Exploration of bacteria isolated from "rusip" fermented tissue of sand sea cucumber Holothuria scabra with fibrinolytic, anticoagulant and antiplatelet activities

Sand sea cucumber (Holothuria scabra) is an important sea commodity containing high levels of protein. Fibrinolytic proteases are extracellularly produced from certain bacteria isolated from fermented, protein-rich substrates. The importance of fibrinolytic proteases is largely attributed to their applicability as anticoagulants and antiplatelets, mainly used in therapy for cardiovascular disease, the number one cause of death in the world. This study aimed to obtain new sources of fibrinolytic protease from bacteria isolated from the fermented tissue of H. scabra captured in Kodek Bay of North Lombok Island, Indonesia. To stimulate the growth of proteolytic bacteria prior to the isolation process, the tissue of the sand sea cucumber was first fermented as "rusip", a fish-based traditional fermentation product from Kalimantan. The selection of fibrinolytic protease producing bacteria was carried out using Skim Milk Agar (SMA) and Fibrin Agar (FA) media followed by crude protease extraction using Skim Milk Broth (SMB). The obtained crude protease extracts were then subjected to anticoagulant and antiplatelet activity tests using Lee-White, Eustrek and Clot-lysis methods, respectively. Bacterial isolates showing fibrinolytic, anticoagulant and antiplatelet activities were identified using Polymerase Chain Reaction (PCR) targeting their 16S rRNA gene. As results, 3 out of 17 bacterial isolates from the fermented tissue of H. scabra identified as Staphylococcus hominis, Bacillus aryabhattai and Staphylococcus saprophyticus could show proteolytic and fibrinolytic activities. Further test results demonstrated specific characteristics of bacterial crude extracts as both antiplatelet and anticoagulant agents. In conclusion, the fermented tissue of H. scabra is a valuable source of bacteria producing fibrinolytic proteases with anticoagulant and antiplatelet activities.