Isolation, Characterization and Purification of α-Galactosidase from Peas
Keywords:
α-galactosidase, Dry peas, DEAE Cellulose column chromatography, SDS-PAGE, DNSA, ImmobilizationAbstract
In India, the peas are cultivated on a large scale. The α-galactosidase enzyme has principle applications in food industries and in the medical field. The α-galactosidase enzyme was purified from dry peas (Pisum sativum). The purification steps include acetone precipitation and DEAE-cellulose column chromatography. The enzyme has maximum activity at 40°C and optimum pH=7.5. The molecular weight of the enzyme was determined by using SDS-PAGE analysis and found to be 110 kDa. The purified enzyme was immobilized by sodium alginate and it was observed that the immobilized enzyme showed an increase in thermal tolerance.
Downloads
References
Mathew, C.D. & Balasubramaniam, K. (1987). Mechanism of action of α-galactosidase. Phytochemistry, 26(5): 1299–1300. https://doi.org/10.1016/S0031-9422(00)81798-7.
Balasubramaniam, K. & Mathew, C.D. (1986). Purification of α-galactosidase from coconut. Phytochemistry, 25(8): 1819–1821. https://doi.org/10.1016/S0031-9422(00)81155-3.
Sozzi, G.O., Cascone, O. & Fraschina, A.A. (1996). Effect of a high-temperature stress on endo-β-mannanase and α- and β-galactosidase activities during tomato fruit ripening. Postharvest Biol. Technol., 9(1): 49–63. https://doi.org/10.1016/0925-5214(96)00014-2.
Kang, H.-C. & Lee, S.-H. (2001). Characteristics of an α-galactosidase associated with grape flesh. Phytochemistry, 58(2): 213–219. https://doi.org/10.1016/S0031-9422(01)00207-2.
Kim, W.D., Kobayashi, O., Kaneko, S., Sakakibara, Y., Park, G.G., Kusakabe, I., Tanaka, H. & Kobayashi, H. (2002). α-Galactosidase from cultured rice (Oryza sativa L. var. Nipponbare) cells. Phytochemistry, 61(6): 621–630. https://doi.org/10.1016/s0031-9422(02)00368-0.
Shivanna, B.D., Ramakrishna, M. & Ramadoss, C.S. (1990). Purification and properties of the anionic form of α-galactosidase from germinating guar (Cyamopsis tetragonolobus). Plant Sci., 72(2): 173–180. https://doi.org/10.1016/0168-9452(90)90080-8.
Plant, A.R. & Moore, K.G. (1982). α-D-mannosidase and α-D-galactosidase from protein bodies of Lupinus angustifolius cotyledons. Phytochemistry, 21(5): 985–989. https://doi.org/10.1016/S0031-9422(00)82402-4.
Plummer, D.T. (1988). An Introduction to Practical Biochemistry. 3rd edition, Tata McGraw Hill Publishing Company Limited, New Delhi. p. 159-160.
Miller, G.L. (1959). Use of Dinitrosalicylic Acid Reagent for Determination of Reducing Sugar. Anal. Chem., 31(3): 426–428. https://doi.org/10.1021/ac60147a030.
de Fátima Viana, S., Guimarães, V.M., José, I.C., e Oliveira, M.G.D.A., Costa, N.M.B., de Barros, E.G., Moreira, M.A. & de Rezende, S.T. (2005). Hydrolysis of oligosaccharides in soybean flour by soybean α-galactosidase. Food Chem., 93(4): 665-670. https://doi.org/10.1016/j.foodchem.2004.09.045.
Laemmli, U.K. (1970). Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4. Nature, 227(5259): 680–685. https://doi.org/10.1038/227680a0.
Kourkoutas, Y., Bekatorou, A., Banat, I.M., Marchant, R. & Koutinas, A.A. (2004). Immobilization technologies and support materials suitable in alcohol beverages production: a review. Food Microbiol., 21(4): 377–397. https://doi.org/10.1016/j.fm.2003.10.005.
Gopalan, C., Rama Sastri, B.V. & Balasubramanian, S.C. (1971). Nutritive value of Indian foods. Hyderabad: National Institute of Nutrition, Indian Council of Medical Research.
Downloads
Published
How to Cite
Issue
Section
License
Copyright (c) 2012 The author(s) retains the copyright of this article.
This work is licensed under a Creative Commons Attribution 4.0 International License.
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.