Metabolic engineering of Corynebacterium glutamicum for the production of pyrone and pyridine dicarboxylic acids

• 成果类型: Article
• 署名作者: Cho, Jae Sung; Luo, Zi Wei; Moon, Cheon Woo; Prabowo, Cindy Pricilia Surya; Lee, Sang Yup
• 署名单位: Korea Advanced Institute of Science & Technology (KAIST); Korea Advanced Institute of Science & Technology (KAIST); Korea Advanced Institute of Science & Technology (KAIST)
• 刊物名称: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
• ISSN/ISSBN: 0027-13648
• DOI: 10.1073/pnas.2415213121
• 发表日期: 2024-11-05
• 关键词: 2-pyrone-4; 6-dicarboxylic acid; bacillus-subtilis; dipicolinic acid; aromatic degradation; efficient production; escherichia-coli; quinolinic acid; polyesters; protocatechuate; intermediate

摘要

Environmental concerns from plastic waste are driving interest in alternative monomers from bio- based sources. Pseudoaromatic dicarboxylic acids are promising alternatives with chemical structures similar to widely used petroleum- based aromatic dicarboxylic acids. However, their use in polyester synthesis has been limited due to production challenges. Here, we report the fermentative production of five pseudoaromatic dicarboxylic acids, including 2- pyrone- 4,6- dicarboxylic acid (PDC) and pyridine dicarboxylic constructed by modulating the expression of nine genes involved in the synthesis and degradation pathways of precursor protocatechuate (PCA) and the glucose- uptake system. Comparative transcriptome analysis of the engineered strain against wild- type fed- batch fermentation conditions enabled the final engineered strain to produce 76.17 +/- 1.24 g/L of PDC. Using this platform strain, we constructed 2,3- , 2,4- , and 2,5- PDCA- producing strains by modulating the expression of key enzymes. Additionally, we demonstrated a previously uncharacterized pathway for 2,3- PDCA biosynthesis. The engineered strains produced 2.79 +/- 0.005 g/L of 2,3- PDCA, 494.26 +/- 2.61 mg/L of 2,4- PDCA, and 1.42 +/- 0.02 g/L of 2,5- PDCA through fed- batch fermentation. To complete the portfolio, we introduced the 2,6- PDCA biosynthetic pathway to an L- aspartate pathway-enhanced C. glutamicum strain, producing 15.01 +/- 0.03 g/L of 2,6- PDCA in fed- batch fermentation. The metabolic engineering strategies developed here will be useful for the production of pseudoaromatic chemicals.