Guanine-ionic liquid derived ordered mesoporous carbon (GIOMC) decorated with gold nanoparticles was used as electrocatalyste for NADH oxidation and electrochemical platform for immobilization of glucose dehydrogenase (GDH) enzyme. The resulting GIOMC/ AuNPs on the glassy carbon electrode can be used as novel redox-mediator free for NADH sensing and this integrated system (GIOMC/AuNPs/GDH) shows excellent electrocatalytic activity toward glucose oxidation. Furthermore, the ionic liquid derived ordered mesoporous carbon derivate with Ph-SO3H (IOMC-PhSO3H) decorated with AuNPs has been developed to bilirubin oxidase enzyme (BOD) immobilization and the GC/IOMC-PhSO3 H/BOD integrated system shows excellent bioelectrocatalytic activity toward oxygen reduction reaction. The proposed mesostructured platforms decorated by AuNPs have been developed to enhance mass transfer and charge transfer from biocatalyst to electrode, leading these bioanode and biocathode used for biofuel cell assembly. Integration designed bioanode and biocathode yielding a membrane-less glucose/O2 biofuel cell with power density of 33 (mW.cm2) at 257 mV. The open circuit voltage of this biofuel cell and maximum produced current density were 508 mVand 0.252 (mA.cm2) respectively.