{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,4]],"date-time":"2026-08-04T16:00:53Z","timestamp":1785859253383,"version":"3.56.0"},"reference-count":55,"publisher":"MDPI AG","issue":"15","license":[{"start":{"date-parts":[[2026,8,4]],"date-time":"2026-08-04T00:00:00Z","timestamp":1785801600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100012166","name":"National Key Research and Development Program of China","doi-asserted-by":"publisher","award":["2025YFA0922800"],"award-info":[{"award-number":["2025YFA0922800"]}],"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":["NSFC-22278313"],"award-info":[{"award-number":["NSFC-22278313"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"award":["NSFC-22278313"],"award-info":[{"award-number":["NSFC-22278313"]}],"id":[{"id":"https:\/\/ror.org\/01h0zpd94","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecules"],"abstract":"<jats:p>2,3-Butanediol (2,3-BD) is a versatile bulk chemical, serving as a high-boiling green solvent, a key monomer for polyesters and polyurethanes, and a crucial intermediate in the production of fine chemicals and cosmetics. Additionally, it is a key intermediate for pharmaceuticals and agrochemicals, supporting green chemistry. Microbial production of 2,3-BD offers a sustainable bioprocess but faces the challenge of high-concentration product inhibition. In an effort to improve 2,3-BD tolerance and production in Corynebacterium glutamicum, the engineered strain CGK4 was subjected to atmospheric and room temperature plasma (ARTP) mutagenesis, from which three superior mutant strains were isolated. Genomic and reverse engineering analyses revealed two mutations beneficial for tolerance and seven mutations beneficial for production. Among these, mutations atpGL250P, cobTV192V, and ctaEL79P increased 2,3-BD titer by 18.0%, 14.5%, and 15.0%, respectively. Furthermore, we found that while certain mutations markedly enhanced stress tolerance, the highest-producing strains achieved a balanced metabolic state rather than maximal tolerance, indicating a non-linear relationship between the two traits. Finally, through mutation combination optimization, the optimal strain K4-cobT-ctaE was constructed. In shake-flask fed-batch fermentation, this strain produced 84.35 \u00b1 3.04 g\/L (2R,3R)-2,3-BD, which was 38.3% higher than that of the parental strain. Our findings provide a promising approach to addressing 2,3-BD tolerance challenges and promote the development of microbial production platforms for industrial application.<\/jats:p>","DOI":"10.3390\/molecules31152714","type":"journal-article","created":{"date-parts":[[2026,8,4]],"date-time":"2026-08-04T15:08:09Z","timestamp":1785856089000},"page":"2714","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Mutagenesis and Analysis for Enhancing (2R,3R)-2,3-Butanediol Tolerance and Production in Corynebacterium glutamicum"],"prefix":"10.3390","volume":"31","author":[{"given":"Zilong","family":"Liu","sequence":"first","affiliation":[{"name":"State Key Laboratory of Synthetic Biology, Tianjin University, Tianjin 300072, China"},{"name":"Frontier Science Center for Synthetic Biology and Key Laboratory of Systems Bioengineering (Ministry of Education), Tianjin University, Tianjin 300072, China"},{"name":"School of Synthetic Biology and Biomanufacturing, Tianjin University, Tianjin 300072, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Haoran","family":"Cui","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Synthetic Biology, Tianjin University, Tianjin 300072, China"},{"name":"Frontier Science Center for Synthetic Biology and Key Laboratory of Systems Bioengineering (Ministry of Education), Tianjin University, Tianjin 300072, China"},{"name":"School of Synthetic Biology and Biomanufacturing, Tianjin University, Tianjin 300072, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuxuan","family":"Jiang","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Synthetic Biology, Tianjin University, Tianjin 300072, China"},{"name":"Frontier Science Center for Synthetic Biology and Key Laboratory of Systems Bioengineering (Ministry of Education), Tianjin University, Tianjin 300072, China"},{"name":"School of Synthetic Biology and Biomanufacturing, Tianjin University, Tianjin 300072, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhiwen","family":"Wang","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Synthetic Biology, Tianjin University, Tianjin 300072, China"},{"name":"School of Synthetic Biology and Biomanufacturing, Tianjin University, Tianjin 300072, China"},{"name":"Key Laboratory of Ministry of Education for Protection and Utilization of Special Biological Resources in Western China, School of Life Sciences, Ningxia University, Yinchuan 750021, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9588-1821","authenticated-orcid":false,"given":"Tao","family":"Chen","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Synthetic Biology, Tianjin University, Tianjin 300072, China"},{"name":"Frontier Science Center for Synthetic Biology and Key Laboratory of Systems Bioengineering (Ministry of Education), Tianjin University, Tianjin 300072, China"},{"name":"School of Synthetic Biology and Biomanufacturing, Tianjin University, Tianjin 300072, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,8,4]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"107783","DOI":"10.1016\/j.biotechadv.2021.107783","article-title":"Prospects on bio-based 2,3-butanediol and acetoin production: Recent progress and advances","volume":"54","author":"Maina","year":"2022","journal-title":"Biotechnol. 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