There has been a dramatic development in the biological production of 2,3-butanediol (2,3-BD) with the help of genetic technology and an advanced fermentation process using various substrates. To produce more effectively, tolerant microorganisms with high productivity should be developed using metabolic engineering and in silico analysis. Fermentation process development and optimization, economical downstream processing for separation and purification is also needed. In this study, fermentative 2,3-BD production system using klebsiella oxytoca, isolation, metabolic engineering of strains, fermentation process, and recovery process, is described.