Summary
Seeds, flowers and leaves of Onopordum turcicum were found to contain proteolytic enzymes able to coagulate milk. Extraction, concentration and identification of the operational parameters affecting the activity of the enzyme complex were followed by partial purification steps involving gel-filtration and ion-exchange chromatography. Milk clotting activity of the enzyme complex was tested in several steps of its purification and an increase of almost 200 fold was obtained. Molecular weight of the proteolytic enzyme fraction having the maximum activity was determined to be about 19000–24000. Isoelectric point (pI) of the enzyme complex with maximum activity was estimated to be in the range 3.3–3.7.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Aworth, O. C., Müller, H. G. (1987) Food Chem., 26: 71.
Barbosa, M., Corradini, C, Battistotti, B. (1981) Scienza e Tecnica Lattiero-Casearia 32: 4, 203–221.
Carlson, A., Hill, C. G., Olson, N.F. (1987.a) Biotech.& Bioeng. 29: 582
Carlson, A., Hill, C.G., Olson, N.F.(1987.b) Biotech.& Bioeng. 29: 590.
Chen, C. H., Zall, R. R. (1986) J. Food Sci. 51: 3, 815.
Carr, P.W., Bowers, L.D., (1980) Immobilized Enzymes in Analytical and Clinical Chemistry, pp. 49, John Wiley and Sons, USA.
Cusack, M., Stephen, G., Powls, R., Beynon, R.L. (1991) Biochem. J., 274: 231.
Dalgeish, D. G. (1983) J. Dairy Res. 50: 531.
Exposito, J.M., Gordillo, C.M., Marino, J.I., Iglesias, J. (1991) Die Nahrung, 35: 2, 139.
Fevereiri, P., Cabral, J. M., Fonseca, M.M.R., Novais, J.M., Pais, M.S. (1986) Biotechnol. Lett. 8: 19–24.
Fox, P. F. (1982) Biochem. Soc. Trans. 10: 282.
Kimmel, J.R., Smith, E. L. (1954) J. Biol. Chem. 207: 515.
Kramer, D., Whitaker, J.R. (1964) J. Biol. Chem. 239: 7, 2178.
Kuwabara, T., Hashimoto, Y. (1990) Plant Cell Physiol, 31: 5, 581.
Löffler, A. (1986) Food Technology Jan: 63.
Prasad, K.M.R., Virupaksha, T.K. (1990) Phytochemistry, 29: 6, 1763.
Rodney, F. B. (1986) Modern Experimental Biochemistry, pp. 75–96. Addison Wesley Pub. Co., Canada.
Rowland, S. J. (1938) J. Dairy Res., 9: 42–46.
Scopes, R. K. (1986) Protein Purification, Principles and Practice, pp. 21–34, 39–60, 67–145, 151–182, Springer, Verlag.
Walstra, P., Jenness, R. (1984) Dairy Chemistry and Physics, pp.107, John Wiley and Sons, New York.
Wieland, H. (1972) Enzymes in Food Processing and Products, Noyes Data Corporation, USA, 102–109.
Whitaker, J.R. (1959) Food Technology 13: 86.
Whitaker, J. R. (1972) Principles of Enzymology for the Food Scientist, pp. 66–98, Marcel Decker Inc., New York.
Whitaker, J. R. (1963) Analytical Chem., 35: 12, 1950–1953.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Tamer, I.M. Identification and partial purification of a novel milk clotting enzyme from Onopordum turcicum . Biotechnol Lett 15, 427–432 (1993). https://doi.org/10.1007/BF00128289
Issue Date:
DOI: https://doi.org/10.1007/BF00128289