Potential of fibrinolytic protease enzyme from tissue of sand sea cucumber (Holothuria scabra) as thrombolysis agent

Abstract

Cardiovascular diseases (CVD) are still the main cause of death in modern society with thrombosis as their most common underlying pathology. The occurrence of thrombosis is characterized by thrombus formation in the circulatory system of the body causing blood clot blockage. Thrombus blocking blood vessels can be destroyed through fibrinolysis by plasmin. Commonly administered fibrinolytic drugs activate plasminogen and convert plasminogen to plasmin which degrades fibrin. However, these agents have limitations such as higher costs, shorter half-lives, allergic reactions and intravenous administration-caused bleeding. Hence, more effective and safer antithrombotic drugs are needed in anti-thrombosis therapy. This paper was intended to assess the ability of Holothuria scabra tissue fibrinolytic proteases as a thrombolytic agent based on literature studies. Summary of recent studies showed that various strong fibrinolytic enzymes could be produced from various animal tissues such as snake and earthworm, yet from sea organism’ tissues are barely reported. In this literature review, the potential of protease enzymes from tissue of H. scabra as thrombolysis agent is summarized and discussed. Among the findings is that H. scabra has the highest protein content among other sea cucumbers in Indonesia, yet the isolation of protease enzymes from H. scabra tissue has not been reported. Hence, there is high possibility in finding novel proteases, which may include serine and metallo-protease known for their thrombolysis activities.

Characteristics and Substrate Specificity of Semi-Purified Bacterial Protease ofBacillusthuringiensisHSFI-12 with Potential as Antithrombotic Agent

“…thuringiensis HSFI-12 Anticoagulant activity assay was carried out by comparing clotting times of the positive control of blood with 10% EDTA solution, negative control of blood with no treatment (as an arti cial thrombus), and samples of blood with treatments of crude and dialysate protease added in various volume variations. The use of this negative control as a thrombus model was previously reported by Hidayati et al (2021a) and Fuad et al (2021) Visual appearances of anticoagulation performance of both crude and dialysate proteases along with controls were displayed in Figure 4. The measured blood clotting time (CT) is displayed in Table 2.…”

“…The bacterial protease, however, did not show direct brinolytic activities. It was proven by the fact that the bacterial protease was unable to utilize brin as a substrate (Hidayati et al, 2021a;Hidayati et al, 2021b). This shows that the thrombolysis activ- ity of protease of strain HSFI-12 likely comes as a plasminogen activator (PA), which action will activate plasmin leading to the cleavage of brins (Akhtar et al, 2017) .…”

“…Therefore, the antithrombotic activity testing included testing of anticoagulant activity using two procedures, namely the clotting time of the Lee and White method as well as the Eustrek method by Giemsa staining is important. Furthermore, testing of brinolytic activity and characterization of the bacterial protease by looking at molecular weight and speci city for certain substrates using the zymography method is required (Hidayati et al, 2021a) .…”

In vivo antithrombotic potential of protease from Bacillus thuringiensis HSFI-12

“…7,8 Plasmin acts in the breakdown of fibrin in blood clots, but when plasmin cannot work normally, enzymatic thrombolytic agent therapy is required. [9][10][11] Commercial thrombolytic agents such as tissue plasminogen activators (tPAs), urokinase (UK), streptokinase (Sk) and nattokinase (Nk) have the disadvantages of causing gastrointestinal bleeding and allergic reactions, are expensive, low specificity and also has short half-life. Therefore, it is necessary to develop new thrombolytic agents.…”

“…Thrombolytic agents can be protease enzymes, which is known to be safe to use, one of which produce by Bacillus spp isolated from fermented products of the digestive organs of sea cucumbers (Holothuria scabra). 9 The protease produced by the strain of Bacillus spp HSFI-12 was then partially purified through ammonium sulphate precipitation followed by dialysis. It was found that in vitro blood lysis activity of the dialysate protease of strain Bacillus spp HSFI-12 was higher than its crude protease and the activity was higher than the commercial Nk as a positive control.…”