Nanozymes have attracted considerable attention as a new type of promising artificial enzyme in recent years. Here, an oxidase-like cobalt-nitrogen-carbon (Co-N/C-900) nanozyme with well-regulated metal atom spatial distribution has been derived from Co-Zn bimetal zeolitic imidazolate framework precursors and used to develop a facile colorimetric sensing method for L-cysteine. With the aid of Co-N/C-900, the colorless 3,3',5,5'-tetramethylbenzidine (TMB) was oxidized to blue oxidized TMB in the absence of H2O2. However, the oxidation was inhibited after the addition of L-cysteine, and the blue color faded to colorless. Thus, Co-N/C-900 exhibited quite good oxidase-like activity with high catalytic efficiency. Therefore, a facile and efficient colorimetric method to sensitively determine L-cysteine with a low detection limit of 33 nM (S/N = 3) has been developed. Furthermore, favorable selectivity and anti-interference ability towards the determination of L-cysteine based on this approach have also been achieved. It is believed that this colorimetric method for the detection of L-cysteine based on Co-N/C-900 will show potential applications in bioscience and bioengineering.