The applications of diesel engines are more as they have more output power and thermal efficiency. But their emissions effect the environment and human being health. Hence there is a lot of scope to search for alternative fuels which promise better performance and emission behaviors of internal combustion (IC) engines. In this regard dairy scum oil methyl ester (DSOME) is a promising sustainable substitute fuel for conventional diesel engines. The standard diesel engine parameters are optimum for diesel but not for DSOME. The present investigation made a effort to use dairy waste scum as a potential source of alternative fuel for diesel engine. In first phase of work, different blends of DSOME (B10, B20, B30 and B100) are prepared and studied their effect on diesel engine performance and emission characteristics. From the study it’s observed that DSOME-B20 exhibits improved performance and emission characteristics among other blends hence it is optimized and carried for further investigation. In second phase of work injector opening pressure (IOP) of the standard engine is modified from 210 bar to 240 bar in step of 10 bar (210, 220, 230 and 240 bars) to study the performance, emission and combustion characteristics of the diesel engine. From the experimental results it’s revealed that increasing the IOP from 210 bar to 230 bar with DSOME-B20, brake thermal efficiency (BTE) is increased while brake specific flue consumption (BSFC), CO, and HC emissions are reduced however, NOx (oxides of nitrogen) is increased marginally. The combustion parameters namely heat release rate (HRR) and cylinder pressure are enhanced significantly with DSOME-B20 at 230 bar IOP. Based on experimental results it’s conclude that 230 bar IOP is the optimum for DSOME-B20 fuel blend operation in diesel engine.