This study aimed to determine the impact of a soccer game on the creatine kinase (Ck) response and recovery and the specific Global Positioning System (GPS)-accelerometry-derived performance analysis during matches and comparing playing positions. A sample composed of 118 observations of 24 professional soccer teams of the Brazil League Serie A was recruited and classified according to playing positions, i.e., Left/Right Defenders (D = 30, age: 25.2 ± 5.8 years, height: 187 ± 5.5 cm, weight: 80 ± 5.8 kg), Offensive Midfielders (OM = 44, age: 25.1 ± 0.2 years, height: 177 ± 0.3 cm, weight: 73 ± 1.2 kg), Forwards (F = 9, age: 25.1 ± 0.2 years, height: 176.9 ± 4.3 cm, weight: 74.5 ± 2.1 kg), Left/Right Wingers (M = 23, age: 24.5 ± 0.5 years, height: 175 ± 1.1 cm, weight: 74 ± 4.4 kg), and Strikers (S = 12, age: 28 ± 0.2 years, height: 184 ± 1.0 cm, weight: 80 ± 1.4 kg). Blood Ck concentration was measured pre-, immediately post-, and 24 h postgame, and the GPS-accelerometry parameters were assessed during games. Findings demonstrated that Ck concentrations were higher at all postgame moments when compared with pregame, with incomplete recovery markers being identified up to 24 h after the game (range: 402–835 U/L). Moreover, Midfielders (108.6 ± 5.6 m/min) and Forwards (109.1 ± 8.3 m/min) had a higher relative distance vs. other positions (100.9 ± 10.1 m/min). Strikers [8.2 (8.1, 9.05) load/min] and Defenders [8.45 (8, 8.8) load/min] demonstrated lower load/min than Wingers [9.5 (9.2, 9.8) load/min], Midfielders [10.6 (9.9, 11.67) load/min], and Forwards [11 (10.65, 11, 15) load/min]. These results could be used to adopt specific training programs and recovery strategies after match according to the playing positions.