Cardiovascular diseases (CVDs) are still the leading mortality causes in the last decades. The ailments are multifactorial characterized by excessive clot (thrombus) formation in the blood vessels. Thrombus could be degraded through thrombolysis mechanism by plasmin activated by various fibrinolysis agents including, urokinase, nattokinase, or streptokinase. However, the use of these agents is restricted by relatively high cost, short half-life, allergic reaction, and bleeding effects. The search for more economical and safer thrombolytic (clot lysis) agents are essential to address the underlying problem in CVD therapy. Among Holothurians, H. scabra has been known to have the highest protein content making it ideal substrate for protease enzymes including fibrinolytic types with clot lysis properties. However, isolation of a proteases with antithrombotic activities either from tissue or from bacteria of H. scabra has not been reported. This study aimed to screen proteolytic and clot lysis activities of crude protease extracts from tissue and bacteria isolated from fermented intestine of H. scabra. Crude protease of tissue of H. scabra was extracted by cold centrifugation, which activity was measured using UV-spectrophotometer. Crude protease of proteolytic bacteria selected by clear zones on skim milk agar (SMA) medium was isolated from nutrient broth (NB). Extracts showing proteolytic activity were subjected to gravimetry-based clot lysis test. As results, crude proteases isolated directly from the H. scabra’s tissue showed low proteolytic activities, thus were no proceed to clot lysis activity test. Crude protease extracted bacteria could show both proteolytic and clot lysis activities. In conclusion, based on this screening study, intestine of H. scabra is a rich source of proteolytic bacteria, some of them could produce crude enzymes showing competitive thrombolysis activities with Nattokinase. Thus, they appeared to have more potentials to be developed as thrombolysis agent than those directly extracted from the organism’s tissue.

Among essential treatment of cardiovascular disorders are fibrinolytic proteases. Most thrombolysis agents are fibrinolytic enzymes from group of bacterial proteases. This work reports a potent bacterium isolated from fermented intestine of H. scabra, which could produce fibrinolytic protease with high thrombolysis activity. Bacterial selection was conducted based on proteolytic and fibrinolytic activities indicated as clear zone on skim milk and fibrin agar media, respectively. Crude proteases from the selected bacterial isolates were subjected to thrombolytic activity test based on gravimetric method, which results were confirmed after 7 repetitions. As result, 4 fibrinolytic protease-producing bacterial isolates HFSI-3, HFSI-4, HFSI-5 and HFSI-8 were obtained. Among them, HFSI-5 isolate identified as Bacillus tequilensis on the basis of the 16S rRNA gene sequencing and morphological properties produced crude protease with the highest thrombolytic activity. The thrombolytic activity of crude protease produced by B. tequilensis HFSI-5 is worthy of comparing to that of standard fibrinolytic enzyme Nattokinase showing its potential as thrombolysis agent.

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.

Cardiovascular disease is among the largest contributors of premature mortality in the world caused by inflammation of blood vessels. The abnormalities provoke thrombus formation or thrombosis blocking blood vessels leading to strokes, heart attacks and coronary artery diseases. Increasing percentage of cardiovascular cases and deaths due to thrombosis has attracted researchers to look for newer thrombolysis agents. Commonly used drugs to treat thrombosis has been limited due to various side effects. Therefore, the search for sources of safer and cheaper fibrinolytic enzymes for handling thrombolysis continues. This study aimed to evaluate potentials of fibrinolytic protease of bacteria isolated from fermented seafood (sea cucumber) products as antithrombotic agents. Information was initially gathered from scientific publications identified using web-based tools including PubMed (National Center for Biotechnology Information), Science Direct (Scopus) and Web of Science (Thomson Reuters) using combinations of search terms including “fibrinolytic enzyme protease”, “endopeptidase”, “fermented food”, “sea cucumber”, “thrombolysis therapy,” “thrombolytic agent,” “fibrinolytic bacteria,” “fibrinolysis,” “protease producing bacteria,” “fibrin degradation,” “holothurians,” etc. We also searched for these terms in national and international organization technical reports and databases. This literature review reveals the prospects of fibrinolytic protease enzymes from bacteria from fermented seafood, particularly sea cucumber as novel antithrombotic agents.

The search of cost-effective bioremediation agent of hospital wastewater is critical since current methods to treat biomedical waste worldwide are still costly and not environmentally friendly. Use of hydrolytic bacteria as bioremediation agent has been known, yet it is important to ensure that they fit safety requirement. This study aimed to establish and implement a simple plate-based pathogenicity selection scoring method to determine the pathogenicity levels of 26 indigenous hydrolytic bacteria isolated from the untreated wastewater of two hospitals in Semarang, Central Java, Indonesia. Bacterial cultivations were carried out in parallel on MacConkey Agar Plate (MAP), Blood Agar Plate (BAP) and Chocolate Agar Plate (CAP) followed by molecular identification. Next, a scoring system was set based on the ability of isolates to produce violet colour on the MA and hemolysis characteristics of the bacteria on both the BAP and CAP media. Based on the scoring system, 6 out of 26 bacterial isolates mostly belong to the members of Bacillus velezensis, B. amyloliquefaciens and B. licheniformis were identified having low pathogenicity, which make them a suitable bioremediation agent of the studied hospital wastewater. As conclusion, the set plate-based pathogenicity scoring system could be a simple, yet useful and reliable tool for selecting non-pathogenic indigenous hydrolytic bacterial strains potential as a bioremediation agent.