<p>Objective: In this study, we aimed to develop a comprehensive health literacy (HL) scale for Japanese workers (CHLS-J) and to investigate the relationship between HL and health-related behaviors.</p><p>Methods: We conducted a cross-sectional study by a questionnaire survey of Japanese workers aged 18 years and older (N=313). The HL scale we previously developed was slightly modified on the basis of the pre-examination and used for developing CHLS-J. Self-reported data on demographic variables, socioeconomic status, health-related behaviors, present illness, past illness, and HL were collected. To determine the factor structure of the HL scale, an exploratory factor analysis was conducted, and the internal consistency of the scale was assessed using Cronbach’s coefficient alpha. The criterion-related validity was evaluated using the Pearson product-moment correlation coefficient.</p><p>Results: From the results of factor analysis, 30 HL items were extracted and the structure was based on three factors (the knowledge and competencies for collecting health information, decision-making and communication, and the motivation for utilizing health information). High CHLS-J scores were significantly associated with food purchasing behaviors based on food labeling and maintaining a well-balanced diet. Furthermore, patients with present or past illness were more likely to have high CHLS-J scores.</p><p>Conclusions: The results show that CHLS-J is mostly a validated and reliable scale, and that the high-HL group had a healthy eating lifestyle. This study suggests that CHLS-J can be used to measure HL and educate Japanese workers on the necessity of health behavioral changes.</p>