A PROCESS FOR THE PREPARATION OF COMMERCIALLY AND PHARMACEUTICALLY IMPORTANT ENZYMES (PECTINASE AND ACID STABLE DISACCHARIDASES) FROM THE PLANT CALOTROPIS GIGANTEA
The present invention relates to a process for the preparation of commercially and pharmaceutically important enzymes from the plant Calotropis gigantea. Calotropis gigantea Linn. (Asclepiadaceae) is a glabrous laticiferous shrub commonly known as the swallow-wort or milkweed. This shrub abounding in milky juice is found chiefly in wastelands in lower Bengal, Himalaya, Assam, Punjab, and south India and also in other south East Asian countries like Srilanka, Singapore, Malay islands, south China. The plant is a rich source of pectinase (polygalacturonase) and acid stable glucosidases (sucrase, maltase, isomaltase). The stem-bark contains (U/ 100 g of fresh tissue) 11000 ± 1000, 850 ± 20, 150 ± 20, and 80 ± 5 of polygalacturonase, sucrase, maltase and isomaltase respectively while the plant latex contains (U/ 100 g of dry latex powder) 1,70,000 ± 5000, 32,500 ± 2500, 5000 ± 500, and 2700 ± 300 of polygalacturonase, sucrase, maltase and isomaltase respectively. Both stem-bark (700 ± 50 U/100 g of fresh tissue) and latex (2000 ± 260 U/ 100 g of dry powder) are a source of acid stable (β-galactosidase (lactase). The enzyme polygalacturonase is active at a pH range of 2 - 7.5 and a temperature range of 25 - 55 C and capable of hydrolyzing crude pectin from guava, citrus peel and apple, α-glucosidases (sucrase, maltase, isomaltase) are stable at a pH range of 2.5 - 6.5 and may be used as an exogenous oral enzyme therapy for disaccharide intolerance. The enzyme composition was also capable of hydrolyzing trisaccharides like raffinose, melezitose and maltotriose. The plant exhibited slight polysaccharidase activities (U/ 100 g latex powder) like amylase (1750 ± 65), xylanase (1730 ± 150), dextranase (1630 ± 20) and inulinase (800 ± 30). Crude pectinase did not require calcium ions for its activity and stability. The milky latex after centrifugation was subjected to ammonium suphate precipitation (80 % saturation) followed by dissolving the protein pellet obtained in 0.1M ac
