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Maximizing the Efficiency of Multi-enzyme Process by Stoichiometry Optimization.
Autoři | |
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Rok publikování | 2014 |
Druh | Článek v odborném periodiku |
Časopis / Zdroj | ChemBioChem |
Fakulta / Pracoviště MU | |
Citace | |
Doi | http://dx.doi.org/10.1002/cbic.201402265 |
Obor | Biochemie |
Klíčová slova | biocatalysis; engineered enzymes |
Popis | Multi-enzyme processes represent an important area of biocatalysis. Their efficiency can be enhanced by optimization of biocatalysts’ stoichiometry. Here we present a workflow for maximizing the efficiency of a three-enzyme system catalysing a five-step chemical conversion. Kinetic models of pathways featuring either wild-type or engineered enzymes were built and the enzyme stoichiometry of each pathway was optimized. Mathematical modelling and one-pot multi-enzyme laboratory experiments provided detailed insights into pathway dynamics, enabled the selection of suitable engineered enzyme and afforded high efficiency while minimizing biocatalyst loadings. The optimizing of stoichiometry in a pathway with engineered enzyme reduced the total biocatalyst load by an impressive 56 %. Our new workflow represents a broadly applicable strategy for optimizing multi-enzyme processes. |
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