IntroductionParacetamol (acetaminophen) is a medicine used for the treatment of fever, pain, and inflammation during pneumococcal infection. ObjectivesTo see how paracetamol affects the transcriptional profile of Streptococcus pneumonia across the genome. MethodsIn this study, microarray analysis was performed for transcriptional profiling. ResultsTranscriptome data showed differential expression of several genes in S. pneumoniae D39 wild-type incorporated with paracetamol in the growth medium. Furthermore, these genes were categorized using Clusters of Orthologous Groups (COG) functional categorization on the basis of the suspected functions of the respective proteins. The majority of differentially expressed genes are in COG categories E (Amino acid transport and metabolism) and I (Lipid transport and metabolism). Analysis of protein–protein interaction networks exhibited compactly connected networks between fatty acid transport/biosynthesis and antibiotic biosynthesis genes. Moreover, pathways enrichment analysis revealed that fatty acid metabolism and biosynthesis genes were significantly affected under the criteria we've established. ConclusionThese results suggest the fatty acid biosynthesis and metabolism genes to be potential target of paracetamol in S. pneumoniae D39 wild-type.