Maximizing the Efficiency of Multienzyme Process by Stoichiometry Optimization

• Published in: Chembiochem : a European journal of chemical biology 2014-09, Vol.15 (13), p.1891-1895
• Main Authors: Dvorak, Pavel, Kurumbang, Nagendra P., Bendl, Jaroslav, Brezovsky, Jan, Prokop, Zbynek, Damborsky, Jiri
• Format: Article
• Language: English
• Subjects: Algorithms; Biocatalysis; Biocatalysts; biotransformations; Efficiency; Enzymes; Enzymes - chemistry; kinetic modeling; Kinetics; Models, Chemical; multienzyme reaction; Optimization; Protein Engineering; stoichiometry optimization; Workflow
• ISSN: 1439-4227; 1439-7633
• DOI: 10.1002/cbic.201402265

Multienzyme processes represent an important area of biocatalysis. Their efficiency can be enhanced by optimization of the stoichiometry of the biocatalysts. Here we present a workflow for maximizing the efficiency of a three‐enzyme system catalyzing a five‐step chemical conversion. Kinetic models of pathways with wild‐type or engineered enzymes were built, and the enzyme stoichiometry of each pathway was optimized. Mathematical modeling and one‐pot multienzyme experiments provided detailed insights into pathway dynamics, enabled the selection of a suitable engineered enzyme, and afforded high efficiency while minimizing biocatalyst loadings. Optimizing the stoichiometry in a pathway with an engineered enzyme reduced the total biocatalyst load by an impressive 56 %. Our new workflow represents a broadly applicable strategy for optimizing multienzyme processes. Recipe for success: We propose a workflow for optimizing complex multienzyme reactions by kinetic modeling and stoichiometry optimization. By using a three‐enzyme system catalyzing a five‐step chemical conversion we show that selection of suitable enzymes and stoichiometry optimization can greatly reduce biocatalyst loadings. This work highlights the potential of kinetic modeling for optimizing industrial biocatalytic processes.