Lignin, derived from agricultural-waste corncob residue, was used to immobilize C. antarctica lipase B (CALB) by taking advantage of the mechanism that the surface hydrophobicity of lignin is positive to the immobilization of lipase by the interfacial activation between the carrier and lipase. The immobilization yield of 93.0 ± 0.2% and enzyme loading of 28.1 ± 0.3 mg/g were obtained under optimized process. The maximum transesterification yield of ethyl palmitate (80.7 ± 1.2%) was obtained within p-nitrophenol palmitate and ethanol as the substrate, transesterification time of 6 h, temperature of 42 ℃, and the organic phase of n-hexane. The recovered activity of immobilized enzyme reached 77.0 ± 0.6% after 20 days of storage, and maintained 88.6 ± 1.4% of its initial activity after 5 cycles application. These results demonstrate that the process presents a useful protocol to immobilize lipase on lignin, which is a fully enzyme-mediated green and sustainable biocatalytic system based exclusively on lignin.