Isolation and Identification Fibrinolytic Protease Endophytic Fungi from Hibiscus Leaves in Shah Alam

Fibrin degradation is an important part in prevention or treatment of intravascular thrombosis and cardiovascular diseases. Plasmin like fibrinolytic enzymes has given new hope to patient with cardiovascular diseases by treating fibrin aggregation related diseases with traditional plasminogen activator which have many side effects. Various researches involving wide range of sources for production of fibrinolytic proteases, from bacteria, fungi, insects and fermented foods. But few have looked into endophytic fungi as a potential source. Sixteen (16) endophytic fungi were isolated from Hibiscus sp. leaves from six different locations in Shah Alam, Selangor. Only two endophytic fungi, FH3 and S13 showed positive fibrinolytic protease activities. FH3 produced 5.78cm and S13 produced 4.48cm on Skim Milk Agar after 4 days of incubation at 27°C. Fibrinolytic activity was observed; 3.87cm and 1.82cm diameter clear zone on fibrin plate of FH3 and S13 respectively. 18srRNA was done for identification of the isolated fungi with positive fibrinolytic protease. S13 had the highest similarity (100%) to that of Penicillium citrinum strain TG2 and FH3 had the highest similarity (99%) to that of Fusarium sp. FW2PhC1, Fusarium sp. 13002, Fusarium sp. 08006, Fusarium equiseti strain Salicorn 8 and Fungal sp. FCASAn-2. Media composition variation showed the effects of carbon nitrogen on protein concentration, where the decrement of 50% of media composition caused drastic decrease in protease of FH3 from 1.081 to 0.056 and also S13 from 2.946 to 0.198.

Identification Characterization and Production of Phytase from Endophytic Fungi

Phytases are acid phosphatase enzymes, which efficiently cleave phosphate moieties from phytic acid, thereby generating myo-inositol and inorganic phosphate. Thirty four isolates of endophytic fungi to produce of phytases were isolated from leaf, stem and root fragments of soybean. Screening of 34 isolates of endophytic fungi identified the phytases produced by Rhizoctonia sp. and Fusarium verticillioides . The phytase production were the best induced by phytic acid and rice bran compared the others inducer in submerged fermentation medium used. The phytase produced by both Rhizoctonia sp. and F. verticillioides have pH optimum at 4.0 and 5.0 respectively. The characterization of phytase from Fusarium verticillioides showed that temperature optimum was 500C and stability until 600C, the pH optimum 5.0 and pH stability was 2.5 – 6.0, and substrate specificity were rice bran>soybean meal>corn> coconut cake, respectively.