Preparation of Bacteria Inoculums
A small loop of fresh bacteria culture was inoculated
in a 250 ml conical flask containing 20 ml medium
[0.5% (w/v) yeast extract, 0.5% (w/v) peptone, 0.1%
(w/v) KH2
PO4
, 0.02% (w/v) MgSO4
.7H2
O and 1% (w/v)
Na2
CO3
(autoclaved separately)] for 18 hours at 37 °C
with shaking at 200 rpm. The cells were then washed
once with normal saline. 10% (v/v) of the cells with
optical density of 0.5 were then inoculated in a 500 ml
conical flask containing 100 ml medium [1% (w/v)
tapioca starch, 0.5% (w/v) peptone, 0.5% (w/v) yeast
extract, 0.02% (w/v) MgSO4
.7H2
O, 0.02% (w/v) FeCl2
and 0.1% (w/v) KH2
PO4
]. The pH of the medium was
adjusted using 1 M Na2
CO3 (autoclaved separately).