<p>Bacterial lipase is of particular interest due to its high activity and potential for genetic modification. The interaction between lipase and detergents has been a subject of inquiry, leading to a research study on the effect of Triton X-100 on Pseudomonas lipase. Both in silico and in vitro methods were employed in this investigation. The in silico results revealed that Triton X-100 can bind to a specific region outside the enzyme’s active site, leading to hydrophobic interactions with Leu278 and Phe240. In the laboratory (in vitro), it was observed that the enzyme exhibited its highest activity in the presence of detergent, particularly at the concentration bordering between the monomer and micelle state. Furthermore, the presence of detergent caused a shift in the optimum temperature of the enzyme activity from 60 to 50oC. The fluorescence spectrum analysis supported the in silico findings, demonstrating a change in the emission spectrum indicating the transfer of aromatic amino acids to a more hydrophilic environment in the presence of detergent. This research confirmed that lipase remains fully active in the presence of the detergent, while also highlighting the potential for the detergent to alter the physico-chemical properties of the enzyme.</p>