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Bacillus subtilis is one of the microbes that is known very potential to degrade heavy oil because it can produce biosurfactants. The presence of biosurfactant makes Bacillus subtilis able to degrade the chains of carbon atoms in heavy oils that are very long and complex. The ability of Bacillus subtilis to produce biosurfactants is caused by the sfp gene which is required to produce Sfp 4-phosphopantetheinyl transferase protein. To know the characteristics and pathway of the sfp gene in producing biosurfactants, molecular analysis was carried out by using bioinformatics. The analysis carried out includes an analysis of the primary structure, secondary structure, and tertiary structure of the protein produced. Further analysis using EcoCyc was conducted to determine the Sfp 4-phosphopantetheinyl transferase protein metabolism pathway. Based on the results of the analysis carried out related to the sfp gene, it has a conserved protein motif associated with a fairly low similarity with some EntD proteins from Escherichia coli. Based on the EcoCyc analysis known that AccS as a group of genes that produce 4-phosphopantetheinyl transferase Sfp and Associated EntD proteins play an important role in maintaining membrane cell function. The expression of the sfp gene is controlled by the Crp and Fur transcription activator. Excessive expression of the SFP gene will help the secretion of secondary metabolites capable of breaking complex carbon atom chains in the heavy oil.