{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,23]],"date-time":"2026-10-23T19:43:23Z","timestamp":1792784603140,"version":"4.5.0"},"reference-count":38,"publisher":"American Chemical Society (ACS)","issue":"39","license":[{"start":{"date-parts":[[2026,9,21]],"date-time":"2026-09-21T00:00:00Z","timestamp":1789948800000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-029"},{"start":{"date-parts":[[2026,9,21]],"date-time":"2026-09-21T00:00:00Z","timestamp":1789948800000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-037"},{"start":{"date-parts":[[2026,9,21]],"date-time":"2026-09-21T00:00:00Z","timestamp":1789948800000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-045"}],"funder":[{"DOI":"10.13039\/501100004028","name":"Jiangnan University","doi-asserted-by":"publisher","award":["KLIB-KF202504"],"award-info":[{"award-number":["KLIB-KF202504"]}],"id":[{"id":"10.13039\/501100004028","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100013314","name":"Higher Education Discipline Innovation Project","doi-asserted-by":"publisher","award":["111-2-06"],"award-info":[{"award-number":["111-2-06"]}],"id":[{"id":"10.13039\/501100013314","id-type":"DOI","asserted-by":"publisher"}]},{"name":"National Basic Research Program of China (973 Program)","award":["2023YFD1300700"],"award-info":[{"award-number":["2023YFD1300700"]}]},{"name":"National Natural Science Foundation of China (NSFC)","award":["32270036"],"award-info":[{"award-number":["32270036"]}],"id":[{"id":"https:\/\/ror.org\/01h0zpd94","id-type":"ROR","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2026,10,7]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Indigoidine is a sustainable natural blue pigment with broad industrial prospects, while its microbial production is hampered by growth inhibition of the producing strain, enzyme-host incompatibility, and insufficient industrial-scale titer. In this study, de novo biosynthesis of indigoidine was achieved in Corynebacterium glutamicum. Efflux pump engineering was employed to alleviate growth inhibition of the production strain, and semi-rational design of the key synthetase BpsA generated the optimal mutant Y433Q with enhanced catalytic activity. Strain CGI23, which integrated the two aforementioned strategies, produced 29.8 g\/L of indigoidine at a mass yield of 0.248 g\/g glucose in shake flasks; when further scaled up in a 5 L fed-batch fermenter, the titer reached 60.7 g\/L with a volumetric productivity of 0.723 g\/L\/h. This work establishes an efficient microbial platform for industrial indigoidine production, providing a reference for integrated engineering of high-performance microbial cell factories.<\/jats:p>","DOI":"10.1021\/acs.jafc.6c05635","type":"journal-article","created":{"date-parts":[[2026,9,22]],"date-time":"2026-09-22T04:02:26Z","timestamp":1790049746000},"page":"30954-30965","source":"Crossref","is-referenced-by-count":0,"title":["Integrated Transporter\nEngineering and Semi-Rational\nEnzyme Design in\n                    <i>Corynebacterium glutamicum<\/i>\n                    for High-Level Natural Blue Pigment Indigoidine Production"],"prefix":"10.1021","volume":"74","author":[{"given":"Junjie","family":"Sun","sequence":"first","affiliation":[{"name":"Jiangnan University , , , ,","place":["Wuxi, Jiangsu, China, 214122"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hui","family":"Gao","sequence":"additional","affiliation":[{"name":"Jiangnan University , , , ,","place":["Wuxi, Jiangsu, China, 214122"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xinyang","family":"Xu","sequence":"additional","affiliation":[{"name":"Jiangnan University , , , ,","place":["Wuxi, Jiangsu, China, 214122"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shougang","family":"Wang","sequence":"additional","affiliation":[{"name":"Jiangnan University , , , ,","place":["Wuxi, Jiangsu, China, 214122"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5602-8262","authenticated-orcid":true,"given":"Meijuan","family":"Xu","sequence":"additional","affiliation":[{"name":"Jiangnan University , , , ,","place":["Wuxi, Jiangsu, China, 214122"]},{"name":"Jiangnan University , , ,","place":["Wuxi, China, 214122"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,9,21]]},"reference":[{"key":"2026100711072411400_cit1","doi-asserted-by":"publisher","first-page":"1342","DOI":"10.3389\/fmicb.2016.01342","article-title":"Leisingera sp. JC1, a bacterial isolate\nfrom Hawaiian Bobtail Squid eggs, produces indigoidine and differentially\ninhibits Vibrios","volume":"7","author":"Gromek","year":"2016","journal-title":"Front. Microbiol"},{"issue":"4","key":"2026100711072411400_cit2","doi-asserted-by":"publisher","first-page":"739","DOI":"10.3390\/microorganisms9040739","article-title":"Natural pigments of bacterial origin\nand their possible\nbiomedical applications","volume":"9","author":"Celedo\u0301n","year":"2021","journal-title":"Microorganisms"},{"key":"2026100711072411400_cit3","doi-asserted-by":"publisher","first-page":"107768","DOI":"10.1016\/j.dyepig.2019.107768","article-title":"Indigoidine \u2013\nBiosynthesized organic semiconductor","volume":"171","author":"Yumusak","year":"2019","journal-title":"Dyes Pigm."},{"issue":"30","key":"2026100711072411400_cit4","doi-asserted-by":"publisher","first-page":"18545","DOI":"10.1021\/acs.jafc.5c05724","article-title":"Recent advances on\nnatural pigment indigoidine: Occurrence, biosynthetic approaches,\nand applications","volume":"73","author":"Li","year":"2025","journal-title":"J. Agric. Food Chem"},{"key":"2026100711072411400_cit5","doi-asserted-by":"publisher","first-page":"113262","DOI":"10.1016\/j.dyepig.2025.113262","article-title":"Advances in the microbial\nsustainable production of pigment indigoidine","volume":"245","author":"Qi","year":"2026","journal-title":"Dyes Pigm"},{"key":"2026100711072411400_cit6","doi-asserted-by":"publisher","first-page":"133908","DOI":"10.1016\/j.foodchem.2022.133908","article-title":"Natural pigments\nof plant origin: Classification, extraction\nand application in foods","volume":"398","author":"Rodri\u0301guez-Mena","year":"2023","journal-title":"Food Chem"},{"issue":"12","key":"2026100711072411400_cit7","doi-asserted-by":"publisher","first-page":"3394","DOI":"10.1039\/C9GC00920E","article-title":"Sustainable\nbioproduction\nof the blue pigment indigoidine: Expanding the range of heterologous\nproducts in R. toruloides to include non-ribosomal peptides","volume":"21","author":"Wehrs","year":"2019","journal-title":"Green Chem."},{"issue":"25","key":"2026100711072411400_cit8","doi-asserted-by":"publisher","first-page":"10931","DOI":"10.1021\/jacs.0c04328","article-title":"Investigation\nof\nindigoidine synthetase reveals a conserved active-site base residue\nof nonribosomal peptide synthetase oxidases","volume":"142","author":"Pang","year":"2020","journal-title":"J. Am. Chem. Soc"},{"issue":"14","key":"2026100711072411400_cit9","doi-asserted-by":"publisher","first-page":"6087","DOI":"10.1007\/s00253-021-11441-5","article-title":"Indigoidine biosynthesis\ntriggered by the heavy metal-responsive\ntranscription regulator: a visual whole-cell biosensor","volume":"105","author":"Hui","year":"2021","journal-title":"Appl. Microbiol. Biotechnol"},{"issue":"24","key":"2026100711072411400_cit10","doi-asserted-by":"publisher","first-page":"10493","DOI":"10.1007\/s00253-020-10990-5","article-title":"Protein scaffold\noptimizes arrangement of constituent enzymes in indigoidine synthetic\npathway to improve the pigment production","volume":"104","author":"Wang","year":"2020","journal-title":"Appl.\nMicrobiol. Biotechnol"},{"key":"2026100711072411400_cit11","doi-asserted-by":"publisher","first-page":"133415","DOI":"10.1016\/j.biortech.2025.133415","article-title":"Pathway optimization\nand membrane engineering for highly efficient\nproduction of indigoidine in engineered Escherichia coli","volume":"440","author":"Li","year":"2026","journal-title":"Bioresour. Technol"},{"issue":"19","key":"2026100711072411400_cit12","doi-asserted-by":"publisher","first-page":"6613","DOI":"10.1021\/acssuschemeng.0c09341","article-title":"High-level production of the natural\nblue pigment indigoidine\nfrom metabolically engineered Corynebacterium glutamicum for sustainable fabric dyes","volume":"9","author":"Ghiffary","year":"2021","journal-title":"ACS Sustainable\nChem. Eng"},{"issue":"1","key":"2026100711072411400_cit13","doi-asserted-by":"publisher","first-page":"2359","DOI":"10.1038\/s41467-023-38119-y","article-title":"Targeted and high-throughput gene knockdown in diverse\nbacteria using synthetic sRNAs","volume":"14","author":"Cho","year":"2023","journal-title":"Nat. Commun."},{"issue":"1","key":"2026100711072411400_cit14","doi-asserted-by":"publisher","first-page":"483","DOI":"10.1021\/acs.jafc.3c05696","article-title":"Establishment of\nan efficient expression and regulation\nsystem in Streptomyces for economical and high-level\nproduction of the natural blue pigment indigoidine","volume":"72","author":"Zhao","year":"2024","journal-title":"J. Agric. Food Chem"},{"issue":"1","key":"2026100711072411400_cit15","doi-asserted-by":"publisher","first-page":"5385","DOI":"10.1038\/s41467-020-19171-4","article-title":"Genome-scale\nmetabolic rewiring improves titers rates and yields\nof the non-native product indigoidine at scale","volume":"11","author":"Banerjee","year":"2020","journal-title":"Nat. Commun."},{"issue":"9","key":"2026100711072411400_cit16","doi-asserted-by":"publisher","first-page":"113087","DOI":"10.1016\/j.celrep.2023.113087","article-title":"Maximizing microbial bioproduction from sustainable\ncarbon sources using iterative systems engineering","volume":"42","author":"Eng","year":"2023","journal-title":"Cell Rep."},{"key":"2026100711072411400_cit17","doi-asserted-by":"publisher","first-page":"229","DOI":"10.1016\/j.ymben.2021.04.015","article-title":"Engineering Pseudomonas putida for efficient aromatic conversion to bioproduct using high throughput\nscreening in a bioreactor","volume":"66","author":"Eng","year":"2021","journal-title":"Metab. Eng"},{"issue":"1","key":"2026100711072411400_cit18","doi-asserted-by":"publisher","first-page":"7758","DOI":"10.1038\/s41467-023-43631-2","article-title":"Discovery\nand remodeling of Vibrio natriegens as a microbial\nplatform for efficient formic acid biorefinery","volume":"14","author":"Tian","year":"2023","journal-title":"Nat. Commun"},{"issue":"23","key":"2026100711072411400_cit19","doi-asserted-by":"publisher","first-page":"9619","DOI":"10.1007\/s00253-019-10121-9","article-title":"Microbial\nsynthesis of the type I polyketide 6-methylsalicylate with Corynebacterium glutamicum","volume":"103","author":"Kallscheuer","year":"2019","journal-title":"Appl.\nMicrobiol. Biotechnol"},{"issue":"8","key":"2026100711072411400_cit20","doi-asserted-by":"publisher","first-page":"1149","DOI":"10.1007\/s10295-015-1642-5","article-title":"Efficient\nproduction of indigoidine\nin Escherichia coli","volume":"42","author":"Xu","year":"2015","journal-title":"J. Ind. Microbiol. Biotechnol"},{"key":"2026100711072411400_cit21","doi-asserted-by":"publisher","first-page":"132808","DOI":"10.1016\/j.biortech.2025.132808","article-title":"Harnessing soybean okara\nfor enhanced microbial production of indigoidine","volume":"434","author":"Chen","year":"2025","journal-title":"Bioresour. Technol."},{"key":"2026100711072411400_cit22","doi-asserted-by":"publisher","first-page":"193","DOI":"10.1186\/s12934-018-1045-1","article-title":"Production\nefficiency of the bacterial non-ribosomal peptide indigoidine relies\non the respiratory metabolic state in S. cerevisiae","volume":"17","author":"Wehrs","year":"2018","journal-title":"Microb. Cell Fact"},{"issue":"6","key":"2026100711072411400_cit23","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0270359","article-title":"Potential of Aspergillus oryzae as a biosynthetic platform for indigoidine, a non-ribosomal\npeptide pigment with antioxidant activity","volume":"17","author":"Panchanawaporn","year":"2022","journal-title":"PLoS\nOne"},{"issue":"14","key":"2026100711072411400_cit24","doi-asserted-by":"publisher","first-page":"8482","DOI":"10.1021\/acs.jafc.5c00934","article-title":"Engineering acidic resistance in\nUDP-galactose 4-epimerase enables efficient UDP-galactose synthesis\nfrom sucrose","volume":"73","author":"Liu","year":"2025","journal-title":"J. Agric. Food Chem"},{"key":"2026100711072411400_cit25","doi-asserted-by":"publisher","first-page":"53","DOI":"10.1007\/978-1-4939-3375-4_3","article-title":"Measurement of nonribosomal peptide\nsynthetase adenylation domain activity using a continuous hydroxylamine\nrelease assay","volume":"1401","author":"Duckworth","year":"2016","journal-title":"Methods Mol. Biol"},{"key":"2026100711072411400_cit26","doi-asserted-by":"publisher","first-page":"106317","DOI":"10.1016\/j.pep.2023.106317","article-title":"Expression of blue\npigment synthetase a from Streptomyces lavenduale reveals insights on the effects\nof refactoring biosynthetic megasynthases for heterologous expression\nin Escherichia coli","volume":"210","author":"Sword","year":"2023","journal-title":"Protein Expression Purif"},{"issue":"5","key":"2026100711072411400_cit27","doi-asserted-by":"publisher","first-page":"1369","DOI":"10.1111\/jam.12141","article-title":"Increasing\nl-isoleucine\nproduction in Corynebacterium glutamicum by overexpressing\nglobal regulator Lrp and two-component export system BrnFE","volume":"114","author":"Yin","year":"2013","journal-title":"J. Appl. Microbiol"},{"issue":"11","key":"2026100711072411400_cit28","doi-asserted-by":"publisher","first-page":"3593","DOI":"10.1007\/s00253-023-12532-1","article-title":"Reduction\nof acetate synthesis, enhanced\narginine export, and supply of precursors, cofactors, and energy for\nimproved synthesis of L-arginine by Escherichia coli","volume":"107","author":"Wang","year":"2023","journal-title":"Appl. Microbiol. Biotechnol"},{"issue":"19","key":"2026100711072411400_cit29","doi-asserted-by":"publisher","first-page":"8465","DOI":"10.1007\/s00253-016-7695-1","article-title":"Roles of export genes cgmA and lysE for the production\nof l-arginine and l-citrulline by Corynebacterium glutamicum","volume":"100","author":"Lubitz","year":"2016","journal-title":"Appl. Microbiol. Biotechnol"},{"issue":"1","key":"2026100711072411400_cit30","doi-asserted-by":"publisher","first-page":"96","DOI":"10.1016\/j.jbiotec.2011.10.002","article-title":"Triggering the production of the cryptic blue pigment\nindigoidine from Photorhabdus luminescens","volume":"157","author":"Brachmann","year":"2012","journal-title":"J. Biotechnol"},{"issue":"12","key":"2026100711072411400_cit31","doi-asserted-by":"publisher","first-page":"9073","DOI":"10.1074\/jbc.M611319200","article-title":"Cloning and characterization\nof a Streptomyces single module type non-ribosomal\npeptide synthetase catalyzing a\nblue pigment synthesis","volume":"282","author":"Takahashi","year":"2007","journal-title":"J. Biol. Chem"},{"issue":"3","key":"2026100711072411400_cit32","doi-asserted-by":"publisher","first-page":"709","DOI":"10.1042\/BJ20110321","article-title":"Rapid and flexible biochemical assays\nfor evaluating 4\u2032-phosphopantetheinyl transferase activity","volume":"436","author":"Owen","year":"2011","journal-title":"Biochem. J"},{"key":"2026100711072411400_cit33","doi-asserted-by":"publisher","first-page":"11893","DOI":"10.3389\/fmicb.2021.711893","article-title":"Growth Response\nand Recovery of Corynebacterium glutamicum Colonies on Single-Cell\nLevel Upon Defined pH Stress Pulses","volume":"12","author":"Ta\u0308uber","year":"2021","journal-title":"Front.\nMicrobiol"},{"issue":"3","key":"2026100711072411400_cit34","doi-asserted-by":"publisher","first-page":"614","DOI":"10.1007\/s12010-008-8493-8","article-title":"Recombinant\nglutamine synthetase\n(GS) from C. glutamicum existed as both hexamers\n& dedocamers and C-terminal His-tag enhanced inclusion bodies\nformation in E. coli","volume":"159","author":"Li","year":"2009","journal-title":"Appl. Biochem. Biotechnol"},{"key":"2026100711072411400_cit35","doi-asserted-by":"publisher","first-page":"149651","DOI":"10.1016\/j.ijbiomac.2025.149651","article-title":"Enhancing the alkaline\ntolerance of Pseudoalteromonas porphyrae \u03ba-carrageenase\nthrough surface charge engineering","volume":"337","author":"Zheng","year":"2026","journal-title":"Int. J.\nBiol. Macromol"},{"issue":"2","key":"2026100711072411400_cit36","doi-asserted-by":"publisher","first-page":"126820","DOI":"10.1016\/j.bmcl.2019.126820","article-title":"Dissecting modular synthases through inhibition: A\ncomplementary chemical and genetic approach","volume":"30","author":"Vickery","year":"2020","journal-title":"Bioorg. Med. Chem. Lett"},{"issue":"1","key":"2026100711072411400_cit37","doi-asserted-by":"publisher","first-page":"4554","DOI":"10.1038\/s41467-020-18365-0","article-title":"Efficient rational\nmodification of\nnon-ribosomal peptides by adenylation domain substitution","volume":"11","author":"Calcott","year":"2020","journal-title":"Nat. Commun"},{"issue":"4","key":"2026100711072411400_cit38","doi-asserted-by":"publisher","first-page":"440","DOI":"10.1016\/j.str.2024.01.010","article-title":"Modular catalytic\nactivity of nonribosomal\npeptide synthetases depends on the dynamic interaction between adenylation\nand condensation domains","volume":"32","author":"Peng","year":"2024","journal-title":"Structure"}],"container-title":["Journal of Agricultural\nand Food Chemistry"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pubs.acs.org\/doi\/pdf\/10.1021\/acs.jafc.6c05635","content-type":"application\/pdf","content-version":"vor","intended-application":"unspecified"},{"URL":"https:\/\/pubs.acs.org\/jafcau\/article-pdf\/74\/39\/30954\/68589245\/acs.jafc.6c05635.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/pubs.acs.org\/jafcau\/article-pdf\/74\/39\/30954\/68589245\/acs.jafc.6c05635.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,10,7]],"date-time":"2026-10-07T15:31:45Z","timestamp":1791387105000},"score":1,"resource":{"primary":{"URL":"https:\/\/pubs.acs.org\/jafcau\/article\/74\/39\/30954\/5434805\/Integrated-Transporter-Engineering-and-Semi"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,9,21]]},"references-count":38,"journal-issue":{"issue":"39","published-online":{"date-parts":[[2026,9,21]]},"published-print":{"date-parts":[[2026,10,7]]}},"URL":"https:\/\/doi.org\/10.1021\/acs.jafc.6c05635","relation":{},"ISSN":["0021-8561","1520-5118"],"issn-type":[{"value":"0021-8561","type":"print"},{"value":"1520-5118","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,9,21]]}}}