Because of the growing high importance of the development of biocatalytic fuel cell (FC) technologies for renewable energy-producing and testing systems for medical or environmental purposes, in this study, we constructed and demonstrated an H2 FC voltammeter working with graphite sample testing micro-strips and based on Escherichia coli microbial cells. Presented H2 FC voltammeter that provides fast and precise testing of bio-electrochemical possible reactions in biosamples for H2 and other gases, is automated with software which works in NI LabVIEW programming environment, has amplifier cascade system with high internal resistance, temperature controlling and resistance cascade. Microbial Hydrogenase (Hyd) enzymes reversibly catalyze the formation and oxidation of H2. Isolation and characterization of O2-tolerant [NiFe]-hydrogenases (Hyds) have given rise to new concepts in H2 FC. Escherichia coli and [NiFe]-Hyds can be applied as a biocatalyst anode in biofuel cells (BFCs). We evaluated the efficiency of applying the 3 µl (1.5 mg cell dry weight) E. coli intact cells or crude extracts on 0.5 cm2 as anode catalyzers in the bio-electrochemical system. The highest electrical potential (up to 0.7 V) was achieved with bacterial whole cells, which were grown on glucose and glycerol.