Vacancy engineering as an effective strategy has been widely employed to regulate the enzyme-mimic activity of nanomaterials by adjusting the surface, electronic structure, and creating more active sites. Herein, we purposed a facile and simple method to acquire transition metal manganese oxide rich in oxygen vacancies (OVs-Mn2O3-400) by pyrolyzing the precursor of the Mn(II)-based metal-organic gel directly. The as-prepared OVs-Mn2O3-400 exhibited superior oxidase-like activity as oxygen vacancies participated in the generation of O2•-. Besides, steady state kinetic constant (Km) and catalytic kinetic constant (Ea) suggested that OVs-Mn2O3-400 had a stronger affinity toward 3,3',5,5'-tetramethylbenzidine and possessed prominent catalytic performance. By taking 2-phospho-l-ascorbic acid as the substrate, which can be converted into reducing substance ascorbic acid in the presence of alkaline phosphatase (ALP), OVs-Mn2O3-400 can be applied as an efficient nanozyme for ALP colorimetric analysis without the help of destructive H2O2. The colorimetric sensor established by OVs-Mn2O3-400 for ALP detection showed a good linearity from 0.1 to 12 U/L and a lower limit of detection of 0.054 U/L. Our work paves the way for designing enhanced enzyme-like activity nanozymes, which is of significance in biosensing.