THE INHIBITORY POTENTIALSOF SEED KERNEL METHANOL EXTRACT OF MangiferaindicaL. AnacardiaceaeAND FRACTIONS ON METALLOPROTEASE OF Echisocellatus.
THE INHIBITORY POTENTIALSOF SEED KERNEL METHANOL EXTRACT OF MangiferaindicaL. AnacardiaceaeAND FRACTIONS ON METALLOPROTEASE OF Echisocellatus.
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Date
2016-02
Authors
OKORO, Peculiar Nwanyibunwa
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Abstract
Mangiferaindica seed kernel methanol extract and its solvent fractions (ethyl acetate, n-butanol, and aqueous) were tested for their anti venom activities against the metalloprotease activity of Echisocellatusvenom in vitro and in vivo.The ethyl acetate fraction was found to be the most active. To determine the mechanism of inhibition, metalloprotease was partially purified from the venom of Echisocellatuswith a yield of 71%, a purification fold of 2.63 and a specific activity of 19.00 μmol/min/mg protein. The enzyme appeared as a band on SDS-PAGE with a molecular weight of 23kDa. The kinetic properties of the enzyme showed a Km of 0.31 mg mL-1 and a Vmax of 9.09 μmol min-1. When the enzyme was incubated with the extract, the kinetic study revealed a mixed non-competitive pattern of inhibition with a Km values of 0.56 and 1.11 mg mL-1 and Vmaxvalues of 6.67 and 4.17μmol min-1 for 5 and 20% inhibitor concentrations respectively. An estimated Ki value of 0.168 mg mL-1 was obtained from a secondary plot demonstrating that the extract has a high affinity for the partially purified enzyme. Thus, could serve as an effective inhibitor. The most active ethyl acetate fraction was used to determinethe in vivo anti venom against metalloprotease induced toxic effect on blood coagulation system (bleeding and clotting time) and some hematological parameters (Packed Cell Volume, Hemoglobin, White Blood Cell and Total Protein count) in mice. The results obtained from the in vivo studies showed a significant increase in the bleeding and clotting time of the mice treated with the venom alone when compared with that of the control. For in the Mice treated with venom and the extract, increase in bleeding and clotting time was observed to be significantly (p<0.05) reduced. Also, the results obtained forhematological parameters showed a significant reduction, except for the WBC
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count which was slightly elevated in the Mice treated with venom alone when compared to the control. Normalcy was significantly restored in the groups treated with the venom and the active fraction. This suggests that the extract has some inhibitory effect in vivo against the action of Echisocellatusvenom. The relevance of these findings would be of importance in the management of snake bite envenomation and the development of anti venom.
Description
A DISSERTATION SUBMITTED TO THE SCHOOL OF POSTGRADUATE STUDIES, AHMADU BELLO UNIVERSITY ZARIA, NIGERIA, IN PARTIAL FULFILLMENT FOR THE AWARD OF MASTER OF SCIENCE DEGREE IN BIOCHEMISTRY
DEPARTMENT OF BIOCHEMISTRY
FACULTY OF SCIENCE
AHMADU BELLO UNIVERSITY,
ZARIA
Keywords
INHIBITORY POTENTIALS,, SEED KERNEL,, METHANOL EXTRACT,, MangiferaindicaL. Anacardiaceae,, FRACTIONS,, METALLOPROTEASE,, Echisocellatus.