Thermo sensitive proteins from W. coagulans Dunal fruits exhibit anti-coagulant, anti-thrombin and anti-trypsin activities
Patil Ajit Babruwahan, Jaiwal Bhimrao Vishwanath, Tak Rajesh Dattatraya
Anti-coagulants are used for the treatment and prophylaxis of thrombotic disorders. The applications of traditional anti-coagulants like warfarin and heparin produce severe complications with bleeding. In present study anti-coagulant, anti-thrombin and anti-trypsin activities were studied from crude aqueous extract (CAE) of Withania coagulans Dunal fruits. Anti-coagulant and trypsin activities of CAE against human plasma were detected by coagulation tests (aPTT, PT and TT) and dot blot assay on X-ray film. Inhibitor assays of CAE against thrombin and trypsin were carried by using synthetic substrate. Prominent delay in clotting time in coagulation tests suggested CAE exhibits an excellent anticoagulant activity. Six trypsin inhibitors (TI) isoforms from CAE were found on X-ray film. Active proteins from CAE of W. coagulans were responsible for activities. CAE showed more anti-trypsin activity (IC50 30.20µg/ml) than anti-thrombin activity (IC50 48.56 µg/ml). Various activities of CAE suggested multifunctional proteins are available in W. coagulans. The proteins of CAE were found to be thermo labial and showed more activity at 400C temperature. The proteins showed activity at wide range of pH and optimum pH was found between 7 and 8. The active proteins from W. coagulans could be applied an alternative anticoagulants for treatment of thrombotic disorders.