All experiments started with overnight pre-cultures in LB medium containing the appropriate antibiotics. For the growth experiments using DBT as a sulfur source, P. azelaica pre-cultures were washed twice with saline solution (0.9% NaCl, w/v) and used to inoculate (initial OD600 of 0.1) 50 ml of sulfur-free BSM medium supplemented with glucose (2 g/l), glycerol (2.45 g/l), 0.1 mM DBT and 1 mM IPTG in 250 ml Erlenmeyer flasks.
For the growth experiments using DBT as a carbon source, P. azelaica pre-cultures were washed twice with saline solution and used to inoculate (initial OD600 of 0.1) 50 ml of pre-sonicated M63 medium supplemented with 6.8 mM glutamic acid, Tween 80 (0.1%) and 2.5 mM DBT or 2.5 mM 2HBP in 250 ml Erlenmeyer flasks.
Optical density at 600 nm (OD600) was measured using a Shimadzu UV–vis spectrophotometer (model UV mini 1,240). Due to the high turbidity of the M63 medium when 2.5 mM DBT was added, growth under these conditions was determined as colony forming units (CFUs) by plating culture samples in LB solid medium.
For the growth experiments using DBT as a carbon source, P. azelaica pre-cultures were washed twice with saline solution and used to inoculate (initial OD600 of 0.1) 50 ml of pre-sonicated M63 medium supplemented with 6.8 mM glutamic acid, Tween 80 (0.1%) and 2.5 mM DBT or 2.5 mM 2HBP in 250 ml Erlenmeyer flasks.
Optical density at 600 nm (OD600) was measured using a Shimadzu UV–vis spectrophotometer (model UV mini 1,240). Due to the high turbidity of the M63 medium when 2.5 mM DBT was added, growth under these conditions was determined as colony forming units (CFUs) by plating culture samples in LB solid medium.