The purpose of this study was to compare markers of glycolytic metabolism in response to the Wingate test and the incremental test in road and mountain bike cyclists, who not different performance level and aerobic capacity. All cyclists executed the Wingate test and incremental test on a cycle ergometer. Maximal power and average power were determined during the Wingate test. During the incremental test the load was increased by 50 W every 3 min, until volitional exhaustion and maximal aerobic power (APmax), maximal oxygen uptake (VO2max), and time of VO2max plateau (Tplateau) were determined. Post-exercise measures of oxygen uptake (VO2post), carbon dioxide excretion, (VCO2post), and the ratio between VCO2/VO2 (RERpost) were collected for 3 min immediately after incremental test completion. Arterialized capillary blood was drawn to measure lactate (La-) and hydrogen (H+) ion concentrations in 3 min after each test. The data demonstrated significant differences between mountain bike and road cyclists for Tplateau, VO2post, VCO2post, La- which was higher-, and RERpost which was lower-, in mountain bike cyclists compare with road cyclists. No differences were observed between mountain bike and road cyclists for APmax, VO2max, H+ and parameters measured in the Wingate test. Increased time of VO2max plateau concomitant to larger post-exercise La- and VO2 values suggests greater anaerobic contribution during incremental testing efforts by mountain bike cyclists compared with road cyclists.