Organizational culture continues to be cited as one of the most important factors for organizations’ success in an increasingly competitive and IT-driven global environment. Given the fact that organizational culture has an influence all over the organization, the complexity of its nature is increased when considering the relationship between business and IT. As a result, different factors that have influence on changing organizational culture were highlighted in literature. These factors are found in the research literature distributed in three main group; micro-environment factors, macro-environment factors and leader’s impact. One of the factors that have not been yet well investigated in researches is concerning business-IT alignment (BITA. Therefore the purpose of this paper is to investigate the impact of BITA maturity on organizational culture. The research process that we have followed is a literature survey followed by an in-depth case study. The result of this research shows a clear interrelation in theories of both BITA and organizational culture, and clear indications of BITA impact on organizational culture and its change. The findings may support both practitioners and researchers in order to understand the insights of the relationships between BITA and organizational culture components and provide a roadmap for improvements or desired changes in organizational culture with highlighted target business area.