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Reverse transcription-PCR experiments revealed that three out of the four genes assigned to lactate dehydrogenase in the genome of\n            <jats:named-content content-type=\"genus-species\">L. lactis<\/jats:named-content>\n            , i.e., the\n            <jats:italic>ldh<\/jats:italic>\n            ,\n            <jats:italic>ldhB<\/jats:italic>\n            , and\n            <jats:italic>ldhX<\/jats:italic>\n            genes, were expressed in the parental strain MG1363. Given that genetic redundancy is often a major cause of metabolic instability in engineered strains, we set out to develop a genetically stable lactococcal host tuned for the production of reduced compounds. Therefore, the\n            <jats:italic>ldhB<\/jats:italic>\n            and\n            <jats:italic>ldhX<\/jats:italic>\n            genes were sequentially deleted in\n            <jats:named-content content-type=\"genus-species\">L. lactis<\/jats:named-content>\n            FI10089, a strain with a deletion of the\n            <jats:italic>ldh<\/jats:italic>\n            gene. The single, double, and triple mutants, FI10089, FI10089\u0394\n            <jats:italic>ldhB<\/jats:italic>\n            , and FI10089\u0394\n            <jats:italic>ldhB<\/jats:italic>\n            \u0394\n            <jats:italic>ldhX<\/jats:italic>\n            , showed similar growth profiles and displayed mixed-acid fermentation, ethanol being the main reduced end product. Hence, the alcohol dehydrogenase-encoding gene, the\n            <jats:italic>adhE<\/jats:italic>\n            gene, was inactivated in FI10089, but the resulting strain reverted to homolactic fermentation due to induction of the\n            <jats:italic>ldhB<\/jats:italic>\n            gene. The three lactate dehydrogenase-deficient mutants were selected as a background for the production of mannitol and 2,3-butanediol. Pathways for the biosynthesis of these compounds were overexpressed under the control of a nisin promoter, and the constructs were analyzed with respect to growth parameters and product yields under anaerobiosis. Glucose was efficiently channeled to mannitol (maximal yield, 42%) or to 2,3-butanediol (maximal yield, 67%). The theoretical yield for 2,3-butanediol was achieved. We show that FI10089\u0394\n            <jats:italic>ldhB<\/jats:italic>\n            is a valuable basis for engineering strategies aiming at the production of reduced compounds.\n          <\/jats:p>","DOI":"10.1128\/aem.05544-11","type":"journal-article","created":{"date-parts":[[2011,8,13]],"date-time":"2011-08-13T02:32:37Z","timestamp":1313202757000},"page":"6826-6835","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":60,"title":["High Yields of 2,3-Butanediol and Mannitol in Lactococcus lactis through Engineering of NAD\n            <sup>+<\/sup>\n            Cofactor Recycling"],"prefix":"10.1128","volume":"77","author":[{"given":"Paula","family":"Gaspar","sequence":"first","affiliation":[{"name":"Instituto de Tecnologia Qu\u00edmica e Biol\u00f3gica, Universidade Nova de Lisboa, Av. 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