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This objective of the research was to examine impact of entrepreneurial knowledge and business practices to entrepreneurship motivation. The sample used was Students of Program Studi Manajemen Fakultas Ekonomi Universitas Sarjanawiyata Yogyakarta. The sampling was conducted by applying a non-probability method with a convenience technique. The questionnaires processed were 288 in number, which were obtained from online survey. The data collected were then analyzed by using a multiple regression analysis technique and two independent sample t-test. The result of the research proved that the family-s background, entrepreneurial character, and business practices have a positive effect on entrepreneurship motivation. There are differences in entrepreneurial motivation of students who have received and have not yet received knowledge about entrepreneurship and business practices.