S.M.M. Mahbubur Rahman
Biotechnology and Genetic Engineering Discipline, Life Science School,
Khulna University, Khulna-9208, Bangladesh
Palash Kumar Sen
Biotechnology and Genetic Engineering Discipline, Life Science School,
Khulna University, Khulna-9208, Bangladesh
M. Fida Hasan
Bangladesh Sugarcane Research Institute, Ishurdi-6620, Pabna, Bangladesh
M.A. Samad Miah
Bangladesh Sugarcane Research Institute, Ishurdi-6620, Pabna, Bangladesh
M. Habibur Rahman
Department of Biochemistry and Molecular Biology, University of Rajshahi, Rajshahi-6205, Bangladesh
ABSTRACT
Sugarcane invertase was purified from mature sugarcane stem tissue to an electrophoretically homogenous state, by successive ion-exchange chromatography on DEAE-Cellulose and CM-Cellulose column chromatography. The molecular weight of purified invertase enzyme was determined 218 kDa on SDS-Polyacrylamide slab gel electrophoresis. When the enzyme was characterized, it was found that the sugarcane invertase is glycoprotein in nature and it contained 7.29% sugar. The height enzyme activity was found at pH 7.2 and 60°C temperature. The Km value of the enzyme was 8.0 mM. NaCl, MnCl2 increased invertase activity remarkably and KCl increased slightly while MgCl2, CaCl2 increased moderately. HgCl2 has completely and FeCl2 showed strong inhibitory effects on enzyme activity while EDTA and acetic acid have remarkable inhibitory effects on invertase activity. CuCl2, ZnCl2 and CdCl2 decreased invertase activity moderately. AgNO3, AlCl3, Glucose and Tris showed less inhibitory effects on invertase activity.
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How to cite this article
S.M.M. Mahbubur Rahman, Palash Kumar Sen, M. Fida Hasan, M.A. Samad Miah and M. Habibur Rahman, 2004. Purification and Characterization of Invertase Enzyme from Sugarcane. Pakistan Journal of Biological Sciences, 7: 340-345.
DOI: 10.3923/pjbs.2004.340.345
URL: https://scialert.net/abstract/?doi=pjbs.2004.340.345
DOI: 10.3923/pjbs.2004.340.345
URL: https://scialert.net/abstract/?doi=pjbs.2004.340.345
REFERENCES
- Krishan, H.B. and S.G. Pueppke, 1990. Cherry fruit invertae: Partial purification, characterization and activity during fruit development. J. Plant Physiol., 135: 662-666.
Direct Link - Lercari, B., 1982. Change in invertase activities during the photoperiodically induced bulb formation of onion (Allium cepa L.). Physiol. Plant., 54: 480-484.
CrossRefDirect Link - Schaffer, A.A., O. Sagee, E.E. Goldschmidt and R. Goren, 1987. Invertase and sucrose synthase activity, carbohydrate status and endogenous IAA levels during citrus leaf development. Physiol. Plant., 69: 151-155.
CrossRefDirect Link - Seitz, K. and A. Lang, 1968. Invertase activity and cell growth in lentil epicotyls. Plant Physiol., 43: 1075-1082.
Direct Link - Ishikawa, N., H. Nakagawa and N. Ogura, 1989. Isoforms of invertase in grape berries. Agric. Biol. Chem., 53: 837-838.
Direct Link - Lowry, O.H., N.J. Rosebrough, A.L. Farr and R.J. Randall, 1951. Protein measurement with the folin phenol reagent. J. Biol. Chem., 193: 265-275.
CrossRefPubMedDirect Link - Miller, G.L., 1959. Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal. Chem., 31: 426-428.
CrossRefDirect Link - Laemmli, U.K., 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, 227: 680-685.
CrossRefDirect Link - Porntaveewat, W., T. Takayanagi and K. Yokotsuka, 1994. Purification and properties of invertase from muscat bailey a grapes. J. Fermenta Bioeng., 78: 288-292.
Direct Link - Kono, Y., T.V. Vedick, L. Fitzmaurice and T.E. Mirkov, 1993. Purification, characterization and sub cellular localization of soluble invertase from tomato fruit. J. Plant Physiol., 141: 385-392.
Direct Link - Geracimo, E., B.R. John and Whitaker, 1990. Purification and partial characterization of potato (Solanum tuberosum invertase and its endogenous proteinaceous inhibitor. Plant Physiol., 92: 386-394.
Direct Link - Krishan, H.B., J.T. Blanchette and T.W. Okita, 1985. Wheat invertase, Characterization of cell wall-bounded and soluble forms. Plant Physiol., 78: 241-245.
Direct Link - Vorster, D.J. and F.C. Botha, 1998. Partial purification and characterization of sugarcane neutral invertase. Phytochemistry, 49: 651-655.
CrossRefDirect Link - Singer, M.B. and R.B. Knox, 1984. Invertase of Lilium pollen: Characterization and activity during in vitro germination. Plant Physiol., 74: 510-515.
Direct Link - Chungling, L., L. Huichiao, W. Aiyu and C. Sung, 1999. Purification and characterization of an alkaline invertase from shoots of etiolated rice seedlings. New Philologist, 142: 427-434.
Direct Link - Hatch, M.D., J.A. Sacher and T.K. Glasziou, 1963. Sugar accumulation cycle in sugar cane. I. Studies on enzymes of the cycle. Plant Physiol., 38: 338-343.
CrossRefDirect Link - Nakanishi, K., W. Wu and K. Yokotsuka, 1991. Purification and some properties of thermostable invertase from wine. J. Fermenta Bioeng., 71: 66-68.
Direct Link - Vitolo, M. and M.T. Yasuda, 1991. Effect of sucrose concentration on the invertase activity of intact yeast cells. Biotechnol. Lett. 13: 53-56.
CrossRefDirect Link - Ishimoto, M. and A. Nakamura, 1997. Purification and properties of β fructofuranosidase from Clostridium perfringens. Bioscience, Biotechno. Biochem., 6: 599-603.
Direct Link