M. Z. Rahman
Department of Biochemistry, Rajshahi University, Rajshahi-6205, Bangladesh.
Z. A. Saud
Department of Biochemistry, Rajshahi University, Rajshahi-6205, Bangladesh.
N. Absar
Department of Biochemistry, Rajshahi University, Rajshahi-6205, Bangladesh.
M. R. Karim
Department of Biochemistry, Rajshahi University, Rajshahi-6205, Bangladesh.
F. Hashinaga
Department of Biochemical Science and Technology,. Faculty of Agriculture,
Kagoshima University, Kagoshima 890-0065, Japan.
ABSTRACT
Two different amylase from the flesh of healthy and fruit-rot disease-affected Moringa fruit were purified by successive chromatography of the crude enzyme extract on DEAE-cellulose followed by CM-cellulose and were purified 46- and 50-fold respectively. Both the enzymes appeared to be homogeneous as judged by polyacrylamide gel electrophoresis. Molecular weights of amylases from healthy and diseased Moringa estimated by gel filtration were 59 kDa and 66.5 kDa respectively. The purified enzymes were classified as -amylase and consist of a single polypeptide chain. The amylase from healthy and diseased Moringa flesh showed the following characteristics: pH optima, 6.8 and 6.4; temperature optima, 38 °C and 40 °C; Km value, 0.28 and 0.22% for starch as substrate respectively.
PDF Citation
How to cite this article
M. Z. Rahman, Z. A. Saud, N. Absar, M. R. Karim and F. Hashinaga, 2000. A Comparative Analysis on the Purification and Characterization of Amylase from the Flesh of Healthy and Disease-affected Moringa Fruit (Moringa oleifera Lam). Pakistan Journal of Biological Sciences, 3: 586-589.
DOI: 10.3923/pjbs.2000.586.589
URL: https://scialert.net/abstract/?doi=pjbs.2000.586.589
DOI: 10.3923/pjbs.2000.586.589
URL: https://scialert.net/abstract/?doi=pjbs.2000.586.589
REFERENCES
- Andrews, P., 1965. The gel-filtration behaviour of proteins related to their molecular weights over a wide range. Biochem. J., 96: 595-606.
CrossRefDirect Link - Beck, E. and P. Ziegler, 1989. Biosynthesis and degradation of starch in higher plants. Ann. Rev. Plant Physiol. Plant Mol. Biol., 40: 95-117.
CrossRefDirect Link - Garcia, E. and F.M. Lajolo, 1988. Starch transformation during banana ripening: The amylase and glucosidase behavior. J. Food Sci., 53: 1181-1186.
CrossRefDirect 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 - MacGregor, E.A., 1993. Relationships between structure and activity in the α‐amylase family of starch‐metabolising enzymes. Starch‐Starke, 45: 232-237.
CrossRefDirect Link - Ornstein, L., 1964. Disc electrophoresis-I: Background and theory. Ann. N. Y. Acad. Sci., 121: 321-349.
CrossRefPubMedDirect Link - Svensson, B., 1994. Protein engineering in the α-amylase family: Catalytic mechanism, substrate specificity and stability. Plant Mol. Biol., 25: 141-157.
PubMedDirect Link - Weber, K. and M. Osborn, 1969. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J. Biol. Chem., 244: 4406-4412.
Direct Link