Maximizing the Efficiency of Multi-enzyme Process by Stoichiometry Optimization.
| Authors | |
|---|---|
| Year of publication | 2014 |
| Type | Article in Periodical |
| Magazine / Source | ChemBioChem |
| MU Faculty or unit | |
| Citation | |
| Doi | http://dx.doi.org/10.1002/cbic.201402265 |
| Field | Biochemistry |
| Keywords | biocatalysis; engineered enzymes |
| Description | 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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