A novel Co(L)(H2O)2 (1) was obtained by hydrothermal method and it exhibited a 1D chain with exposed carboxyl groups, the unique coordination mode made it have unusual physical and chemical stability. Meanwhile, 1 showed peroxidase-like and weak oxidase-like activity. 1 as a peroxidase mimic enzyme had an excellent affinity for the substrates luminol and H2O2. Compared with HRP, 1 had catalytic activity in a wide pH range and showed the best catalytic activity at pH 7.4. Meanwhile, the catalysis process of 1 was reversible and recyclable, and the catalytic activity remained stable after different pH and temperatures and long-time storage. Based on the inhibition of glutathione on luminol-H2O2-MOF 1 chemiluminescence signal, a chemiluminescence method for the determination of glutathione has been proposed with high sensitivity and selectivity and had been applied for detecting glutathione in cell lysate with satisfactory results. Display omitted •A novel Co (Ⅱ)-based MOF material with double enzyme mimetic activity was synthesized.•The unique coordination mode made Co-MOF have unusual physical and chemical stability.•The Co-MOF had favorable catalytic activity in a wide pH range.•A CL sensing method for glutathione was developed using the luminol-H2O2-Co-MOF system under neutral condition.