Abstract: Extracellular metabolites in the supernatants 52 Bacillus-like strains were analyzed by HPLC along with the quadrupole time-of-flight mass spectrometry (HPLC-Q-TOF-MS). Subsequently, the molecular feature extraction (MFE) was performed on the data using the software MassHunter followed by submitting the information for comparison against the mass spectrometry database, Metlin. Free oleic acid was thereby detected in the cultural supernatant of Bacillus plakortidis, FJAT-14858. Its matching score with oleic acid was 95.61, accurate mass was 284.2693, and relative content in the metabolites collected was up to 1.62%. To date, B. plakortidis FJAT-14858 was the first Bacillus-like bacterium known to produce free oleic acid with no cytotoxicity to the bacteria per se.