Long-chain dicarboxylic acids (LCDAs) are important chemicals used in a wide range of industries, including pharmaceuticals, plastics, and lubricants. However, current production methods are inefficient and non-sustainable, relying on biotransformation strategies that involve expensive downstream processing and pathogenic production strains. In this proposal, I aim to overcome these limitations by focusing on four different aspects of LCDA production. First, I will screen for novel, non-pathogenic Wickerhamiella strains that can produce LCDAs from vegetable oils to create a more sustainable production process while avoiding the usage of pathogenic strains. Secondly, I will optimize the best Wickerhamiella strain for C16-C18 LCDA synthesis using genetic engineering techniques, thereby increasing the efficiency of LCDA production. Thirdly, I will computationally explore the transportome of Wickerhamiella as discovering LCDA transporters and their mechanisms will help develop new strategies for improving production. Finally, I will engineer LCDA transporters to export LCDAs at low pH through, among others, directed evolution. This will reduce the downstream processing cost and enable a continuous recovery process. In conclusion, by screening for novel Wickerhamiella strains, optimizing LCDA production, exploring the transportome, and engineering LCDA transporters, I hope to revolutionize the LCDA industry and pave the way for more sustainable industrial practices.