Two-dimensional gel electrophoresis (2-DE) was employed to assess the thermo-tolerance characteristics of Bifrdobacterium infantis ATCC 27920 to mild heat adaptation. When exposed to various heat levels, pH, and hydrogen peroxide ($H_2O_2$) stress conditions, B. infantis ATCC 27920 exhibited high level of stress resistance. Under mild-heat treatment ($46^{circ}C$), no significant change in viability level was observed after 2 hr. Interestingly, improved viability was observed in mild-heat adapted ($46^{circ}C$ for 1 hr) cultures exposed to $55^{circ}C$, in comparison to control experiments. Viability was not affected by pH, bile, and $H_2O_2$ stress conditions. 2-DE analysis revealed those mild-heat adaptation up-regulated 4 proteins and down-regulated 3 proteins. Among these protein spots, isopropyhnalate dehydratase (leuD), glycosyltransferase (glgA), and ribosomal protein L5 (rp1E) were identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALD1-TOF/MS).