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Apart from an engineered glycerol utilization pathway that generates NADH, the strain was equipped with the NADH-dependent reductive branch of the TCA cycle (rTCA) and a heterologous SA exporter. However, the results indicated that a significant amount of carbon still entered the CO2-releasing oxidative TCA cycle. The current study aimed to tune down the flux through the oxidative TCA cycle by targeting the mitochondrial uptake of pyruvate and cytosolic intermediates of the rTCA pathway, as well as the succinate dehydrogenase complex. Thus, we tested the effects of deletions of MPC1, MPC3, OAC1, DIC1, SFC1, and SDH1 on SA production. The highest improvement was achieved by the combined deletion of MPC3 and SDH1. The respective strain produced up to 45.5\u00a0g\/L of SA, reached a maximum SA yield of 0.66 gSA\/gglycerol, and accumulated the lowest amounts of byproducts when cultivated in shake-flasks. Based on the obtained data, we consider a further reduction of mitochondrial import of pyruvate and rTCA intermediates highly attractive. Moreover, the approaches presented in the current study might also be valuable for improving SA production when sugars (instead of glycerol) are the source of carbon.<\/jats:p>","DOI":"10.1093\/femsyr\/foae009","type":"journal-article","created":{"date-parts":[[2024,4,8]],"date-time":"2024-04-08T17:24:00Z","timestamp":1712597040000},"source":"Crossref","is-referenced-by-count":12,"title":["Mitochondrial membrane transporters as attractive targets for the fermentative production of succinic acid from glycerol in <i>Saccharomyces cerevisiae<\/i>"],"prefix":"10.1093","volume":"24","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-8203-4575","authenticated-orcid":false,"given":"Toni","family":"Renduli\u0107","sequence":"first","affiliation":[{"name":"School of Science, Constructor University , Campus Ring 1, 28759 Bremen ,","place":["Germany"]},{"name":"Centre of Molecular and Environmental Biology, Department of Biology, University of Minho , Campus de Gualtar, 4710-057 Braga ,","place":["Portugal"]},{"name":"Institute of Science and Innovation for Bio-Sustainability (IB-S), University of Minho , Campus de Gualtar, 4710-057 Braga ,","place":["Portugal"]}]},{"given":"Andreea","family":"Perpelea","sequence":"additional","affiliation":[{"name":"School of Science, Constructor University , Campus Ring 1, 28759 Bremen ,","place":["Germany"]}]},{"given":"Juan Paulo Ragas","family":"Ortiz","sequence":"additional","affiliation":[{"name":"School of Science, Constructor University , Campus Ring 1, 28759 Bremen ,","place":["Germany"]}]},{"given":"Margarida","family":"Casal","sequence":"additional","affiliation":[{"name":"Centre of Molecular and Environmental Biology, Department of Biology, University of Minho , Campus de Gualtar, 4710-057 Braga ,","place":["Portugal"]},{"name":"Institute of Science and Innovation for Bio-Sustainability (IB-S), University of Minho , Campus de Gualtar, 4710-057 Braga ,","place":["Portugal"]}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7947-3000","authenticated-orcid":false,"given":"Elke","family":"Nevoigt","sequence":"additional","affiliation":[{"name":"School of Science, Constructor University , Campus Ring 1, 28759 Bremen ,","place":["Germany"]}]}],"member":"286","published-online":{"date-parts":[[2024,4,8]]},"reference":[{"key":"2024121916364332200_bib1","doi-asserted-by":"crossref","first-page":"1123","DOI":"10.1111\/j.1567-1364.2009.00537.x","article-title":"Metabolic engineering of Saccharomyces cerevisiae for production of carboxylic acids: current status and challenges","volume":"9","author":"Abbott","year":"2009","journal-title":"FEMS Yeast Res"},{"key":"2024121916364332200_bib2","doi-asserted-by":"crossref","first-page":"54","DOI":"10.1016\/j.copbio.2016.02.034","article-title":"Production of succinic acid by metabolically engineered microorganisms","volume":"42","author":"Ahn","year":"2016","journal-title":"Curr Opin Biotechnol"},{"key":"2024121916364332200_bib3","doi-asserted-by":"crossref","first-page":"15","DOI":"10.1111\/j.1574-6976.2001.tb00570.x","article-title":"Stoichiometry and compartmentation of NADH metabolism in Saccharomyces cerevisiae","volume":"25","author":"Bakker","year":"2001","journal-title":"FEMS Microbiol Rev"},{"key":"2024121916364332200_bib4","doi-asserted-by":"crossref","first-page":"911","DOI":"10.15252\/embj.201490197","article-title":"Regulation of mitochondrial pyruvate uptake by alternative pyruvate carrier complexes","volume":"34","author":"Bender","year":"2015","journal-title":"EMBO J"},{"key":"2024121916364332200_bib5","doi-asserted-by":"crossref","first-page":"293","DOI":"10.1128\/jb.62.3.293-300.1951","article-title":"Studies on Lysogenesis. 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