Nitrogen (N) content in algae hydrothermal liquefaction (HTL) biocrude is high (5–8 wt%) and generally presumed to result in high NOx emissions during combustion. However, to our knowledge a very limited previous work on diesel engine performance and emissions of N-containing fuels. In order to investigate this issue, pyridine, an N-heterocyclic compound commonly found in algae biocrude, was blended with diesel fuel. This study investigated the influence of N in fuels, using a surrogate fuel to simulate algal biocrude, to determine the combustion behavior and emissions profile of an industrial multi-cylinder diesel engine. The presence of N in the fuel affected its physical properties. Density was slightly higher than neat diesel, while the viscosity, the flash point and the higher heating value (HHV) of the N-containing fuels reduced with increasing N content. The flash point of N-containing fuels were reduced, which affects the storage and transportation of the fuel. The engine load between 25 and 75% was observed to have an effect on engine performance parameters. Compared to diesel, N-containing fuels emitted both lower carbon monoxide (CO) and unburned hydrocarbons (HC). Increasing nitrogen oxides (NOx) emissions were observed with increasing N content in the fuels. At 50% and 75% loads, NOx emissions from N0.1 (0.1 wt% N), N0.5 (0.5 wt% N) and N2 (2 wt% N) were lower than for EUROIII. Particulate matter (PM) was lower for N-containing fuels compared to diesel fuel except for N0.1.