Lipoxygenase is an enzyme, mainly produced by plants, capable of converting unsaturated fatty acids to fatty acids. It has vast application potential in the food, pharmaceutical and agricultural industries. The aim of this study was to isolate novel lipoxygenase-producing bacteria from the environment and to investigate the lipoxygenase enzymatic properties for industrial production. The strain, NC1, isolated from cultivation soils, was identified as Bacillus subtilis based on the phenotypic characteristics and 16S rRNA gene sequencing. This strain formed a pink color around the colony when cultured on indamine dye formation plates. The production of lipoxygenase by B. subtilis NC1 was influenced by the composition of the medium and linoleic acid concentrations. The optimum temperature and pH for lipoxygenase activity was determined to be 40 °C and pH 6, respectively. The enzyme showed relatively high stability at temperatures ranging from 20–50 °C and acid-neutral regions. In addition, the lipoxygenase produced by B. subtilis NC1 was able to degrade commercially available oils including sunflower seed oil and Perilla oil. In this study, a useful indigenous bacterium was isolated, and the fundamental physicochemical data of bacterial lipoxygenase giving it industrial potential are presented.