Engineering a Seven Enzyme Biotransformation using Mathematical Modelling and Characterized Enzyme Parts

Multi‐step enzyme reactions offer considerable cost and productivity benefits. Process models offer a route to understanding the complexity of these reactions, and allow for their optimization. Despite the increasing prevalence of multi‐step biotransformations, there are few examples of process mode...

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Bibliographic Details
Published in:ChemCatChem 2019-08, Vol.11 (15), p.3474-3489
Main Authors: Finnigan, William, Cutlan, Rhys, Snajdrova, Radka, Adams, Joseph P., Littlechild, Jennifer A., Harmer, Nicholas J.
Format: Article
Language:English
Subjects:
Biotransformation
Complexity
Economic models
Enzyme cascade
Enzymes
Genetic algorithms
Impact analysis
Isolated enzymes
Kinetics
Optimization
Pathway optimization
Reaction engineering
Regeneration
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Summary:Multi‐step enzyme reactions offer considerable cost and productivity benefits. Process models offer a route to understanding the complexity of these reactions, and allow for their optimization. Despite the increasing prevalence of multi‐step biotransformations, there are few examples of process models for enzyme reactions. From a toolbox of characterized enzyme parts, we demonstrate the construction of a process model for a seven enzyme, three step biotransformation using isolated enzymes. Enzymes for cofactor regeneration were employed to make this in vitro reaction economical. Good modelling practice was critical in evaluating the impact of approximations and experimental error. We show that the use and validation of process models was instrumental in realizing and removing process bottlenecks, identifying divergent behavior, and for the optimization of the entire reaction using a genetic algorithm. We validated the optimized reaction to demonstrate that complex multi‐step reactions with cofactor recycling involving at least seven enzymes can be reliably modelled and optimized. What a (bio)transformation! We demonstrate the construction and validation of a kinetic model for a seven enzyme, three step biotransformation (including cofactor regeneration) using isolated enzymes. We employed the model to optimize the entire reaction using a genetic algorithm. The validated reaction demonstrates that complex multi‐step reactions can be reliably modelled and optimized.
ISSN:1867-3880
1867-3899
DOI:10.1002/cctc.201900646