Abstract In the present study, hybrid bio-support catalyst (BSC) has been prepared for the production of biodiesel from Castor oil. Lipase from Pseudomonas cepacia has been immobilized on a hybrid bio-support material (BSM) by entrapment method and characterized. The highest immobilized lipase activity and loading efficiency were 87.28 (U/g) and 55.2%, respectively. The composition of Castor oil expelled from Castor seeds evaluated through Gas Chromatograph-Mass Spectrometer (GC-MS) technique. GCMS spectra show the percentage composition of ricinoleic, linoleic, oleic, stearic, palmitic, and nonanoic acid, as 78.67, 9.62, 6.65, 1.53, 2.19, and 0.08, respectively. Optimized conditions for the transesterification of Castor oil with propan-2-ol as acyl acceptor in solvent free medium were, 10% immobilized lipase relying on oil-weight, molar ratio of alcohol to oil (6:1) at 50 ± 1 °C for the period of 24 h. The product yield was estimated as 75% while the calculated conversion of biodiesel was approximately 78%. The reusability of hybrid BSC for 12 runs was found to be 70% of its initial biodiesel yield after 6 cycles. The properties of oil and biodiesel were calculated using American Standards for Testing Material (ASTM) D6751; formation of biodiesel confirmed by Proton Nuclear Magnetic Resonance (1H NMR) and Fourier Transform Infrared (FTIR) analysis.