Extended Data Fig. 1: MAR Purification.
A) Anaerobic phototrophic growth of wild type R. rubrum in 2 L roux bottles. B) Lysate, soluble fraction, and DEAE fraction containing MAR natively purified from wild type R. rubrum. Experiments were conducted n = 2 times with similar results. C) Activity and functional requirements of native MAR activity in the DEAE fraction (B) for n = 1 sample. All reactions unless otherwise indicated contained 4 mM ATP, 6 mM dithionite, 1 mM MT-EtOH, and ATP regeneration system. D) Plasmid-based MAR gene expression system. Red stars indicate location of poly-histidine sequences. E-G) Total hydrocarbons produced when MAR gene constructs in (D) are expressed in the marBHDK, nflDK, nifB, and nifEN deletion strain to verify histidine tags do not alter in vivo activity. Previously it was shown that the nflDK genes of unknown function are not involved in MAR activity5. Plasmid 1, pPUF-MCS3; 2, pPUF-M4; 3, pPUF-MarBHis6-MarHHis6-MarDHis6-MarK (Supplementary Table 2). In E-G averages and standard deviation error bars are for n = 3 independent experiments.
