Assessment of enzyme producing efficiencies of endophytic fungi
Extracellular enzymes assay was conducted to investigate the production of enzymes by the endophytic fungi
Amylase activity
Amylase enzyme was assessed by growing the fungi on glucose yeast extract peptone (GYP) agar medium (glucose- 1g, yeast extract- 0.1g, agar- 15 g, distilled water 1000mL and pH 6) containing 1 % soluble starch after 5 days incubation , the plates with fungal colony were flooded with 1% iodine and 2% potassium iodide. The appearance of clear zone surrounding the colony was considered positive for amylase enzyme. Protease activity
Protease assay was performed by growing the endophytic fungi on GYP gar media amended with 1% skim milk and pH was adjusted to 6.5 …show more content…
To GYP agar media, CMC(carboxy methyl cellulose) was added which acted as a substrate. The plates were then kept for incubation at 28 °C for 5 days. The plates were then flooded with 1 % Congo red dye solution for 20 minutes and destained with NaCl solution for 15 minutes. Appearance of light yellow area around the fungal colonies indicated the presence of cellulase enzyme.
Optimization of culture conditions for protease production
The procedure adopted for optimization of various parameters influencing protease production was to evaluate the effect of each parameter independently keeping others as constant. The optimized parameters were incorporated in subsequent experiments. . The fungal culture was grown in 25 ml of production media g/L- (skim milk: 10.0, yeast extract: 0.5, NaCl: 0.5, K2HPO4: 2.0, MgSO4.2 H2O: 0.05, CaCO3: 0.02, FeSO4.7H2O: 0.01) in 100 ml flasks and autoclaved at 121° C, 15psi for 15 minutes.
Effect of temperature, pH, carbon and nitrogen sources on protease activity
The experiment was carried out at different temperature (27° C, 32° C, 37° C) and pH (4, 6 and 8). Different sources of carbon such as maltose, sucrose and glucose at 1.5 % (w/v) were used and nitrogen sources such as beef extract, peptone and yeast extract at 0.3 % (w/v) were used to determine their effect on the production of protease
Extracellular enzymes assay was conducted to investigate the production of enzymes by the endophytic fungi
Amylase activity
Amylase enzyme was assessed by growing the fungi on glucose yeast extract peptone (GYP) agar medium (glucose- 1g, yeast extract- 0.1g, agar- 15 g, distilled water 1000mL and pH 6) containing 1 % soluble starch after 5 days incubation , the plates with fungal colony were flooded with 1% iodine and 2% potassium iodide. The appearance of clear zone surrounding the colony was considered positive for amylase enzyme. Protease activity
Protease assay was performed by growing the endophytic fungi on GYP gar media amended with 1% skim milk and pH was adjusted to 6.5 …show more content…
To GYP agar media, CMC(carboxy methyl cellulose) was added which acted as a substrate. The plates were then kept for incubation at 28 °C for 5 days. The plates were then flooded with 1 % Congo red dye solution for 20 minutes and destained with NaCl solution for 15 minutes. Appearance of light yellow area around the fungal colonies indicated the presence of cellulase enzyme.
Optimization of culture conditions for protease production
The procedure adopted for optimization of various parameters influencing protease production was to evaluate the effect of each parameter independently keeping others as constant. The optimized parameters were incorporated in subsequent experiments. . The fungal culture was grown in 25 ml of production media g/L- (skim milk: 10.0, yeast extract: 0.5, NaCl: 0.5, K2HPO4: 2.0, MgSO4.2 H2O: 0.05, CaCO3: 0.02, FeSO4.7H2O: 0.01) in 100 ml flasks and autoclaved at 121° C, 15psi for 15 minutes.
Effect of temperature, pH, carbon and nitrogen sources on protease activity
The experiment was carried out at different temperature (27° C, 32° C, 37° C) and pH (4, 6 and 8). Different sources of carbon such as maltose, sucrose and glucose at 1.5 % (w/v) were used and nitrogen sources such as beef extract, peptone and yeast extract at 0.3 % (w/v) were used to determine their effect on the production of protease