Production of Bioactive Peptides in Tomato Seed Protein Isolate Fermented by Water Kefir Culture: Optimization of the Fermentation Conditions

  • Manel MechmecheEmail author
  • Hamida Ksontini
  • Moktar Hamdi
  • Faten Kachouri


The aim of this work was to explore the use of protein isolate from tomato seed as a medium for the growth of water kefir mixture culture. Response surface methodology (RSM) and a central composite rotatable design were used to optimize the fermentation conditions of tomato seed isolate by water kefir mixture culture, including inoculum level, sucrose concentration and ascorbic acid concentrations, in order to improve cell growth and exopolysaccharides production. The models established showed that the values of R2 were high and the p-values < 0.0001 were also suitable for this experiment, which confirmed the effectiveness of these models. The evaluation of the radical scavenging activity of the isolate after 24 h of fermentation showed an improvement of about 74%. HPLC analysis showed a significant decrease of the concentration of total amino-acids exceeding 155%, especially for glutamic acid and aspartic acid. After 24 h of fermentation, the protein isolate contains respectively by about 41.27 and 20.29 mg/100 g of glutamic acid and aspartic acid. FTIR results showed that the fermentation favors the production of new amides and aromatic compounds. The results of these experiments indicated that RSM design is promising approach for the optimization of tomato isolate fermentation conditions, and that water kefir culture could degrade and convert the proteins into bioactive peptides that contribute positively in the improvement of antioxidant activity.


Tomato waste Water kefir culture Response surface methodology Bioactive peptides Antioxidant activity 


Compliance with Ethical Standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical Approval

This article does not contain any studies with human participants or animals performed by any of the authors.


  1. Chandrasekara A, Shahidi F (2012) Bioaccessibility and antioxidant potential of millet grain phenolics as affected by simulated in vitro digestion and microbial fermentation. J Funct Foods 4(1):226–237. CrossRefGoogle Scholar
  2. Corona O, Randazzo W, Alessandro M, Guarcello R, Nicola F, Erten H, Moschetti G, Settanni L (2016) Characterization of kefir-like beverages produced from vegetable juices. LWT Food Sci Technol 66:572–581. CrossRefGoogle Scholar
  3. Garcia-Mora P, Frias J, Peñas E, Zielinski H, Giménez-Bastida JA, Wiczkowski W, Zielinska D, Martínez-Villaluenga C (2015) Simultaneous release of peptides and phenolics with antioxidant, ACE-inhibitory and anti-inflammatory activities from pinto bean (Phaseolus vulgaris L. var. pinto) proteins by subtilisins. J Funct Foods 18:319–332. CrossRefGoogle Scholar
  4. Grobben GJ, Sikkema J, Smith MR, De Bont JAM (1995) Production of extracellular polysaccharides by Lactobacillus delbrueckii ssp. bulgaricus NCFB 2772 grown in a chemically defined medium. J Appl Bacteriol 79:103–107. CrossRefGoogle Scholar
  5. Hati S, Sakure A, Mandal S (2017) Synthesis and functional identification of oligopeptides derived from the α3/5-conotoxins. Int J Pept Res Ther 23:297. CrossRefGoogle Scholar
  6. Iskandar MM, Lands LC, Sabally K, Azadi B, Meehan B, Mawji N, Skinner CD, Kubow S (2015) High hydrostatic pressure pretreatment of whey protein isolates improves their digestibility and antioxidant capacity. Foods 4:184–207. CrossRefPubMedPubMedCentralGoogle Scholar
  7. Joglekar AM, May AT (1987) Product excellence through design of experiments. Cereal Foods World 32:857–868Google Scholar
  8. Juan MY, Chou CC (2010) Enhancement of antioxidant activity, total phenolic and flavonoid content of black soybeans by solid state fermentation with Bacillus subtilis BCRC 14715. Food Microbiol 27(5):586–591. CrossRefPubMedGoogle Scholar
  9. Kagkli DM, Corich V, Bovo B, Lante A, Giacomini A (2016) Antiradical and antimicrobial properties of fermented red chicory (Cichorium intybus L.) by-products. Ann Microbiol 66(4):1377–1386. CrossRefGoogle Scholar
  10. Kembhavi AA, Kulkarni A, Pant A (1993) Salt-tolerant and thermostable alkaline protease from Bacillus subtilis NCIM No. 64. Appl Biochem Biotechnol 38:83–92. CrossRefPubMedGoogle Scholar
  11. Kiosseoglou V, Theodorakis K, Doxastakis G (1989) The rheology of tomato seed protein isolate films at the corn oil-water interface. Colloid Polym Sci 267:834–838. CrossRefGoogle Scholar
  12. Kumar V, Sheoran P, Gupta A, Yadav J, Tiwari SK (2016) Antibacterial property of bacteriocin produced by Lactobacillus plantarum LD4 isolated from a fermented food. Ann Microbiol. CrossRefGoogle Scholar
  13. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of the bacteriophage T4. Nature 227:680–686. CrossRefPubMedPubMedCentralGoogle Scholar
  14. Lee JI, Song KY, Chon JW, Hyeon JY, Seo KH (2011) Inhibitory effect of polysaccharide from kefir grain on the infection of MA-104 cell by human rotavirus. Korean J Food Sci Anim Resour 31(1):81–85. CrossRefGoogle Scholar
  15. Li T, Wang X, Wang Y, Fan T, Xu Y, Fan Z (2017) Characterization of antimicrobial peptides isolated from the processing by-products of African catfish Clarias gariepinus. Int J Pept Res Ther 23:227. CrossRefGoogle Scholar
  16. Liu X, Li T, Liu B, Zhao H, Zhou F, Zhang B (2016) An external addition of soy protein isolate hydrolysate to sourdough as a new strategy to improve quality of Chinese steamed bread. J Food Qual 39(1):3–12. CrossRefGoogle Scholar
  17. Marazza JA, Nazareno MA, de Giori GS, Garro MS (2012) Enhancement of the antioxidant capacity of soymilk by fermentation with Lactobacillus rhamnosus. J Funct Foods 4(3):594–601. CrossRefGoogle Scholar
  18. Mechmeche M, Kachouri F, Chouaibi M, Ksontini H, Setti K, Hamdi M (2017a) Optimization of extraction parameters of protein isolate from tomato seed using response surface methodology. Food Anal Methods. 10(3):809–819. CrossRefGoogle Scholar
  19. Mechmeche M, Kachouri F, Yaghlane H, Ksontini H, Setti K, Hamdi M (2017b) Kinetic analysis and mathematical modeling of growth parameters of Lactobacillus plantarum in protein-rich isolates from tomato seed. Food Sci Technol Int 23(2):128–141. CrossRefPubMedGoogle Scholar
  20. Moayedi A, Hashemi M, Safari M (2016) Valorization of tomato waste proteins through production of antioxidant and antibacterial hydrolysates by proteolytic Bacillus subtilis: optimization of fermentation conditions. J Food Sci Technol 53(1):391–400. CrossRefPubMedGoogle Scholar
  21. Neuzil J, Gebicki JM, Stocker R (1993) Radical-induced chain oxidation of proteins and its inhibition by chain-breaking antioxidants. Biochem J 293:601–610CrossRefGoogle Scholar
  22. Piermaria JA, Pinotti A, Garcia MA, Abraham AG (2009) Films based on kefiran, an expopolysaccharide obtained from kefir grain: development and characterization. Food Hydrocoll 23:684–690CrossRefGoogle Scholar
  23. Qiao D, Hua B, Gan D, Sun Y, Ye H, Zeng X (2009) Extraction optimized by using response surface methodology, purification and preliminary characterization of polysaccharides from Hyriopsis cumingii. Carbohydr Polym 76:422–429. CrossRefGoogle Scholar
  24. Rathore S, Salmerón I, Pandiella SS (2012) Production of potentially probiotic beverages using single and mixed cereal substrates fermented with lactic acid bacteria cultures. Food Microbiol 30:239–244. CrossRefPubMedGoogle Scholar
  25. Re R, Pellegrini N, Proteggente A, Pannala A, Yang M, Rice-Evans C (1999) Antioxidant activity applying an improved ABTS radical cation decolorization assay. Free Radic Bio Med 26:1231–1237. CrossRefGoogle Scholar
  26. Rodrigues KL, Caputo LRG, Carvalho JCT, Evangelista J, Schneedorf JM (2005) Antimicrobial and healing activity of kefir and kefiran extract. Int J Antimicrob Agents 25(5):404–408. CrossRefPubMedGoogle Scholar
  27. Savadkoohi S, Farahnaky A (2012) Dynamic rheological and thermal study of the heat-induced gelation of tomato-seed proteins. J Food Eng 113:479–485. CrossRefGoogle Scholar
  28. Sogi DS, Garg SK, Bawa AS (2002) Functional properties of seed meals and protein concentrates from tomato-processing waste. J Food Sci 67:2997–3001. CrossRefGoogle Scholar
  29. Sun T, Ho CT (2005) Antioxidant activities of buckwheat extracts. Food Chem 90:743–749. CrossRefGoogle Scholar
  30. Tayuan C, Tannock GW, Rodtong S (2011) Growth and exopolysaccharide production by Weissella sp. from low-cost substitutes for sucrose. Afr J Microbiol Res 5(22):3693–3701. CrossRefGoogle Scholar
  31. Wouters D, Bernaert N, Anno N, Van Droogenbroeck B, De Loose M, Van Bockstaele E, De Vuyst L (2013) Application’ and validation of autochthonous lactic acid bacteria starter cultures for controlled leek fermentations and their influence on the antioxidant properties of leek. Int J Food Microbiol 165:121–133. CrossRefPubMedGoogle Scholar
  32. Xiao Y, Xing G, Rui X, Li W, Chen X, Jiang M, Dong M (2014) Enhancement of the antioxidant capacity of chickpeas by solid state fermentation with Cordyceps militaris SN-18. J Funct Foods 10:210–222. CrossRefGoogle Scholar

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© Springer Science+Business Media, LLC, part of Springer Nature 2017

Authors and Affiliations

  • Manel Mechmeche
    • 1
    • 2
    Email author
  • Hamida Ksontini
    • 1
    • 2
  • Moktar Hamdi
    • 1
    • 2
  • Faten Kachouri
    • 1
    • 2
  1. 1.Laboratory of Microbial Ecology and Technology (LETMI)National Institute of Applied Sciences and Technology (INSAT)TunisTunisia
  2. 2.Superior School of Food Industry at Tunis (ESIAT)TunisTunisia

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