2.2 Growth of bacterial cultures
Cultivations were carried out at 30 °C and 200 rpm in a Multitron shaker
(Infors, Bottmingen, Switzerland). Microorganisms were cultivated in an
LB pre-culture for ca. 20 h, from which an M9* pre-culture (1% v/v) was
inoculated and incubated for another 12-16 h. From this culture, an M9*
main culture was inoculated to a starting OD450 of 0.2.
Heterologous gene expression was induced with 1 mM isopropyl
β-d-1-thiogalactopyranoside (IPTG) when the cultures reached an
OD450 of ~0.5. Incubation was continued
for another 4-6 h, and cells were harvested for SDS-PAGE analyses, CO
spectra analyses, and/or activity or toxicity assays (see below).