{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,29]],"date-time":"2025-10-29T13:42:33Z","timestamp":1761745353879,"version":"build-2065373602"},"reference-count":27,"publisher":"MDPI AG","issue":"22","license":[{"start":{"date-parts":[[2020,11,12]],"date-time":"2020-11-12T00:00:00Z","timestamp":1605139200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100002878","name":"Consejer\u00eda de Econom\u00eda, Innovaci\u00f3n, Ciencia y Empleo, Junta de Andaluc\u00eda","doi-asserted-by":"publisher","award":["D.P12-TEP-725","PAIDI TEP-105","PAIDI CTS-569"],"award-info":[{"award-number":["D.P12-TEP-725","PAIDI TEP-105","PAIDI CTS-569"]}],"id":[{"id":"10.13039\/501100002878","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>D-DIBOA (4-hydroxy-(2H)-1,4-benzoxazin-3-(4H)-one) is an allelopathic-derived compound with interesting herbicidal, fungicidal, and insecticide properties whose production has been successfully achieved by biocatalysis using a genetically engineered Escherichia coli strain. However, improvement and scaling-up of this process are hampered by the current methodology for D-DIBOA quantification, which is based on high-performance liquid chromatographic (HPLC), a time-consuming technique that requires expensive equipment and the use of environmentally unsafe solvents. In this work, we established and validated a rapid, simple, and sensitive spectrophotometric method for the quantification of the D-DIBOA produced by whole-cell biocatalysis, with limits of detection and quantification of 0.0165 and 0.0501 \u00b5mol\u00b7mL\u22121 respectively. This analysis takes place in only a few seconds and can be carried out using 100 \u00b5L of the sample in a microtiter plate reader. We performed several whole-cell biocatalysis strategies to optimize the process by monitoring D-DIBOA production every hour to keep control of both precursor and D-DIBOA concentrations in the bioreactor. These experiments allowed increasing the D-DIBOA production from the previously reported 5.01 mM up to 7.17 mM (43% increase). This methodology will facilitate processes such as the optimization of the biocatalyst, the scaling up, and the downstream purification.<\/jats:p>","DOI":"10.3390\/ijms21228523","type":"journal-article","created":{"date-parts":[[2020,11,12]],"date-time":"2020-11-12T10:00:32Z","timestamp":1605175232000},"page":"8523","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Optimization of the Biocatalysis for D-DIBOA Synthesis Using a Quick and Sensitive New Spectrophotometric Quantification Method"],"prefix":"10.3390","volume":"21","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-0196-6443","authenticated-orcid":false,"given":"Gema","family":"Cabrera","sequence":"first","affiliation":[{"name":"Department of Chemical Engineering and Food Technology, Campus Universitario de Puerto Real, University of Cadiz, 11510 Puerto Real, Spain"},{"name":"Institute of Viticulture and Agri-Food Research (IVAGRO)\u2014International Campus of Excellence (ceiA3), University of Cadiz, 11510 Puerto Real, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Teresa","family":"Linares","sequence":"additional","affiliation":[{"name":"Department of Biomedicine, Biotechnology and Public Health-Biochemistry and Molecular Biology, Campus Universitario de Puerto Real, University of Cadiz, 11510 Puerto Real, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Maria Elena","family":"de la Calle","sequence":"additional","affiliation":[{"name":"Department of Chemical Engineering and Food Technology, Campus Universitario de Puerto Real, University of Cadiz, 11510 Puerto Real, Spain"},{"name":"Institute of Viticulture and Agri-Food Research (IVAGRO)\u2014International Campus of Excellence (ceiA3), University of Cadiz, 11510 Puerto Real, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1643-2831","authenticated-orcid":false,"given":"Domingo","family":"Cantero","sequence":"additional","affiliation":[{"name":"Department of Chemical Engineering and Food Technology, Campus Universitario de Puerto Real, University of Cadiz, 11510 Puerto Real, Spain"},{"name":"Institute of Viticulture and Agri-Food Research (IVAGRO)\u2014International Campus of Excellence (ceiA3), University of Cadiz, 11510 Puerto Real, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4459-4627","authenticated-orcid":false,"given":"Antonio","family":"Valle","sequence":"additional","affiliation":[{"name":"Institute of Viticulture and Agri-Food Research (IVAGRO)\u2014International Campus of Excellence (ceiA3), University of Cadiz, 11510 Puerto Real, Spain"},{"name":"Department of Biomedicine, Biotechnology and Public Health-Biochemistry and Molecular Biology, Campus Universitario de Puerto Real, University of Cadiz, 11510 Puerto Real, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2645-9528","authenticated-orcid":false,"given":"Jorge","family":"Bolivar","sequence":"additional","affiliation":[{"name":"Department of Biomedicine, Biotechnology and Public Health-Biochemistry and Molecular Biology, Campus Universitario de Puerto Real, University of Cadiz, 11510 Puerto Real, Spain"},{"name":"Institute of Biomolecules (INBIO), University of Cadiz, 11510 Puerto Real, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2020,11,12]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Rimando, A.M., and Duke, S.O. 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