Aims: The main objective of this study was to examine the influence of individualism on entrepreneurial opportunity exploitation among small and medium enterprise owners in Tanzania through the mediation effect of innovativeness. Specific objectives were to examine the influence of individualism on entrepreneurial opportunity exploitation and to examine the mediating effect of innovativeness on the influence of individualism on entrepreneurial opportunity exploitation. Methodology: The study employed an explanatory research design. 370 small and medium enterprise owners were studied using a cross-section survey questionnaire. Partial least square structural equation modelling was used to test the instruments’ validity and reliability and testing the direct and indirect hypotheses on the effects of individualism on entrepreneurial opportunity exploitation. Results: After testing the hypotheses, the direct effects showed that individualism positively and significantly influences entrepreneurial opportunity exploitation among small and medium enterprise owners. Testing mediation effects, it was found that innovativeness partially mediates the relationship between individualism and entrepreneurial opportunity. Therefore, innovativeness has positive significant indirect effects on the relationship between individualism and entrepreneurial opportunity exploitation.  Conclusion: Individualism directly and indirectly influences small and medium enterprise owners to exploit entrepreneurial opportunities. The study helps policymakers to understand that individualistic values, if well addressed in entrepreneurship policy, can help to promote innovativeness and entrepreneurial opportunity exploitation. Practically, the study implies that well-structured culture influences small and medium enterprises owners’ behavior to exploit opportunities.