<P><B>Abstract</B></P> <P>Mycelia pellets were employed as biological carrier in a continuous stirred tank reactor to reduce biomass washout and enhance hydrogen production from cornstalk hydrolysate. Hydraulic retention time (HRT) and influent substrate concentration played critical roles on hydrogen production of the bioreactor. The maximum hydrogen production rate of 14.2mmol H<SUB>2</SUB> L<SUP>−1</SUP> h<SUP>−1</SUP> was obtained at optimized HRT of 6h and influent concentration of 20g/L, 2.6 times higher than the counterpart without mycelia pellets. With excellent immobilization ability, biomass accumulated in the reactor and reached 1.6g/L under the optimum conditions. Upon further energy conversion analysis, continuous hydrogen production with mycelia pellets gave the maximum energy conversion efficiency of 17.8%. These results indicate mycelia pellet is an ideal biological carrier to improve biomass retention capacity of the reactor and enhance hydrogen recovery efficiency from lignocellulosic biomass, and meanwhile provides a new direction for economic and efficient hydrogen production process.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Mycelia pellets were employed to improve H<SUB>2</SUB> production from cornstalk hydrolysate. </LI> <LI> Continuous process demonstrated the immobilization efficiency of mycelia pellets. </LI> <LI> Mycelia pellets immobilized CSTR gave a high energy conversion efficiency. </LI> <LI> This study provides an efficient way for H<SUB>2</SUB> production from cornstalk hydrolysate. </LI> </UL> </P>