Abstract: The wholecell biocatalyst is a novel technology for biodiesel production. It can not only overcome the needs of highenergy and environmental pollution in traditional chemical process, but also avoid the high cost and short lifetime of enzymes in immobilizedenzyme catalyzing system. Therefore, the wholecell biocatalyst attracts extensive attention in recent years. In this paper the production conditions of lipase from Rhizopus oryzae were optimized and its enzymatic properties were studied. The results indicated that the suitable conditions for lipase production from R.oryzae were as the follows: nitrogen source was 2% tryptone and 0.1% NaNO3, carbon source was 0.3% soybean, initial pH was 5.5, incubation temperature was 28℃, and incubation time was 58 h, respectively. Furthermore, the properties of the lipase were also characterized. The optimal pH and temperature of lipase for catalysis were 7.5 and 35℃. The lipase was relatively stable in pH range of 7-9 and at 40℃, and Mg2+ had a stimulative effect on the lipase activity.