<p>T he sulforaphane production of wild-type <i>Lactobacillus plantarum</i> <i>CR12</i> (<i>L. plantarum CR12</i>) is limited. Researchers found that adaptive laboratory evolution (ALE) technology can enhance the production of microorganism by altering the growth environment of microorganisms. However, the effectiveness of enhancing the transformation function of <i>L. plantarum</i> <i>CR12</i> and the safety of the adaptive strain are unknown. To clarify the above problem, ALE technology was used to enhance the sulforaphane conversion rate of <i>L. plantarum</i> <i>CR12</i>. The results showed that the glucoraphanin conversion rate, sulforaphane production rate, and sulforaphane conversion efficiency of adaptive strain were significantly higher than those of the wild-type strain. The adaptive strain could produce sulforaphane in vivo and regulate the gut microbiota structure. The genomes showed that the adaptive strain consists of a 4,165,885&#xa0;bp chromosome with a GC content of 36.8%, encoding 4172 protein-coding genes. Compared with the wild-type strain, the adaptive strain expresses more glycoside hydrolases and glycosyl transferases, indicating that the adaptive strain possesses higher carbohydrate metabolic capability. Genes related to glucoraphanin metabolism (<i>cliT</i>, <i>bglF</i>, and <i>bglA</i>) were detected in both the genomes of the wild-type strain and adaptive strain, indicating that both the wild-type strain and adaptive strain possess sulforaphane production capabilities. Therefore, the adaptive strain can be safely used to increase the in vivo production of sulforaphane, ultimately improving human health. This research is aimed at providing genetic evidence for further formulating rational genetic engineering strategies to transform the wild-type strain into an overproducer.</p>