Abstract
A number of glycosidase enzymes have been isolated and identified in healthy fresh leaves of turnip. Myrosinase (β-thioglucosidase, EC. 3.2.3.1), disaccharase (β-fructofuranosidase, EC. 3.2.1.26), andβ-amylase (α-1,4-glucan maltohydrolase, EC 3.2.1.2) have been isolated and purified in powder form. The purification methods were salting out with ammonium sulfate, DEAE-cellulose column chromatography, hydroxylapatite batch chromatography, and gel filtration through Sephadex G-200. Four isoenzymes of myrosinase enzyme could be isolated. The most active one was purified (131.3 times) and found to have an SA of 19.7 U/mg. An enzyme causing hydrolysis of amylose and glycogen has been isolated and partially purified. It had an SA of 22.6 U/mg and 49.1-fold purification. Seven isoenzymes of disaccharase were isolated, but only one was purified (C) with SA of 1448.5 U/mg and 1316.8-fold purification.
The yield of the purified myrosinase and disaccharase enzymes was 3.68 and 0.5 mg, respectively, from 100 gm dry wt of turnip leaves. Confirmation of purity with disk electrophoresis was performed. A single sharp band was obtained for each pure enzyme by disk electrophoresis. The chromatographic analysis of the hydrolytic end product of theβ-amylase indicated the presence of maltose. This results confirm theβ-form of amylase enzyme.
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References
Jwanny, E. W., Montanari, L., and Fantozzi, P. (1993),Bioresource Technol. 43, 67.
Gaines, R. D. and Goering, K. J. (1962),Arch. Biochem. Biophys. 96, 13.
Snowden, D. R. and Gaines, R. D. (1969),Phytochemistry 8, 1649.
Chapman, G. W., Jr., Pallas, J. E., Jr., and Mendicino, J. (1972),Biochem. Biophys. Acta 276, 491.
Jwanny, E. W. and El-Sayed, S. T. (1993), in Proceeding of the 13th Egyptian Chemical Conference, April 10–13 (9th Arab Chemical Conference).
Thies, W. (1988),Fat Sci. Technol. 90, 311.
Somogyi, M. (1952),J. Biol. Chem. 195, 19.
Nelson, N. (1944),J. Biol. Chem. 153, 375.
Lowry, O. L., Rosebrough, N. J., Farr, A. L., and Randall, R. J. (1951),J. Biol. Chem. 193, 265.
Layne, E. (1957), inMethods in Enzymology, Colowick, S. P. and Kaplan, N. O., eds., vol. 111, Academic, New York, p. 147.
Wrede, F. (1941), inDie Methoden der Fermentforschung, Bamann E. and Myrback, K., eds., vol. 2, Thieme, Leipzig, p. 835.
Schwimmer, S. (1961),Acta Chem. Scand. 15, 535.
Peterson, E. A. and Soben, H. A. (1956),J. Am. Chem. Soc. 78, 751.
Tiselius, A., Jenten, S. H., and Levin, O. (1956),Arch. Biochem. Biophys. 65, 132.
Davis, B. J. (1964),Ann. NY Acad. Sci. 121, 404.
Wilson, C. M. (1959),Anal. Chem. 131, 1199.
Jorgensen, B. B. and Jorgensen, O. B. (1963),Acta Chem. Scand. 17, 1765.
Vasiléna, Z. V. (1956),Lenia 133.
MacGibbon, D. B. and Allison, R. M. (1970),Phytochemistry 9, 541.
Spencer, R. and Weston, T. J. (1966),Tobacco 163, 28.
Myrback, K. (1960). inThe Enzyme, Boyer, P. D., Lardy, H., and Myrbach, K., eds., vol. 4, Academic, New York, p. 384.
Thoma, J. A., Sprodlin, J. E., and Dygert, S. (1971),The Enzymes, Boyer, P. D., ed., Academic, New York, p. 115.
Shi-Ching, H., Jong-Ching, S., and Hsien-Yi, S. (1977), Chung-Kuo Nung Yeh Hua Hsuch Hui Chin, 15, 3. cf. C. A. 87(19), 150405 w (1977).
Okamoto, K., and Akazawa, T. (1978),Agric. Biol. Chem. 42, 1379.
Green Wood, C. T. and Milne, E. A. (1968),Adv. Carbohydr. Chem. 23, 281.
Subbaramaiah, K. and Sharma, R. (1990),Phytochemistry 29, 1417.
Abdullah, M. and French, D. (1996),Nature 210, 200.
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Jwanny, E.W., El-Sayed, S.T. Glycosidases of turnip leaf tissues. Appl Biochem Biotechnol 49, 23–34 (1994). https://doi.org/10.1007/BF02888844
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DOI: https://doi.org/10.1007/BF02888844