Systems‐wide analysis and engineering of metabolic pathway fluxes in bio‐succinate producing Basfia succiniciproducens. Issue 11 (9th July 2013)

• Record Type: Journal Article
• Authors: Becker, Judith; Reinefeld, Jasper; Stellmacher, René; Schäfer, Rudolf; Lange, Anna; Meyer, Hanna; Lalk, Michael; Zelder, Oskar; von Abendroth, Gregory; Schröder, Hartwig; Haefner, Stefan; Wittmann, Christoph
• Is Part Of: Biotechnology and bioengineering. Volume 110:Issue 11(2013:Nov.)
• Page Start: 3013
• Page End: 3023
• Publication Date: 2013-07-09
• Subjects: Biotechnology -- Periodicals; Bioengineering -- Periodicals; 660.6
• DOI: 10.1002/bit.24963
• Languages: English
• ISSNs: 0006-3592

Abstract

<abstract abstract-type="main" xml:lang="en"> <title>ABSTRACT</title> <sec id="bit24963-sec-0001" sec-type="section"> <p> <italic>Basfia succiniciproducens</italic> has been recently isolated as novel producer for succinate, an important platform chemical. In batch culture, the wild type exhibited a high natural yield of 0.75 mol succinate (mol glucose)<sup>−1</sup>. Systems‐wide <sup>13</sup>C metabolic flux analysis identified undesired fluxes through pyruvate‐formate lyase (PflD) and lactate dehydrogenase (LdhA). The double deletion strain <italic>B. succiniciproducens</italic> Δ<italic>ldhA</italic> Δ<italic>pflD</italic> revealed a 45% improved product yield of 1.08 mol mol<sup>−1</sup>. In addition, metabolic flux analysis unraveled the parallel in vivo activity of the oxidative and reductive branch of the TCA cycle in <italic>B. succiniciproducens</italic>, whereby the oxidative part mainly served for anabolism. The wild type re‐directed surplus NADH via a cycle involving malic enzyme or via transhydrogenase, respectively, to supply NADPH for anabolism, because the fluxes through the oxidative PPP and isocitrate dehydrogenase, that also provide this cofactor, were not sufficient. This was not observed for the deletion mutants, <italic>B. succiniciproducens</italic> Δ<italic>pflD</italic> and Δ<italic>ldhA</italic> Δ<italic>pflD</italic>, where PPP and isocitrate dehydrogenase flux alone matched with the reduced anabolic NADPH demand. The integration of the production performance into the theoretical flux space, computed by elementary flux mode analysis, revealed that <italic>B. succiniciproducens</italic> Δ<italic>ldhA</italic> Δ<italic>pflD</italic> reached 62% of the theoretical maximum yield. Biotechnol. Bioeng. 2013;110: 3013–3023. © 2013 Wiley Periodicals, Inc.</p> </sec> </abstract>