Living cells oxidize a wide variety of fuels by employing enzymes as catalysts for energy conversion. It has been shown that many of these cellular metabolic enzymes exist in sequential and proximal organization within living organisms. This organization, called a metabolon, results in channeling of substrates between enzymes. Biofuel cell researchers have studied multienzyme systems, but they have not investigated the need for proximal three-dimension organization in efficient energy conversion. This work outlines the use of a Krebs cycle metabolon catalyst obtained through the in situ cross-Linking of protein within the mitochondria of Saccharomyces cervisea. This cross-linking maintains the integrity of the sequential complexation that exists naturally in the intact biological system. These complexes channel substrate through the enzyme cascade and improve mass transport, thus increasing the current and power density of pyruvate/air enzymatic biofuel cells.