Microbial electrode catalysis such as microbial fuel cells or electrosynthesis involves electron exchange with the electrodes located at the cell exterior; i.e., extracellular electron transport (EET). Despite the vast amount of research on the kinetics of EET to optimize the catalysis rate, the relevance of other factors, including upstream metabolic reactions, has scarcely been investigated. Herein, we report an in vivo electrochemical assay to confirm whether EET limits anodic current production (j) for the lactate oxidation of Shewanella oneidensis MR-1. Addition of riboflavin, which specifically enhances the EET rate, increased j only in the early phase before j saturation. In contrast, when we removed a trace metal ion necessary for upstream reactions from the electrolyte, a significant decrease in j and the lactate consumption rate was observed only after j saturation. These data suggest that the limiting factor for j shifted from EET to upstream reactions, highlighting the general importance of enhancing, for example, microbial metabolism, especially for long-standing practical applications. Our concept to specifically control the rate of EET could be applicable to other bioelectrode catalysis systems as a strategy to monitor their rate-limiting factors. (C) 2016 Elsevier Ltd. All rights reserved.