The controlled enhancement of antibacterial properties of biomedical materials can be achieved by using laser-induced micro-texturing of the surfaces. Laser-induced periodic surface structures (LIPSS) play an important role in the formation of antibacterial surfaces. The development of the antibacterial properties of a surface depends not only on the parameters of obtained microstructure but also on certain types of bacteria. Therefore, the aim of this study is to obtain enhanced antibacterial properties of titanium surfaces with LIPSS against Gram-positive Staphylococcus aureus and Gram-negative Escherichia coli. Titanium samples were irradiated by femtosecond (fs) laser by varying hatch spacing. The experimental results confirmed that morphological and surface chemical composition changes of the titanium surface induced by fs laser radiation lead to enhanced antibacterial properties against S.aureus by more than 99 % and E.coli more than 80 % compared with non-irradiated titanium samples.