Potential Anticoagulant Activity of Trypsin Inhibitor Purified from an Isolated Marine Bacterium Oceanimonas Sp. BPMS22 and its Kinetics

Abstract

Protease inhibitors control major biological protease activities to maintain physiological homeostasis. Marine bacteria isolated from oligotrophic conditions could be taxonomically distinct, metabolically unique, and offers a wide variety of biochemicals. In the present investigation, marine sediments were screened for the potential bacteria that can produce trypsin inhibitors. A moderate halotolerant novel marine bacterial strain of Oceanimonas sp. BPMS22 was isolated, identified, and characterized. The effect of various process parameters like salt concentration, temperature, and pH was studied on the growth of the bacteria and production of trypsin inhibitor. Further, the trypsin inhibitor was purified to near homogeneity using anion exchange, size exclusion, and affinity chromatography. The purified trypsin inhibitor was found to competitively inhibit trypsin activity with an inhibition coefficient, K, of 3.44 ± 0.13 μM and second-order association rate constant, k, of 1.08 × 10 M S. The proteinaceous trypsin inhibitor had a molecular weight of approximately 30 kDa. The purified trypsin inhibitor showed anticoagulant activity on the human blood samples.

Novel Amphibian Bowman–Birk-Like Inhibitor with Antioxidant and Anticoagulant Effects Ameliorates Pancreatitis Symptoms in Mice

“…Consistently, the resonance successively increased with increasing concentrations of trypsin, indicating the direct interactions between trypsin and Ranacin, and the binding constant K D value is about 1.93 μM (Figure C). Considering that some trypsin inhibitors also inhibit the proteases in coagulation cascade system, its effect on blood coagulation was measured. , It appeared that Ranacin bound to components of plasma and prolonged the plasma recalcification time (PRT) in a dose-dependent manner (Figure D,E). Moreover, Ranacin inhibited thrombin-induced platelet aggregation and delayed the activated partial thromboplastin time (APTT) in a concentration-dependent manner as well (Figure F,G).…”

“…Male BALB/c mice at the age of 5–6 weeks ( n = 4) were injected intravenously with 0.9% saline (control), heparin (200 U/kg), or Ranacin (5 mg/kg). After 1 h, all animals were administrated with 2.0% isoflurane to anesthetize and expose their left carotid artery through a midline incision in the neck as previously reported . A 5 mm filter paper saturated with 10% (w/v) FeCl 3 solution was then applied onto the exposed carotid artery for 5 min to induce thrombus formation .…”

“…After 1 h, all animals were administrated with 2.0% isoflurane to anesthetize and expose their left carotid artery through a midline incision in the neck as previously reported. 34 A 5 mm filter paper saturated with 10% (w/v) FeCl 3 solution was then applied onto the exposed carotid artery for 5 min to induce thrombus formation. 36 A laser speckle perfusion imaging system (RWD RFLSI Pro + , Shenzhen, China) was used to monitor blood flow in animals for 20 min and the time of complete vessel occlusion was recorded.…”

Association Mechanism and Conformational Changes in Trypsin on Its Interaction with Atrazine: A Multi- Spectroscopic and Biochemical Study with Computational Approach

“…The activity of TYP was measured using BApNA as a substrate according to the reported literature [ 28 , 29 , 30 ]. The absorbance at 400 nm was recorded to evaluate the TYP activity.…”

Marine Antithrombotics

“…BPMS22 was shown to increase the clotting time from 7 to 20 min in an in vitro analysis. The extent of the observed coagulation was 33% less than the control [ 54 ]. Another study reported that a serine protease isolated from Sipunculus nudus (a marine worm) exhibited in vitro fibrinolytic activity by activating plasminogen.…”