Designing rational nanostructure to promote the oxygen reduction reaction (ORR) catalytic activity of microbial fuel cells (MFCs) is desirable but still remains a huge challenge. In this work, an elaborately designed strategy is proposed to deposit layered double hydroxides (LDHs) on the surface of ZIF-67 grown along nanofibers, thereby obtaining nanoflower-branch composites ([email protected]) with a rich cavity structure supported by electrospinning nanofibers. During the pyrolysis process, the variable cobalt in ZIFs is captured by LDHs nanosheets to generate CoNi alloy. As expected, [email protected] exhibits brilliant ORR catalytic activity. The as-prepared catalyst is an outstanding cathode in MFC, with a maximum power density of 1390.37 mW/m2, superior to Pt/C corresponding MFC (843.67 mW/m2). Impressively, the nanofiber-derived catalyst exhibits long-term durability in single-chamber MFCs. This work provides a new perspective for the combination of LDHs and nanofiber-derived materials, and gives promising performance in realistic MFCs applications.