A combined experimental and modelling approach for the Weimberg pathway optimisation

Lu Shen, Martha Kohlhaas, Junichi Enoki, Roland Meier, Bernhard Schönenberger, Roland Wohlgemuth, Robert Kourist, Felix Niemeyer, David van Niekerk, Christopher Bräsen, Jochen Niemeyer, Jacky Snoep, Bettina Siebers*

*Corresponding author for this work

Research output: Contribution to JournalArticleAcademicpeer-review

Abstract

The oxidative Weimberg pathway for the five-step pentose degradation to α-ketoglutarate is a key route for sustainable bioconversion of lignocellulosic biomass to added-value products and biofuels. The oxidative pathway from Caulobacter crescentus has been employed in in-vivo metabolic engineering with intact cells and in in-vitro enzyme cascades. The performance of such engineering approaches is often hampered by systems complexity, caused by non-linear kinetics and allosteric regulatory mechanisms. Here we report an iterative approach to construct and validate a quantitative model for the Weimberg pathway. Two sensitive points in pathway performance have been identified as follows: (1) product inhibition of the dehydrogenases (particularly in the absence of an efficient NAD+ recycling mechanism) and (2) balancing the activities of the dehydratases. The resulting model is utilized to design enzyme cascades for optimized conversion and to analyse pathway performance in C. cresensus cell-free extracts.

Original languageEnglish
Article number1098
Pages (from-to)1-13
Number of pages13
JournalNature Communications
Volume11
Issue number1
DOIs
Publication statusPublished - 27 Feb 2020

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Shen, L., Kohlhaas, M., Enoki, J., Meier, R., Schönenberger, B., Wohlgemuth, R., ... Siebers, B. (2020). A combined experimental and modelling approach for the Weimberg pathway optimisation. Nature Communications, 11(1), 1-13. [1098]. https://doi.org/10.1038/s41467-020-14830-y