Understanding pro-environmental attitudes is critical to encouraging pollution-minimizing behaviors. Therefore, identifying associated factors is essential for understanding different types of pro-environmental attitudes. We aimed to investigate the associations among individuals’ college-level science course enrollment and their perceptions of the level of spending to improve and protect the environment, as well as their pro-environmental attitudes. We used nationwide population-based cross-sectional survey data from 2,348 individuals obtained from the General Social Survey in the United States. An ordered logistic model was used to examine the associations among college-level science course enrollment, environmental perception, and pro-environmental attitude. We found that science course enrollment was positively associated (OR:1.80, 95% CI: 1.17–2.75) with individuals’ pro-environmental attitudes. We also found that the perception that “too little” is spent on improving and protecting the environment was positively associated (OR:6.68, 95% CI: 2.46–18.12) with a pro-environmental attitude. Understanding how people’s college-level science education and positive environmental perceptions are associated with their positive pro-environmental attitudes could facilitate national environmental policy and the allocation of necessary funds.