The demand for alternative sources of clean, sustainable, and renewable energy has been a focus of research around the world for the past few decades. Microbial/enzymatic biofuel cells are one of the popular technologies for generating electricity from organic substrates. Currently, one of the promising fuel options is based on glucose due to its multiple advantages: high energy intensity, environmental friendliness, low cost, etc. The effectiveness of biofuel cells is largely determined by the activity of biocatalytic systems applied to accelerate electrode reactions. For this work with aerobic granular sludge as a basis, a nitrogen-fixing community of microorganisms has been selected. The microorganisms were immobilized on a carbon material (graphite foam, carbon nanotubes). The bioanode was developed from a selected biological material. A membraneless biofuel cell glucose/oxygen, with abiotic metal catalysts and biocatalysts based on a microorganism community and enzymes, has been developed. Using methods of laboratory electrochemical studies and mathematical modeling, the physicochemical phenomena and processes occurring in the cell has been studied. The mathematical model includes equations for the kinetics of electrochemical reactions and the growth of microbiological population, the material balance of the components, and charge balance. The results of calculations of the distribution of component concentrations over the thickness of the active layer and over time are presented. The data obtained from the model calculations correspond to the experimental ones. Optimization for fuel concentration has been carried out.