摘要
Dodecanedioic acid (DDA) is an important monomer with wide applications in industry, but its chemical synthesis is energy-intensive and contributes to greenhouse gas emissions. To address these sustainability concerns, we previously engineered an alkane-assimilating yeast Candida viswanathii using a split CRISPR system for multi-gene integration, achieving record DDA titers and high conversion efficiency. Building on this genetic foundation, in this study we further developed and optimized the bioprocess for high-yield DDA production in a 3-L fermenter, followed by efficient purification. By identifying low-cost nutrient combinations to replace the expensive Yeast Nitrogen Base (YNB), we reduced the medium cost by 28.8-fold. We further evaluated the process parameters including surfactant supplementation, nutrient feeding and cell density at the onset of DDA production. Such strategic process engineering improved the DDA titer from 90 g/L to 162 g/L and enhanced the total DDA yield from 240 g to 453 g. Importantly, the optimized process achieved a molar conversion efficiency of ≈99%. Furthermore, we developed a simple purification method, yielding 95% DDA recovery and >99% product purity. This study demonstrates the potential of integrated bioprocess engineering to enable sustainable and economically viable microbial production of DDA.