Fig. 2

Model system for evaluating (a) short chain alkane uptake in E. Coli. b Knockout strains for candidate transporters OmpW and FadL were selected from the Keio collection. c The AlkBGT alkane monooxygenase complex from P. putida, encoded on pGEc41, was used as a reporter for alkane import, allowing quantification of intracellular alkane oxidation to alcohols, aldehydes, and fatty acids by gas chromatography. d Complementation of fadL in the ΔfadL knockout strain to restore phenotype was performed with IPTG inducible vector pASKAfadL, whereas the rhamnose inducible vectors: pRHA67KfadL, pRHA67KompW, and pRHA67KalkL permitted fine tuning of overexpression to optimize alkane import via FadL, OmpW, and known alkane importer AlkL. e pSB50C7, harboring an alkane inducible AlkS-GFP fluorescence biosensor, was used as a independent reporter system to confirm the results from AlkBGT alkane oxidation in the ΔfadL strain