OBTAINING OF β-LACTOGLOBULIN BY GEL FILTRATION OF COW MILK WHEY

Volodymyr Yukalo, Kateryna Datsyshyn, Liudmyla Storozh

Abstract


Milk whey proteins carry out a number of important biological functions and also they are precursors of many biologically active peptides (antihypertensive peptides, antagonists of opioid receptors, regulators of intestinal motility, immunomodulatory, anti-microbial and anti-cancer peptides, appetite regulators and so on.). An important stage in natural bioactive peptides obtaining from milk whey proteins is the isolation of homogeneous proteins-precursors. Considering the significant difference in the molecular masses of whey proteins, a promising method for their selection is gel filtration. The purpose of the research was the fractionation of bioactive peptides precursors from milk whey using gel filtration on Sephadex G-150. The whey was obtained from fresh skimmed milk after isoelectric precipitation of casein. Gel filtration was carried out on the columns from a liquid chromatography kit by the “Reanal” company. The fractional composition and the degree of homogeneity of milk whey proteins were determined by disc-electrophoresis in the plates of a polyacrylamide gel. A repeated gel filtration of fractions from the chromatographic peaks, separated into sections, was performed to increase the fractionation efficiency. While choosing a dextran gel for gel filtration of precursors of biologically active peptides from milk whey proteins, we have taken into account the range of their molecular weights (from 10000 to 150000 Da), the ability to form supramolecular structures (β-LG), as well as the previously obtained results of gel filtration. As a result, it was shown that repeated gel filtration of milk whey on Sephadex G-150 allows efficiently fractionate the proteins-precursors of bioactive peptides. The range of peptides and proteins molecular weights that can be fractionated on this Sephadex is from 5000 to 300 000 Da. The usage of repeated gel filtration on Sephadex G-150 with the chromatogram separation into sectors allows to effectively fractionate proteins-precursors of bioactive peptides from milk whey. In particular, homogeneous β-lactoglobulin (degree of homogeneity > 95 %) and partially purified α-lactalbumin, as well as a group of immunoglobulins and a proteose-peptone fraction were obtained.


Keywords


milk whey proteins; β-lactoglobulin; gel filtration; disc-electrophoresis

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References


Park, Y. W. (Ed.) (2009). Bioactive components in milk and dairy products. Wiley-Blackwell, 426. doi: https://doi.org/10.1002/9780813821504

Fox, P., Uniacke-Lowe, T., McSweeney, P., O’Mahony, J. (2015). Dairy Chemistry and Biochemistry. Springer, 584. doi: https://doi.org/10.1007/978-3-319-14892-2

Oo, T. Z., Cole, N., Garthwaite, L., Willcox, M. D. P., Zhu, H. (2010). Evaluation of synergistic activity of bovine lactoferricin with antibiotics in corneal infection. Journal of Antimicrobial Chemotherapy, 65 (6), 1243–1251. doi: https://doi.org/10.1093/jac/dkq106

McSweeney, P. L. H., Fox P. F. (Eds.) (2013) Advanced Dairy Chemistry. Springer, 548. doi: https://doi.org/10.1007/978-1-4614-4714-6

Bonnaillie, L. M., Tomasula, P. M. (2012). Fractionation of Whey Protein Isolate with Supercritical Carbon Dioxide To Produce Enriched α-Lactalbumin and β-Lactoglobulin Food Ingredients. Journal of Agricultural and Food Chemistry, 60 (20), 5257–5266. doi: https://doi.org/10.1021/jf3011036

Konrad, G., Kleinschmidt, T. (2008). A new method for isolation of native α-lactalbumin from sweet whey. International Dairy Journal, 18 (1), 47–54. doi: https://doi.org/10.1016/j.idairyj.2007.06.004

Lozano, J. M., Giraldo, G. I., Romero, C. M. (2008). An improved method for isolation of β-lactoglobulin. International Dairy Journal, 18 (1), 55–63. doi: https://doi.org/10.1016/j.idairyj.2007.05.003

Osterman, L. A. (1985). Khromatografiya belkov i nukleinovykh kislot. Moscow: Nauka, 536.

Yukalo, V. G., Datsyshyn, K. Ye. (2018). Dextran gels for the exclusive chromatography of milk serum proteins. Scientific Messenger of LNU of Veterinary Medicine and Biotechnologies, 20 (85), 3–8. doi: https://doi.org/10.15421/nvlvet8501

Yukalo, V. G., Storozh, L. A. (2007). Vydilennia elektroforetychno gomogennoi frakcii β-CN-5P kazeinu korovіachogo moloka. Medychna khimiya, 2, 91–95.

Yukalo, V. G., Datsyshyn, K. Ye. (2018). Gel filtration of cow milk whey proteins. Food Science and Technology, 12 (4), 72–78. doi: https://doi.org/10.15673/fst.v12i4.1183

Osterman, L. A. (1981). Metody issledovaniya belkov i nukleinovykh kislot: Elektroforez i ultratsentrifugirovaniye. Moscow: Nauka.

Studier, F. W. (1973). Analysis of bacteriophage T7 early RNAs and proteins on slab gels. Journal of Molecular Biology, 79 (2), 237–248. doi: https://doi.org/10.1016/0022-2836(73)90003-x

Yukalo, V. G., Yavorskyi, B. I., Storozh, L. A., Solovodzinska, I. Ye. (2007). Kilkisnyi elektroforetychnyi analiz bilkiv kazeinovoho kompleksu. Biolohiya tvaryn, 9 (12), 295–298.




DOI: http://dx.doi.org/10.21303/2504-5695.2019.00859

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ISSN 2504-5695 (Online), ISSN 2504-5687 (Print)