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• W2065603832 abstract "The filamentous fungus Aspergillus niger is an efficient host for the recombinant production of the glycosylated enzyme fructofuranosidase, a biocatalyst of commercial interest for the synthesis of pre-biotic sugars. In batch culture on a minimal glucose medium, the recombinant strain A. niger SKAn1015, expressing the fructofuranosidase encoding suc1 gene secreted 45 U/mL of the target enzyme, whereas the parent wild type SKANip8 did not exhibit production. The production of the recombinant enzyme induced a significant change of in vivo fluxes in central carbon metabolism , as assessed by 13 C metabolic flux ratio analysis. Most notably, the flux redistribution enabled an elevated supply of NADPH via activation of the cytosolic pentose phosphate pathway (PPP) and mitochondrial malic enzyme, whereas the flux through energy generating TCA cycle was reduced. In addition, the overall possible flux space of fructofuranosidase producing A. niger was investigated in silico by elementary flux mode analysis. This provided theoretical flux distributions for multiple scenarios with differing production capacities. Subsequently, the measured flux changes linked to improved production performance were projected into the in silico flux space. This provided a quantitative evaluation of the achieved optimization and a priority ranked target list for further strain engineering. Interestingly, the metabolism was shifted largely towards the optimum flux pattern by sole expression of the recombinant enzyme, which seems an inherent attractive property of A. niger. Selected fluxes, however, changed contrary to the predicted optimum and thus revealed novel targets—including reactions linked to NADPH metabolism and gluconate formation. ► Coupling of 13 C flux and elementary flux mode analysis for systems analysis. ► Novel approach integrating in vivo and in silico flux analysis for strain design. ► Optimization of recombinant enzyme production in filamentous fungi. ► Identification of NADPH metabolism as key target towards improved protein production." @default.
• W2065603832 created "2016-06-24" @default.
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• W2065603832 date "2012-01-01" @default.
• W2065603832 modified "2023-09-24" @default.
• W2065603832 title "Integration of in vivo and in silico metabolic fluxes for improvement of recombinant protein production" @default.
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• W2065603832 doi "https://doi.org/10.1016/j.ymben.2011.11.002" @default.
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