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-free pathway for heme biosynthesis, knockout studies on Gram-positive model organism Bacillus subtilis have been performed. These reveal that ΔchdC strains exhibit heme auxotrophic behavior during aerobic and anaerobic growth, highlighting that ChdC has likely no anaerobic alternative. Free and protein bound FMN (in the form of flavodoxins YkuN and YkuP from B. subtilis) were subsequently characterized as alternative co-substrates. Their reaction with heterologously expressed ChdC from B. subtilis was characterized in different settings. By polarographic dioxygen level determination, liquid chromatography, mass spectrometry and time-resolved spectroscopy, these reactions were shown to be possible and promoted under anaerobic conditions and at elevated pH-values. Overall, the results presented in this study confirm the necessity and the capability of ChdC to react anaerobically in a Gram-positive model organism.
Falb et al. (Fri,) studied this question.