AUTHOR=Zhang Xiaoxue , Ning Guohui , Qi Mingyue , Li Jiahui , Zhang Xuena , Hao Rui , Zhao Xuefeng , Wang Xiaomin , Yang Zhixin TITLE=Study on the degradation pathway of benzo[a]pyrene by Bacillus sp. strain M1 JOURNAL=Frontiers in Microbiology VOLUME=Volume 16 - 2025 YEAR=2025 URL=https://www.frontiersin.org/journals/microbiology/articles/10.3389/fmicb.2025.1633648 DOI=10.3389/fmicb.2025.1633648 ISSN=1664-302X ABSTRACT=Bacillus sp. strain M1, originally isolated from soil contaminated with polyaromatic hydrocarbons during earlier work of our group, has a strong ability to degrade benzo[a]pyrene (BaP). Here we show that the bacteria can utilize phthalic acid, protocatechuic acid, salicylic acid, catechol or gentisic acid as the sole carbon source, with utilization of 65.50–92.28% within 2 days. The metabolites formed during culture of Bacillus sp. strain M1 in mineral salt medium with BaP as the sole carbon source were detected by GC–MS. Intermediate metabolites were identified that included 4,5-dimethylphenanthrene, 9-ethyl-10-methylanthracene, 2-ethylacridine, phthalate derivatives, silylated catechol, and salicylic acid. The involvement of six relevant intracellular enzymes, namely phthalate dioxygenase, salicylate hydroxylase, catechol-1,2-dioxygenase, catechol-2,3-dioxygenase, gentisic acid dioxygenase and dehydrogenases, was demonstrated, and their activities correlated with the level of BaP degradation. Incorporating all findings, we propose a degradation pathway of BaP by strain M1 that starts with an enzymic attack of the C-9/C-10 or the C-7/C-8 bond. The pathway is summarized as BaP → cis-4,5-pyrene dihydrodiol → 4,5-dimethyl phenanthrene → 1-hydroxy-2-naphthalic acid. In addition, two new products, 9-ethyl-10-methylanthracene and 2-ethylacridine, were discovered.