{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,18]],"date-time":"2026-10-18T11:59:06Z","timestamp":1792324746256,"version":"4.4.0"},"reference-count":55,"publisher":"American Chemical Society (ACS)","issue":"29","license":[{"start":{"date-parts":[[2026,7,17]],"date-time":"2026-07-17T00:00:00Z","timestamp":1784246400000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-029"},{"start":{"date-parts":[[2026,7,17]],"date-time":"2026-07-17T00:00:00Z","timestamp":1784246400000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-037"},{"start":{"date-parts":[[2026,7,17]],"date-time":"2026-07-17T00:00:00Z","timestamp":1784246400000},"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":["22178372"],"award-info":[{"award-number":["22178372"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["22208368"],"award-info":[{"award-number":["22208368"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100002367","name":"Chinese Academy of Sciences","doi-asserted-by":"publisher","award":["306GJHZ2023019GC"],"award-info":[{"award-number":["306GJHZ2023019GC"]}],"id":[{"id":"10.13039\/501100002367","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100002367","name":"Chinese Academy of Sciences","doi-asserted-by":"publisher","award":["306GJHZ2025003BS"],"award-info":[{"award-number":["306GJHZ2025003BS"]}],"id":[{"id":"10.13039\/501100002367","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100012166","name":"National Key Research and Development Program of China","doi-asserted-by":"publisher","award":["2022YFC2106100"],"award-info":[{"award-number":["2022YFC2106100"]}],"id":[{"id":"10.13039\/501100012166","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100014219","name":"National Science Fund for Distinguished Young Scholars","doi-asserted-by":"publisher","award":["22325807"],"award-info":[{"award-number":["22325807"]}],"id":[{"id":"10.13039\/501100014219","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/na","name":"Tianjin Major Science and Technology Projects and Engineering Programs","doi-asserted-by":"publisher","award":["25ZXZSSS00190"],"award-info":[{"award-number":["25ZXZSSS00190"]}],"id":[{"id":"10.13039\/na","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2026,7,29]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Riboflavin is an essential nutrient widely used in food, feed, and pharmaceutical industries, yet efficient screening of high-yield microbial strains remains a major bottleneck in industrial production. Here, we developed an integrated strategy combining dual selective pressure, optimized microplate fermentation, and stepwise high-throughput screening to accelerate the isolation of riboflavin-overproducing Bacillus subtilis mutants. A dual selective pressure system employing phosphonoarginine-5-amino-6-(d-ribosylamino)uracil (Arp) and the GTP analog 8-azaguanine (8-AZG) enhanced the positive mutation rate. Optimization of microplate culture enabled strong correlation across 96-well, 24-well, and shake-flask fermentations. Stepwise screening further enriched stable high producers, achieving a final positive rate of 21.4%. The best mutant, Hs-G87, produced 4.1 g\/L riboflavin in shake flasks and 32.1 g\/L in a 5 L bioreactor (yield of 0.11 g\/g glucose; productivity of 0.594 g\/L\/h), demonstrating strong industrial potential.<\/jats:p>","DOI":"10.1021\/acs.jafc.5c13930","type":"journal-article","created":{"date-parts":[[2026,7,17]],"date-time":"2026-07-17T07:22:36Z","timestamp":1784272956000},"page":"22952-22964","source":"Crossref","is-referenced-by-count":1,"title":["Dual Selective\nPressure and Intelligent Screening\nof Mutagenized\n                    <i>Bacillus subtilis<\/i>\n                    for\nEnhanced Riboflavin Production"],"prefix":"10.1021","volume":"74","author":[{"given":"Fangyu","family":"Zhang","sequence":"first","affiliation":[{"name":"China Three Gorges University , , , ,","place":["Yichang, Hubei, China, 443002"]},{"name":"Chinese Academy of Sciences , , ,","place":["Tianjin, China, 300308"]},{"name":"National Center of Technology Innovation for Synthetic Biology , ,","place":["Tianjin, China, 300308"]},{"name":"Chinese Academy of Sciences , , ,","place":["Tianjin, China, 300308"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Guangqing","family":"Du","sequence":"additional","affiliation":[{"name":"Chinese Academy of Sciences , , ,","place":["Tianjin, China, 300308"]},{"name":"National Center of Technology Innovation for Synthetic Biology , ,","place":["Tianjin, China, 300308"]},{"name":"Chinese Academy of Sciences , , ,","place":["Tianjin, China, 300308"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Miaomiao","family":"Xia","sequence":"additional","affiliation":[{"name":"Chinese Academy of Sciences , , ,","place":["Tianjin, China, 300308"]},{"name":"National Center of Technology Innovation for Synthetic Biology , ,","place":["Tianjin, China, 300308"]},{"name":"Chinese Academy of Sciences , , ,","place":["Tianjin, China, 300308"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dongqin","family":"Ding","sequence":"additional","affiliation":[{"name":"Chinese Academy of Sciences , , ,","place":["Tianjin, China, 300308"]},{"name":"National Center of Technology Innovation for Synthetic Biology , ,","place":["Tianjin, China, 300308"]},{"name":"Chinese Academy of Sciences , , ,","place":["Tianjin, China, 300308"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhengzi","family":"Ji","sequence":"additional","affiliation":[{"name":"China Three Gorges University , , , ,","place":["Yichang, Hubei, China, 443002"]},{"name":"Chinese Academy of Sciences , , ,","place":["Tianjin, China, 300308"]},{"name":"National Center of Technology Innovation for Synthetic Biology , ,","place":["Tianjin, China, 300308"]},{"name":"Chinese Academy of Sciences , , ,","place":["Tianjin, China, 300308"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ziyu","family":"Yang","sequence":"additional","affiliation":[{"name":"China Three Gorges University , , , ,","place":["Yichang, Hubei, China, 443002"]},{"name":"Chinese Academy of Sciences , , ,","place":["Tianjin, China, 300308"]},{"name":"National Center of Technology Innovation for Synthetic Biology , ,","place":["Tianjin, China, 300308"]},{"name":"Chinese Academy of Sciences , , ,","place":["Tianjin, China, 300308"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Junlin","family":"Yang","sequence":"additional","affiliation":[{"name":"Yu Xing Biotechnology (Group) Co., Ltd , , ,","place":["Xingtai, Hebei, China, 055550"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yufeng","family":"Wang","sequence":"additional","affiliation":[{"name":"Yu Xing Biotechnology (Group) Co., Ltd , , ,","place":["Xingtai, Hebei, China, 055550"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cuigai","family":"Cheng","sequence":"additional","affiliation":[{"name":"Yu Xing Biotechnology (Group) Co., Ltd , , ,","place":["Xingtai, Hebei, China, 055550"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dachun","family":"Gong","sequence":"additional","affiliation":[{"name":"China Three Gorges University , , , ,","place":["Yichang, Hubei, China, 443002"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5857-394X","authenticated-orcid":true,"given":"Dawei","family":"Zhang","sequence":"additional","affiliation":[{"name":"Chinese Academy of Sciences , , ,","place":["Tianjin, China, 300308"]},{"name":"National Center of Technology Innovation for Synthetic Biology , ,","place":["Tianjin, China, 300308"]},{"name":"Chinese Academy of Sciences , , ,","place":["Tianjin, China, 300308"]},{"name":"University of Chinese Academy of Sciences , ,","place":["Beijing, China, 100190"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,7,17]]},"reference":[{"issue":"3","key":"2026072910071553000_cit1","doi-asserted-by":"publisher","first-page":"950","DOI":"10.3390\/ijms21030950","article-title":"Riboflavin:\nThe health benefits of a forgotten natural vitamin","volume":"21","author":"Suwannasom","year":"2020","journal-title":"Int. J. Mol. Sci."},{"key":"2026072910071553000_cit2","doi-asserted-by":"publisher","first-page":"115","DOI":"10.1016\/j.eng.2021.03.018","article-title":"Biotechnological\nstrategies of riboflavin biosynthesis in microbes","volume":"12","author":"Zhang","year":"2022","journal-title":"Eng."},{"issue":"4\u20135","key":"2026072910071553000_cit3","doi-asserted-by":"publisher","first-page":"659","DOI":"10.1007\/s10295-016-1842-7","article-title":"Bioproduction of riboflavin: A bright\nyellow history","volume":"44","author":"Revuelta","year":"2017","journal-title":"J. Ind. Microbiol. Biotechnol."},{"issue":"1","key":"2026072910071553000_cit4","doi-asserted-by":"publisher","first-page":"164","DOI":"10.3390\/microorganisms11010164","article-title":"Metabolic engineering\nof Bacillus\nsubtilis for riboflavin production: A review","volume":"11","author":"Liu","year":"2023","journal-title":"Microorganisms"},{"key":"2026072910071553000_cit5","doi-asserted-by":"publisher","first-page":"173","DOI":"10.1186\/s12934-020-01436-8","article-title":"Bacillus subtilis: A universal cell\nfactory for industry, agriculture, biomaterials and medicine","volume":"19","author":"Su","year":"2020","journal-title":"Microb. Cell Fact."},{"key":"2026072910071553000_cit6","doi-asserted-by":"publisher","first-page":"125228","DOI":"10.1016\/j.biortech.2021.125228","article-title":"Metabolic\nengineering\nof Bacillus subtilis for enhancing riboflavin production by alleviating\ndissolved oxygen limitation","volume":"333","author":"You","year":"2021","journal-title":"Bioresour. Technol."},{"key":"2026072910071553000_cit7","doi-asserted-by":"publisher","first-page":"58","DOI":"10.1016\/j.ymben.2020.05.002","article-title":"Rational engineering\nof transcriptional riboswitches leads to enhanced metabolite levels\nin Bacillus subtilis","volume":"61","author":"Boumezbeur","year":"2020","journal-title":"Metab. Eng."},{"issue":"41","key":"2026072910071553000_cit8","doi-asserted-by":"publisher","first-page":"12241","DOI":"10.1021\/acs.jafc.1c04471","article-title":"Rational engineering\nof Escherichia\ncoli for high-level production of riboflavin","volume":"69","author":"Liu","year":"2021","journal-title":"J. Agric. Food Chem."},{"issue":"14","key":"2026072910071553000_cit9","doi-asserted-by":"publisher","first-page":"gkaf693","DOI":"10.1093\/nar\/gkaf693","article-title":"A quorum sensing-controlled\ntype I Crispi toolkit for dynamically regulating metabolic flux","volume":"53","author":"Zheng","year":"2025","journal-title":"Nucleic Acids Res."},{"issue":"5","key":"2026072910071553000_cit10","doi-asserted-by":"publisher","first-page":"1801","DOI":"10.1021\/acssynbio.1c00640","article-title":"Increased\nproduction of riboflavin by coordinated expression of multiple genes\nin operons in Bacillus subtilis","volume":"11","author":"You","year":"2022","journal-title":"ACS Synth.\nBiol."},{"issue":"4","key":"2026072910071553000_cit11","doi-asserted-by":"publisher","first-page":"3934","DOI":"10.1016\/j.biortech.2010.11.120","article-title":"Enhancement\nof riboflavin production with Bacillus subtilis by expression and\nsite-directed mutagenesis of zwf and gnd gene from Corynebacterium\nglutamicum","volume":"102","author":"Wang","year":"2011","journal-title":"Bioresour. Technol."},{"issue":"4","key":"2026072910071553000_cit12","doi-asserted-by":"publisher","first-page":"1264","DOI":"10.1021\/acssynbio.4c00884","article-title":"Advancing VB12 production:\nInsights into enhancing VB12 titer in Ensifer\nadhaerens Casida A through ARTP mutagenesis and multiomics analysis","volume":"14","author":"Wang","year":"2025","journal-title":"ACS Synth. Biol."},{"key":"2026072910071553000_cit13","doi-asserted-by":"publisher","first-page":"5639","DOI":"10.1007\/s00253-015-6678-y","article-title":"Quantitative\nevaluation of DNA damage and mutation rate by atmospheric and room-temperature\nplasma (ARTP) and conventional mutagenesis","volume":"99","author":"Zhang","year":"2015","journal-title":"Appl. Microbiol. Biotechnol."},{"key":"2026072910071553000_cit14","doi-asserted-by":"publisher","first-page":"63","DOI":"10.1186\/s40643-025-00907-3","article-title":"Current breakthroughs and advances\nin atmospheric room temperature plasma (ARTP) technology for biomanufacturing","volume":"12","author":"Li","year":"2025","journal-title":"Bioresour. Bioprocess"},{"key":"2026072910071553000_cit15","doi-asserted-by":"publisher","first-page":"16","DOI":"10.1007\/s11274-016-2156-7","article-title":"Highly improved\nacarbose production\nof Actinomyces through the combination of ARTP and penicillin susceptible\nmutant screening","volume":"33","author":"Ren","year":"2017","journal-title":"World J. Microbiol. Biotechnol."},{"issue":"14","key":"2026072910071553000_cit16","doi-asserted-by":"publisher","first-page":"6363","DOI":"10.1007\/s00253-020-10703-y","article-title":"ARTP\nmutation and adaptive\nlaboratory evolution improve probiotic performance of Bacillus coagulans","volume":"104","author":"Liu","year":"2020","journal-title":"Appl. Microbiol. Biotechnol."},{"issue":"1","key":"2026072910071553000_cit17","doi-asserted-by":"publisher","first-page":"27","DOI":"10.1080\/07391102.2012.745821","article-title":"Solution structures\nof purine base analogues 6-chloroguanine,\n8-azaguanine and allopurinol","volume":"32","author":"Gogia","year":"2014","journal-title":"J. Biomol. Struct.\nDyn."},{"key":"2026072910071553000_cit18","doi-asserted-by":"publisher","first-page":"139","DOI":"10.1016\/j.procbio.2024.11.020","article-title":"Enhancing\nriboflavin production by modulating the expression of the rib operon\nin Bacillus subtilis","volume":"148","author":"Yao","year":"2025","journal-title":"Process Biochem."},{"key":"2026072910071553000_cit19","doi-asserted-by":"publisher","first-page":"123856","DOI":"10.1016\/j.biortech.2020.123856","article-title":"Target-site directed\nrational high-throughput\nscreening system for high sophorolipids production by Candida bombicola","volume":"315","author":"Chen","year":"2020","journal-title":"Bioresour. Technol."},{"key":"2026072910071553000_cit20","doi-asserted-by":"publisher","first-page":"1748","DOI":"10.3389\/fmicb.2019.01748","article-title":"Real-time detection of riboflavin\nproduction by Lactobacillus\nplantarum strains and tracking of their gastrointestinal survival\nand functionality in vitro and in vivo using mCherry labeling","volume":"10","author":"Mohedano","year":"2019","journal-title":"Front. Microbiol."},{"key":"2026072910071553000_cit21","doi-asserted-by":"publisher","first-page":"D1670","DOI":"10.1093\/nar\/gkae973","article-title":"Generic Diagramming Platform (GDP):\nA Comprehensive Database of High-Quality Biomedical Graphics","volume":"53","author":"Jiang","year":"2025","journal-title":"Nucleic Acids Res."},{"key":"2026072910071553000_cit22","doi-asserted-by":"publisher","first-page":"159","DOI":"10.1186\/s12934-024-02426-w","article-title":"De novo engineering\nriboflavin production in Bacillus subtilis by overexpressing the downstream\ngenes in the purine biosynthesis pathway","volume":"23","author":"Liu","year":"2024","journal-title":"Microb.\nCell Fact."},{"key":"2026072910071553000_cit23","doi-asserted-by":"publisher","first-page":"142072","DOI":"10.1016\/j.foodchem.2024.142072","article-title":"Combined fermentation\nand ARTP mutagenesis to enhance lipase activity\nof Penicillium camembertii and its application for high-purity 1,3-diacylglycerol\npreparation","volume":"465","author":"Ji","year":"2025","journal-title":"Food Chem."},{"key":"2026072910071553000_cit24","doi-asserted-by":"publisher","first-page":"1051425","DOI":"10.3389\/fmicb.2022.1051425","article-title":"Toxic Effect\nand Inability of L-Homoserine to Be a Nitrogen Source for Growth of\nEscherichia coli Resolved by a Combination of In Vivo Evolution Engineering\nand Omics Analyses","volume":"13","author":"Alkim","year":"2022","journal-title":"Front. Microbiol."},{"issue":"7","key":"2026072910071553000_cit25","doi-asserted-by":"publisher","first-page":"1813","DOI":"10.1128\/JB.01166-09","article-title":"Alternative Route\nfor Glyoxylate Consumption during Growth on Two-Carbon Compounds by\nMethylobacterium extorquens AM1","volume":"192","author":"Okubo","year":"2010","journal-title":"J. Bacteriol."},{"key":"2026072910071553000_cit26","doi-asserted-by":"publisher","first-page":"82","DOI":"10.1016\/j.ymben.2018.05.012","article-title":"Adaptation to the Coupling of Glycolysis\nto Toxic Methylglyoxal Production in tpiA Deletion Strains of Escherichia\ncoli Requires Synchronized and Counterintuitive Genetic Changes","volume":"48","author":"McCloskey","year":"2018","journal-title":"Metab. Eng."},{"key":"2026072910071553000_cit27","doi-asserted-by":"publisher","first-page":"33","DOI":"10.3390\/genes7070033","article-title":"Stationary-phase mutagenesis in stressed Bacillus subtilis\ncells operates by Mfd-dependent mutagenic pathways","volume":"7","author":"Go\u0301mez-Marroqui\u0301n","year":"2016","journal-title":"Genes"},{"issue":"5","key":"2026072910071553000_cit28","doi-asserted-by":"publisher","DOI":"10.1128\/aac.01462-23","article-title":"Exploring the links between SOS response,\nmutagenesis,\nand resistance during the recovery period","volume":"68","author":"Ghosh","year":"2024","journal-title":"Antimicrob.\nAgents Chemother."},{"key":"2026072910071553000_cit29","doi-asserted-by":"publisher","first-page":"484","DOI":"10.1186\/s12866-025-04120-2","article-title":"Enhancement of carrimycin\nproduction in Streptomyces spiramyceticus 54-IA by the reporter-guided\nselection method with traditional mutagenesis","volume":"25","author":"Li","year":"2025","journal-title":"BMC Microbiol."},{"key":"2026072910071553000_cit30","doi-asserted-by":"publisher","first-page":"603514","DOI":"10.3389\/fbioe.2020.603514","article-title":"Identification of 8-azaguanine\nbiosynthesis-related genes provides insight into the enzymatic and\nnon-enzymatic biosynthetic pathway for 1,2,3-triazole","volume":"8","author":"Hou","year":"2020","journal-title":"Front. Bioeng. Biotechnol."},{"key":"2026072910071553000_cit31","doi-asserted-by":"publisher","first-page":"fnab154","DOI":"10.1093\/femsle\/fnab154","article-title":"Iterative mutagenesis\ninduced by atmospheric and room temperature plasma treatment under\nmultiple selection pressures for the improvement of coenzyme Q10 production\nby Rhodobacter sphaeroides","volume":"368","author":"Wang","year":"2021","journal-title":"FEMS Microbiol.\nLett."},{"key":"2026072910071553000_cit32","doi-asserted-by":"publisher","DOI":"10.3791\/63849","article-title":"Enhanced\nreproducibility\nand precision of high-throughput quantification of bacterial growth\ndata using a microplate reader","volume":"185","author":"Abkar","year":"2022","journal-title":"J. Vis. Exp."},{"issue":"4","key":"2026072910071553000_cit33","doi-asserted-by":"publisher","first-page":"565","DOI":"10.1002\/mlf2.12147","article-title":"Measurement of optical density of microbes by multi-light path transmission\nmethod","volume":"3","author":"Wang","year":"2024","journal-title":"mLife"},{"key":"2026072910071553000_cit34","doi-asserted-by":"publisher","first-page":"1831","DOI":"10.1007\/s00217-022-04009-8","article-title":"Evaluation of microtiter plate as\na high-throughput screening platform for beer fermentation","volume":"248","author":"Zhao","year":"2022","journal-title":"Eur. Food Res. Technol."},{"key":"2026072910071553000_cit35","doi-asserted-by":"publisher","first-page":"147","DOI":"10.1186\/s12934-016-0546-z","article-title":"High-throughput fermentation screening for the yeast Yarrowia lipolytica\nwith real-time monitoring of biomass and lipid production","volume":"15","author":"Back","year":"2016","journal-title":"Microb. Cell Fact."},{"key":"2026072910071553000_cit36","doi-asserted-by":"publisher","first-page":"661","DOI":"10.1007\/s11274-013-1492-0","article-title":"Enhanced\nriboflavin production by\nrecombinant Bacillus subtilis RF1 through the optimization of agitation\nspeed","volume":"30","author":"Man","year":"2014","journal-title":"World J. Microbiol. Biotechnol."},{"issue":"6","key":"2026072910071553000_cit37","doi-asserted-by":"publisher","first-page":"1118","DOI":"10.1002\/bit.22341","article-title":"The baffled\nmicrotiter plate: increased oxygen transfer\nand improved online monitoring in small scale fermentations","volume":"103","author":"Funke","year":"2009","journal-title":"Biotechnol. Bioeng."},{"key":"2026072910071553000_cit38","doi-asserted-by":"publisher","first-page":"1202836","DOI":"10.3389\/fbioe.2023.1202836","article-title":"Technical upgrade of an open-source liquid handler\nto support bacterial colony screening","volume":"11","author":"del Olmo Lianes","year":"2023","journal-title":"Front.\nBioeng. Biotechnol."},{"key":"2026072910071553000_cit39","doi-asserted-by":"publisher","first-page":"119","DOI":"10.1016\/j.procbio.2022.05.018","article-title":"Enhancing protease\nactivity of Bacillus subtilis using UV-laser random mutagenesis and\nhigh-throughput screening","volume":"119","author":"Lu","year":"2022","journal-title":"Process Biochem."},{"key":"2026072910071553000_cit40","doi-asserted-by":"publisher","first-page":"117376","DOI":"10.1016\/j.bios.2025.117376","article-title":"Biosensor-based dual-color\ndroplet microfluidic platform for precise high-throughput screening\nof erythromycin hyperproducers","volume":"278","author":"Zhao","year":"2025","journal-title":"Biosens. Bioelectron."},{"key":"2026072910071553000_cit41","doi-asserted-by":"publisher","first-page":"119","DOI":"10.1007\/s11274-020-02901-7","article-title":"Degeneration of industrial bacteria\ncaused by genetic instability","volume":"36","author":"Peng","year":"2020","journal-title":"World J. Microbiol.\nBiotechnol."},{"issue":"1","key":"2026072910071553000_cit42","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1038\/sj.jim.2900590","article-title":"GTP cyclohydrolase\nII and 3,4-dihydroxy-2-butanone\n4-phosphate synthase are rate-limiting enzymes in riboflavin synthesis\nof an industrial Bacillus subtilis strain used for riboflavin production","volume":"22","author":"Hu\u0308mbelin","year":"1999","journal-title":"J. Ind. Microbiol. Biotechnol."},{"issue":"1","key":"2026072910071553000_cit43","doi-asserted-by":"publisher","first-page":"70","DOI":"10.1016\/j.jsb.2017.12.006","article-title":"Structural\ncharacterization of ribT from Bacillus subtilis reveals it as a GCN5-related\nN-acetyltransferase","volume":"202","author":"Srivastava","year":"2018","journal-title":"J. Struct. Biol."},{"key":"2026072910071553000_cit44","doi-asserted-by":"publisher","first-page":"138","DOI":"10.1016\/j.ymben.2018.05.022","article-title":"Integrated\nwhole-genome and transcriptome sequence analysis reveals the genetic\ncharacteristics of a riboflavin-overproducing Bacillus subtilis","volume":"48","author":"Wang","year":"2018","journal-title":"Metab. Eng."},{"key":"2026072910071553000_cit45","doi-asserted-by":"publisher","first-page":"1893","DOI":"10.1007\/s11274-014-1611-6","article-title":"Enhancement\nof riboflavin production by deregulating gluconeogenesis in Bacillus\nsubtilis","volume":"30","author":"Wang","year":"2014","journal-title":"World J. Microbiol. Biotechnol."},{"issue":"45","key":"2026072910071553000_cit46","doi-asserted-by":"publisher","first-page":"29140","DOI":"10.1021\/acsomega.0c03867","article-title":"Manipulation of purine metabolic\nnetworks for riboflavin production in Bacillus subtilis","volume":"5","author":"Sun","year":"2020","journal-title":"ACS Omega"},{"issue":"1","key":"2026072910071553000_cit47","doi-asserted-by":"publisher","first-page":"49","DOI":"10.1016\/S1096-7176(03)00007-7","article-title":"Reducing maintenance\nmetabolism by metabolic engineering of respiration improves riboflavin\nproduction by Bacillus subtilis","volume":"5","author":"Zamboni","year":"2003","journal-title":"Metab. Eng."},{"issue":"1","key":"2026072910071553000_cit48","doi-asserted-by":"publisher","first-page":"150","DOI":"10.1186\/s12934-017-0764-z","article-title":"Mixomics analysis of\nBacillus subtilis: effect of oxygen availability on riboflavin production","volume":"16","author":"Hu","year":"2017","journal-title":"Microb. Cell Fact."},{"issue":"4","key":"2026072910071553000_cit49","doi-asserted-by":"publisher","first-page":"531","DOI":"10.1007\/s13205-014-0252-7","article-title":"Optimization\nof fermentation upstream parameters and\nimmobilization of Corynebacterium glutamicum MH 20\u201322 B Cells\nto enhance the production of L-Lysine","volume":"5","author":"Razak","year":"2015","journal-title":"3 Biotech"},{"key":"2026072910071553000_cit50","doi-asserted-by":"publisher","first-page":"1070","DOI":"10.3390\/microorganisms11041070","article-title":"Characterization\nof a riboflavin-producing mutant of\nBacillus subtilis isolated by droplet-based microfluidics screening","volume":"11","author":"Xu","year":"2023","journal-title":"Microorganisms"},{"key":"2026072910071553000_cit51","doi-asserted-by":"publisher","first-page":"783","DOI":"10.1007\/s00253-007-1049-y","article-title":"Optimization\nof riboflavin production by recombinant Bacillus subtilis RH44 using\nstatistical designs","volume":"76","author":"Wu","year":"2007","journal-title":"Appl. Microbiol. Biotechnol."},{"issue":"4","key":"2026072910071553000_cit52","doi-asserted-by":"publisher","first-page":"045012","DOI":"10.1088\/1758-5090\/ad628e","article-title":"Riboflavin\noverproduction from diverse feedstocks with engineered Corynebacterium\nglutamicum","volume":"16","author":"Pe\u0301rez-Garci\u0301a","year":"2024","journal-title":"Biofabrication"},{"key":"2026072910071553000_cit53","doi-asserted-by":"publisher","first-page":"1315","DOI":"10.1007\/s00253-011-3325-0","article-title":"The improvement\nof riboflavin production in Ashbya gossypii via disparity mutagenesis\nand DNA microarray analysis","volume":"91","author":"Park","year":"2011","journal-title":"Appl. Microbiol.\nBiotechnol."},{"key":"2026072910071553000_cit54","doi-asserted-by":"publisher","first-page":"67","DOI":"10.1016\/j.copbio.2017.06.007","article-title":"Recent advances in systems\nmetabolic engineering tools and strategies","volume":"47","author":"Chae","year":"2017","journal-title":"Curr. Opin. Biotechnol."},{"issue":"1","key":"2026072910071553000_cit55","doi-asserted-by":"publisher","first-page":"253","DOI":"10.1186\/s12934-025-02873-z","article-title":"Advances in high-throughput mutation breeding\nsystems integrating atmospheric and room-temperature plasma (ARTP)\nwith droplet-based microfluidics","volume":"24","author":"Xu","year":"2025","journal-title":"Microb. Cell\nFact."}],"container-title":["Journal of Agricultural\nand Food Chemistry"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pubs.acs.org\/doi\/pdf\/10.1021\/acs.jafc.5c13930","content-type":"application\/pdf","content-version":"vor","intended-application":"unspecified"},{"URL":"https:\/\/pubs.acs.org\/jafcau\/article-pdf\/74\/29\/22952\/65987647\/acs.jafc.5c13930.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/pubs.acs.org\/jafcau\/article-pdf\/74\/29\/22952\/65987647\/acs.jafc.5c13930.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,7,29]],"date-time":"2026-07-29T14:16:59Z","timestamp":1785334619000},"score":1,"resource":{"primary":{"URL":"https:\/\/pubs.acs.org\/jafcau\/article\/74\/29\/22952\/5212686\/Dual-Selective-Pressure-and-Intelligent-Screening"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,7,17]]},"references-count":55,"journal-issue":{"issue":"29","published-online":{"date-parts":[[2026,7,17]]},"published-print":{"date-parts":[[2026,7,29]]}},"URL":"https:\/\/doi.org\/10.1021\/acs.jafc.5c13930","relation":{},"ISSN":["0021-8561","1520-5118"],"issn-type":[{"value":"0021-8561","type":"print"},{"value":"1520-5118","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,7,17]]}}}