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Recently, the de novo synthesis of 1,5\u2010PDO has been established, but the accumulation of intermediates and low yield of product limit its further application. In this study, based on the\n                    <jats:sc>l<\/jats:sc>\n                    \u2010lysine high\u2010producing\n                    <jats:italic>Escherichia coli<\/jats:italic>\n                    , an efficient microbial cell factory containing a 5\u2010hydroxyvalerate synthesis module (5\u2010HV) and a 1,5\u2010PDO synthesis module was designed. By screening the enzymes of different modules, a 1,5\u2010PDO\u2010synthesizing recombinant strain with the best combination of MmCAR, Yahk, and GabT was obtained. The amino acid residues in the adenosine domain of CAR were rationally mutated to glutamic acid to obtain the variant MmCAR\n                    <jats:sup>Q302E<\/jats:sup>\n                    , which had enhanced activity against 5\u2010HV and reduced its accumulation. Subsequently, the accumulation of 5\u2010HV was further reduced by enhancing the expression of CAR through RBS engineering and fixing CAR with the help of EutM protein scaffold. In addition, the endogenous gene\n                    <jats:italic>ycjQ<\/jats:italic>\n                    of\n                    <jats:italic>E. coli<\/jats:italic>\n                    was deleted to reduce the reoxidation of 1,5\u2010PDO. Finally, the 1,5\u2010PDO yield reached 12.9\u2009g\/L under the optimized fermentation conditions, achieving efficient biosynthesis of 1,5\u2010PDO and lower accumulation of 5\u2010HV, which is the highest yield reported in\n                    <jats:italic>E. coli<\/jats:italic>\n                    so far.\n                  <\/jats:p>","DOI":"10.1002\/bit.70167","type":"journal-article","created":{"date-parts":[[2026,2,5]],"date-time":"2026-02-05T08:00:23Z","timestamp":1770278423000},"page":"1166-1180","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["De Novo Biosynthesis of 1,5\u2010Pentanediol by Metabolically Engineered\n                    <i>Escherichia coli<\/i>"],"prefix":"10.1002","volume":"123","author":[{"given":"Chen","family":"Ma","sequence":"first","affiliation":[{"name":"State Key Laboratory of Materials\u2010Oriented Chemical Engineering, College of Biotechnology and Pharmaceutical Engineering 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