For the transfer experiments (Section 3.5), the cells were initially
grown in 3 L Erlenmeyer flasks containing 2 L of the modified
M9 medium with 1000 mM (92.1 g/L) glycerol and 25.0 mg/L
CoCl24H2O in 100 mM phosphate buffer at 37 C and 100 rpm
for 24 h. For the second stage (bioconversion), cells were harvested
at the 24th h when the biomass concentration reached 4.6 g/L
CDW. Cells were centrifuged (4 C, 5000 rpm and 10 min), washed
once with ice-cold 100 mM phosphate buffer (pH 7.0), and re-suspended
to be 4.6 g/L CDW (14.8 OD600) in bioconversion medium
(0.5 L in 1.0 L Erlenmeyer flask). Bioconversion medium contained
only glycerol (700 mM or 64.5 g/L) and CoCl24H2O (35 mg/L) in