CoFe-layered double oxide derived from Hydrotalcite-like combining biochar (CoFe-LDO-BC) was prepared and applied to catalytic ozonation process for removing methylene blue (MB). The catalyst has not only greater specific surface area (about 26 times and 2 times more than that of BC and LDO respectively) and more abundant oxygen vacancies, but inhibited metal agglomeration and accelerated the electron transfer between Co (III)/Co (II) effectively. TOC analysis revealed mineralization rate of methylene blue was 3.7 × 10−2 min−1 (R2 = 0.996, pH = 9, catalyst dosage = 10 mg, ozone = 0.35 mg/min) in the catalytic ozonation system, much higher than that of single ozone (1.5 ×10−2 min−1, R2 = 0.907). Accordingly, the TOC degradation efficiency increased from 31.6% to 68.25%. Moreover, the lower metal ion leaching made the catalyst still maintain higher mineralization ability, and its mineralization efficiency decreased at only 7.9% point after five cycles. The results from XPS, EPR, NH3-TPD and Py-IR confirmed the oxygen vacancies and metal acid sites on catalyst surface could synergistically activate ozone to produce·OH, O2·- and 1O2, and·OH was dominating in mineralization process. The degradation pathway of MB in CoFe-LDO-BC+O3 catalytic system was proposed. This work is helpful for exploring and preparing stable and efficient ozone catalysts.