Numerous countries are investigating alternative fuel sources in response to the escalating issue of energy inadequacy. Using environmentally sustainable biodiesel as a potential alternative to fossil fuels, particularly from waste sources, is a developing prospect. This study aims to examine the feasibility of utilizing industry leather waste as a diesel fuel substitute. Traditional transesterification was used to obtain methyl ester out of leather waste. After processing, 81.93% of methyl ester was produced. Bio-silica (Bio-Si) is used as a fuel additive to enhance combustion and decrease emissions. This work utilized a leather industry waste fat biodiesel (LIWFB), LIWFB blend (B50), LIWFB blend with Bio-Si nanoparticles (B50Bio-Si50, B50Bio-Si75, and B50Bio-Si100 ppm) to analyze the engine outcome parameters at standard operating conditions. Experimental results revealed that adding Bio-Si in the biodiesel blend increased thermal brake efficiency (BTE) but was lower in diesel fuel. The biodiesel blends reduced NOx emissions more than Bio-Si nanoparticle blends. Furthermore, the smoke opacity was reduced by 31.87%, hydrocarbon (HC) emissions were reduced by 34.14%, carbon monoxide (CO) emissions were decreased by 43.97%, and oxides of nitrogen (NOx) emissions were slightly increased by 4.45% for B50Bio-Si100 blend compared to neat diesel. This investigation determined that all the emissions remained lower for all combinations than neat diesel, with a small increase in NOx emissions. Therefore, the LIWFB blend with Bio-Si nanoparticles was a viable diesel fuel alternative in diesel engines.