title: Autodisplay of glucose-6-phosphate dehydrogenase for redox cofactor regeneration at the cell surface.
abstract: Inherent cofactor regeneration is a pivotal feature of whole cell biocatalysis. For specific biotechnological applications, surface display of enzymes is emerging as a tool to circumvent mass transfer limitations or enzyme stability problems. Even complex reactions can be accomplished applying displayed enzymes. Yet, industrial utilization of the technique is still impeded by lacking cofactor regeneration at the cell surface. Here, we report on the surface display of a glucose-6-phoshate dehydrogenase (G6PDH) via Autodisplay to address this limitation and regenerate NADPH directly at the cell surface. The obtained whole cell biocatalyst demonstrated similar kinetic parameters compared to the purified enzyme, more precisely KMvalues of 0.2 mM for NADP+and calculated total turnover numbers of 107. However, the KMfor the substrate G6P increased by a factor of 7 to yield 1.5 mM. The whole cell biocatalyst was cheaper to produce, easy to separate from the reaction mixture and reusable in consecutive reaction cycles. Furthermore, lyophilization allowed storage at room temperature. The whole cell biocatalyst displaying G6PDH was applicable for NADPH regeneration in combination with soluble as well as surface displayed enzymes and model reactions in combination with bacterial CYP102A1 and human CYP1A2 were realized. Biotechnol. Bioeng. 2017;114: 1658-1669. © 2017 Wiley Periodicals, Inc.
authors.affiliation: Institut für Pharmazeutische und Medizinische Chemie, PharmaCampus, Westfälische Wilhelms-Universität Münster, Corrensstraße 48, 48149 Münster, Germany.
authors.forename: Jan
authors.initials: J
authors.lastname: Schüürmann
authors.affiliation: Institut für Pharmazeutische und Medizinische Chemie, PharmaCampus, Westfälische Wilhelms-Universität Münster, Corrensstraße 48, 48149 Münster, Germany.
authors.forename: Paul
authors.initials: P
authors.lastname: Quehl
authors.affiliation: Institut für Pharmazeutische und Medizinische Chemie, PharmaCampus, Westfälische Wilhelms-Universität Münster, Corrensstraße 48, 48149 Münster, Germany.
authors.forename: Fabian
authors.initials: F
authors.lastname: Lindhorst
authors.affiliation: Institut für Pharmazeutische und Medizinische Chemie, PharmaCampus, Westfälische Wilhelms-Universität Münster, Corrensstraße 48, 48149 Münster, Germany.
authors.forename: Kristina
authors.initials: K
authors.lastname: Lang
authors.affiliation: Institut für Pharmazeutische und Medizinische Chemie, PharmaCampus, Westfälische Wilhelms-Universität Münster, Corrensstraße 48, 48149 Münster, Germany.
authors.forename: Joachim
authors.identifier: 0000-0002-0666-2676
authors.initials: J
authors.lastname: Jose
mesh_terms: D002462:Cell Membrane / Q000235:genetics / Q000378:metabolism*; D003067:Coenzymes / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli / Q000502:physiology*; D015971:Gene Expression Regulation, Enzymologic / Q000235:genetics; D005954:Glucosephosphate Dehydrogenase / Q000235:genetics / Q000378:metabolism*; D009249:NADP / Q000235:genetics / Q000378:metabolism*; D010084:Oxidation-Reduction; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D003067:Coenzymes; D011994:Recombinant Proteins; D009249:NADP; D005954:Glucosephosphate Dehydrogenase
keywords: NADPH; autodisplay; cofactor regeneration; glucose-6-phosphate dehydrogenase; surface display; whole cell biocatalysis