Lipase produced from microorganisms can be used in many industrial applications, such as detergent formulation, oil/fat degradation, pharmaceutical synthesis, biodiesel and cosmetics production. Pseudomonas sp. AB2 was isolated and identified by 16SrRNA from waste cooking oil (WCO). The lipase showed a molecular weight of 28.6 KDa by SDS–PAGE. The enzyme activity was highest after 24 h at 200 rpm agitation when (4X10⁴ CFU) was used as inoculums. The maximum lipase production was observed at 35°C and pH 7. As the Tributyrine increase the enzyme activity decrease, while the yeast extract increase the activity with concentration higher than 0.5 g/l. The GC analysis of the WCO showed that the fatty acid profile of the oil was Palmetic (31.79%), Stearic (2.41 %), Oleic (65.8 %); while the total saturated acid was (34.2 %) and unsaturated acid was (65.8%). Pseudomonas sp. AB2 lipase, which exhibited a potential for catalyzing the biodiesel production, was further purified and immobilized on 20.4 mmol of Tetramethoxyorthosilicate (TMOS). The immobilized lipase from Pseudomonas sp. AB2 could be used as a biocatalyst for the application of enzyme-catalyzed biodiesel synthesis