Laccase and borate–fructose complex were investigated by coincidence in a solid-state fermentation of Edenia sp. TS-76 under fructose oxidase screening. Laccase was purified to homogeneity with a 34-fold purification and 32% yield. Fructose had no significant effect on laccase activity, whereas borate reduced laccase activity by 60–90%; conversely, the borate–fructose complex increased laccase activity by nearly fourfold at pH 7.5. The complex caused a shift in the optimal pH for laccase from 5.0 to 7.5 and served as a highly efficient mediator. Borate complexed with fructose provides an alternative, time-saving, and specific method for serum fructose determination.