Phosvitin phosphopeptide preparation using immobilised trypsin and enhancing calcium absorption in growing rats

https://doi.org/10.17221/425/2015-CJFSCitation:Zhong Q., Li X., Hu W., Zeng B., Liang R., Liu H., Li Z., Zhang Z. (2016): Phosvitin phosphopeptide preparation using immobilised trypsin and enhancing calcium absorption in growing rats. Czech J. Food Sci., 34: 325-331.
download PDF
We demonstrate a new, efficient method for the preparation of phosvitin phosphopeptides using immobilised-trypsin enzymolysis technology. Immobilised trypsin was prepared using a covalent binding method, and then was added to degrade egg yolk phosvitin for the production of phosphopeptides. In our results, the prepared immobilised trypsin demonstrated a higher hydrolysing activity toward phosvitin than free trypsin, and the hydrolysing activity was retained well even after trypsin was repeatedly used up to five times. Interestingly, the phosvitin phosphopeptides prepared with immobilised trypsin demonstrated a lower N/P ratio and a higher calcium-binding efficiency than those prepared with free trypsin. Furthermore, phosphopeptides significantly increased the rate of calcium absorption and serum calcium content in vivo. Based on these results, we conclude that trypsin immobilised onto chitosan has a greater phosvitin hydrolysing activity than free trypsin, and the prepared phosphopeptides can be used as a new calcium supplement to significantly increase calcium absorption in growing rats.
References:
ABE YUTAKA, ITOH TAKATOSHI, ADACHI SUSUMU (1982): Fractionation and Characterization of Hen's Egg Yolk Phosvitin. Journal of Food Science, 47, 1903-1907  https://doi.org/10.1111/j.1365-2621.1982.tb12910.x
 
Allen Russell James Laurence (1940): The estimation of phosphorus. Biochemical Journal, 34, 858-865  https://doi.org/10.1042/bj0340858
 
Ali Khan Amjad, Alzohairy Mohammad A. (2010): Recent Advances and Applications of Immobilized Enzyme Technologies: A Review. Research Journal of Biological Sciences, 5, 565-575  https://doi.org/10.3923/rjbsci.2010.565.575
 
Berrocal Rafael, Chanton Serge, Juillerat Marcel A., Favillare Blaise, Scherz Jean-Claude, Jost Rolf (1989): Tryptic phosphopeptides from whole casein. II. Physicochemical properties related to the solubilization of calcium. Journal of Dairy Research, 56, 335-  https://doi.org/10.1017/S0022029900028776
 
Castellani O, Guérin-Dubiard C, David-Briand E, Anton M (2004): Influence of physicochemical conditions and technological treatments on the iron binding capacity of egg yolk phosvitin. Food Chemistry, 85, 569-577  https://doi.org/10.1016/j.foodchem.2003.08.002
 
Grizzuti K., Perlmann G.E. (1970): Conformation of the phosphoprotein, phosvitin. Journal of Biological Chemistry, 245: 2573–2578.
 
Hansen Marianne, Sandström Brittmarie, Lönnerdal Bo (1996): The Effect of Casein Phosphopetides on Zinc and Calcium Absorption from High Phytate Infant Diets Assessed in Rat Pups and Caco-2 Cells. Pediatric Research, 40, 547-552  https://doi.org/10.1203/00006450-199610000-00006
 
Homaei Ahmad Abolpour, Sariri Reyhaneh, Vianello Fabio, Stevanato Roberto (2013): Enzyme immobilization: an update. Journal of Chemical Biology, 6, 185-205  https://doi.org/10.1007/s12154-013-0102-9
 
CHOI I, JUNG C, CHOI H, KIM C, HA H (2005): Effectiveness of phosvitin peptides on enhancing bioavailability of calcium and its accumulation in bones. Food Chemistry, 93, 577-583  https://doi.org/10.1016/j.foodchem.2004.10.028
 
Ishikawa S.-I., Tamaki S., Arihara K., Itoh M. (2007): Egg Yolk Protein and Egg Yolk Phosvitin Inhibit Calcium, Magnesium, and Iron Absorptions in Rats. Journal of Food Science, 72, S412-S419  https://doi.org/10.1111/j.1750-3841.2007.00417.x
 
Jiang Bo, Mine Yoshinori (2000): Preparation of Novel Functional Oligophosphopeptides from Hen Egg Yolk Phosvitin. Journal of Agricultural and Food Chemistry, 48, 990-994  https://doi.org/10.1021/jf990600l
 
JIANG Bo, MINE Yoshinori (2001): Phosphopeptides Derived from Hen Egg Yolk Phosvitin: Effect of Molecular Size on the Calcium-binding Properties. Bioscience, Biotechnology and Biochemistry, 65, 1187-1190  https://doi.org/10.1271/bbb.65.1187
 
Kapoor Mukesh, Kuhad Ramesh Chander (2007): Immobilization of Xylanase from Bacillus pumilus Strain MK001 and its Application in Production of Xylo-oligosaccharides. Applied Biochemistry and Biotechnology, 142, 125-138  https://doi.org/10.1007/s12010-007-0013-8
 
Kitts D.D. (2005): Antioxidant properties of casein-phosphopeptides. Trends in Food Science & Technology, 16: 549–554.
 
Klomklao Sappasith, Benjakul Soottawat, Kishimura Hideki, Osako Kazufumi, Tanaka Munehiko (2010): A heat-stable trypsin inhibitor in adzuki bean (Vigna angularis): effect of extraction media, purification and biochemical characteristics. International Journal of Food Science & Technology, 45, 163-169  https://doi.org/10.1111/j.1365-2621.2009.02117.x
 
Ko K. Y., Nam K. C., Jo C., Lee E. J., Ahn D. U. (2011): A simple and efficient method for preparing partially purified phosvitin from egg yolk using ethanol and salts. Poultry Science, 90, 1096-1104  https://doi.org/10.3382/ps.2010-01138
 
Korhonen Hannu, Pihlanto Anne (2006): Bioactive peptides: Production and functionality. International Dairy Journal, 16, 945-960  https://doi.org/10.1016/j.idairyj.2005.10.012
 
Lei Bo, Wu Jianping (2012): Purification of egg yolk phosvitin by anion exchange chromatography. Journal of Chromatography A, 1223, 41-46  https://doi.org/10.1016/j.chroma.2011.12.023
 
Li Y., Tomé D., Desjeux J. F. (1989): Indirect effect of casein phosphopeptides on calcium absorption in rat ileum in vitro. Reproduction Nutrition Développement, 29, 227-233  https://doi.org/10.1051/rnd:19890210
 
Ling Y.X., Sheng K.H., Jiang S.W. (2007): Study on enzymatic characteristics of immobilizing trypsin with difference carrier. Biotechnology, 17: 71–73. (in Chinese)
 
Maqueda C., Morillo E. (1990): Determination of calcium by atomic-absorption spectrometry in samples dissolved by acid mixtures. Fresenius' Journal of Analytical Chemistry, 338, 253-254  https://doi.org/10.1007/BF00323018
 
Park O., Allen J.C. (1998): Preparation of Phosphopeptides Derived from αs-Casein and β-Casein Using Immobilized Glutamic Acid-Specific Endopeptidase and Characterization of Their Calcium Binding. Journal of Dairy Science, 81, 2858-2865  https://doi.org/10.3168/jds.S0022-0302(98)75845-X
 
Park O., Swaisgood H.E., Allen J.C. (1998): Calcium Binding of Phosphopeptides Derived from Hydrolysis of a αs-Casein or β-Casein Using Immobilized Trypsin. Journal of Dairy Science, 81, 2850-2857  https://doi.org/10.3168/jds.S0022-0302(98)75844-8
 
Samaraweera Himali, Zhang Wan-gang, Lee Eun Joo, Ahn Dong U (2011): Egg Yolk Phosvitin and Functional Phosphopeptides-Review. Journal of Food Science, 76, R143-R150  https://doi.org/10.1111/j.1750-3841.2011.02291.x
 
download PDF

© 2021 Czech Academy of Agricultural Sciences | Prohlášení o přístupnosti