Effect of Temperature, pH, Enzyme to Substrate Ratio, Substrate Concentration and Time on the Antioxidative Activity of Hydrolysates from Goat Milk Casein by Alcalase

Open access


The effect of hydrolysis temperature (45, 50, 55, 60 and 65°C), pH (7.0, 7.5, 8.0, 8.5 and 9.0), enzyme to substrate (E/S) ratio (1.0, 1.5, 2.0, 2.5 and 3.0%), substrate concentration (2, 3, 4, 5 and 6%) and hydrolysis time (30-240min) on antioxidant peptides hydrolysated from goat’s milk casein by Alcalase was investigated using single factor experiment. In order to obtain high DPPH radical-scavenging activity, metal-chelating activity and superoxide radical scavenging activity, the optimal conditions were hydrolysis time of 150 min, temperature of 50°C, pH 8.0, E/S ratio of 2.0% and substrate concentration of 4.0%. The hydrolysis time, hydrolysis temperature, pH, E/S ratio and substrate concentration had a significant influence on degree of hydrolysis, metal-chelating activity, DPPH and superoxide radical scavenging activity on casein hydrolysate of goat milk by Alcalase, the results were beneficial for further provide theoretical basis for production of antioxidant peptides.

1. Adler-Nissen, J. (1986). Enzymatic hydrolysis of food protein. London: Elsevier Applied Science Publishers, 12–14.

2. Alferez, M. J. M., Lopez-Aliaga, I., Nestares, T., et al. (2006). Dietary goat milk improves iron bioavailability in rats with induced ferropenic anaemia in comparison with cow milk. International Dairy Journal, 16 (7), 813-821.

3. Decker, E. A., Welch, B. (1990). Role of ferritin as a lipid oxidation catalyst in muscle food. Journal Agricultural Food Chemistry, 38, 674–677.

4. Dröge, W. (2002). Free radicals in the physiological control of cell function. Physiological Reviews, 82(1), 47-95.

5. Ferreira, N.G., Hultin, H.O., (1994). Liquefying cod fish frames under acidic conditions with a fungal enzyme. J. Food Proc. Pres., 18, 87.

6. Haenlein, G. F. W. (2004). Goat milk in human nutrition. Small Ruminant Research, 51(2), 155-163.

7. Han Ji-fu, Ren Jian-bo. (2003). Study on the optimum conditions of the preparation of soluble peptides from corn gluten meal hydrolysated with Alcalase, Journal of Jilin Agricultural. 28(3), 46-49.

8. He Ting. (2008). Hydrolysates derived from Decapterus maruadsi protein with controlled enzymatic modification and its antioxidant effect. Wuxi: Jiangnan university, 25-28.

9. Head, E. (2009). Oxidative damage and cognitive dysfunction: antioxidant treatments to promote healthy brain aging. Neurochemical Research, 34(4), 670-8.

10. Hodgkinson, A. J., Mcdonald, N. A., Kivits, L. J., et al. (2012). Allergic responses induced by goat milk α s1-casein in a murine model of gastrointestinal atopy. Journal of Dairy Science, 95(1), 83-90.

11. Kim S.K., Kim Y.T., Byun H.G., et al. (2001). Isolation and characterization of antioxidative peptides from gelatin hydrolysate of Alaska pollack skin. J. Agric. Food Chem, 49, 1984-1989.

12. Lobo, V., Patil, A., Phatak, A., et al. (2010). Free radicals, antioxidants and functional foods: impact on human health. Pharmacognosy Reviews, 4(8), 118-26.

13. Marklund, S., Marklund, G. (1974). Involvement of the superoxide anion radical in the autoxidation of pyrogallal and a convenient assay for superoxide dismutase. Eur J Biochem, 47 (3), 469-474.

14. Millάn F., Clemente A., Vioque J., et al. (1999). Production of extensive chickpea (Cicer arietinum L.) protein hydrolysates with reduced antigenic activity. J Agric Food Chem, 47(9), 3776-3781.

15. Murphy, M. P., Holmgren, A., Larsson, N. G., et al. (2011). Unraveling the biological roles of reactive oxygen species. Cell Metabolism, 13(4), 361-6.

16. Nagpal, R., Behare, P., Rana, R. et al. (2011). Bioactive peptides derived from milk proteins and their health beneficial potentials: an update. Food & Function, 2(1), 18-27.

17. Park, Y. W. (2007). Impact of goat milk and milk products on human nutrition. Cab Reviews Perspectives in Agriculture Veterinary Science Nutrition & Natural Resources, 2(81).

18. Peng Zhi-ying. (2002). An introduction to food enzymology. Beijing: China light industry press, 163-164.

19. Rafter, J. (2003). Probiotics and colon cancer. Best Prac. Res. Cli. Gastroentrol, 17 (5), 849-859.

20. Ramos A., Visozo A., Piloto, J. et al. (2003). Screening of antimutagenicity via antioxidant activity in Cuban medicinal plants. Journal of Ethnopharmacology, 87, 241–246.

21. Shu Guowei, Zhang Qian, Chen He, et al. (2015). Effect of five proteases including alcalase, flavourzyme, papain, proteinase k and trypsin on antioxidative activities of casein hydrolysate from goat milk. Acta Universitatis Cibiniensis Series E: FOOD TECHNOLOGY, 19(2),65-74

22. Udenigwe, C. C. (2014). Bioinformatics approaches, prospects and challenges of food bioactive peptide research. Trends in Food Science & Technology, 36(2), 137-143.

23. Wu H.C., Chen H.M., Shiau C.Y. (2003). Free amino acids and peptides as related to antioxidant properties in protein hydrolysates of mackerel (Scomber austriasicus). Food Res. Int., 36, 949–957.

24. Xiao Hong, Sun Yu-min, Duan Yu-feng. (2006). Study on the Hydrolytic Condition of Oxya chinensis (Thunberg) Protein with Alcalase. Nat Prod Res Dev, 18, 822-824.

25. Yu, L. (2001). Freer adical scavenging properties of conjugated linoleic acids. Journal of Agricultural and Food Chemistry, 49, 3452–3456.

26. Zhang Han-jun, Han jun-mei, Liu Da-chuan. (2007). Study on double low rapeseed protein isolated and its antioxidant properties by Alcalase hydrolysis. Grain and oil processing. 12, 77-80.

Acta Universitatis Cibiniensis. Series E: Food Technology

The Journal of „Lucian Blaga“ University of Sibiu

Journal Information


All Time Past Year Past 30 Days
Abstract Views 0 0 0
Full Text Views 230 230 78
PDF Downloads 105 105 46