Pushing the boundaries of phosphorylase cascade reaction for cellobiose production II: Model-based multiobjective optimization

Alexander Sigg, Mario Klimacek, Bernd Nidetzky*

*Korrespondierende/r Autor/-in für diese Arbeit

Publikation: Beitrag in einer FachzeitschriftArtikelBegutachtung

Abstract

The inherent complexity of coupled biocatalytic reactions presents a major challenge for process development with one-pot multienzyme cascade transformations. Kinetic models are powerful engineering tools to guide the optimization of cascade reactions towards a performance suitable for scale up to an actual production. Here, we report kinetic model-based window of operation analysis for cellobiose production (≥100 g/L) from sucrose and glucose by indirect transglycosylation via glucose 1-phosphate as intermediate. The two-step cascade transformation is catalyzed by sucrose and cellobiose phosphorylase in the presence of substoichiometric amounts of phosphate (≤27 mol% of substrate). Kinetic modeling was instrumental to uncover the hidden effect of bulk microviscosity due to high sugar concentrations on decreasing the rate of cellobiose phosphorylase specifically. The mechanistic-empirical hybrid model thus developed gives a comprehensive description of the cascade reaction at industrially relevant substrate conditions. Model simulations serve to unravel opposed relationships between efficient utilization of the enzymes and maximized concentration (or yield) of the product within a given process time, in dependence of the initial concentrations of substrate and phosphate used. Optimum balance of these competing key metrics of process performance is suggested from the model-calculated window of operation and is verified experimentally. The evidence shown highlights the important use of kinetic modeling for the characterization and optimization of cascade reactions in ways that appear to be inaccessible to purely data-driven approaches.
Originalspracheenglisch
Seiten (von - bis)566-579
Seitenumfang14
FachzeitschriftBiotechnology and Bioengineering
Jahrgang121
Ausgabenummer2
Frühes Online-Datum21 Nov. 2023
DOIs
PublikationsstatusVeröffentlicht - Feb. 2024

ASJC Scopus subject areas

  • Angewandte Mikrobiologie und Biotechnologie
  • Bioengineering
  • Biotechnology

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