Figure 2.

Expression of 2SL genes enables a B. subtilis ΔyxeK mutant to grow on 2SC.B. subtilis 168 harboring an empty pHCMC04 vector (EV) and B. subtilis ΔyxeK mutants harboring an empty pHCMC04 vector or pHCMC04 containing B. subtilis yxeK, D. dadantii (Dd) 2SL, or E. italicus (Ei) 2SL coding sequences were grown overnight at 37 °C in 5 ml LB medium with 5 μg ml⁻¹ chloramphenicol. Cells were washed twice with ED minimal medium lacking sulfur, diluted to absorbances (600 nm) of 1.0, 0.2, 0.04, and 0.008, and 5 μl and was spotted on ED minimal medium plates containing 0.2% xylose and either no added sulfur (−) or 2 mM sulfate (SO₄) or 0.25 mM 2SC as the sole sulfur source. Plates were incubated for 16 h at 37 °C. 2SC, S-(2-succino)cysteine; 2SL, S-(2-succino) lyase.