In this study, the effect of reaction conditions on the transesterification of soybean oil and methanol was investigated in a high-pressure-fixed-bed-reactor-system with zinc aluminate catalysts. Without catalysts, high-pressurereaction at 300 ℃ and 1,200 psi brought 19% yields of methyl esters, which was caused by the approach of reaction condition to supercritical point of methanol. However, except the specific reaction condition, the yields in the reaction with no catalyst were very low below 4.5%. The zinc aluminate was prepared as catalyst by coprecipitation and characterized with N2 gas adsorption/desorption and X-ray diffraction. With catalyst, the effect of the reaction parameters such as temperature, pressure, and molar ratio of reactants on biodiesel production was demonstrated. The higher temperature, pressure, and methanol molar ratio to soybean oil, the more yields of methyl esters. It was proved that among the reaction parameters, the reaction temperature be the most influential variable on methyl ester yields.