PREPARATION OF PEPTIDE HYDROLYSATES FROM BETA CASEIN ISOLATED AND PURIFIED FROM IRAQI CAMEL MILK
DOI:
https://doi.org/10.36103/bnwe4t23Keywords:
active peptides, casein, enzymatic hydrolysates, gel filtration, ion exchangeAbstract
The current study aimed to isolate and purify the Beta Casein (β-CN) protein from Iraqi Camel milk by urea, and ion exchange chromatography using a DEAE-Cellulose column, and gel filtration by using a Sephadex-G100 column, determining the molecular weight (M.wt) of protein isolate by polyacrylamide gel electrophoresis in the presence of the denaturant Sodium Dodecyl Sulphate (SDS). Enzymatic hydrolyzates were prepared using both pepsin and trypsin enzymes and their mixture (1:1), then the degree of hydrolysis (DH) was estimated after incubation for 8 h. and compared with bovine protein and standard proteins, the elution of β-CN using a DEAE-Cellulose column showed the appearance of two separate peaks, and their molecular weight was 24 KDa and 20 KDa Respectively. The Sephadex-G100 gel filtration technique also showed the appearance of a single peak whose molecular weight was 24 KDa compared to β-CN, which weighed 23 KDa. The results showed that the highest degree of decomposition was 52.32% by using a mixture of pepsin and trypsin. The results indicate that the protein β-CN separated from camel milk can be highly purified using ion exchange chromatography and gel filtration techniques, and peptide hydrolysates can be prepared from it.
Received: 12/2/2024
Accepted: 26/5/2024
Published: 2026/4/30
References
Abbes, F. B., R. Belhattab., M. Seghier., and, D. L. Anes-Boulahbal. 2021. In vitro antioxidant and antiviral activity of camel milk casein hydrolysates. Journal of Applied Biological Sciences, 15(1):101-112. https://jabsonline.org/index.php/jabs/article/view/797.
Abd El-Aziz, M., J.M. Kassem., F.M. Aasem., and, H.M. Abbas. 2022. Physicochemical properties and health benefits of camel milk and its applications in dairy products: A Review. Egyptian Journal of Chemistry, 65(5):101.118.
https://doi.org/10.21608/EJCHEM.2021.92589.4383
Abderrahmane, F. A., A.B. Mezmaze Chekroun., D. J. A. Saidi., and, O. M. Kheroua. 2015. In vitro digestibility of the dromedary whey proteins: potential uses in infant milk allergies. International Journal of Pharmacology and Pharmaceutical Science, 7(2): 115-120. Google Scholar
Abdullahi, A. 2019. Camel milk-A Review. Journal of Animal Sciences and Livestock Production, 3(1):13-18.
Google Scholar
Ahmed, S.A., and A.J. Al-Mousawi. 2021. Preparation of lysozyme enzyme coated with chitosan nanoparticles. Plant Arch, (21): 1079-1087.
https://doi.org/10.51470/plantarchives.2021.v21.s1.169.
Akan, E. 2021. An evaluation of the in vitro antioxidant and antidiabetic potentials of camel and donkey milk peptides released from casein and whey proteins. Journal of Food Science and Technology, 58(10): 3743-3751.
https://doi.org/10.1007/s13197-021-05076-7.
Akindykova, A., C. Cakir-Kiefer., A. Baubekova., and, S. Jurjanz. 2019. Isolation and characterization of camel milk proteins. International Journal of Biology and Chemistry, 12(1): 4-10.
https://doi.org/10.26577/ijbch-2019-i1-2.
Al-easawi, M. A. H., E. Z. Naji., and S. B. Mohammed. 2020. Extraction and purification of Beta-galactosidase from local almond and itsuse for lactose intolerance treatment. Iraqi Journal of Agricultural Sciences, 51(3): 767-776. https://doi.org/10.36103/ijas.v51i3.1032.
Alhaj, O. A., R. Lajnaf., Z. Jrad., M. A. Alshuniaber., H. A. Jahrami., and, M. F. Serag El-Din. 2022. Comparison of ethanol stability and chemical composition of camel milk from five samples. Animals, 12(5): 615. https://doi.org/10.3390/ani12050615.
AlKhalidy SJ, and K.S. Dosh 2023 Isolation and Purification of αs-CN from Sheep Milk and Measuring the Effectiveness of Its Enzymatic Hydrolysis in Inhibiting ACE1. Chemical Methodologies, 7(2):156166. doi.org/10.22034/chemm.2023.366342.1618
AlKhalidy, S. J., and, K. S. Dosh. 2022. Isolation and purification of Β-CN from sheep milk and measuring the effectiveness of Its enzymatic hydrolysis in inhibiting ACE1. Ann. For. Res, 2022, 65.1: 8584-8596.
https://doi.org/10.22034/chemm.2023.366342.1618.
Al-Saleh, A. A., A. A.Metwalli., E. A. Ismail., and, O. A. Alhaj. 2014. Antioxidative activity of camel milk casein hydrolysates. Journal of Camel Practice and Research, 21(2): 229-237.
https://doi.org/10.5958/2277 8934.2014.00041.1.
Al-Shaikh, S. A. H., and Doosh, K. S. 2024. Evaluation of the Toxicological Efficacy of Protein Hydrolysates from Camel Milk β-Casein Against Different Types of Cancer Cell Lines in Vitro. In IOP Conference Series. Earth and Environmental Science (Vol. 1371, No. 6, p. 062027). IOP Publishing.
https:// 10.1088/1755-1315/1371/6/062027.
AOAC 2016. International Official Methods of Analysis of A.O.A.C International-20th Edition, 2016; AOAC: Gaithersburg, MD, USA, 2016.
Darani, K. K., M. Jahadi., A. D. Tripathi., V. Paul., A. Nandan, Chakraborty, and A. Agarwal. 2023. Probiotic dairy dessert from camel milk–A Review. Indian Journal of Dairy Science, 76(3):203-2015.
https://doi.org/10.33785/IJDS.2023.v76i03.001.
El‐Agamy, E. I. 2009. Bioactive components in camel milk. Bioactive components in milk and dairy products, 159-194.
DOI:10.1002/9780813821504.
Ellouze, M., C. Vial., H. Attia., and, M. A. Ayadi. 2021. Effect of pH and heat treatment on structure, surface characteristics and emulsifying properties of purified camel β-casein. Food Chemistry, 365(2):130421. https://doi.org/10.1016/j.foodchem.2021.130421
Felfoul, I., J. Jardin., F. Gaucheron., H. Attia., and, M. A. Ayadi. 2017. Proteomic profiling of camel and cow milk proteins under heat treatment. Food Chemistry, 100 (216):161-169.https://doi.org/10.1016/j.foodchem.2016.08.007.
Ganzorig, K., T. Urashima., and, K. Fukuda. 2020. Exploring potential bioactive peptides in fermented bactrian camel’s milk and mare’s milk made by mongolian nomads. Foods, 9(12):1817.
https://doi.org/10.3390/foods9121817.
Gaytan, A.Q., R A. Ali, Z.M. Ali, M.A. Majid, H.H. Alwan, and A.R.A. Abdullah, 2019. Separation of protein and lactose from whey disposed from abo-ghraab dairy factory by using membrane technology. Iraq journal of market research and consumer protection, 11(1):37-47. http://dx.doi.org/10.28936/jmracpc11.1.(4)
Haddad, Z. A., and K. S. Doosh. 2023. Isolation, purification and characterization of a-Lactalbumin from camel milk and study its antioxidant activity in oils. In IOP Conference Series: Earth and Environmental Science. 1262, (6): 062042. IOP Publishing.
doi:10.1088/1755-1315/1262/6/062042
Ho, T. M., Z. Zou., and, N. Bansal. 2022. Camel milk: A Review of its nutritional value, heat stability, and potential food products. Food ResearchInternational,153(2):110870. https://doi.org/10.1016/j.foodres.2021.110870.
Hoseini, S. M., M. Anushiravani., M. J. Mojahedi., M. Hami., S. Zibaee., H.Rakhshandeh., and, A. P. Tavassoli. 2020. The efficacy of camel milk and tarangabin (manna of alhagi maurorum (combination therapy on glomerular filtration rate in patients with chronic kidney disease: a randomized controlled trial. Avicenna Journal of Phytomedicine, 10(2):170.
PMCID: PMC7103430
Kappeler, S., Z. Farah., and, Z. Puhan. 1998. Sequence analysis of camelus dromedarius milk caseins. Journal of Dairy Research, 65(2): 209-222. https://doi.org/10.1017/S0022029997002847.
Karaman, A. D., F. Yildiz Akgul., S. Ögut., H. Secilmis Canbay., and, V. Alvarez. 2021. Gross composition of raw camel’s milk produced in Turkey. Food Science and Technology, 42(4): e59820.
https://doi.org/10.1590/fst.59820.
Khalil, A. H., and S. S. Lafta. 2023. Studying the physiochemical, sensory and microbiological properties of yoghurt which are fortified by encapsulated folic acid. In IOP Conference Series: Earth and Environmental Science. 1259(1): 012067. IOP Publishing.
DOI 10.1088/1755-1315/1259/1/012067.
Kim, W., Y. Wang., and, C. Selomulya. 2020. Dairy and plant proteins as natural food emulsifiers. Trends in Food Science and, Technology, 105: 261-272.
https://doi.org/10.1016/j.tifs.2020.09.012.
Kumar, D., Chatli, M. K., Singh, R., Mehta, N., and Kumar, P. 2016. Enzymatic hydrolysis of camel milk casein and its antioxidant properties. Dairy Science & Technology, 96(3), 391-404.
https://doi.org/10.1007/s13594-015-0275-9.
Lajnaf, R., H. Gharsallah, H. Attia, and M. A. Ayadi, 2021. Comparative study on antioxidant, antimicrobial, emulsifying and physico-chemical properties of purified bovine and camel β-casein. LWT, 140(1):110842. https://doi.org/10.1016/j.lwt.2020.110842.
Lajnaf, R., L. Picart-Palmade., H.Attia, S. Marchesseau., and, M. A. Ayadi. 2022. Foaming and air-water interfacial properties of camel milk proteins compared to bovine milk proteins. Food Hydrocolloids, 126(12): 107470. https://doi.org/10.1016/j.foodhyd.2021.107470
Liang, L. I., and, Y. Luo. 2020. Casein and pectin: Structures, interactions, and applications. Trends in Food Science and, Technology, 97(1): 391-403.
https://doi.org/10.1016/j.tifs.2020.01.027
Maehre, H.K., L. Dalheim., G.K. Edvinsen., E.O. Elvevoll., and, I.J. Jensen. 2018. Protein determination—method matters. Foods, 7(1):5.
https://doi.org/10.3390/foods7010005.
Maree, S., J.L. Du Preez, L.H. Du Plessis, J. Plessis,, and, M. Gerber, 2020. A novel HPLC method developed and validated for the detection and quantification of atorvastatin, fluvastatin, pitavastatin and pravastatin during transdermal delivery studies. Die Pharmazie-An International Journal of Pharmaceutical Sciences, 75(5):164-166. https://doi.org/10.1691/ph.2020.0007.
Mati, A., C. Senoussi-Ghezali, S.A. Zennia, D. Almi-Sebbane, H. El-Hatmi, and J.M. Girardet, 2017. Dromedary camel milk proteins, a source of peptides having biological activities–A Review. International Dairy Journal, 73: 25-37.
DOI: 10.1016/j.idairyj.2016.12.001.
Mohamed, H., M. Ayyash., and, A. Kamal-Eldin, 2022. Effect of heat treatments on camel milk proteins–A Review. International Dairy Journal, 133: 105404. https://doi.org/10.1016/j.idairyj.2022.105404.
Mohamed, H., P. Nagy., J. Agbaba., and, A. Kamal-Eldin. 2021. Use of near and mid infra-red spectroscopy for analysis of protein, fat, lactose and total solids in raw cow and camel milk. Food Chemistry, 334: 127436.
https://doi.org/10.1016/j.foodchem.2020.127436
Mudgil, P., A.A. Redha., N.P. Nirmal., and S. Maqsood. 2023. In vitro antidiabetic and antihypercholesterolemic activities of camel milk protein hydrolysates derived upon simulated gastrointestinal digestion of milk from different camel breeds. Journal of Dairy Science, 106(5):3098-3108. https://doi.org/10.3168/jds.2022-22701.
Ptiček Siročić, A., L.Kratofil Krehula., Z. Katančić., and Z. Hrnjak-Murgić. 2016. Characterization of casein fractions–Comparison of commercial casein and casein extracted from cow’s milk. Chemical and Biochemical Engineering Quarterly, 30(4): 501-509.
https://doi.org/10.15255/CABEQ.2015.2311
Redha, A. A., H. Valizadenia., S.A. Siddiqui., and S. Maqsood. 2022. A state-of-art review on camel milk proteins as an emerging source of bioactive peptides with diverse nutraceutical properties. Food Chemistry, 373(11):131444.
https://doi.org/10.1016/j.foodchem.2021.131444
Saliha, S. A. Z., A. Dalila., S. Chahra, B. H. Saliha., and M. Abderrahmane. 2013. Separation and characterization of major milk protein from Algerian dromedary (camel US dromedaries). Emirates Journal of Food and Agriculture, 25(4):283-290. https://doi.org/10.9755/ejfa.v25i4.15496.
Swelum, A. A., M.T. El-Saadony., M. Abdo., R.A. Ombarak., E. O. Hussein., G. Suliman., and Abd M. E. El-Hack. 2021. Nutritional, antimicrobial and medicinal properties of camel’s milk: A Review. Saudi Journal of Biological Sciences, 28(5): 3126-3136.
https://doi.org/10.1016/j.sjbs.2021.02.057.
Taghipour, M. J., H. Ezzatpanah, and M. Ghahderijani. 2023. In vitro and in silico studies for the identification of anti-cancer and antibacterial peptides from camel milk protein hydrolysates. Plos one, 18(7): e0288260.
https://doi.org/10.1371/journal.pone.0288260.
Tagliazucchi, D., S. Shamsia., and, A. Conte. 2016. Release of angiotensin converting enzyme-inhibitory peptides during in vitro gastro-intestinal digestion of camel milk. International Dairy Journal, 100(56): 119-128.
https://doi.org/10.1016/j.idairyj.2016.01.009
Vorob’ev, M. M. 2022. Modeling of proteolysis of β-lactoglobulin and β-casein by trypsin with consideration of secondary masking of intermediate polypeptides.
International Journal of Molecular Sciences, 23(15):8089.
https://doi.org/10.3390/ijms23158089
Wang, R., Z. Han., R. Ji., Y. Xiao., R.Si., F. Guo., and L. Yi. 2020. Antibacterial activity of trypsin-hydrolyzed camel and cow whey and their fractions. Animals, 10(2): 337. https://doi.org/10.3390/ani10020337.
Downloads
Published
Issue
Section
License
Copyright (c) 2026 Sharaf. Ali. Hadi. Al-Shaikh, Kifah. Saed. Doosh
This work is licensed under a Creative Commons Attribution 4.0 International License.
2.jpg)
