The aim of this study was to evaluate polyhydroxybutyrate (PHB) production and productivity with supplements under fed-batch cultivation at bioreactor scale (1.3 L). In this study, multiple supplements including oxidative enzyme, mediators, surfactants and silicon nanoparticles were added to Cupriavidus necator culture growing on alkaline pretreatment liquor (APL). At 1.3 L bioreactor scale, PHB production reached 3.3 g/L. To further enhance PHB production, fed-batch cultivation with two different feeding strategies were applied. Under single pulse feeding of 300 mL medium, PHB production reached 4.0 g/L. Under 4 pulses feeding of 75 mL medium each time, PHB production reached 4.5 g/L. This is the highest PHB production from lignin that the authors are aware of in literature.