In order to optimize the use of residues of enzymatic hydrolysis of corn stalk (REHCS) and explore the low‐cost and sustainable raw material substitute for carbon fibers, three types of lignin samples were extracted from REHCS by various extraction methods, and then they were converted into carbon fibers (CFs) by electrospinning, thermostabilization, and carbonization under the same process conditions. The microstructure and mechanical properties of the three types of carbonized fibers were different. The CFs from the ethanol organosolv lignin were actually smooth and brittle carbon films. The CFs from the formic acid/acetic acid organosolv lignin had microscopic pores, causing poor mechanical properties. Comparatively, the CFs from the alkaline lignin demonstrated preferable microstructure and mechanical properties. The reasons for the differences were analyzed by characterizing the lignin samples, precursor fibers, and resultant CFs. © 2017 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2018, 135, 45580.