In this study, we construct a green and high-performance platform using Pickering emulsions for biphasic catalysis. The oil-in-water Pickering emulsions stabilized by the lignin/chitosan nanoparticles (Lig/Chi NPs) have great stability and alkali resistance, showing pH-responsive reversible emulsification and demulsification which can be recycled at least three times. The Pickering emulsion also has fluorescence and wide availability to different oil-to-water volume ratios, types of oil, storage times, temperatures, and ion concentrations. When this system is applied to the lipase-catalyzed reaction for the hydrolysis of p-nitrophenol palmitate, it will provide stable and large oil-water reaction interface areas, and the negatively charged lipase will enrich at the emulsion interface by electrostatic adsorption of the positively charged Lig/Chi NPs to achieve immobilization (lipase-Lig/Chi NPs). The reaction conversion rate can reach nearly 100% in 30 min, which is nearly three times higher than that of the conventional two-phase system. Moreover, the lipases in Pickering emulsion stabilized by Lig/Chi NPs exhibit great recyclability because of the protection of Lig/Chi NPs.