Recently, renewable energy is very important use for industrial development, transport because it is environmental friendly and can reduce high cost of imported fossil fuel. The evolution of biodiesel production has been reported in many researchers. Transesterification process which is mainly reaction for biodiesel is required catalysts such as acid, base catalyst or biocatalyst. In this study, 134 isolates were selected from soil contaminated with oil in Khon Kaen region by using culture medium containing 1% olive oil and 0.0001% rhodamine B with incubation at 30° C for 48 h. These lipase-producing bacteria were also determined their activities on agar medium with 1% tributyrin. The isolate NA37 showed high lipase activity (190 mU/ml) detecting with p-nitrophenyl palmitate as a substrate. Cooking-palm oil was contributed lipase production of the NA37 more than other oils. The optimum temperature and pH for lipase production is 30° C and pH 9, resulting in lipase activity of 481 mU/ml.