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The use of biofuel is essential for the reduction of greenhouse gas emission. This paper highlights the use of biodiesel as a means of reducing greenhouse gas emission from the diesel engine of heavy-duty vehicles. Biodiesel is fatty acid methyl ester (FAME) obtained through ester exchange reaction by adding methanol to oil, such as rapeseed oil, soybean oil, palm oil, etc. The CO₂ emission from combustion of biodiesel is defined to be equivalent to the CO₂ volume absorbed by its raw materials or plants in their course of growth. On the other hand, however, biodiesel is known to increase the NOx emission when compared with operating with conventional diesel fuel, then suppressing this increase is regarded as a critical issue. This study is intended to identify the fuel properties of biodiesel free from increase in the NOx emission.