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.
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