Abstract. The aim of this study was to investigate the influence of newly proposed bicycle's crank to crank angle setting on the Vastus Lateralis (VL) and Bicep Femoris (BF) muscle activity during cycling. Procedures of Conconi Test were used throughout the experiment for the data collection purpose. The muscles activities were recorded using surface electromyography and software LabChart7. The raw data were further processed in time (Root-Mean-Square, RMS) and frequency (Mean Power Frequency, MPF) domain. It was found that 0° crank to crank setting (similar to conventional crank to crank angle setting) caused the prime mover VL (Normalized RMS = 0.119) to fatigue more than BF (Normalized RMS = 0.102). This setting is expected to decrease the cycling performance. In addition, -5° is the best crank to crank angle setting that causes least fatigue to both VL and BF. In short, to increase the cycling performance by avoiding the fatigue to the main muscles, -5° is the suggested as setting angle for the proposed crank design.