OPTIMIZATION OF SCLEROTIUM FORMATION OF SCLEROTIUM ROLFSII RELATED TO LECTIN’S BINDING ACTIVITY TOWARDS CARBOHYDRATE EPITOPE OF ERYTHROCYTE

• Main Author: Zahra, Muhandinni
• Format: Theses
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/46981
• Institution: Institut Teknologi Bandung

Sclerotium rolfsii lectin (SRL) is known to be able to detect glycan-based cancer biomarkers. The binding activity of SRL in sclerotial bodies is reported higher compared to mycelia. Exposing mycelia to stress conditions induces oxidative stress in cells, triggering higher sclerotia formation. In this research, optimization of sclerotia formation in S. rolfsii was performed by introducing physical and chemical of stress conditions to fungal mycelia. S. rolfsii was grown in liquid media containing potato extract 20%, peptone 0,5% and dextrose 2% for 11 days with 150 rpm shaking condition at room temperature. The strategy of stress exposure was done by obtaining optimum biomass of mycelia in liquid media, to be later stress-induced. S. rolfsii was then exposed to physical (fragmentation, cold shock, electric current) and chemical stress (H2O2, garlic extract, and Mycostatin®), while control was not exposed to any stress condition. In fragmentation stress, the culture was crushed in a blender with 10.000 rpm blender with time variation of 30, 45, and 60 seconds. In cold-shock stress, S. rolfsii was incubated in 8°C with time variation 6, 12, 24 hours. In electric current stress, S. rolfsii was shocked for 10 seconds in voltage variation of 5, 50, and 100 V. S. rolfsii was subsequently incubated without shaking for 4 weeks at room temperature. In chemical stress, H2O2, was added to S. rolfsii with dose variation of 0,001%, 0,01%, and 0,1%, garlic extract 0,01%, 0,1%, and 1%, also Mycostatin® 0,001%, 0,01%, and 0,1%. All was done in different flasks separately. Results showed that 0,01% Mycostatin® yielded the highest (4,877 mg/ml) dry weight of sclerotia significantly, compared to control, while appeared to be insignificant to some other treatments. Unexpectedly, 45 seconds of fragmentation showed the highest protein content compared to control but turned out to be insignificant to some other treatments. For hemagglutination activity, 50 V electric current indicated significantly higher activity (1381,18 U/mL) than control but insignificant to some other treatments as well. PCA analysis indicates that protein content positively interacts with hemagglutination activity, while both appears to have negative interaction with dry weight of sclerotium. It is assumed that stress conditions affect the lectin surface structure to somehow increases the binding activity of the lectin with ligands.