TY - JOUR
T1 - A combined experimental and modelling approach for the Weimberg pathway optimisation
AU - Shen, Lu
AU - Kohlhaas, Martha
AU - Enoki, Junichi
AU - Meier, Roland
AU - Schönenberger, Bernhard
AU - Wohlgemuth, Roland
AU - Kourist, Robert
AU - Niemeyer, Felix
AU - van Niekerk, David
AU - Bräsen, Christopher
AU - Niemeyer, Jochen
AU - Snoep, Jacky
AU - Siebers, Bettina
PY - 2020/2/27
Y1 - 2020/2/27
N2 - 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.
AB - 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.
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U2 - 10.1038/s41467-020-14830-y
DO - 10.1038/s41467-020-14830-y
M3 - Article
C2 - 32107375
AN - SCOPUS:85080142082
VL - 11
SP - 1
EP - 13
JO - Nature Communications
JF - Nature Communications
SN - 2041-1723
IS - 1
M1 - 1098
ER -