Cell pellets were washed to remove residual LBB by
resuspending in M9 minimal salts medium containing no carbon
source and pelleting again. The M9 medium was discarded after
centrifugation and the washed pellets were resuspended in M9
minimal salts medium containing [Cnmim]Cl (n = C2–C10). All
samples were incubated at 28uC and 100 rpm, and 1 ml samples
were removed periodically and analysed by HPLC. Samples
containing cells were cleaned by centrifugation at 12000 g for15 min and then 0.22 mm filtered prior to analysis. The HPLC
system used was an Agilent 1260 Infinity equipped with a G1311C
quat pump, G1329B autosampler, G1316A column compartment
and G1314C variable wavelength detector. The column was a
Phenomenex Jupiter 5u C18 300A with dimensions of 250 6
4.6 mm65 mm. For analysis of cell suspensions the mobile phase
was 75% acetonitrile: 25% 10 mM K2HPO, and for supernatants
the mobile phase was 80% acetonitrile: 20% 5 mM K2HPO4/
5 mM H2SO4. All samples were run isocratically with a flow rate
of 1 ml/min. The sample size was 5 mL and all measurements
were made at a wavelength of 210 nm, with degradation expressed
as a percentage of the control. All analysis was carried out in
duplicate and was conducted at least weekly for up to 63 days.