Bio-based succinate from sucrose: High-resolution 13 C metabolic flux analysis and metabolic engineering of the rumen bacterium Basfia succiniciproducens - CNRS - Centre national de la recherche scientifique Accéder directement au contenu
Article Dans Une Revue Metabolic Engineering Année : 2017

Bio-based succinate from sucrose: High-resolution 13 C metabolic flux analysis and metabolic engineering of the rumen bacterium Basfia succiniciproducens

Résumé

Succinic acid is a platform chemical of recognized industrial value and accordingly faces a continuous challenge to enable manufacturing from most attractive raw materials. It is mainly produced from glucose, using microbial fermentation. Here, we explore and optimize succinate production from sucrose, a globally applied substrate in biotechnology, using the rumen bacterium Basfia succiniciproducens DD1. As basis of the strain optimization, the yet unknown sucrose metabolism of the microbe was studied, using C-13 metabolic flux analyses. When grown in batch culture on sucrose, the bacterium exhibited a high succinate yield of 1 mol mol(-1) and a by-product spectrum, which did not match the expected PTS-mediated sucrose catabolism. This led to the discovery of a fructokinase, involved in sucrose catabolism. The flux approach unraveled that the fructokinase and the fructose PTS both contribute to phosphorylation of the fructose part of sucrose. The contribution of the fructokinase reduces the undesired loss of the succinate precursor PEP into pyruvate and into pyruvate-derived by-products and enables increased succinate production, exclusively via the reductive TCA cycle branch. These findings were used to design superior producers. Mutants, which (i) overexpress the beneficial fructokinase, (II) lack the competing fructose PTS, and (iii) combine both traits, produce significantly more succinate. In a fed-batch process, B. succiniciproducens Delta fruA achieved a titer of 71 g L-1 succinate and a yield of 2.5 mol mol(-1) from sucrose.
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Dates et versions

hal-01886444 , version 1 (23-05-2019)

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Paternité - Pas d'utilisation commerciale - Pas de modification

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Anna Lange, Judith Becker, Dennis Schulze, Edern Cahoreau, Jean-Charles Portais, et al.. Bio-based succinate from sucrose: High-resolution 13 C metabolic flux analysis and metabolic engineering of the rumen bacterium Basfia succiniciproducens. Metabolic Engineering, 2017, 44, pp.198-212. ⟨10.1016/j.ymben.2017.10.003⟩. ⟨hal-01886444⟩
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