Single-photon subtraction plays important roles in optical quantum information processing as it provides a non-Gaussian characteristic in continuous-variable quantum information. While the conventional way of implementing single-photon subtraction based on a low-reflectance beam splitter works properly for a single-mode quantum state, it is unsuitable for a multimode quantum state because a single photon is subtracted from all multiple modes without maintaining their mode coherence. Here, we experimentally implement and characterize a mode-tunable coherent single-photon subtractor based on sum-frequency generation. It can subtract a single photon exclusively from one desired time-frequency mode of light or from a coherent superposition of multiple time-frequency modes. To experimentally characterize the time-frequency modes of the single-photon subtractor, we employ quantum process tomography based on coherent states. The mode-tunable coherent single-photon subtractor will be an essential element for realizing non-Gaussian quantum networks necessary to get a quantum advantage in information processing.