title: Maximizing the efficiency of multienzyme process by stoichiometry optimization.
abstract: 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.
authors.affiliation: Loschmidt Laboratories, Department of Experimental Biology and Research Centre for Toxic Compounds in the Environment RECETOX, Faculty of Science, Masaryk University, Kamenice 5/A13, 625 00 Brno (Czech Republic); International Clinical Research Center, St. Anne's University Hospital Brno, Pekarska 53, 656 91 Brno (Czech Republic).
authors.forename: Pavel
authors.initials: P
authors.lastname: Dvorak
authors.forename: Nagendra P
authors.initials: NP
authors.lastname: Kurumbang
authors.forename: Jaroslav
authors.initials: J
authors.lastname: Bendl
authors.forename: Jan
authors.initials: J
authors.lastname: Brezovsky
authors.forename: Zbynek
authors.initials: Z
authors.lastname: Prokop
authors.forename: Jiri
authors.initials: J
authors.lastname: Damborsky
mesh_terms: D000465:Algorithms; D055162:Biocatalysis; D004798:Enzymes / Q000737:chemistry*; D007700:Kinetics; D008956:Models, Chemical; D015202:Protein Engineering; D057188:Workflow
chemical_list: D004798:Enzymes
keywords: biocatalysis; biotransformations; kinetic modeling; multienzyme reaction; stoichiometry optimization