COMPARISON OF DIFFERENT METHODS FOR BETA LACTOGLOBULIN ISOLATION

The aim of this study was to introduce a simple, high efficient and less expensive method for isolation of βLg from whey. Anion exchange chromatography, pepsin enzyme treatment and ultrafiltration tecniques were preferred for isolation process to compare differences. In addition to, centrifuge ultrafiltration techniques were using for the first time for isolation of β-Lg from whey. Physicochemical analysis of whey samples indicated that protein and β-Lg content in whey samples changed from 0.07 to 0.8% and 0.24 to 0.29 g/L, respectively. Treatment with the use of pepsin enzyme, anion exchanges chromatography and ultrafiltration techniques, resulted to β-Lg of 1.43, 6.56 and 43.59 folds respectively. Our results showed that ultrafiltration techniques are rapid and efficient that allows high protein yield and has advantages over other methods since it preserves the native structure of β-Lg. Additionally, when the enzymatic hydrolysis was used together with ultrafiltration technique, it was found efficient and pure than the enzymatic hydrolysis together with dialyse membrane. Also this study concluded that pepsin enzyme treatment and anion exchange chromatography are economic methods but they are not efficient enough and very time consuming. However isolation efficiency can be increased the use of isolation methods together

___

Aich, R., Batabyal, S. & Joardar, S.N. (2015). Isolation and purification of betalactoglobulin from cow milk. Veterinary World, 8, 621-624.

Bhattacharjee, S., Bhattacharjee, C. & Datta, S. (2006). Studies on the fractionation of β- lactoglobulin from casein whey using ultrafiltration and ion-exchange membrane chromatography. Journal of Membrane Science, 275(1), 141-150.

Bottomley, R. (1991). Process for obtaining concentrates having a high α-lactalbumin content from whey. US Patent, Patent number: 5,008, 376.

Chatterton, D.E., Smithers, G., Roupas, P. & Brodkorb, A. (2006). Bioactivity of β- lactoglobulin and α-lactalbumin— Technological implications for processing. International Dairy Journal, 16(11), 1229-1240.

Cheang, B., & Zydney, A. L. (2003). Separation of α‐lactalbumin and β‐lactoglobulin using membrane ultrafiltration. Biotechnology and Bioengineering, 83(2), 201-209.

Cheang, B., & Zydney, A. L. (2004). A two-stage ultrafiltration process for fractionation of whey protein isolate. Journal of Membrane Science, 231(1), 159-167.

De Wit, J. (1998). Nutritional and functional characteristics of whey proteins in food products. Journal of Dairy Science, 81(3), 597-608.

Elgar, D.F., Norris, C.S., Ayers, J.S., Pritchard, M., Otter, D.E., & Palmano, K.P. (2000). Simultaneous separation and quantitation of the major bovine whey proteins including proteose peptone and caseinomacropeptide by reversed-phase high-performance liquid chromatography on polystyrene–divinylbenzene. Journal of Chromatography A, 878(2), 183-196.

Fox, P. (2003). Milk proteins: general and historical aspects Advanced Dairy Chemistry—1 Proteins (pp. 1-48): Springer. ISBN 978-1-4419-8602-3

Guo, M., Fox, P., Flynn, A., & Kindstedt, P. (1995). Susceptibility of β-Lactoglobulin and Sodium Caseinate to Proteolysis by Pepsin and Trypsin. Journal of Dairy Science, 78(11), 2336-2344.

Hernández-Ledesma, B., Recio, I., & Amigo, L. (2008). β-Lactoglobulin as source of bioactive peptides. Amino Acids, 35(2), 257-265.

Jang, H.D., & Swaisgood, H.E. (1990). Disulfide bond formation between thermally denatured β-lactoglobulin and κ-casein in casein micelles. Journal of Dairy Science, 73(4), 900-904.

Karagözlü, C., & Bayarer, M. (2004). Peyniraltı suyu proteinlerinin fonksiyonel özellikleri ve sağlık üzerine etkileri. Ege Üniv. Ziraat Fakültesi Dergisi, 41, 197-207. Kinekawa, Y.-I., & Kitabatake, N. (1996).

Purification of β-Lactoglobulin from Whey Protein Concentrate by Pepsin Treatment. Journal of Dairy Science, 79(3), 350-356.

Laemmli, U.K. (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, 227(5259), 680-685.

Lieske, B., Konrad, G., & Faber, W. (1997). A new spectrophotometric assay for native β-lactoglobulin in raw and processed bovine milk. International Dairy Journal, 7(12), 805-812.

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.

Morr, C., & Ha, E. (1993). Whey protein concentrates and isolates: processing and functional properties. Critical Reviews in Food Science & Nutrition, 33(6), 431- 476.

Muller, A., Chaufer, B., Merin, U., & Daufin, G. (2003). Purification of alpha-lactalbumin from a prepurified acid whey: Ultrafiltration or precipitation. Le Lait, 83(6), 439-451.

Outinen, M. (2010). Effect of pre-treatment of cheese milk on the composition and characteristics of whey and whey products. TKK Dissertation 212, Aalto University, p. 44. ISBN 978-9-5260- 3006-7

Reddy, I.M., Kella, N.K., & Kinsella, J.E. (1988). Structural and conformational basis of the resistance of beta-lactoglobulin to peptic and chymotryptic digestion. Journal of Agricultural and Food Chemistry, 36(4), 737-741.

Sannier, F., Bordenave, S., & Piot, J. (2000). Purification of goat β-lactoglobulin from whey by an ultrafiltration membrane enzymic reactor. Journal of Dairy Research, 67(01), 43-51.

Şanlidere Aloğlu, H. (2013). The effect of various heat treatments on the antioxidant capacity of milk before and after simulated gastrointestinal digestion. International Journal of Dairy Technology, 66(2), 170-174.

Stojadinovic, M.,Burazer, L., Ercili-Cura,D., Sancho, A.,Buchert, J.,Velickovica T.C. & Stanic-Vucinica, D. (2012). One-step method for isolation and purification of native β-lactoglobulin. Journal of the Science of Food and Agriculture, 92, 1432-1440

Tunçtürk, Y., Andiç, S., & Ocak, E. (2010). Homojenizasyon ve pastörizasyonun Beyaz Peynir ve Peyniraltı Suyu Bileşimine Etkisi. Gıda/The Journal of Food, 35(5), 339-345.

Ye, X., Yoshida, S., & Ng, T. (2000). Isolation of lactoperoxidase, lactoferrin, α- lactalbumin, β-lactoglobulin B and β- lactoglobulin A from bovine rennet whey using ion exchange chromatography. The International Journal of Biochemistry & Cell Biology, 32(11), 1143-1150.

Yerlikaya, O., Kınık, Ö., & Akbulut, N. (2010). Peyniraltı Suyunun Fonksiyonel Özellikleri ve Peyniraltı Suyu Kullanılarak Üretilen Yeni Nesil Süt Ürünleri. Gıda/The Journal of Food, 35(4), 289-296.

Young, V. R. (1994). Adult amino acid requirements: the case for a major revision in current recommendations. The Journal of Nutrition, 124(8 Suppl), 1517S-1523S.