{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,4]],"date-time":"2026-09-04T06:12:02Z","timestamp":1788502322113,"version":"build-2803163510"},"reference-count":54,"publisher":"Wiley","license":[{"start":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T00:00:00Z","timestamp":1788393600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"},{"start":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T00:00:00Z","timestamp":1788393600000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/501100012166","name":"National Basic Research Program of China","doi-asserted-by":"publisher","award":["2024YFA0917900"],"award-info":[{"award-number":["2024YFA0917900"]}],"id":[{"id":"10.13039\/501100012166","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["32101170"],"award-info":[{"award-number":["32101170"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["analyticalsciencejournals.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Biotech &amp;amp; Bioengineering"],"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>\n                    <jats:sc>d<\/jats:sc>\n                    \u2010Pantothenic acid (DPA) is an essential vitamin with broad applications. In this study, we engineered\n                    <jats:italic>Corynebacterium glutamicum<\/jats:italic>\n                    for high\u2010titer DPA production by integrating dynamic pathway regulation with structure\u2010guided protein engineering. Functional characterization revealed that endogenous CgPanE and BsPanE2 function as \u03b1\u2010hydroxy acid dehydrogenases rather than ketopantoate reductases, whereas heterologous EcPanE and BsPanE, bifunctional activity toward both ketoisovalerate and ketopantoate, possess larger active cavities. A stationary\u2010phase promoter (\n                    <jats:italic>P<\/jats:italic>\n                    <jats:sub>\n                      <jats:italic>4\u2010N14<\/jats:italic>\n                    <\/jats:sub>\n                    ) was employed to delay ketopantoate reductase expression, reducing precursor consumption. Multiple screening strategies were employed to identify candidate residues for alanine scanning and saturation mutagenesis, which yielded two beneficial mutants, T119I and I183S. Notably, I183S exhibited the most prominent improvements, with a 2.25\u2010fold increase in specific activity and a 1.67\u2010fold higher\n                    <jats:italic>k<\/jats:italic>\n                    <jats:sub>cat<\/jats:sub>\n                    <jats:italic>\/K<\/jats:italic>\n                    <jats:sub>m<\/jats:sub>\n                    ratio than the wild\u2010type. Molecular dynamics simulations indicated that the mutations enhanced catalytic efficiency by providing a more stable catalytic environment, tighter binding with the catalytic units, expanding the active cavity, and shortening the substrate tunnel length. The final engineered strain achieved a DPA titer of 36.12\u2009g\/L in a 5\u2009L bioreactor. This study establishes a robust chassis for sustainable DPA production and provides a generalizable framework for engineering other valuable biochemicals.\n                  <\/jats:p>","DOI":"10.1002\/bit.70371","type":"journal-article","created":{"date-parts":[[2026,9,4]],"date-time":"2026-09-04T05:16:40Z","timestamp":1788499000000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Enhancing\n                    <scp>d<\/scp>\n                    \u2010Pantothenic Acid Production in\n                    <i>Corynebacterium glutamicum<\/i>\n                    via Integrating Dynamic Regulation With Structure\u2010Guided Protein Engineering of Ketopantoate Reductase"],"prefix":"10.1002","author":[{"given":"Qing","family":"Yang","sequence":"first","affiliation":[{"name":"State Key Laboratory of Green Chemical Synthesis and Conversion Zhejiang University of Technology Hangzhou China"},{"name":"The National and Local 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