As one of the main oxygen-containing organic products generated from the hydrothermal conversion of biomass, levulinic acid (LA) has the potential to be further upgraded. This work investigated the steam reforming (SR) process of biomass-derived LA to produce H2. A series of Ni catalysts supported by various spinels were synthesized via co-precipitation and impregnation. The Ni active metal dispersed well on the NiAl2O4 catalyst with high specific surface area, thereby exhibiting high catalytic activity. Among all the catalysts tested, 15Ni/NiAl2O4 showed the best performance for SR of LA, resulting in high carbon conversion of 96.3%, H2 yield of 92.8%, and H2 concentration of 67.9% at a reaction temperature of 800 & DEG;C. Additionally, the influences of reaction temperature, steam-to-carbon ratio (S/C), and liquid hourly space velocity (LHSV) were also examined. Moreover, during the 40-h continuous SR process of LA, the 15Ni/NiAl2O4 catalyst maintained its outstanding catalytic activity. This study provides an encouraging route for upgrading biomass-derived LA into eco-friendly and high-value fuels, thereby advancing the sustainability of the biomass refining process.