| Summary: | Zearalenone (ZEN) is a widespread mycotoxin found in grain and feed, presenting a serious threat to animal and human health. This study investigated the ability of the novel strain B73, isolated from petroleum-contaminated soil, to detoxify ZEN. B73 was identified as <i>Bacillus spizizenii</i> through physiological and biochemical tests, and further confirmed based on the 16S rRNA gene sequence and the complete genome sequence. <i>B. spizizenii</i> B73 was capable of degrading up to 99.3% of ZEN at a concentration of 10 μg/mL in a minimal medium (pH = 7.0) within 8 h at 37 °C via HPLC-UV. In addition, <i>B. spizizenii</i> B73 was used to treat ZEN-contaminated wheat bran, dried distillers grains (DDGS), and corn meal, whereby the respective degradation rates reached 96.32%, 98.73%, and 80.31% after 36 h of treatment. HPLC-Q-Exactive-MS/MS analysis revealed one of the degradation products to have the formula C<sub>17</sub>H<sub>24</sub>O<sub>4</sub>. <i>B. spizizenii</i> B73 is a novel strain isolated from petroleum-contaminated soil, and the extracellular enzymes secreted by this strain show a remarkable ability to degrade ZEN.
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