{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,23]],"date-time":"2026-08-23T04:22:11Z","timestamp":1787458931443,"version":"build-2736575974"},"reference-count":36,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2026,8,21]],"date-time":"2026-08-21T00:00:00Z","timestamp":1787270400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100007807","name":"Japan Science Society","doi-asserted-by":"crossref","id":[{"id":"10.13039\/501100007807","id-type":"DOI","asserted-by":"crossref"}]},{"id":[{"id":"https:\/\/ror.org\/03aeyg892","id-type":"ROR","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100002241","name":"Japan Science and Technology Agency","doi-asserted-by":"publisher","award":["JPMJSP2124"],"award-info":[{"award-number":["JPMJSP2124"]}],"id":[{"id":"10.13039\/501100002241","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100002241","name":"Japan Science and Technology Agency","doi-asserted-by":"publisher","award":["JPMJAN23D6"],"award-info":[{"award-number":["JPMJAN23D6"]}],"id":[{"id":"10.13039\/501100002241","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100002241","name":"Japan Science and Technology Agency","doi-asserted-by":"publisher","award":["JPMJGX23B4"],"award-info":[{"award-number":["JPMJGX23B4"]}],"id":[{"id":"10.13039\/501100002241","id-type":"DOI","asserted-by":"publisher"}]},{"award":["JPMJSP2124"],"award-info":[{"award-number":["JPMJSP2124"]}],"id":[{"id":"https:\/\/ror.org\/00097mb19","id-type":"ROR","asserted-by":"publisher"}]},{"award":["JPMJAN23D6"],"award-info":[{"award-number":["JPMJAN23D6"]}],"id":[{"id":"https:\/\/ror.org\/00097mb19","id-type":"ROR","asserted-by":"publisher"}]},{"award":["JPMJGX23B4"],"award-info":[{"award-number":["JPMJGX23B4"]}],"id":[{"id":"https:\/\/ror.org\/00097mb19","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["BioTech"],"abstract":"<jats:p>Raspberry ketone (RK) is a valuable natural flavor compound, but its extraction from plants is inefficient because of its low natural abundance. Microbial production of RK offers a promising alternative; however, insufficient availability of malonyl-CoA can limit RK biosynthesis. In this study, we introduced a malonate-dependent malonyl-CoA supply module into an engineered Escherichia coli strain designed for de novo RK production through heterologous expression of malonyl-CoA synthetase MatB and malonate transporter MatC. In the presence of malonate, the introduction of the MatB\u2013MatC module increased RK production by 2.3-fold. To optimize malonate supplementation, RK pathway metabolites, including intracellular acyl-CoA intermediates, were quantified by liquid chromatography\u2013mass spectrometry, and the resulting metabolite profiles were analyzed by principal component analysis, hierarchical clustering, and correlation analysis. This study indicated that 50-mM malonate was optimal, yielding 24 mg\/L RK during 96-deep-well plate cultivation. Fed-batch optimization increased RK production to 301 mg\/L in a 100-mL jar fermenter, and scale-up cultivation in a 2-L jar fermenter produced 340 mg\/L RK. In this study, optimizing MatB\u2013MatC-dependent malonyl-CoA supply, combined with pathway-level metabolic profiling and controlled fed-batch cultivation, enhanced de novo RK production in E. coli.<\/jats:p>","DOI":"10.3390\/biotech15030071","type":"journal-article","created":{"date-parts":[[2026,8,21]],"date-time":"2026-08-21T15:51:05Z","timestamp":1787327465000},"page":"71","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Optimizing MatB\u2013MatC-Dependent Malonyl-CoA Supply for Enhanced Raspberry Ketone Production in Escherichia coli"],"prefix":"10.3390","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0009-0006-0484-4165","authenticated-orcid":false,"given":"Kurumi","family":"Usui","sequence":"first","affiliation":[{"name":"Degree Programs in Life and Earth Sciences, Microbiology Research Center for Sustainability, University of Tsukuba, 1-1-1 Tennodai, Tsukuba 305-8572, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Naoki","family":"Takaya","sequence":"additional","affiliation":[{"name":"Faculty of Life and Environmental Sciences, Microbiology Research Center for Sustainability, University of Tsukuba, 1-1-1 Tennodai, Tsukuba 305-8572, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0454-3315","authenticated-orcid":false,"given":"Shunsuke","family":"Masuo","sequence":"additional","affiliation":[{"name":"Faculty of Life and Environmental Sciences, Microbiology Research Center for Sustainability, University of Tsukuba, 1-1-1 Tennodai, Tsukuba 305-8572, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,8,21]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1270","DOI":"10.1002\/biot.200700076","article-title":"Microbial production of natural raspberry ketone","volume":"2","author":"Beekwilder","year":"2007","journal-title":"Biotechnol. 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