{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,8]],"date-time":"2026-10-08T01:53:10Z","timestamp":1791424390041,"version":"4.3.3"},"reference-count":217,"publisher":"MDPI AG","issue":"13","license":[{"start":{"date-parts":[[2025,6,28]],"date-time":"2025-06-28T00:00:00Z","timestamp":1751068800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Riboflavin (vitamin B2) is an essential micronutrient required for all living organisms. It is naturally synthesized by plants and most microorganisms, including the bacterium Bacillus subtilis, the filamentous fungus Ashbya gossypii, and the yeast Candida famata\u2014all of which are known to be riboflavin overproducers. The choice of production organism in industrial applications depends on factors such as yield, ease of cultivation, and the availability of genetic tools. As a result, several microorganisms are commonly used, and their relative prominence can shift over time with advances in metabolic engineering and process optimization. This review presents a comparative analysis of riboflavin biosynthesis across prokaryotic and eukaryotic systems, with a particular focus on regulatory mechanisms governing flavinogenesis. Special attention is given to recent advances in metabolic engineering strategies, including the application of CRISPR\/Cas9 genome editing in Bacillus subtilis and Ashbya gossypii. In yeast systems, significant improvements in riboflavin production have been achieved primarily through the manipulation of transcriptional regulators (e.g., SEF1, SFU1, TUP1) and metabolic genes. The role of other important genes (PRS3, ADE4, ZWF1, GND1, RFE1, VMA1, etc.) in riboflavin overproduction in C. famata is described. The review also explores the use of alternative, low-cost feedstocks\u2014including lignocellulosic hydrolysates and dairy by-products\u2014to support more sustainable and economically viable riboflavin production. Although considerable progress has been achieved in genetic optimization and bioprocess development, further work is required to fine-tune metabolic flux and maximize riboflavin synthesis, particularly under industrial conditions. This review highlights key opportunities for future research aimed at refining metabolic interventions and expanding the use of renewable substrates for environmentally sustainable riboflavin production.<\/jats:p>","DOI":"10.3390\/ijms26136243","type":"journal-article","created":{"date-parts":[[2025,7,1]],"date-time":"2025-07-01T05:28:28Z","timestamp":1751347708000},"page":"6243","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":19,"title":["Regulation of Riboflavin Biosynthesis in Microorganisms and Construction of the Advanced Overproducers of This Vitamin"],"prefix":"10.3390","volume":"26","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-2850-6073","authenticated-orcid":false,"given":"Justyna","family":"Ruchala","sequence":"first","affiliation":[{"name":"Faculty of Biotechnology, Medical College, University of Rzesz\u00f3w, \u0106wikli\u0144skiej 2D, 35-601 Rzesz\u00f3w, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alicja","family":"Najdecka","sequence":"additional","affiliation":[{"name":"Faculty of Biotechnology, Medical College, University of Rzesz\u00f3w, \u0106wikli\u0144skiej 2D, 35-601 Rzesz\u00f3w, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0557-3476","authenticated-orcid":false,"given":"Dominik","family":"Wojdyla","sequence":"additional","affiliation":[{"name":"Faculty of Biotechnology, Medical College, University of Rzesz\u00f3w, \u0106wikli\u0144skiej 2D, 35-601 Rzesz\u00f3w, Poland"},{"name":"The Doctoral School of the University of Rzesz\u00f3w, University of Rzesz\u00f3w, 35-959 Rzesz\u00f3w, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wen","family":"Liu","sequence":"additional","affiliation":[{"name":"Department of Molecular Genetics and Biotechnology, Institute of Cell Biology, NAS of Ukraine, Drahomanov Street, 14\/16, 79005 Lviv, Ukraine"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andriy","family":"Sibirny","sequence":"additional","affiliation":[{"name":"Faculty of Biotechnology, Medical College, University of Rzesz\u00f3w, \u0106wikli\u0144skiej 2D, 35-601 Rzesz\u00f3w, Poland"},{"name":"Department of Molecular Genetics and Biotechnology, Institute of Cell Biology, NAS of Ukraine, Drahomanov Street, 14\/16, 79005 Lviv, Ukraine"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,6,28]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Mosegaard, S., Dipace, G., Bross, P., Carlsen, J., Gregersen, N., and Olsen, R.K.J. 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