{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,21]],"date-time":"2026-10-21T14:39:48Z","timestamp":1792593588333,"version":"4.5.0"},"reference-count":50,"publisher":"American Chemical Society (ACS)","issue":"41","license":[{"start":{"date-parts":[[2026,10,6]],"date-time":"2026-10-06T00:00:00Z","timestamp":1791244800000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-029"},{"start":{"date-parts":[[2026,10,6]],"date-time":"2026-10-06T00:00:00Z","timestamp":1791244800000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-037"},{"start":{"date-parts":[[2026,10,6]],"date-time":"2026-10-06T00:00:00Z","timestamp":1791244800000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-045"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["32572534"],"award-info":[{"award-number":["32572534"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Key Research and Development Program of Wuhan City","award":["2025021202030436"],"award-info":[{"award-number":["2025021202030436"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2026,10,21]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Monascus azaphilones (MAs) are bioactive compounds that have been utilized in food for over a millennium. These molecules are classified into orange (OMAs), yellow (YMAs), and red MAs (RMAs), with compositional ratios determining the product quality. Cofactors critically influence component ratios, but their individual roles in controlling MA ratios remain unclear. This study established a metabolic library by chemically perturbing the cofactor levels in M. purpureus HJ11. Machine learning analysis of cofactor concentrations and MA components identified NADH as the regulator driving YMAs biosynthesis while suppressing OMAs and RMAs. To validate this, we engineered a cofactor supply system by replacing native isocitrate dehydrogenase with Pseudomonas stutzeri phosphite dehydrogenase, enabling NADH control via an isomaltose adjustment. The engineered strain produced azaphilones with purities of 91% (YMA), 93% (OMA), and 87% (RMA) under optimized protocols. These findings reveal the regulatory role of cellular redox balance in azaphilone diversity and establish a programmable framework for metabolite composition control in Monascus species.<\/jats:p>","DOI":"10.1021\/acs.jafc.5c12726","type":"journal-article","created":{"date-parts":[[2026,10,6]],"date-time":"2026-10-06T22:01:36Z","timestamp":1791324096000},"page":"32219-32228","source":"Crossref","is-referenced-by-count":0,"title":["Machine Learning-Guided Cofactor Engineering for Tunable\nAzaphilone Component Ratios in\n                    <i>Monascus<\/i>\n                    spp."],"prefix":"10.1021","volume":"74","author":[{"given":"Wenrui","family":"Huang","sequence":"first","affiliation":[{"name":"Huazhong Agricultural University , , , ,","place":["Wuhan, Hubei Province, China, 430070"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jiajia","family":"Li","sequence":"additional","affiliation":[{"name":"Huazhong Agricultural University , , , ,","place":["Wuhan, Hubei Province, China, 430070"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yali","family":"Duan","sequence":"additional","affiliation":[{"name":"Huazhong Agricultural University , , , ,","place":["Wuhan, Hubei Province, China, 430070"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuting","family":"Liu","sequence":"additional","affiliation":[{"name":"Huazhong Agricultural University , , , ,","place":["Wuhan, Hubei Province, China, 430070"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nanwei","family":"Wang","sequence":"additional","affiliation":[{"name":"Huazhong Agricultural University , , , ,","place":["Wuhan, Hubei Province, China, 430070"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6264-929X","authenticated-orcid":true,"given":"Mu","family":"Li","sequence":"additional","affiliation":[{"name":"Huazhong Agricultural University , , , ,","place":["Wuhan, Hubei Province, China, 430070"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,10,6]]},"reference":[{"issue":"5","key":"2026102109071201500_cit1","doi-asserted-by":"publisher","first-page":"1589","DOI":"10.1021\/acs.jafc.1c07588","article-title":"Systematic metabolic engineering for the production of azaphilones\nin Monascus purpureus HJ11","volume":"70","author":"Duan","year":"2022","journal-title":"J. Agric. Food Chem."},{"issue":"1","key":"2026102109071201500_cit2","doi-asserted-by":"publisher","first-page":"114053","DOI":"10.1016\/j.lwt.2022.114053","article-title":"An efficient\nmicrobial-based method for production of high-purity Monascus azaphilones pigments","volume":"170","author":"Duan","year":"2022","journal-title":"LWT-food Sci. Technol."},{"issue":"5","key":"2026102109071201500_cit3","doi-asserted-by":"publisher","first-page":"555","DOI":"10.1111\/1541-4337.12145","article-title":"Edible filamentous\nfungi from the species Monascus: Early traditional\nfermentations, modern molecular biology, and\nfuture genomics","volume":"14","author":"Chen","year":"2015","journal-title":"Comp. Rev. Food Sci. Food Safe"},{"key":"2026102109071201500_cit4","doi-asserted-by":"publisher","DOI":"10.1038\/srep08331","article-title":"Complete genome sequence\nand transcriptomics analyses reveal pigment biosynthesis and regulatory\nmechanisms in an industrial strain, Monascus purpureus YY-1","volume":"5","author":"Yang","year":"2015","journal-title":"Sci. Rep."},{"issue":"7","key":"2026102109071201500_cit5","doi-asserted-by":"publisher","first-page":"4917","DOI":"10.1039\/C7SC00475C","article-title":"Orange, red, yellow: Biosynthesis\nof azaphilone pigments in Monascus fungi","volume":"8","author":"Chen","year":"2017","journal-title":"Chem. Sci."},{"issue":"4","key":"2026102109071201500_cit6","doi-asserted-by":"publisher","first-page":"561","DOI":"10.1039\/C8NP00060C","article-title":"Nature and nurture:\nconfluence of pathway determinism with metabolic and chemical serendipity\ndiversifies Monascus azaphilone pigments","volume":"36","author":"Chen","year":"2019","journal-title":"Nat. Prod. Rep."},{"issue":"6","key":"2026102109071201500_cit7","doi-asserted-by":"publisher","first-page":"2102","DOI":"10.1039\/C7FO00406K","article-title":"Alleviation\nof metabolic syndrome\nby monascin and ankaflavin: the perspective of Monascus functional foods","volume":"8","author":"Lin","year":"2017","journal-title":"Food Funct."},{"issue":"3","key":"2026102109071201500_cit8","doi-asserted-by":"publisher","first-page":"1078","DOI":"10.1016\/j.foodchem.2007.09.021","article-title":"Effect of the Monascus pigment threonine\nderivative on regulation of the cholesterol level in mice","volume":"107","author":"Jeun","year":"2008","journal-title":"Food Chem."},{"issue":"4\u20135","key":"2026102109071201500_cit9","doi-asserted-by":"publisher","first-page":"406","DOI":"10.1002\/mnfr.200500238","article-title":"Cytotoxic and antimitotic\neffects of N-containing Monascus metabolites studied using immortalized\nhuman kidney epithelial cells","volume":"50","author":"Knecht","year":"2006","journal-title":"Mol. 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Agents\nChemother."},{"issue":"11","key":"2026102109071201500_cit29","doi-asserted-by":"publisher","first-page":"1938","DOI":"10.1177\/09603271211014573","article-title":"Nicotinamide improves NAD+ levels to protect against acetaminophen-induced acute liver injury\nin mice","volume":"40","author":"Xu","year":"2021","journal-title":"Hum. Exp. Toxicol."},{"key":"2026102109071201500_cit30","doi-asserted-by":"publisher","first-page":"287","DOI":"10.1016\/j.neuropharm.2017.05.009","article-title":"Methylene blue stimulates\nsubstrate-level phosphorylation\ncatalysed by succinyl-CoA ligase in the citric acid cycle","volume":"123","author":"Komlo\u0301di","year":"2017","journal-title":"Neuropharmacology"},{"issue":"1","key":"2026102109071201500_cit31","doi-asserted-by":"publisher","first-page":"807","DOI":"10.3390\/ijms14010807","article-title":"A model of interaction\nbetween nicotinamide adenine dinucleotide\nphosphate (NADPH) oxidase and apocynin analogues by docking method","volume":"14","author":"Jiang","year":"2013","journal-title":"Int. J. Molecular Sci."},{"issue":"7","key":"2026102109071201500_cit32","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0131663","article-title":"Dichloroacetate,\nthe pyruvate dehydrogenase\ncomplex and the modulation of mESC pluripotency","volume":"10","author":"Rodrigues","year":"2015","journal-title":"PLoS One"},{"issue":"12","key":"2026102109071201500_cit33","doi-asserted-by":"publisher","DOI":"10.1016\/j.heliyon.2020.e05741","article-title":"The efficacy\nof the anticancer 3-bromopyruvate is potentiated by antimycin and\nmenadione by unbalancing mitochondrial ROS production and disposal\nin U118 glioblastoma cells","volume":"6","author":"Petricciuolo","year":"2020","journal-title":"Heliyon"},{"issue":"2","key":"2026102109071201500_cit34","doi-asserted-by":"publisher","first-page":"66","DOI":"10.1016\/j.trechm.2024.12.007","article-title":"Driving non-enzymatic\nreactions towards biocompatible\nchemistry","volume":"7","author":"Xue","year":"2025","journal-title":"Trends Chem."},{"issue":"1","key":"2026102109071201500_cit35","doi-asserted-by":"publisher","first-page":"213","DOI":"10.1021\/acssynbio.2c00500","article-title":"Versatile strategy\nfor the construction of a transcription factor-based orthogonal gene\nexpression toolbox in Monascus spp","volume":"12","author":"Duan","year":"2023","journal-title":"Acs Synth. 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Adv."},{"issue":"2","key":"2026102109071201500_cit39","doi-asserted-by":"publisher","first-page":"e70151","DOI":"10.1002\/jobm.70151","article-title":"Enhanced Monacolin\nK production in Monascus purpureus via mutagenesis\nand nitrogen source optimization","volume":"66","author":"Yang","year":"2026","journal-title":"J. Basic Microbiol."},{"key":"2026102109071201500_cit40","doi-asserted-by":"publisher","first-page":"151844","DOI":"10.1016\/j.ijbiomac.2026.151844","article-title":"Glucose-6-phosphate\ndehydrogenase facilitates the synthesis of yellow Monascus pigment by providing energy","volume":"358","author":"Zhang","year":"2026","journal-title":"Int. J. Mol.\nSci."},{"key":"2026102109071201500_cit41","doi-asserted-by":"publisher","first-page":"490","DOI":"10.1016\/j.synbio.2026.01.002","article-title":"Metabolic modulation\nof TCA cycle by S-nitrosylation in Monascus spp","volume":"12","author":"Meng","year":"2026","journal-title":"Synth. Syst. 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Biotechnol."},{"key":"2026102109071201500_cit45","doi-asserted-by":"publisher","first-page":"227","DOI":"10.1016\/j.ymben.2021.07.003","article-title":"Integrated knowledge\nmining, genome-scale\nmodeling, and machine learning for predicting Yarrowia lipolytica bioproduction","volume":"67","author":"Czajka","year":"2021","journal-title":"Metab. Eng."},{"key":"2026102109071201500_cit46","doi-asserted-by":"publisher","first-page":"417","DOI":"10.1007\/978-1-0716-2617-7_18","article-title":"Machine learning and hybrid methods\nfor metabolic pathway modeling","volume":"2553","author":"Cuperlovic-Culf","year":"2023","journal-title":"Methods Mol.\nBiol."},{"issue":"1","key":"2026102109071201500_cit47","doi-asserted-by":"publisher","first-page":"127","DOI":"10.1021\/acssynbio.8b00398","article-title":"Machine\nlearning of designed translational control allows predictive pathway\noptimization in Escherichia coli","volume":"8","author":"Jervis","year":"2019","journal-title":"ACS Synth. 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Eng."}],"container-title":["Journal of Agricultural\nand Food Chemistry"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pubs.acs.org\/doi\/pdf\/10.1021\/acs.jafc.5c12726","content-type":"application\/pdf","content-version":"vor","intended-application":"unspecified"},{"URL":"https:\/\/pubs.acs.org\/jafcau\/article-pdf\/74\/41\/32219\/68764552\/acs.jafc.5c12726.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/pubs.acs.org\/jafcau\/article-pdf\/74\/41\/32219\/68764552\/acs.jafc.5c12726.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,10,21]],"date-time":"2026-10-21T13:49:57Z","timestamp":1792590597000},"score":1,"resource":{"primary":{"URL":"https:\/\/pubs.acs.org\/jafcau\/article\/74\/41\/32219\/5445675\/Machine-Learning-Guided-Cofactor-Engineering-for"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,10,6]]},"references-count":50,"journal-issue":{"issue":"41","published-online":{"date-parts":[[2026,10,6]]},"published-print":{"date-parts":[[2026,10,21]]}},"URL":"https:\/\/doi.org\/10.1021\/acs.jafc.5c12726","relation":{},"ISSN":["0021-8561","1520-5118"],"issn-type":[{"value":"0021-8561","type":"print"},{"value":"1520-5118","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,10,6]]}}}