Induction of ACC deaminase activity:
One litre culture of Pseudomonas pulida was grown on DF were minimal medium
plus (NH4
)2 SO4
for 48hr. The cells were pelleted by centrifugation in a
sorval RC 5C centrifuge at 8300 Xg for l0min, at room temperature, washed
with 500ml of 0.1M phosphate buffer, pll 7.0 and then centrifuged again
under the same condition. The cell pellet was resuspended in 500ml of DF
salts minimal medium and 90ml was distributed into 5 separate sterile 250ml
conical flasks. 10ml of ACC in DF salts minimal medium was added to flask
to give final ACC concentration of 1 .mM, l00µM. l0µM, lµM and l00nM. The
cell suspensions were then incubated at room temperature on an orbital shaker
at 200 - 250 rpm for 24h. The cells were subsequently pelleted by high speed
centrifugation and dialyzed cell free extracts were prepared and assayed for
ACC deaminase activity.