12.5g of nutrient broth was dissolved in a 1000ml conical flask containing 500ml of distilled water. 40 ml nutrient broth was measured by volumetric cylinder and poured into ten conical flasks of 100ml. These flasks were then plugged with cotton and covered with aluminum foil. Flasks were autoclaved at 121°C for 15minutes and cooled at room temperature. Media in the flasks were inoculated with bacterial culture and flasks were placed on water bath shaker over night 30°C for further studies.