Identification of Pseudomonas protegens and Bacillus subtilis Antimicrobials for Mitigation of Fuel Biocontamination

Hydrocarbon fuel biofouling and biocorrosion require expensive cleanup of aviation infrastructures unless appropriate sustainment measures are applied. The identification of novel biological control agents offers promising alternatives to the current chemical biocides used in fuel sustainment. In this study, 496 microbial fuel isolates from our in-house repository were screened to identify new endogenously produced antimicrobial compounds. Using agar plug screening, liquid culture growth testing, and Jet A fuel culture assays, the two fuel-isolate strains <i>Pseudomonas protegens</i> #133, and <i>Bacillus subtilis</i> #232 demonstrated promising biocontrol activity against bacteria, yeast, and filamentous fungi. Liquid chromatography-quadrupole time of flight tandem mass spectrometry (LC-QTOF-MS/MS) of #232 culture filtrate identified several common lipopeptide antimicrobials including gageostatin C, gageopeptin B, and miscellaneous macrolactins. In contrast, LC-QTOF-MS/MS identified the siderophore pyochelin as one of the predominant compounds in #133 culture filtrate with previously demonstrated antimicrobial effect. Jet fuel microbial consortium culture testing of #133 culture filtrate including flow-cytometry live/dead cell mechanism determination demonstrated antimicrobial action against Gram-positive bacteria. The study concludes that antimicrobial compounds secreted by #133 have bactericidal effects against <i>Gordonia</i> sp. and cause cell death through bacterial lysis and membrane damage with potential applications in the biocidal treatment of hydrocarbon-based aviation fuels.