Blue light irradiation (BLI) is an FDA‐approved method for treating certain types of infections, like acne, and is becoming increasingly attractive as an antimicrobial strategy as the prevalence of antibiotic‐resistant “superbugs” rises. However, no study has delineated the effectiveness of BLI throughout different bacterial growth phases, especially in more BLI‐tolerant organisms such as Escherichia coli. While the vast majority of E. coli strains are nonpathogenic, several E. coli pathotypes exist that cause infection within and outside the gastrointestinal tract. Here, we compared the response of E. coli strains from five phylogenetic groups to BLI with a 455 nm wavelength (BLI 455), using colony‐forming unit and ATP measurement assays. Our results revealed that BLI 455 is not bactericidal, but can retard E. coli growth in a manner that is dependent on culture age and strain background. This observation is critical, given that bacteria on and within mammalian hosts are found in different phases of growth.