Little is known yet which character strengths matter the most at the workplace. The aim of the present investigation was thus to assess the relationship between the level of the 24 character strengths with overall job satisfaction in a general working population (N = 12,499) as well as in eight occupational subgroups (nurses, physicians, supervisors, office workers, clinical psychologists, social workers/educators, economists, and secondary-school teachers) and to compare the overall level of character strengths across the occupational subgroups. Results showed that the character strengths zest, hope, curiosity, love, and gratitude, and emotional strengths in general, related most strongly to overall job satisfaction. These relationships differed between the occupational subgroups. For example, persistence emerged as a top-five strength in three of the eight occupational subgroups, teamwork in two, and bravery, leadership, forgiveness, and self-regulation in one subgroup each. Furthermore, mean differences in character strengths between the subgroups were small and significant for 21 of the 24 strengths. Knowing which individual strengths as well as strengths factors are more important for specific working populations can help to develop and apply more effective strength-based interventions in the workplace, thus improving positive and reducing negative work-related outcomes.