The organoleptic properties of plant-based meat alternatives do not meet consumer expectations due to the lack of characteristic flavors resembling meat. To address this challenge, a fermentation system utilizing Laetiporussulphureus was developed to generate a meat-like and fatty flavor from a vegetable source, onion. By means of multiple stir bar sorptive extraction and gas chromatography-mass spectrometry-olfactometry, an unsaturated aldehyde, (E,Z)-2,4-decadienal, which imparts a tallow-like and fatty odor, and a sulfurous compound benzothiazole, with a broth-like odor were identified, which well contributed to the characteristic odor of the supernatant. (E,Z)-2,4-Decadienal as the most important odorant (odor activity value = 206) was biosynthesized by transformation of linoleic acid with L.sulphureus, as revealed by isotopic tracing experiments. For the first time in Basidiomycota, the biogenetic pathway of (E,Z)-2,4-decadienal from linoleic acid was proposed.