrates (CPC1 and CPC2). These hydrolysates were evaluated for their antimicrobial activity against Gram-positive (G+) and Gram-negative (G) microorganisms. The protein hydrolysates were purified by ultrafiltration through a membrane with 3 kDa nominal molecular weight, for evaluation of hypocholesterolemic activity. An inhibition zone was observed when the hydrolysate was tested against S. aureus, and minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) values were obtained. Peptides from chia protein with molecular mass lower than 3 kDa reduced up to 80.7% of 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMG-CoA reductase) enzymatic reaction velocity. It was also observed that, independent of the method used to obtain chia proteins, the fractions showed relevant bioactivity. Moreover, the intensity of the bioactivity varied with the method for obtaining the protein and with the enzyme used in the hydrolysis process. This is the first report to demonstrate that chia peptides are able to inhibit cholesterol homeostasis.


Antibacterial By-product Hypocholesterolemic Bioactive peptides Salvia hispanica





Analysis of variance


Brain heart infusion agar


Brain heart infusion


Partially defatted chia flour


Chia protein concentrate 1


Chia protein concentrate 2


Degree of hydrolysis







HMG-CoA reductase

3-hydroxy-3-methylglutaryl coenzyme A reductase


Minimal bactericidal concentration


Minimal inhibitory concentration


Molecular weight


Nominal molecular weight limit


Protein-rich fraction


Sequential Alcalase-Flavourzyme


Very low density lipoprotein.


Compliance with Ethical Standards

Conflict of Interest

The authors declare that they have no conflict of interest.

Supplementary material

11130_2018_668_MOESM1_ESM.pdf (356 kb)
ESM 1 (PDF 356 kb)
11130_2018_668_MOESM2_ESM.pdf (123 kb)
ESM 2 (PDF 123 kb)
11130_2018_668_MOESM3_ESM.pdf (238 kb)
ESM 3 (PDF 237 kb)


  1. 1.
    Grundy SM, Veja GL (1988) Plasma cholesterol responsiveness to saturated fatty acids. Am J Clin Nutr 47:822–824CrossRefPubMedGoogle Scholar
  2. 2.
    Rader DJ (2003) Regulation of reverse cholesterol transport and clinical implications. Am J Cardiol 92:42–49CrossRefGoogle Scholar
  3. 3.
    Pak VV, Koo MS, Kasymova TD, Kwon DY (2005) Isolation and identification of peptides from soy 11S-globulin with hypocholesterolemic activity. Chem Nat Comp 41(6):710–714CrossRefGoogle Scholar
  4. 4.
    Marques MR, Freitas RAMS, Carlos ACCC, Siguemoto ES, Fontanari GG, Arêas JAG (2015) Peptides from cowpea present antioxidant activity, inhibit cholesterol synthesis and its solubilisation into micelles. Food Chem 168:288–293CrossRefPubMedGoogle Scholar
  5. 5.
    Soares RAM, Mendonça S, de Castro LIA, Menezes ACCCC, Arêas JAG (2015) Major peptides from amaranth (Amaranthus cruentus) protein inhibit HMG-CoA reductase activity. Int J Mol Sci 16(2):4150–4160CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Osman A, Goda HA, Abdel-Hamid M, Badran SM, Otte J (2016) Antibacterial peptides generated by alcalase hydrolysis of goat whey. LWT - Food Sci Technol 65:480–486CrossRefGoogle Scholar
  7. 7.
    Abdel-Hamid M, Goda HA, De Gobba C, Jenssen H, Osman A (2016) Antibacterial activity of papain hydrolysed camel whey and its fractions. Int Dairy J 61:91–98CrossRefGoogle Scholar
  8. 8.
    Hwang CF, Chen YA, Luo C, Chiang WD (2016) Antioxidant and antibacterial activities of peptide fractions from flaxseed protein hydrolysed by protease from Bacillus altitudinis HK02. Int J Food Sci Technol 51:681–689CrossRefGoogle Scholar
  9. 9.
    Segura-Campos MR, Salazar-Vega IM, Chel-Guerrero LA, Betancur-Ancona DA (2013) Biological potential of chia (Salvia hispanica L.) protein hydrolysates and their incorporation into functional foods. LWT - Food Sci Technol 50:723–731CrossRefGoogle Scholar
  10. 10.
    Coelho MS, Salas-Mellado MM (2016) Biological properties of chia (Salvia hispanica L.) proteins. In: Betancur-Ancona D, Segura-Campos MR (Eds.). Salvia Hispanica L.: properties, applications and health. Nova Publishers, New York, pp 1–345Google Scholar
  11. 11.
    Ambigaipalan P, Al-Khalifa AS, Shahidi F (2015) Antioxidant and angiotensin I converting enzyme (ACE) inhibitory activities of date seed protein hydrolysates prepared using Alcalase, Flavourzyme and Thermolysin. J Funct Foods 18:1125–1137CrossRefGoogle Scholar
  12. 12.
    Betancur-Ancona D, Sosa-Espinoza T, Ruiz-Ruiz J, Segura-Campos M, Chel-Guerrero L (2014) Enzymatic hydrolysis of hard-to-cook bean (Phaseolus vulgaris L.) protein concentrates and its effects on biological and functional properties. Int J Food Sci Technol 49:2–8CrossRefGoogle Scholar
  13. 13.
    Pedroche J, Yust MM, Girón-Calle J, Alaiz M, Millán F, Vioque J (2002) Utilization of chickpea protein isolates for production of peptides with angiotensin I-converting enzyme (ACE) inhibitory activity. J Sci Food Agric 82:960–965CrossRefGoogle Scholar
  14. 14.
    Haney EF, Petersen AP, Lau CK, Jing W, Storey DG, Vogel HJ (2013) Mechanism of action of puroindoline derived tryptophan-rich antimicrobial peptides. Biochim Biophys Acta 1828(8):1802–1181CrossRefPubMedGoogle Scholar
  15. 15.
    Adler-Nissen J (1986) Enzymatic hydrolysis of food proteins. Elsevier Applied Science Publishers Ltd., New YorkGoogle Scholar
  16. 16.
    Zhao Q, Xiong H, Selomulya C, Chen XD, Zhong H, Sun W, Zhou Q, Wang S (2012) Enzymatic hydrolysis of rice dreg protein: effects of enzyme type on the functional properties and antioxidant activities of recovered proteins. Food Chem 134:1360–1367CrossRefPubMedGoogle Scholar
  17. 17.
    García-Moreno PJ, Batista I, Pires C, Bandarra NM, Espejo-Carpio FJ, Guadix A, Guadix EM (2014) Antioxidant activity of protein hydrolysates obtained from discarded Mediterranean fish species. Food Res Int 65:469–476CrossRefGoogle Scholar
  18. 18.
    Shi Y, Kovacs-Nolan J, Jiang B, Tsao R, Mine Y (2014) Antioxidant activity of enzymatic hydrolysates from eggshell membrane proteins and its protective capacity in human intestinal epithelial Caco-2 cells. J Funct Foods 10:35–45CrossRefGoogle Scholar
  19. 19.
    Ketnawa S, Benjakul S, Martínez-Alvarez O, Rawdkuen S (2017) Fish skin gelatin hydrolysates produced by visceral peptidase and bovine trypsin: bioactivity and stability. Food Chem 215:383–390CrossRefPubMedGoogle Scholar
  20. 20.
    Sun Y, Chang R, Li Q, Li B (2016) Isolation and characterization of an antibacterial peptide from protein hydrolysates of Spirulina platensis. Eur Food Res Technol 242:685–692CrossRefGoogle Scholar
  21. 21.
    Nazzaro F, Fratianni F, De Martino L, Copolla R, de Feo V (2013) Effect of essential oils on pathogenic bacteria. Pharmaceuticals 6(12):1451–1474CrossRefPubMedPubMedCentralGoogle Scholar
  22. 22.
    Sila A, Nedjar-Arroume N, Hedhili K, Chataigné G, Balti R, Nasri M, Dhulster B, Bougatef A (2014) Antibacterial peptides from barbel muscle protein hydrolysates: activity against some pathogenic bacteria. LWT – Food Sci Technol 55(1):183–188CrossRefGoogle Scholar
  23. 23.
    Wald M, Schwarz K, Rehbein H, Bußmann B, Beermann C (2016) Detection of antibacterial activity of an enzymatic hydrolysate generated by processing rainbow trout by-products with trout pepsin. Food Chem 205:221–228CrossRefPubMedGoogle Scholar
  24. 24.
    Yokota S, Fujii N (2007) Contributions of the lipopolysaccharide outer core oligosaccharide region on the cell surface properties of Pseudomonas aeruginosa. Comp Immunol Microbiol Infect Dis 30:97–109CrossRefPubMedGoogle Scholar
  25. 25.
    Pak VV, Koo M, Kwon DY, Shakhidoyatov KM, Yun L (2010) Peptide fragmentation as an approach in modeling of an active peptide and designing a competitive inhibitory peptide for HMG-CoA reductase. Bioorganic Medic Chem 18(12):4300–4309CrossRefGoogle Scholar
  26. 26.
    Hammershøj M, Nebel C, Carstens JH (2008) Enzymatic hydrolysis of ovomucin and effect on foaming properties. Food Res Int 41:522–531CrossRefGoogle Scholar
  27. 27.
    Pak VV, Koo M, Lee N, Kim MS, Kwon DY (2005) Structure-activity relationships of the peptide Ile-Ala-Val-Pro and its derivatives revealed using the semi-empirical AM1 method. Chem Nat Compd 41(4):454–460CrossRefGoogle Scholar
  28. 28.
    Sigma quality control test procedure (1999) Product information, SSCASE01.001Google Scholar
  29. 29.
    Otto T, Baik B, Czuchajowska Z (1997) Wet fractionation of garbanzo bean and pea flours. Cereal Chem 74:141–146CrossRefGoogle Scholar
  30. 30.
    Nolsoe H, Undeland I (2009) The acid and alkaline solubilization process for the isolation of muscle proteins: state of the art. Food Bioprocess Technol 2:1–27CrossRefGoogle Scholar
  31. 31.
    Ragab DM, Babiker EE, Eltinay AH (2004) Fractionation, solubility and functional properties of cowpea (Vigna unguiculata) proteins as affected by pH and/or salt concentration. Food Chem 84:207–212CrossRefGoogle Scholar
  32. 32.
    Nielsen PM, Petersen D, Dambmann C (2001) Improved method for determining food protein degree of hydrolysis. J Food Sci 66(5):642–646CrossRefGoogle Scholar
  33. 33.
    CLSI (2012) M02-A11: Performance standards for antimicrobial disk susceptibility tests; approved standard, 11th Ed. CLSI (Clinical and Laboratory Standards Institute), vol 32(1). CLSI, WayneGoogle Scholar
  34. 34.
    Valgas C, Machado de Souza S, Elza FA, Smânia EFA, Smânia AJ (2007) Screening methods to determine antibacterial activity of natural products. Braz J Microbiol 38:369–380CrossRefGoogle Scholar
  35. 35.
    CLSI (2012) M07-A9: Methods for dilution antimicrobial susceptibility tests for bacteria that grow aerobically; Aprpoved Standard, 9th Ed. CLSI (Clinical and Laboratory Standards Institute), vol 32(2). CLSI, WayneGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Michele Silveira Coelho
    • 1
  • Rosana Aparecida Manólio Soares-Freitas
    • 2
  • José Alfredo Gomes Arêas
    • 2
  • Eliezer Avila Gandra
    • 3
  • Myriam de las Mercedes Salas-Mellado
    • 1
  1. 1.Laboratory of Food Technology, School of Chemistry and FoodFederal University of Rio GrandeRio GrandeBrazil
  2. 2.Faculty of Public HealthUniversity of São PauloSão PauloBrazil
  3. 3.Center of Chemical, Pharmaceutical and Food SciencesFederal University of PelotasPelotasBrazil

Personalised recommendations