Isolation, Purification and Characterization of Antimicrobial Peptides Produced from Saccharomyces boulardii

  • Alaa Kareem Naimah
  • Alaa Jabbar Abd Al-Manhel
  • Manar Jabbar Al-Shawi


Saccharomyces boulardii was used for antimicrobial peptides production. Separation process of produced antimicrobial peptides was conducted using ultrafiltration technique through dialysis membranes with porous 10 (MWCO) kDa. The inhibition activity was determined against four bacterial isolates. As a result, higher inhibition zone against Bacillus cereus were 26, 29 and 33 mm after adding 50, 75 and 100 µL of concentrated peptide, respectively. After that, peptide passed through the Sephadex G-50 column to achieve purified peptide using gel filtration. The high activity of purified peptide was confirmed based on the second peak reaching to 37 mm of bacterial inhibition zone while other peaks did not show any inhibition against tested bacteria. Some of the important characteristics of purified bioactive peptide were applied. Antimicrobial peptides stability was studied and found to be stable at pH range from 5 to 7 values studied in addition to its inhibition activity reached to 100%. Regarding thermal stability, it was observed that the peptide was fully activity at a both 60–80 °C for 30 min. Moreover, molecular weight of a peptide was identified using electrophoresis technique with SDS measured at 5792 Dalton.


Saccharomyces boulardii Antimicrobial peptides Ultrafiltration 


  1. Agyei D, Danquah MK (2011) Industrial-scale manufacturing of pharmaceutical-grade bioactive peptides. Biotechnol Adv 29(3):272–277CrossRefPubMedGoogle Scholar
  2. Ali MAE, Abdel-Fatah OM, Janson JC, Elshafei AM (2012) Antimicrobial potential of Saccharomyces boulardii extracts and fractions. J Appl Sci Res 8(8):4537–4543Google Scholar
  3. Bahar AA, Ren D (2013) Antimicrobial Peptides Pharmaceuticals. 6(12):1543–1575. doi: 10.3390/ph6121543 PubMedGoogle Scholar
  4. Berni Canani R, Cucchiara S, Cuomo R, Pace F, Papale F (2011) Saccharomyces boulardii: a summary of the evidence for gastroenterology clinical practice in adults and children. Eur Rev Med Pharmacol Sci 15(7):809–822PubMedGoogle Scholar
  5. Bhunia AK, Johnson MC, Ray B (1988) Purification, characterization and antimicrobial spectrum of a bacteriocin produced by Pediococcus acidilactici. J Appl Microbiol 65(4):261–268Google Scholar
  6. Biswas SR, Ray P, Johnson MC, Ray B (1991) Influence of growth conditions on the production of a bacteriocin pediocin AcH, by Pediococcus acidilactici H. Appl Environ Microbiol 57:1265-1 267Google Scholar
  7. Bussey H (1991) K1 killer toxin, a pore forming protein from yeast. Mol Microbiol 5(10):2339–2343CrossRefPubMedGoogle Scholar
  8. Buts J-P, Dekeyser N, Stilmant C, Delem E, Smets F, Sokal E (2006) Saccharomyces boulardii produces in rat small intestine a novel protein phosphatase that inhibits Escherichia coli endotoxin by dephosphorylation. Pediatric Res, 60:24–29. doi: 10.1203/01.pdr.0000220322.31940.29 CrossRefGoogle Scholar
  9. Buyuksirit T, Kuleasan H (2014) Antimicrobial agents produced by yeasts. Int J Biol Biomol Agric Food Biotechnol Eng 8(10):1114–1117Google Scholar
  10. Dean A, Voss D (1999) Design and analysis of experiments, Springer, New York, p 740CrossRefGoogle Scholar
  11. Demain AL, Phaff HJ, Kurtzman CP (1998) The industrial and agricultural significance of yeasts. In: Kurtzman CP, Fell JW (eds) The yeasts, a taxonomic study, 4th edn., Elsevier Science, AmsterdamGoogle Scholar
  12. Ge J, Sun Y, Xin X, Wang Y, Ping W (2016) Purification and partial characterization of a novel bacteriocin synthesized by Lactobacillus paracasei HD1-7 isolated from Chinese sauerkraut juice. Scientific reports, p 6Google Scholar
  13. Hatoum R, Labrie S, Fliss I (2012) Antimicrobial and probiotic properties of yeasts: from fundamental to novel applications. Front Microbiol 3:421. doi: 10.3389/fmicb.2012.00421 CrossRefPubMedPubMedCentralGoogle Scholar
  14. He R, Alashi A, Malomo SA, Girgih AT, Chao D, Ju X, Aluko RE (2013) Antihypertensive and free radical scavenging properties of enzymatic rapeseed protein hydrolysates. Food Chem 141:153–159. doi: 10.1016/j.foodchem.2013.02.087 CrossRefPubMedGoogle Scholar
  15. Hedstrom L (2002) Serine protease mechanism and specificity. Chem Rev 102:4501–4524. doi: 10.1021/cr000033x CrossRefPubMedGoogle Scholar
  16. Jin Z, Shinde PL, Yang YX, Choi JY, Yoon SY, Hahn T-W, Lim HT, Park YK, Hahm KS, Joo JW (2009) Use of refined potato (Solanum tuberosum L. cv. Gogu valley) protein as an alternative to antibiotics in weanling pigs. Livestock Sci 124:26–32. doi: 10.1016/j.livsci.2008.12.003 CrossRefGoogle Scholar
  17. Kelesidis T, Pothoulakis C (2012) Efficacy and safety of the probiotic Saccharomyces boulardii for the prevention and therapy of gastrointestinal disorders. Therapeutic Adv Gastroenterol 5(2):111–125. doi: 10.1177/1756283X11428502 CrossRefGoogle Scholar
  18. Lee JK, Gopal R, Seo CH, Cheong HS, Park YK (2012) Isolation and purification of a novel deca-antifungal peptide from potato (Solanum tuberosum L. cv. Jopung) against. Int J Mol Sci 13:4021–4032. doi: 10.3390/ijms13044021 CrossRefPubMedPubMedCentralGoogle Scholar
  19. Magaña MD, Segura-Campos M, Dávila-Ortiz G, Betancur-Ancona D, Chel-Guerrero L (2015) ACE-I inhibitory properties of hydrolysates from germinated and ungerminated Phaseolus lunatus proteins. Food Sci Technol (Campinas) 35(1):167–174CrossRefGoogle Scholar
  20. Minh NP (2015) Alcalase and protamex hydrolysis of bioactive peptides from soybean. Bull Environ Pharmacol Life Sci 4(7):132–143Google Scholar
  21. Murzyn A, Krasowska A, Stefanowicz P, Dziadkowiec D, Łukaszewicz M (2010) Capric acid secreted by S. boulardii inhibits C. albicans filamentous growth, adhesion and biofilm formation. PLoS ONE 5(8):e12050. doi: 10.1371/journal.pone.0012050 CrossRefPubMedPubMedCentralGoogle Scholar
  22. Niamah AK (2010) Production of pediocin like bacteriocin from a local isolate of Pediococcus acidilactici and using it as foods preservative. Ph.D. thesis, College of Agriculture, University of Basrah, 177p. doi: 10.13140/RG.2.2.31314.35529
  23. Niamah AK (2014) Determination, identification of bioactive compounds extracts from yellow banana peels and used in vitro as antimicrobial. Int J Phytomed 6(4):625–632Google Scholar
  24. Niamah AK (2017) Physicochemical and microbial characteristics of yogurt added with Saccharomyces boulardii. Curr Res Nutr Food Sci.
  25. Niamah AK, Al-Manhel AJA, Al-Shawi MJ (2017) Study of inhibitory spectrum of metabolic extract from Saccharomyces boulardii yeast against some food related bacteria. Pak J Food Sci 27(1):26–32Google Scholar
  26. Palfree RG, Bussey H (1979) Yeast killer toxin: purification and characterisation of the protein toxin from Saccharomyces cerevisiae. Eur J Biochem 93(3):487–493CrossRefPubMedGoogle Scholar
  27. Pfeiffer P, Radler F (1982) Purification and characterization of extracellular and intracellular killer toxins of Saccharomyces cerevisiae- strain 28. J Gen Microbiol 128:2699–2706Google Scholar
  28. Sah BNP (2016) Identification of bioactive peptides produced in synbiotic yogurt having anticancer properties. Ph.D. thesis, Victoria University, Melbourne, AustraliaGoogle Scholar
  29. Sah BNP, Vasiljevic T, McKechnie S, Donkor ON (2016) Antibacterial and antiproliferative peptides in synbiotic yogurt—Release and stability during refrigerated storage. J Dairy Sci 99(6):4233–4242. doi: 10.3168/jds.2015-10499 CrossRefPubMedGoogle Scholar
  30. Smith BJ (1984) SDS poly acrylamide gel electrophoresis of proteins In: Walker JM (ed) Methods in molecular biology, The Humana press, New York, pp 41–55Google Scholar
  31. Schved F, Lalazar A, Henis Y, Juven BJ (1993) Purification, partial characterization and plasmid-linkage of pediocin SJ-1, a bacteriocin produced by Pediococcus acidilactici. J Appl Microbiol 74(1):67–77Google Scholar
  32. Yazawa K, Numata K (2014) Recent advances in chemoenzymatic peptide syntheses. Molecules, 19(9):13755–13774. doi: 10.3390/molecules190913755 CrossRefPubMedGoogle Scholar
  33. Zaouche A, Loukil C, De Lagausie P, Peuchmaur M, Macry J, Fitoussi F, Bernasconi P, Bingen E, Cezard J (2000) Effects of oral Saccharomyces boulardii on bacterial overgrowth, translocation, and intestinal adaptation after small-bowel resection in rats. Scand J Gastroenterol 35:160–165. doi: 10.1080/003655200750024326 CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2017

Authors and Affiliations

  • Alaa Kareem Naimah
    • 1
  • Alaa Jabbar Abd Al-Manhel
    • 1
  • Manar Jabbar Al-Shawi
    • 1
  1. 1.Department of Food Science, Agriculture CollegeBasrah UniversityBasraIraq

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