Measurement of Heat and Pressure Induced Denaturation of Whey Protein Isolate Using Reversed-Phase HPLC and FTIR-Spectroscopy
Trends in Peptide and Protein Sciences,
Vol. 4 (2019),
1 January 2019
,
Page 1-6 (e11)
https://doi.org/10.22037/tpps.v4i0.26676
Abstract
The protein molecules experience various external stresses leading to denaturation of protein during the process of transforming original whey to the concentrated whey proteins or while the concentrated proteins are used in the protein-enriched food formulation. This study was designed for a comparative assessment of the denaturation of whey protein isolate (WPI) under an important thermal stress, isothermal heat treatment (IHT), and high hydrostatic pressure treatment (HPT). The type and extent of denaturation were determined using reversed-phase HPLC and FTIR spectroscopy. The HPLC results demonstrated that the isothermal heat treatment caused higher denaturation of protein due to IHT at 80oC for 600s (88.38%) than that of HPT (58.5%). However, the infra-red spectroscopic analyses suggested that the HPT caused severe destruction of the structural conformation of WPI. The state of protein has a great impact on food formation; hence, the findings of this study would alert the concentrate protein producers and protein-enriched food manufacturers to prepare more active functional foods.
HIGHLIGHTS
- Heat (IHT) and pressure (HPT) stresses caused 88% and 58% WPI to denature, respectively.
- Reversed phase-HPLC determined the denatured protein through aggregation.
- FTIR together with HPLC is required for better characterization of denatured protein.
- Conformation of protein
- Denaturation of protein
- FTIR spectroscopy
- Reversed-phase HPLC
- Whey protein.
How to Cite
References
Anandharamakrishnan, C., Rielly, C. D. and A. G. F. Stapley, (2008). ″Loss of solubility of α-lactalbumin and β-lactoglobulin during the spray drying of whey proteins.″ Swiss Society of Food Science and Technology, 41: 270-277.
Anandharamakrishnan, C., Riellya, C. D. and A. G. F. Stapleya, (2007). ″Effects of process variables on the denaturation of whey proteins during spray drying.″ Drying Technology, 25(5): 799-807.
Byler, D. M. and H. Susi, (1986). ″Examination of the secondary structure of proteins by deconvolved FTIR spectra.″ Biopolymers, 25: 469−487.
Felipe, X., Capellas, M. and A. J. R. Law, (1997). ″Comparison of the effects of high-pressure treatments and heat pasteurization on the whey proteins in goat’s milk.″ Journal of Agricultural and Food Chemistry, 45: 627-631.
Ferreira, I. M. P. L. V. O., Mendes, E., and M. A. Ferreira, (2001). ″HPLC/UV analysis of proteins in dairy products using a hydrophobic interaction chromatographic column.″ Analytical Sciences, 17: 499-501.
Fu, F., Deoliveira, D. B., Trumble, W. R., Sarkar, H. K. and B.R. Singh, (1994). ″Secondary structure estimation of proteins using the amide III region of Fourier Transform Infrared Spectroscopy: application to analyze calcium binding-induced structural changes in calsequestrin.″ Applied Spectroscopy, 48: 1432-1441.
Godfrey, K., Robinson, S., Barker, D.J.P., Osmond, C. and V. Cox, (1996). ″Maternal nutrition in early and late pregnancy in relation to placental and fetal growth.″ British Medical Journal 312:, 410-414.
Grossi, A., Olsen, K., Bolumar, T., Rinnan, A., Øgendal, L. H. and V. Orlien, (2016). ″The effect of high pressure on the functional properties of pork myofibrillar proteins.″ Food Chemistry, 196: 1005–1015.
Hastings, G., Thomas, J., Witt, K., Giarikos, D. and R. Razeghifard, (2018). ″Starch, lipid, and protein accumulation in nutrient-stressed microalgal cells studied using Fourier Transform Infrared Microscopy.″ Frontiers in Science, Technology, Engineering and Mathematics, 2(2): 1-8.
Haque, M. A., Aldred, P., Chen, J., Barrow, C. J., and B. Adhikari, (2013). ″Comparative study of denaturation of whey protein isolate (WPI) in convective air drying and isothermal heat treatment processes.″ Food Chemistry, 141: 702–711.
Haque, M. A., Aldred, P., Chen, J., Barrow, C. and B. Adhikari, (2014). ″Drying and denaturation characteristics of α-lactalbumin, β-lactoglobulin and bovine serum albumin in a convective drying process.″ Journal of Agricultural and Food Chemistry, 62 (20):, 4695–4706.
Haque, M. A., Aldred, P., Chen, J. and B. Adhikari, (2015). ″Denaturation and physical characteristics of spray dried whey protein isolate powders produced in the presence and absence of lactose, trehalose and polysorbate- 80″. Drying Technology, 33: 1243–1254.
Hoffman, J. R. and M. J. Falvo, (2004). ″Protein – which is best?″ Journal of Sports Science and Medicine 3: 118-130.
Jimenez, M., Garcia, H.S. and C. I. Beristain, (2004). ″Spray-drying microencapsulation and oxidative stability of conjugated linoleic acid.″ European Food Research and Technology, 219 (6): 588–592.
Kim, Y. D., Morr, C. V. and T. W. Schenz, (1996). ″Microencapsulation properties of gum Arabic and several food proteins: Liquid orange oil emulsion particles.″ Journal of Agricultural and Food Chemistry, 44(5): 1308–1313.
Kong, J. and S. Yu, (2007). ″Fourier transform infrared spectroscopic analysis of protein secondary structures.″ Acta Biochimica Et Biophysica Sinica, 39: 549–559.
Mosharaf, M. K., Tanvir, M. Z. H., Haque, M. M., Haque, M. A., Khan, M. A. A., Molla, A. H., Alam, M. Z., Islam, M. S. and M. R. Talukder, (2018). ″Metal-adapted bacteria isolated from wastewaters produce biofilms by expressing proteinaceous curli fimbriae and cellulose nanofibers.″ Frontiers in Microbiology, 9: Article 13314.
Navarro, F., Harouna, S., Calvo, M., Pérez, M. D. and Sánchez, L (2015). ″Kinetic and thermodynamic parameters for thermal denaturation of ovine milk lactoferrin determined by its loss of immunoreactivity.″ Journal of Dairy Science, 98(7):4328-37.
Ng-Kwai-Hang, K. F. (2011). Milk proteins heterogeneity, fractionation, and isolation. McGill University, Montreal, QC, Canada, Elsevier Ltd., pp. 751-764.
Parris, N. and M. A. Baginski, (1991). ″A rapid method for the determination of whey protein denaturation.″ Journal of Dairy Science, 74: 58-64.
Silverstein, R. M., Bassler, G. C. and Morrill, T.C. (1981). Spectrometric identification of organic compounds. 4th Edn., John Wiley and Sons, New York, USA. pp. 25-35.
Siwatt, P. (2014). ″FTIR spectra of organic functional group compositions in pm2.5 collected at Chiang-Mai city, Thailand during the haze episode in March 2012.″ Journal of Applied Sciences, 14: 2967-2977.
Vadim, V. M., Karel, H., Johannes, F., Patrick, M. and B. Claude, (1996). ″high pressure effects on protein structure and function.″ PROTEINS: Structure, Function, and Genetics, 24: 81-91.
van de Weert, M., Haris, P. I., Hennink, W. E. and D. J. A. Crommelin, (2001). ″Fourier Transform Infrared Spectrometric Analysis of Protein Conformation: Effect of sampling method and stress factors.″ Analytical Biochemistry, 297: 160-169.
- Abstract Viewed: 328 times
- PDF Downloaded: 169 times