Abstract

COMPARATIVE STUDIES ON ANTICOAGULANT ENZYME FROM PLANT LATEX SAMPLES

Ashwin Mishra, Anusha P., Lakshmikantha M. R., Nagamani T. S. and Mahesh M.*

ABSTRACT

Plant latex is composed of organic, inorganic and hydrolytical enzymes and is a source of proteases which belongs to cysteine or serine super family. It also exhibits many pharmacological properties used in folk medicines and are involved in haemostasis. In present study, latex collected from eleven plant samples such as Calotropis gigantea, Croton bonplandianum, Ficus benghalensis, Ficus carica, Synadenium umbellatum, Nerium oleander, Artocarpus heterophyllus, Manilkara zapota, Ficus religiosa, Carica papaya and Plumeria acutifolia to evaluate the best source for anticoagulant enzyme. Among the eleven samples three samples gave the highest anticoagulant activity confirmed by blood clotting and milk clotting activity. These three samples were subjected for purification by 3 step method such as salt precipitation & dialysis, ion exchange chromatography and gel filtration chromatography. The fold purification and percentage yield of plants are 6.33 and 34.04 for Croton banplandianum, 54.83 and 86.69 for Carica papaya, 22.35 and 66.77 for plumeria acutifolia respectively. These enzymes show optimum pH at 7, Temperature at 40°C, Incubation time for 10 min and Substrate concentration activity increases till 9 mM. The molecular weight of purified protease enzyme of Croton bonplandianum, Carica papaya and Plumeria acutifolia are ~35kDa, ~25kDa and ~34kDa respectively detected by SDS PAGE. Purified enzymes subjected for immobolization, the stability of enzymes of Croton bonplandiaum and Plumeria acutifolia found to be high in Polyurethane method and that of Carica papaya found to be high in Polyacrylamide method. In APTT test, clotting time of citrated plasma for control is 32±2 sec and there was no clotting for Croton bonplandiaum, Carica papaya and Plumeria acutifolia.

Keywords: Plant latex, Anticoagulant, Chromatography, SDS PAGE, Blood clotting.