Clotting Activities of Partially Purified Extracts of Moringa oleifera L. on Dromedary Camel Milk
June 1, 2017
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Abstract: Processing camel milk into shelf stable value added milk products is not yet well developed.
Unlike the milk of cows and small ruminants, camel milk does not readily coagulate by rennet due to its
inherent properties. Therfore, experiments were conducted to evaluate the clotting activities of partially
purified Moringa oleifera extract on camel milk and to identify the optimum pH, temperature and
concentration of the partially purified extract that would result in strong curd of camel milk. These
included three temperature points (55, 60 and 65°C), three pH values(4.5, 5 and 5.5) and five levels of
partially purified Moringa oleifera extracts obtained from seed and leaf samples (0, 10, 20, 30 and 40%). The
results revealed that temperature, pH, and concentrations of partially purified seed and leaf extracts of
Moringa oleifera had significant (p<0.05) effect on the clotting activities of camel milk. The highest camel
milk clotting activity and curd firmness were observed at pH 5, temperature of 65°C and partially purified
extract concentration of 10% for both seeds and leaves, while the lowest values were recorded at pH 5.5,
temperature of 55°C and a partially purified extract concentration of 40%, respectively. An increase in
camel milk clotting activity was observed with a decrease in milk pH from 5.5 to 4.5. Camel milk clotting
activities increased with increasing temperature. However, it decreased with increase in partially purified
extract concentration for both seed and leaf extracts. Therefore, the capability of the partially purified
extract of Moringa oleifera seeds to coagulate camel milk and to form firm curd combined with its high
ratio of milk clotting to proteolytic activity could make it a useful rennet substitute in the dairy industry.
Keywords: Ammonium sulfate fractionation; Curd firmness; Milk-clotting activity; Partially purified
extract
Terefe, M. A., A. Kebede, and M. Kebede. “Clotting Activities of Partially Purified Extracts of Moringa Oleifera L. On Dromedary Camel Milk”. East African Journal of Sciences, vol. 11, no. 2, June 2017, pp. 117-28, doi:10.20372/eajs.v11i2.407.
References
Adedeji, B. A. and Nwanekezi, E. C. 1987. Effect of partial substitution of cow's milk with soymilk on Yield and Quality attribute of West African cheese. International Journal of Food Science and Technology, 22: 135-138.
Ahmed, N. A. A. and El Zubeir, I. E. M. 2011. Effect of salt level on some physical and chemical properties and acceptability of camel milk cheese. Journal of Camelid Science, 4: 40-48.
Amna E. T., Ebraheem A. S. A., , Abdelmoez M. A., Fatima J. Khider, F.J., Mohamed, O. E. and Ahmed, M. I. A. 2014. Partial purification of milk-clotting enzyme from the seeds of Moringa oleifera. Journal microbiology,bitechnoloy and food science, 4(1): 58-62.
Antao, C. M. and Malcata, F. X. 2005. Plant serine proteases: biochemical, physiological and molecular features. Plant Physiology and Biochemistry, 43: 637–650.
Anusha, R., Singh, M. K. and Bindhu O. S. 2014. Characterization of potential milk coagulants from Calotropis gigantean plant parts and their hydrolytic pattern of bovine casein. European Food Research and technology, 238: 997-1006.
Attia, H., Kherouatou, N., Nasri M, Khorchani T. 2000. Characterization of the dromedary milk casein micelle and study of its changes during acidification. Dairy Science Technology, 80 (5): 503-515.
Beka, R. G., Krier, F., Botquin, M., Guiama, V. D., Donn, P., Libouga, D. G., Mbofung, C. M., Dimitrov, K., Slomianny, M.-C., Guillochon, D. and Vercaigne-Marko, D. 2014. Characterisation of a milk-clotting extract from Balanites aegyptiaca fruit pulp. International Dairy Journal, 34: 25-31.
Egito, A. S., Girardet, J. M., Laguna, L. E., Poirson, C., Mollé, D., Miclo, L. 2007. Milk-clotting activity of enzyme extracts from sunflower and Albizia seeds and specific hydrolysis of bovine k-casein. International Dairy Journal, 17: 816-825.
Fahey, J. W. 2005. Moringa oleifera: A Review of the Medical Evidence for Its Nutritional, Therapeutic, and Prophylactic Properties. Part 1.Trees for Life International Journal of Crude Drug Research, 26 (1): 29-32.
FAO (Food and Agriculture Organization of the United Nations). 1990. The technology of traditional milk products in developing countries. FAO Animal Production and Health Paper 85. FAO, Rome, Italy. 333pp.
FAO (Food and Agriculture Organization of the United Nations). 2001. The Technology of Making Cheese from Camel Milk (Camelus dromedarius). Animal Production and Health Paper, Vol. 113. Rome, Italy.
Farah, Z. and Bachmann, M. R. 1987. Rennet coagulation properties of camel milk. Milchwissenschaft, 42: 689-692.
Foltmann, B. and Quist, B. K. 1998. On the enzymatic and the coagulation stages of the renneting process. In Proceedings of the 15th, International Dairy Congress, 2: 655-661 London
Gomez., K. A. and Gomez, A. A 1984. Statistical Procedures for Agricultural Research. 2nd ed. John Wiley and Sons Inc., New York.
Guiama, V. D., Libouga, D. G., Ngah, E. and Mbofung, C. M. 2010. Milk clotting activity of berries extracts from nine Solanum plants. African Journal Biotechnology, 9(25): 3911-3918.
Hashim, M. M., Mingsheng, D., Iqbal, M. F. and Xiaohong, C. 2011. Ginger rhizome as a potential source of milk coagulating cysteine protease. Phytochemistry, 72: 458-464. http://dx.doi.org/10.1016/j.phytochem.2010.12.002.
IDP. 2006. Integrated development plan of Dire Dawa Administration, 2006/7-2010/2011. Berhanena Selam Printing press. 92 pp.
Jacob, M., Jaros, D. and Rohm, H. 2011. Recent advances in milk clotting enzymes. International Journal of Dairy Technology, 64 (1): 14 33.
Khan, B. B. and Iqbal, A. 2001. Production and composition of camel milk: Review. Pakistan Journal of Agricultural Sciences, 38: 3-4.
Khan, H., Athar, I. H. and Aslam, M. 2004. Evaluation of cheese prepared by processing camel milk. Pakistan Journal Zoology, 36: 323-326.
Kherouatou, N., Moncef, N. and Attia, H. 2003. A study of the dromedary casein micelle and its changes during acidification. Brazil Journal Food, 34: 32-38.
Lowry, O. H., Rosebrough, N. J., Farr, A. L. and Randall, R. J. 1951. Protein measurement with the Folin phenol reagent. Journal of Biological Chemistry, 193: 265-275
Macedo, I. Q., Faro, C. J. and Pires, E. M. 1996. Caseinolytic Specificity Of Cardosin, an Aspartic Protease from the Cardoon Cynaracardunculus L. Action on Bovine β- and α -Casein and Comparison with Chymosin. Journal of Agricultural and Food Chemistry, 44: 42-47.
Maryam Salami, Ali Akbar Moosavi-Movahedi, Faezeh Moosavi-Movahedi, Mohammad Reza Ehsani, Reza Yousefi, Mohammad Farhadi, Amir Niasari-Naslaji, Ali Akbar Saboury, Jean-Marc Chobert and Thomas Haertlé. 2011. Biological activity of camel milk casein following enzymatic digestion. Journal of Dairy Research, 78: 471–478. .
Mazorra-Manzano, M. A., Perea-Gutiérrez, T. C., Lugo-Sánchez, M. E., Ramirez-Suarez, J. C., Torres-Llanez, M. J., González-Córdova, A. F. and Vallejo-Cordoba, B. 2013. Comparison of the milk-clotting properties of three plant extracts. Food Chemistry, 141: 1902–1907.
Mehaia, M. A. 2006. Manufacture of fresh soft white cheese (Domiati-type) from dromedary camels milk using ultra filtration process. Journal of Food Technology, 4: 206-212.
Mohamed Ahmed, I. A., Morishima, I., Babiker, E. E. and Mori, N. 2009a. Characterization of partially purified milk-clotting enzyme from Solanumdubium Fresen seeds. Food Chemistry, 116 (2): 395–400.
Mohamed Ahmed, I. A., Morishima, I., Babiker, E. E. and Mori, N. 2009b. Dubiumin, a chymotrypsin-like serine protease from the seeds of Solanum dubium Fresen. Phytochemistry, 70 (4):483-491.
Mohamed Ahmed, I.A., Babiker, E.E. and MORI N. 2010. pH stability and influence of salts on activity of a milk-clotting enzyme from Solanumdubium seeds and its enzymatic action on bovine caseins. LWT of Food Science and Technology, 43: 759-764.
Najera, A. I., De Renobales, M. and Barron, L. J. R. 2003. Effects of pH, temperature, CaCl2 and enzyme concentrations on the rennet-clotting properties of milk: a multi factorial study. Food Chemistry, 80: 345-352.
Nestor, G. -M., Rubi, C.-G. D. and Hector, J. -C. 2012. Exploring the Milk-Clotting Properties of a Plant Coagulant from the Berries of S. elaeagnifolium var. Cavanilles. Journal of Food Science, 71 (1): C89-C94.
Pontual, E. V., Carvalho, B. E. A., Bezerra, R. S., Coelho, L. C. B. B., Napoleao, T. H. and Paiva, P. M. G. 2012. Caseinolytic and milk-clotting activities from Moringa oleifera flowers. Food Chemistry, 135: 1848-1854.
Roseiro, L. B., Barbosa, M. M., Ames, J. and Wilbey, R. 2003. Cheese making with vegetable coagulants; the use of Cynaracardunculus L. for the production of ovine milk cheeses. International Journal of Dairy Technology, 56: 76 - 85.
Salvador, A. and Fiszman, S. M. 2004. Textural and sensory characteristics of whole and skimmed flavoured set-type yoghurt during long storage.Journal of dairy Science, 87 (12): 4033-4041.
Silva, S. V. and Malcata, F. X. 2005. Studies pertaining to coagulant and proteolytic activities of plant proteases from Cynara cardunculus. Food Chemistry, 89: 19-26.
Soledad, C., Sidrach, L., Lo´pez-Molin, D. and Rodrıguez-Lopez, J. N. 2007. Characterization of the milk clotting properties of extracts from Animal Production and Health Paper Vol. 113.Artichoke (Cynarascolymus L.) flowers. International Dairy Journal, 17: 1393-1400.
Verissimo, P., Esteves, C., Faro, C. and Pires, E. 1995. The vegetable rennet of Cynaracardunculus L. contains two proteinases with chymosin and pepsin-like specificities. Biotechnology Letters, 17: 614–645.
Yonas Hailu, Eyassu Seifu and Zelalem Yilma. 2014. Clotting activity of camel milk using crude extracts of ginger (Zingiber officinale) rhizome. African Journal of Food Science Technology, 5 (3): 90-95.
Adedeji, B. A. and Nwanekezi, E. C. 1987. Effect of partial substitution of cow's milk with soymilk on Yield and Quality attribute of West African cheese. International Journal of Food Science and Technology, 22: 135-138.
Ahmed, N. A. A. and El Zubeir, I. E. M. 2011. Effect of salt level on some physical and chemical properties and acceptability of camel milk cheese. Journal of Camelid Science, 4: 40-48.
Amna E. T., Ebraheem A. S. A., , Abdelmoez M. A., Fatima J. Khider, F.J., Mohamed, O. E. and Ahmed, M. I. A. 2014. Partial purification of milk-clotting enzyme from the seeds of Moringa oleifera. Journal microbiology,bitechnoloy and food science, 4(1): 58-62.
Antao, C. M. and Malcata, F. X. 2005. Plant serine proteases: biochemical, physiological and molecular features. Plant Physiology and Biochemistry, 43: 637–650.
Anusha, R., Singh, M. K. and Bindhu O. S. 2014. Characterization of potential milk coagulants from Calotropis gigantean plant parts and their hydrolytic pattern of bovine casein. European Food Research and technology, 238: 997-1006.
Attia, H., Kherouatou, N., Nasri M, Khorchani T. 2000. Characterization of the dromedary milk casein micelle and study of its changes during acidification. Dairy Science Technology, 80 (5): 503-515.
Beka, R. G., Krier, F., Botquin, M., Guiama, V. D., Donn, P., Libouga, D. G., Mbofung, C. M., Dimitrov, K., Slomianny, M.-C., Guillochon, D. and Vercaigne-Marko, D. 2014. Characterisation of a milk-clotting extract from Balanites aegyptiaca fruit pulp. International Dairy Journal, 34: 25-31.
Egito, A. S., Girardet, J. M., Laguna, L. E., Poirson, C., Mollé, D., Miclo, L. 2007. Milk-clotting activity of enzyme extracts from sunflower and Albizia seeds and specific hydrolysis of bovine k-casein. International Dairy Journal, 17: 816-825.
Fahey, J. W. 2005. Moringa oleifera: A Review of the Medical Evidence for Its Nutritional, Therapeutic, and Prophylactic Properties. Part 1.Trees for Life International Journal of Crude Drug Research, 26 (1): 29-32.
FAO (Food and Agriculture Organization of the United Nations). 1990. The technology of traditional milk products in developing countries. FAO Animal Production and Health Paper 85. FAO, Rome, Italy. 333pp.
FAO (Food and Agriculture Organization of the United Nations). 2001. The Technology of Making Cheese from Camel Milk (Camelus dromedarius). Animal Production and Health Paper, Vol. 113. Rome, Italy.
Farah, Z. and Bachmann, M. R. 1987. Rennet coagulation properties of camel milk. Milchwissenschaft, 42: 689-692.
Foltmann, B. and Quist, B. K. 1998. On the enzymatic and the coagulation stages of the renneting process. In Proceedings of the 15th, International Dairy Congress, 2: 655-661 London
Gomez., K. A. and Gomez, A. A 1984. Statistical Procedures for Agricultural Research. 2nd ed. John Wiley and Sons Inc., New York.
Guiama, V. D., Libouga, D. G., Ngah, E. and Mbofung, C. M. 2010. Milk clotting activity of berries extracts from nine Solanum plants. African Journal Biotechnology, 9(25): 3911-3918.
Hashim, M. M., Mingsheng, D., Iqbal, M. F. and Xiaohong, C. 2011. Ginger rhizome as a potential source of milk coagulating cysteine protease. Phytochemistry, 72: 458-464. http://dx.doi.org/10.1016/j.phytochem.2010.12.002.
IDP. 2006. Integrated development plan of Dire Dawa Administration, 2006/7-2010/2011. Berhanena Selam Printing press. 92 pp.
Jacob, M., Jaros, D. and Rohm, H. 2011. Recent advances in milk clotting enzymes. International Journal of Dairy Technology, 64 (1): 14 33.
Khan, B. B. and Iqbal, A. 2001. Production and composition of camel milk: Review. Pakistan Journal of Agricultural Sciences, 38: 3-4.
Khan, H., Athar, I. H. and Aslam, M. 2004. Evaluation of cheese prepared by processing camel milk. Pakistan Journal Zoology, 36: 323-326.
Kherouatou, N., Moncef, N. and Attia, H. 2003. A study of the dromedary casein micelle and its changes during acidification. Brazil Journal Food, 34: 32-38.
Lowry, O. H., Rosebrough, N. J., Farr, A. L. and Randall, R. J. 1951. Protein measurement with the Folin phenol reagent. Journal of Biological Chemistry, 193: 265-275
Macedo, I. Q., Faro, C. J. and Pires, E. M. 1996. Caseinolytic Specificity Of Cardosin, an Aspartic Protease from the Cardoon Cynaracardunculus L. Action on Bovine β- and α -Casein and Comparison with Chymosin. Journal of Agricultural and Food Chemistry, 44: 42-47.
Maryam Salami, Ali Akbar Moosavi-Movahedi, Faezeh Moosavi-Movahedi, Mohammad Reza Ehsani, Reza Yousefi, Mohammad Farhadi, Amir Niasari-Naslaji, Ali Akbar Saboury, Jean-Marc Chobert and Thomas Haertlé. 2011. Biological activity of camel milk casein following enzymatic digestion. Journal of Dairy Research, 78: 471–478. .
Mazorra-Manzano, M. A., Perea-Gutiérrez, T. C., Lugo-Sánchez, M. E., Ramirez-Suarez, J. C., Torres-Llanez, M. J., González-Córdova, A. F. and Vallejo-Cordoba, B. 2013. Comparison of the milk-clotting properties of three plant extracts. Food Chemistry, 141: 1902–1907.
Mehaia, M. A. 2006. Manufacture of fresh soft white cheese (Domiati-type) from dromedary camels milk using ultra filtration process. Journal of Food Technology, 4: 206-212.
Mohamed Ahmed, I. A., Morishima, I., Babiker, E. E. and Mori, N. 2009a. Characterization of partially purified milk-clotting enzyme from Solanumdubium Fresen seeds. Food Chemistry, 116 (2): 395–400.
Mohamed Ahmed, I. A., Morishima, I., Babiker, E. E. and Mori, N. 2009b. Dubiumin, a chymotrypsin-like serine protease from the seeds of Solanum dubium Fresen. Phytochemistry, 70 (4):483-491.
Mohamed Ahmed, I.A., Babiker, E.E. and MORI N. 2010. pH stability and influence of salts on activity of a milk-clotting enzyme from Solanumdubium seeds and its enzymatic action on bovine caseins. LWT of Food Science and Technology, 43: 759-764.
Najera, A. I., De Renobales, M. and Barron, L. J. R. 2003. Effects of pH, temperature, CaCl2 and enzyme concentrations on the rennet-clotting properties of milk: a multi factorial study. Food Chemistry, 80: 345-352.
Nestor, G. -M., Rubi, C.-G. D. and Hector, J. -C. 2012. Exploring the Milk-Clotting Properties of a Plant Coagulant from the Berries of S. elaeagnifolium var. Cavanilles. Journal of Food Science, 71 (1): C89-C94.
Pontual, E. V., Carvalho, B. E. A., Bezerra, R. S., Coelho, L. C. B. B., Napoleao, T. H. and Paiva, P. M. G. 2012. Caseinolytic and milk-clotting activities from Moringa oleifera flowers. Food Chemistry, 135: 1848-1854.
Roseiro, L. B., Barbosa, M. M., Ames, J. and Wilbey, R. 2003. Cheese making with vegetable coagulants; the use of Cynaracardunculus L. for the production of ovine milk cheeses. International Journal of Dairy Technology, 56: 76 - 85.
Salvador, A. and Fiszman, S. M. 2004. Textural and sensory characteristics of whole and skimmed flavoured set-type yoghurt during long storage.Journal of dairy Science, 87 (12): 4033-4041.
Silva, S. V. and Malcata, F. X. 2005. Studies pertaining to coagulant and proteolytic activities of plant proteases from Cynara cardunculus. Food Chemistry, 89: 19-26.
Soledad, C., Sidrach, L., Lo´pez-Molin, D. and Rodrıguez-Lopez, J. N. 2007. Characterization of the milk clotting properties of extracts from Animal Production and Health Paper Vol. 113.Artichoke (Cynarascolymus L.) flowers. International Dairy Journal, 17: 1393-1400.
Verissimo, P., Esteves, C., Faro, C. and Pires, E. 1995. The vegetable rennet of Cynaracardunculus L. contains two proteinases with chymosin and pepsin-like specificities. Biotechnology Letters, 17: 614–645.
Yonas Hailu, Eyassu Seifu and Zelalem Yilma. 2014. Clotting activity of camel milk using crude extracts of ginger (Zingiber officinale) rhizome. African Journal of Food Science Technology, 5 (3): 90-95.
Copyright (c) 2018 Mezgebu Abate Terefe, Ameha Kebede, Misrak Kebede
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