Back
 AJPS  Vol.3 No.5 , May 2012
Kinetic Studies on β-Galactosidase Isolated from Apricots (Prunus armeniaca kaisa)
Abstract: β-galactosidase was extracted from apricots (Prunus armeniaca kaisa) and characterized biochemically. Three isoenzymes (β-gal I, β-gal II and β-gal III) were obtained by salt fractionation and ionexchange and Sephadex G-100 column chromatography. β-galactosidase II showed a high ability to hy-drolyze the substrate p-nitrophenyl β-D-galactopyranoside than that of β-galactosidase I and III. The individual peaks showed charge homogeneity as revealed by single band on polyacrylamide gel. The molecular weight of β-gal I, β-gal II and β-gal III as determined by gel filtration was found to be 44.15, 34.70 and 23.71 KDa respectively. The optimum pH for the activity different isozymes was found between 4 and 6. The isoenzymes were determined to be thermally stable upto 40?C. The Km value for β-gal I was 1.85 mM which was higher than that of β-gal II (Km = 1.7), and β-gal III (Km = 1.19). The Vmax value for β-gal I, β-gal II and β-gal III was found to be 0.52, 0.70 and 0.38 μmole/min respectively.
Cite this paper: S. Gulzar and S. Amin, "Kinetic Studies on β-Galactosidase Isolated from Apricots (Prunus armeniaca kaisa)," American Journal of Plant Sciences, Vol. 3 No. 5, 2012, pp. 636-645. doi: 10.4236/ajps.2012.35077.
References

[1]   J. Gray, S. Picton, J. Shabbeer, W. Schuch and D. Grierson, “Molecular Biology of Fruit Ripening and Its Manipulation with Antisense Genes,” Plant Molecualr Biology, Vol. 19, No. 1, 1992, pp. 69-87. doi:10.1007/BF00015607

[2]   A. P. Ranwala, C. Suematsu and H. Masuda, “The Role of β-Galactosidases in the Modification of Cell Wall Components during Muskmelon Fruit Ripening,” Plant Physiology, Vol. 100, No. 3, 1992, pp. 1318-1325. doi:10.1104/pp.100.3.1318

[3]   C. S. Ross, S. Cavin, T. Wegrzyn, E. A. MacRae and R. J. Redgwell, “Apple β-Galactosidase Activity against Cell Wall Polysaccharides and Characterization of a Related cDNA Clone,” Plant Physiology, Vol. 106, No. 2, 1994, pp. 521-528. doi:10.1104/pp.106.2.521

[4]   D. A. Brummell and M. H. Harpster, “Cell Wall Metabolism in Fruit Softening and Quality and Its Manipulation in Transgenic Plants,” Plant Molecualr Biology, Vol. 47, No. 1-2, 2001, pp. 311-340. doi:10.1023/A:1010656104304

[5]   E. J. DeVeau, K. C. Gross, D. J. Huber and A. E. Watada, “Degradation and Solubilization of Pectin by β-Galactosidases Purified from Avocado Mesocarp,” Physiologia Plantarum, Vol. 87, 1993, pp. 279-285

[6]   A. T. Carey, K. Holt, S. Picard, R. Wilde, G. A. Tucker, C. R. Bird, W. Schuch and G. R. Seymour, “Tomato Exo-1 → 4-β-D-Galactanase: Isolation, Changes during Ripening in Normal and Mutant Tomato Fruit and Characterization of a Related cDNA Clone,” Plant Physiology, Vol. 108, No. 3, 1995, pp. 1099-1107. doi:10.1104/pp.108.3.1099

[7]   C. M. Carrington and R. Pressey, “β-Galactosidase II Activity in Relation to Changes in Cell Wall Galactosyl Composition during Tomato Ripening,” Journal of the American Society for Horticultural Science, Vol. 121, 1996, pp. 132-136.

[8]   Y. Hirano, Y. Tsumuraya and Y. Hashimoto, “Characterization of Spinach leaf α-L-Arabinofuranosidases and β-Galactosidases and Their Synergistic Action on an Endogenous Arabinogalactan-Protein,” Physiologia Plantarum, Vol. 92, No. 2, 1994, pp. 286-296. doi:10.1111/j.1399-3054.1994.tb05339.x

[9]   S. C. Li, J. W. Han, K. C. Chen and C. S. Chen, “Purification and Characterization of Isoforms of β-Galactosidases in Mung Bean Seedlings,” Phytochemistry, Vol. 57, No. 3, 2001, pp. 349-359. doi:10.1016/S0031-9422(01)00022-X

[10]   T. Kotake, S. Dina, T. Konishi, S. Kaneko, K. Igarashi, et al., “Molecular Cloning of a β-Galactosidase from Radish That Specifically Hydrolyzes β-1 → 3- and β-1 → 6-Galactosyl Residues of Arabinogalactan Protein,” Plant Physiology, Vol. 138, No. 3, 2005, pp. 1563-1576. doi:10.1104/pp.105.062562

[11]   P. Hrubá, D. Honys, D. Twell, V. Capková and J. Tupy, “Expression of β-Galactosidase and β-Xylosidase Genes during Microspore and Pollen Development,” Planta, Vol. 220, No. 6, 2005, pp. 931-940. doi:10.1007/s00425-004-1409-0

[12]   A. Wu and J. Liu, “Isolation of the Promoter of a Cotton Beta-Galactosidase Gene GhGal1 and Its Expression in Transgenic Tobacco Plants,” Science in China Series C: Life Sciences, Vol. 49, No. 2, 2006, pp. 105-114. doi:10.1007/s11427-006-0105-7

[13]   K. D. Golden, M. A. John and E. A. Kean, “β-Galactosidase from Coffea arabica and Its Role in Fruit Ripening,” Phytochemistry, Vol. 34, No. 2, 1993, pp. 355-360. doi:10.1016/0031-9422(93)80008-G

[14]   I. K. Kang, S. G. Suh, K. C. Gross and J. K. Byun, “N-Terminal Amino Acid Sequence of Persimmon Fruit β-Galactosidase,” Plant Physiology, Vol. 105, No. 3, 1994, pp. 975-979. doi:10.1104/pp.105.3.975

[15]   P. K. Andrews and S. Li, “Partial Purification and Characterization of β-D-Galactosidase from Sweet Cherry, a Non-Climacteric fruit,” Journal of Agricultural and Food Chemistry, Vol. 42, No. 10, 1994, pp. 2177-2182. doi:10.1021/jf00046a019

[16]   Z. M. Ali, S. Armugam and H. Lazan, “β-Galactosidase and Its Significance in Ripening Mango Fruit,” Phytochemistry, Vol. 38, No. 5, 1995, pp. 1109-1114. doi:10.1016/0031-9422(94)00804-3

[17]   D. H. Lee, S.-G. Kang, S.-G. Suh and J. K. Byun, “Purification and Characterization of a β-Galactosidase from Peach Prunus persica,” Molecules and Cells, Vol. 15, 2003, pp. 68-74.

[18]   H. Lazan, S. Y. Ng, L. Y. Goh And Z. M. Ali, “Papaya beta-Galactosidase/Galactanase Isoforms in Differential Cell Wall Hydrolysis and Fruit Softening During Ripening,” Plant Physiology and Biochemistry, Vol. 42, No. 11, 2004, pp. 847-853. doi:10.1016/j.plaphy.2004.10.007

[19]   G. S. Ross, E. A. MacRae and R. J. Redgwell, “Isolation and Activity against Specific Fruit Cell-Wall Polysaccharides,” Planta, Vol. 189, No. 4, 1993, pp. 499-506. doi:10.1007/BF00198212

[20]   D. L. Smith, D. A. Starrett and K. C. Gross, “A Gene Coding for Tomato Fruit β-Galactosidase II Is Expressed during Fruit Ripening Cloning, Characterization and Expression Pattern,” Plant Physiology, Vol. 117, No. 2, 1998, pp. 417-423. doi:10.1104/pp.117.2.417

[21]   D. L. Smith and K. C. Gross, “A Family of at Least Seven Beta-Galactosidase Genes Is Expressed during Tomato Fruit Development,” Plant Physiology, Vol. 123, No. 3, 2000, pp. 1173-1183. doi:10.1104/pp.123.3.1173

[22]   L. Trainotti, R. Spinello, A. Piovan, S. P. Spolaore and G. Casadoro, “β-Galactosidases with a Lectin-Like Domain Are Expressed in Strawberry,” Journal of Experimental Botany, Vol. 52, No. 361, 2001, pp. 1635-1645. doi:10.1093/jexbot/52.361.1635

[23]   Z. Wu and J. K. Burns, “A β-Galactosidase Gene Is Expressed during Mature Fruit Abscission of ‘Valencia’ Orange Citrus sinensis,” Journal of Experimental Botany, Vol. 55, No. 402, 2004, pp. 1483-1490. doi:10.1093/jxb/erh163

[24]   J. Zhuang, J. Su, X. Li and W. Chen, “Cloning and Expression Analysis of β-Galactosidase Gene Related to Softening of Banana Musa sp. Fruit,” Journal of Plant Physiology and Molecular Biology, Vol. 32, No. 4, 2006, pp. 411-419.

[25]   O. H. Lowry, N. J. Rosebrough, A. L. Farr and R. J. Randall, “Protein Measurement with Folin Phenol Reagent,” Journal of Biological Chemistry, Vol. 193, 1951, pp. 265-275.

[26]   R. Pressey, “β-Galactosidase in Ripening Tomatoes,” Plant Physiology, Vol. 71, No. 1, 1983, pp. 132-135. doi:10.1104/pp.71.1.132

 
 
Top