Partial Purification and Characterization of Invertase from the Flowers of Madhuca longifolia (Mi)
M.K.B. Weerasooriya and H.P. Yatawara
Department of Chemistry
University of Kelaniya, Sri Lanka
ABSTRACT
Enzyme Invertase (b-fructofuranosidase, EC 3.2.1.26), present in the flowers of Madhuca longifolia plays an important role during the preparation of fermented Ayurvedic drugs known as 'Arishta'. This enzyme was partially purified with a yield of 11.6%, using (NH4)2SO4 fractionation, followed by gel filtration through Sepharose 4B and DEAE cellulose chromatography at pH 6.5 and 4.2. The molecular mass of the enzyme as determined by elution through Sepharose 4B gel column was found to be about 210 kDa . The enzyme showed a broad pH optimum between 4 -7. Optimum assay temperature was 37ºC and above 45ºC enzyme activity slowly declined and inactivated around 80ºC. Approximate Km value for the enzyme for sucrose is 125 mM.
INTRODUCTION
In ayurveda, fermentation processes have been used for ages to modify the potency of drugs and to extract and to preserve active constituents from medicinal plants. Therapeutic application of these fermended drugs are based on the properties of ingredients and the method of preparation. In general, these fermended ayurvedic preparations possess a moderate potency and administered in the convalescent phase of a disease (Namijoshi 1979; Caraka Samhita 1949). Once such fermented preparations known as 'Arishtas' are widely used in Sri Lanka as well as in other South Asian countries. 'Arishtas' are weak alcoholic preparations used for the treatment of a wide variety of diseases and specially, prescribed for diseases which require long term treatments. . . . . .
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