Metabolic Engineering of Corynebacterium glutamicum for High‐Level Production of 1,5‐Pentanediol, a C5 Diol Platform Chemical

Abstract The biobased production of chemicals is essential for advancing a sustainable chemical industry. 1,5‐Pentanediol (1,5‐PDO), a five‐carbon diol with considerable industrial relevance, has shown limited microbial production efficiency until now. This study presents the development and optimization of a microbial system to produce 1,5‐PDO from glucose in Corynebacterium glutamicum via the l‐lysine‐derived pathway. Engineering began with creating a strain capable of producing 5‐hydroxyvaleric acid (5‐HV), a key precursor to 1,5‐PDO, by incorporating enzymes from Pseudomonas putida (DavB, DavA, and DavT) and Escherichia coli (YahK). Two conversion pathways for further converting 5‐HV to 1,5‐PDO are evaluated, with the CoA‐independent pathway—utilizing Mycobacterium marinum carboxylic acid reductase (CAR) and E. coli YqhD—proving greater efficiency. Further optimization continues with chromosomal integration of the 5‐HV module, increasing 1,5‐PDO production to 5.48 g L⁻¹. An additional screening of 13 CARs identifies Mycobacterium avium K‐10 (MAP1040) as the most effective, and its engineered M296E mutant further increases production to 23.5 g L⁻¹. A deep‐learning analysis reveals that Gluconobacter oxydans GOX1801 resolves the limitations of NADPH, allowing the final strain to produce 43.4 g L⁻¹ 1,5‐PDO without 5‐HV accumulation in fed‐batch fermentation. This study demonstrates systematic approaches to optimizing microbial biosynthesis, positioning C. glutamicum as a promising platform for sustainable 1,5‐PDO production.