Abstract
This study aims to identify antioxidant and antimicrobial peptides from sheep milk produced using Lactobacillus plantarum (KGL3A). It was inferred that antioxidative and antimicrobial activities increased with increasing incubation time, and antioxidative properties (ABTS assay, superoxide free radical & hydroxyl free radical scavenging activity were 34.5, 34.7, and 29.2% respectively) and antimicrobial properties against Escherichia coli, S. typhimurium, E. faecalis, & B. cereus were 11.3, 12.7, 13.3, & 12.3 mm. However, inoculation of culture at a level of 2.5% and 48 h fermentation give the highest proteolysis activities. Fermented sheep milk fractions of 3 & 10 kDa were analysed for antioxidative and antimicrobial activity, and the 10 kDa permeate showed the highest ABTS assay. The hydroxyl free radical scavenging activity was greatest in 10 kDa retentate and superoxide free radical scavenging activity was observed in 3 kDa permeate (34.7, 43.4, and 34.6%, respectively). Antimicrobial activity of 10 kDa retentate against B. cereus & E. coli (13.3 mm) was greater than 3 and 10 kDa retentate against S. typhimurium (13 mm) and 3 kDa retentate against E. faecalis (13.7 mm). The molecular weight of the protein was estimated using SDS-PAGE. On electrophoresis on a 2-D gel, 6 peptides were identified using RP-LC/MS. BIOPEP, a database for antioxidative and antimicrobial peptides, validated the antioxidative & antimicrobial activities of several peptides in sheep's milk that has been fermented. Sheep milk fermented using Lactobacillus could be considered a novel source of antioxidative and antimicrobial proteins.
Similar content being viewed by others
Data availability
All data generated or analysed during this study are included in this published article.
References
Bulet P, Cociancich S, Dimarcq JL, Lambert J, Reichhart JM, Hoffmann D, Hetru C, Hoffmann JA (1991) Insect immunity. Isolation from a coleopteran insect of a novel inducible antibacterial peptide and of new members of the insect defensin family. J Biol Chem 266(36):24520–24525. https://doi.org/10.1016/S0021-9258(18)54260-5
Burrow K, Young W, Carne A, McConnell M, Hammer N, Scholzeg M, El-Din Bekhit A (2018) Consumption of sheep milk compared to cow milk can affect trabecular bone ultrastructure in a rat model. Food Funct 10(1):163–171. https://doi.org/10.1039/C8FO01598H
Carrasco-Castilla J, Hernandez A, Lvarez AJ, Jimenez-Martinez C, Jacinto-Hernandez C, Alaiz M, Giron-Calle J, Vioque J, Davila-Ortiz G (2012) Antioxidant and metal chelating activities of Phaseolus vulgaris L. var. Jamapa protein isolates, phaseolin and lectin hydrolysates. Food Chem 131(2):1157–1164. https://doi.org/10.1016/j.foodchem.2011.09.084
Chang CY, Wu KC, Chiang SH (2007) Antioxidant properties and protein compositions of porcine haemoglobin hydrolysates. Food Chem 100:1537–1543
Chifíriuc MC, Cioaca AB, Lazar V (2011) In vitro assay of the antimicrobial activity of kephir against bacterial and fungal strains. Anaerobe 17:433–435. https://doi.org/10.1016/j.anaerobe.2011.04.020
Corrêa AP, Daroit DJ, Fontoura R, Meira SM, Segalin J, Brandelli A (2014) Hydrolysates of sheep cheese whey as a source of bioactive peptides with antioxidant and angiotensin-converting enzyme inhibitory activities. Peptides 61:48–55. https://doi.org/10.1016/j.peptides.2014.09.001
de Lima MD, da Silva RA, da Silva MF, da Silva PA, Costa RM, Teixeira JA, Porto AL (2018) Brazilian kefir-fermented sheep’s milk, a source of antimicrobial and antioxidant peptides. Probiotics Antimicrob Proteins 10(3):446–455. https://doi.org/10.1007/s12602-017-9365-8
Delgado MC, Nardo A, Pavlovic M, Rogniaux H, Añón MC, Tironi VA (2016) Identification and characterization of antioxidant peptides obtained by gastrointestinal digestion of amaranth proteins. Food Chem 197:1160–1167. https://doi.org/10.1016/j.foodchem.2015.11.092
Gavino VC, Gavino G, Leblanc MJ, Tuchweber B (2000) An isomeric mixture of conjugated linoleic acids but not pure cis-9, trans-11- octadecadienoic acid affects body weight gain and plasma lipids in hamsters. J Nutr 130(1):27–29. https://doi.org/10.1093/jn/130.1.27
Harwig SS, Kokryakov VN, Swiderek KM, Aleshina GM, Zhao C, Lehrer RI (1995) Prophenin-1, an exceptionally proline-rich antimicrobial peptide from porcine leukocytes. FEBS Lett 362(1):65–69. https://doi.org/10.1016/0014-5793(95)00210-Z
Hashemi SMB, Gholamhosseinpour A (2020) Effect of ultrasonication treatment and fermentation by probiotic Lactobacillus plantarum strains on goat milk bioactivities. Int J Food Sci Technol 55(6):2642–2649
Hati S, Shilpa V, Brij S, Vandna K, Surajit M (2013) Antioxidative activity and polyphenol content in fermented soy milk supplemented with WPC-70 by probiotic Lactobacilli. Int Food Res J 20(5):2125
Hoess A, Watson S, Siber GR, Liddington R (1993) Crystal structure of an endotoxin-neutralizing protein from the horseshoe crab, Limulus anti-LPS factor, at 1.5 A resolution. The EMBO J 12(9):3351–3356. https://doi.org/10.1002/j.1460-2075.1993.tb06008.x
Houston ME Jr, Kondejewski LH, Karunaratne DN, Gough M, Fidai S, Hodges RS, Hancock RE (1998) Influence of preformed α-helix and α-helix induction on the activity of cationic antimicrobial peptides. The J Peptide Res 52(2):81–88. https://doi.org/10.1111/j.1399-3011.1998.tb01361.x
Je JY, Qian ZJ, Lee SH, Byun HG, Kim SK (2008) Purification and antioxidant properties of bigeye tuna (Thunnus obesus) dark muscle peptide on free radical-mediated oxidative systems. J Med Food 11(4):629–637. https://doi.org/10.1089/jmf.2007.0114
Khaled E (2009) Antimicrobial activity of sheep yoghurt prepared by different commercial starter strains. Alexandria Science Exchange Journal 30(2):188–196
Li Y, Wei H, Hsieh TC, Pallas DC (2008) Cdc55p-mediated E4orf4 growth inhibition in Saccharomyces cerevisiae is mediated only in part via the catalytic subunit of protein phosphatase 2A. J Virol 82(7):3612–3623. https://doi.org/10.1128/JVI.02435-07
Li Y, Sadiq FA, Liu T et al (2015) Purification and identification of novel peptides with inhibitory effect against angiotensin I converting enzyme and optimization of process conditions in milk fermented with the yeast Kluyveromyces marxianus. J Funct Foods 16:278–288. https://doi.org/10.1016/j.jff.2015.04.043
Liu JR, Chen MJ, Lin CW (2005) Antimutagenic and antioxidant properties of milk−kefir and soymilk−kefir. J Agric Food Chem 53(7):2467–2474. https://doi.org/10.1021/jf048934k
Liu R, Wang M, Duan JA, Guo JM, Tang YP (2010) Purification and identification of three novel antioxidant peptides from Cornu Bubali (water buffalo horn). Peptides 31(5):786–793. https://doi.org/10.1016/j.peptides.2010.02.016
Lopez-Exposito I, Gomez-Ruiz JA, Amigo L, Recio I (2006a) Identification of antibacterial peptides from ovine αs2-casein. Int Dairy J 16(9):1072–1080. https://doi.org/10.1016/j.idairyj.2005.10.006
Lopez-Exposito I, Minervini F, Amigo L, Recio I (2006b) Identification of antibacterial peptides from bovine κ-casein. J Food Prot 69(12):2992–2997. https://doi.org/10.4315/0362-028X-69.12.2992
Mohanty D, Jena R, Choudhury PK, Pattnaik R, Mohapatra S, Saini MR (2016) Milk derived antimicrobial bioactive peptides: a review. Int J Food Prop 19(4):837–846. https://doi.org/10.1080/10942912.2015.1048356
Moslehishad M, Ehsani MR, Salami M et al (2013) The comparative assessment of ACE-inhibitory and antioxidant activities of peptide fractions obtained from fermented camel and bovine milk by Lactobacillus rhamnosus PTCC 1637. Int Dairy J 29(2):82–87
O’Shea M, Bassaganya-Riera J, Mohede ICM (2004) Immunomodulatory properties of conjugated linoleic acid. Am J Clin Nutr 79(8):1199–1206. https://doi.org/10.1093/ajcn/79.6.1199S
Panchal G, Hati S, Sakure A (2020) Characterization and production of novel antioxidative peptides derived from fermented goat milk by L. fermentum. LWT-Food Sci Technol 119:108887. https://doi.org/10.1016/j.lwt.2019.108887
Panchal G, Hati S, Darji V, Prajapati J (2020b) Antioxidant activities, proteolytic activity and growth behavior of Lactobacillus cultures during fermentation of goat milk. Ind J Dairy Sci 73(1):57–66
Papadimitriou CG, Vafopoulou-Mastrojiannaki A, Silva SV (2007) Identification of peptides in traditional and probiotic sheep milk yoghurt with angiotensin I-converting enzyme (ACE)-inhibitory activity. Food Chem 105(2):647–656. https://doi.org/10.1016/j.foodchem.2007.04.028
Parmar H (2017) Isolation and purification of ACE-inhibitory peptides derived from fermented surti goat milk, Dissertation, Anand Agricultural University, AAU, Anand, Gujarat, India
Rana S, Bajaj R, Mann B (2018) Characterization of antimicrobial and antioxidative peptides synthesized by L. rhamnosus C6 fermentation of milk. Int J Peptide Res Therap 24(2):309–21. https://doi.org/10.1007/s10989-017-9616-2
Rival SG, Fornaroli S, Boeriu CG, Wichers HJ (2001) Caseins and casein hydrolysates. 1. Lipoxygenase inhibitory properties. J Agric Food Chem 49(1):287–294. https://doi.org/10.1021/jf000392t
Rodríguez-Pazo N, Vázquez-Araújo L, Pérez-Rodríguez N, Cortés-Diéguez S, Domínguez JM (2013) Cell-free supernatants obtained from fermentation of cheese whey hydrolyzates and phenylpyruvic acid by Lactobacillus plantarum as a source of antimicrobial compounds, bacteriocins, and natural aromas. Appl Biochm Biotechnol 171:1042–1060. https://doi.org/10.1007/s12010-013-0408-7
Schillinger U (1989) Antibacterial activity of Lactobacillus’s sake isolated from meat. Appl Environ Microbiol 55(8):1901–1906. https://doi.org/10.1128/aem.55.8.1901-1906.1989
Selsted ME, Brown DM, DeLange RJ, Harwig SS, Lehrer RI (1985) Primary structures of six antimicrobial peptides of rabbit peritoneal neutrophils. J Biol Chem 260(8):4579–4584. https://doi.org/10.1016/S0021-9258(18)89110-4
Sheng J, Yang X, Chen J, Peng T, Yin X, Liu W, Yang X (2019) Antioxidative effects and mechanism study of bioactive peptides from defatted walnut (Juglans regia L.) meal hydrolysate. J Agric Food Chem 67(12):3305–3312. https://doi.org/10.1021/acs.jafc.8b05722
Siow HL, Gan CY (2016) Extraction, identification, and structure–activity relationship of antioxidative and α-amylase inhibitory peptides from cumin seeds (Cuminum cyminum). J Funct Foods 22:1–12. https://doi.org/10.1016/j.jff.2016.01.011
Solanki D, Hati S, Sakure A (2017) in silico and in vitro analysis of novel angiotensin I-converting enzyme (ace) inhibitory bioactive peptides derived from fermented camel milk (Camelus dromedarius). Int J Pept Res Ther 19(4):275–380. https://doi.org/10.1007/s10989-017-9577-5
Storici P, Tossi A, Lenarčič B, Romeo D (1996) Purification and structural characterization of bovine cathelicidins, precursors of antimicrobial peptides. Eur J Biochem 238(3):769–776. https://doi.org/10.1111/j.1432-1033.1996.0769w.x
Sugiyama K, Ogino T, Ogata K (1990) Rapid purification and characterization of histatins (histidine-rich polypeptides) from human whole saliva. Arch Oral Biol 35(6):415–419. https://doi.org/10.1016/0003-9969(90)90202-L
Tian M, Fang B, Jiang L, Guo H, Cui J, Ren F (2015) Structure-activity relationship of a series of antioxidant tripeptides derived from β-Lactoglobulin using QSAR modeling. Dairy Sci Technol 95(4):451–463. https://doi.org/10.1007/s13594-015-0226-5
Tonolo F, Fiorese F, Moretto L, Folda A, Scalcon V, Grinzato A, Ferro S, Arrigoni G, Bindoli A, Feller E, Bellamio M (2020) Identification of new peptides from fermented milk showing antioxidant properties: mechanism of action. Antioxidants 9(2):117. https://doi.org/10.3390/antiox9020117
Wang W, Smith DK, Moulding K, Chen HM (1998) The dependence of membrane permeability by the antibacterial peptide cecropin B and its analogs, CB-1 and CB-3, on liposomes of different composition. J Biol Chem 273(42):27438–27448. https://doi.org/10.1074/jbc.273.42.27438
Yang Y, Zheng N, Yang J, Bu D, Wang J, Ma L, Sun P (2014) Animal species milk identification by comparison of two-dimensional gel map profile and mass spectrometry approach. Int Dairy J 35(1):15–20. https://doi.org/10.1016/j.idairyj.2013.09.008
Ye Y, Barange M, Beveridge M, Garibaldi L, Gutierrez N, Anganuzzi A, Taconet M (2017) FAO’s statistic data and sustainability of fisheries and aquaculture: Comments on Pauly and Zeller. Mar Policy 81:401–405. https://doi.org/10.1016/j.marpol.2017.03.012
Yilmaz-Ersan L, Ozcan T, Akpinar-Bayizit A, Sahin S (2016) The antioxidative capacity of Kefir produced from goat milk. Int J Chem Eng Appl 7(1):22–26
Yu D, Feng MQ, Sun J, Xu XL, Zhou GH (2020) Protein degradation and peptide formation with antioxidant activity in pork protein extracts inoculated with Lactobacillus plantarum and Staphylococcus simulans. Meat Sci 160:107958
Acknowledgements
We are very much thankful to the Dr. Rank, Animal Breeding Department, College of Veterinary Science, Anand for helping in sheep breed selection.
Funding
None.
Author information
Authors and Affiliations
Contributions
Conceptualization: AS, SH, GBP, MM, GK; Methodology: SD, JK, BB; Formal analysis and investigation: JK, SD, BB; Writing: JK, SD, BB; Original draft preparation: JK, SD, BB; Review and editing: SD, SH, AS, GBP, MM; Supervision: AS, GBP, MM; All the authors viewed and approved the final version of the manuscript. SH, AS and JK contributed equally to this study.
Corresponding author
Ethics declarations
Conflict of interest
All the authors declare that there is no conflict of interest.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Ashokbhai, J.K., Basaiawmoit, B., Sakure, A. et al. Purification and characterization of antioxidative and antimicrobial peptides from lactic-fermented sheep milk. J Food Sci Technol 59, 4262–4272 (2022). https://doi.org/10.1007/s13197-022-05493-2
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s13197-022-05493-2