This study aimed to develop a maker spirit problem-based learning (PBL) innovation and entrepreneurship course and explore its impact on students' learning effectiveness. A mixed research method was adopted for this study, in which qualitative and quantitative data were collected for inductive analysis. The research subjects comprised 49 students who took the 'Innovation and Entrepreneurship Course' at a university of science and technology. Further, an 18-week experimental teaching activity was implemented, in which the 'Maker Spirit -- PBL Innovation and Entrepreneurship Course' was innovatively developed. Results revealed that most students showed significant positive learning effectiveness in two levels, namely 'innovation and entrepreneurship', and 'maker practice', after the experimental teaching. Therefore, five major implementation points of the 'Maker Spirit -- PBL Innovation and Entrepreneurship Course' were proposed to provide references for future implementations and applications in the teaching field.