The purpose of this research is to find out the influence of implementation occupational health and safety to toll gatherer employee’s working motivation at PT Trans Marga Jateng. The data were collected by doing observation, literature studies and questionnaire. The techniques of taking sample are probability sampling. The population of this research are the toll gatherer employees, there are 67 respondents with 41 are taken as samples. SPSS version 20 used to process the data. The result shows that the simple linier regression is Y = 6,242 + 0,664X. It means that occupational health and safety has a positive and significant effect to toll gatherer employees. The correlation value is 0,799. It means that the correlation between occupational health and safety to working motivation is strong or high. Then, the coefficient determination (R Square) is 0,639, it means that the working motivation can be explained by occupational health and safety of 63,9% while 36,1% is explained by other variables not examined in this research. The t value is 8,307 (bigger than t table 2,0227) with the significant value is 0,000 (less than 0,05), it means that occupational health and safety has a positive and significant effect to the working motivation.