Enzymes are used to make a production of biomass based process more efficient and more selective. Lipase is a biocatalyst in the breakdown of fat reaction that is widely used in the industry, including the industry in Indonesia. Lipase production can be carried out by bacterial solid state fermentation by using agro-industry waste substrate containing carbon and nitrogen. The developments of various industrial sectors are demanding the use and application of lipase commercially in industrial processes as biocatalysts chosen by its ability to work in a friendly environment and have high specificity. Fermentation of Aspergillus niger are able to produce enzyme lipase that can be done by using solid-state fermentation method. In this study palm oil empty fruit bunches (POEFB), bagasse, and palm oil sludge are used as fermentation substrates and will be treated variations of inducer concentration and fermentation time. The results of solid state fermentation of solid substrates POEFB with inducer concentration of 8% for 7 days showed the highest activity value of 2.2 U/mL and 8.2 U/mL in the form of dry extract lipase. The result of the dry lipase enzyme immobilized so that enzyme was stable in repetitive use with cross linking adsorption method using macroporous resin as a support. Experiments showed that empty fruit bunches of oil palm fermentation substrate could produce lipase enzyme with enzyme loading of 56.6% wt. Enzyme activity test carried out in the synthesis of biodiesel through interesterification reaction mole ratio of reactants palm oil and metal acetate 1:12 at 40o C operating temperature conditions for 50 hours in 4 reaction cycles. Biodiesel synthesis results were analyzed using High Perfomance Liquid Chromatography (HPLC) showed biodiesel yield values of 48.6% and the enzyme was able to move up to 68.60% initial yield of 4 cycles of biodiesel synthesis.