Deep eutectic solvents (DESs) as a novel class of ionic liquid analogues has been widely used in various fields. Herein, a multifunctional biomass-based DES composed of choline chloride (ChCl) and glucose (GLU) was synthesized and used for the modification of vanadium phosphorus oxide (VPO) catalyst, which is the sole catalyst for the selective oxidation of n-butane to maleic anhydride (MA) in industry. The DES is more green, cost-effective and efficient compared with traditional modifier (metal inorganics and organic polymers). A combination of structure and catalytic performance was investigated in detail to deeply understand its functions. Initially, this DES plays the role of structure directing agent at the stage of precursor synthesis, which can induce the formation of active plane and obtain highly crystalline precursors. Accordingly, the active phase with highly crystalline was obtained after the activation of precursors. Additionally, the DES was decomposed during the precursors activation, resulting in the formation of porous structure around 20 nm on the active plane. Besides this, the surface chemical state including the valence state and acid-base property was changed significantly due to the function of DES. All of these lead to about an 11% increase of MA mass yield, which has great significance to the high value utilization of low carbon alkanes.