The aims of the presented study are to compare submerged, static, and solid-state fermentation in the production of gibberellic acid (GA3), indole acetic acid (IAA), and abscisic acid (ABA) by Inonotus hispidus, to optimize with a statistical approach, and to determine the kinetic parameters under flask and reactor conditions. The maximum concentrations of GA3, (2478.85 ± 68.53 mg/L), ABA, (273.26 ± 6.17 mg/L) and IAA (30.67 ± 0.19 mg/L) were obtained in submerged conditions. After optimization, these values reached 2998.85 ± 28.85, 339.47 ± 5.50, and 34.56 ± 0.25 mg/L, respectively. Immobilization of fungal cells on synthetic fiber, polyurethane foam, and alginate beads resulted in an increase in plant growth regulators (PGR) production by 5.53%− 5.79% under optimized conditions. At the reactor scale, a significant increase was observed for GA3 concentration, 5441.54 mg/L, which was 2.14 and 1.45 times higher than non-optimized and optimized conditions in the flask scale, respectively. The maximum values for ABA and IAA were 390.39 and 44.79 mg/L, respectively. Although the specific growth rate (µ) decreases relatively from non-optimized flask conditions to optimized reactor conditions, it was observed that the PGR amounts produced per liter medium (rp) and per gram biomass (Qp) increased significantly. This is the first report on the synthesis of PGR by Inonotus hispidus which could be crucial for sustainable agriculture.