A metabolic pathway for the biodesulfurization of model organosulfur compounds e.g., dibenzothiophene (DBT), is proposed. This pathway, defined as extended 4S pathway, incorporates the traditional 4S pathway with the methoxylation pathway from 2-hydroxybiphenyl (HBP) to 2-methoxybiphenyl (2-MBP). The formation of 2-MBP was confirmed by the gas chromatography-mass spectrometry (GC-MS) analysis. A similar pathway was also obtained in the desulfurization of 4,6-dimethyldibenzothiophene (4,6-DMDBT), confirming the methoxylation reaction in the desulfurization process by the Mycobacterium sp. strain. Compared with 2-HBP, 2-MBP has much slighter inhibition effect on the cell growth and desulfurization activity. Thus, the methoxylation pathway from 2-HBP to 2-MBP would make less inhibitory effect on the microbe. The new pathway with 2-MBP as the end product may be an alternative for the further desulfuration of the fossil fuels.