ABSTRACTStreptococcus mitisis a normal member of the human oral microbiota and a leading opportunistic pathogen causing infective endocarditis (IE). Despite the complex interactions betweenS. mitisand the human host, understanding ofS. mitisphysiology and its mechanisms of adaptation to host-associated environments is inadequate, especially compared with other IE bacterial pathogens. This study reports the growth-promoting effects of human serum onS. mitisand other pathogenic streptococci, includingS. oralis,S. pneumoniae, andS. agalactiae. Using transcriptomic analyses, we identified that, with the addition of human serum,S. mitisdownregulates uptake systems for metal ions and sugars, fatty acid biosynthetic genes, and genes involved in stress response and other processes related with growth and replication.S. mitisupregulates uptake systems for amino acids and short peptides in response to human serum. Zinc availability and environmental signals sensed by the induced short peptide binding proteins were not sufficient to confer the growth-promoting effects. More investigation is required to establish the mechanism for growth promotion. Overall, our study contributes to the fundamental understanding ofS. mitisphysiology under host-associated conditions.IMPORTANCES. mitisis exposed to human serum components during commensalism in the human mouth and bloodstream pathogenesis. However, the physiological effects of serum components on this bacterium remain unclear. Using transcriptomic analyses,S. mitisbiological processes that respond to the presence of human serum were revealed, improving the fundamental understanding ofS. mitisphysiology in human host conditions.