title: One-step synthesis of 2-keto-3-deoxy-d-gluconate by biocatalytic dehydration of d-gluconate.
abstract: 2-Keto-3-deoxy-sugar acids are key intermediates of central metabolism and integral constituents of bacterial (lipo)polysaccharides and cell wall components and are therefore continuously and highly demanded in related research fields. The stereospecific chemical synthesis of chiral 2-keto-deoxy-sugar acids involves a multitude of reaction steps, while in metabolic pathways only few conversions lead to the same 2-keto-3-deoxy sugar acids from easily available carbohydrate precursors. Here we present a straightforward and highly economic one-step biocatalytic synthesis procedure of 2-keto-3-deoxy-d-gluconate (KDG) from d-gluconate using recombinant gluconate dehydratase (GAD) from the hyperthermophilic crenarchaeon Thermoproteus tenax. This method is highly advantageous to KDG production schemes described so far for several reasons: (i) the d-gluconate is completely converted to stereochemically pure D-KDG without side-product formation, (ii) the final KDG yield is approximately 90%, (iii) the newly developed quantitative and qualitative LC-MS analysis method enabled the simultaneous detection of d-gluconate and KDG and (iv) the T. tenax GAD as biocatalyst can be provided by a simple and rapid procedure involving only two precipitation steps. The described utilization of dehydratases for 2-keto-3-deoxy sugar acid syntheses represents a highly resource-efficient one-step preparation and offers potential short synthetic routes toward a broad range of 2-keto-3-deoxy sugar acids and their derivatives.
authors.affiliation: Molecular Enzyme Technology and Biochemistry, University of Duisburg-Essen, Universitätsstrasse 5, 45117 Essen, Germany.
authors.forename: Kohei
authors.initials: K
authors.lastname: Matsubara
authors.affiliation: Sigma-Aldrich, Industriestrasse 25, CH-9470 Buchs, Switzerland.
authors.forename: Rudi
authors.initials: R
authors.lastname: Köhling
authors.affiliation: Sigma-Aldrich, Industriestrasse 25, CH-9470 Buchs, Switzerland.
authors.forename: Bernhard
authors.initials: B
authors.lastname: Schönenberger
authors.affiliation: Molecular Enzyme Technology and Biochemistry, University of Duisburg-Essen, Universitätsstrasse 5, 45117 Essen, Germany.
authors.forename: Theresa
authors.initials: T
authors.lastname: Kouril
authors.affiliation: Molecular Enzyme Technology and Biochemistry, University of Duisburg-Essen, Universitätsstrasse 5, 45117 Essen, Germany.
authors.forename: Dominik
authors.initials: D
authors.lastname: Esser
authors.affiliation: Molecular Enzyme Technology and Biochemistry, University of Duisburg-Essen, Universitätsstrasse 5, 45117 Essen, Germany.
authors.forename: Christopher
authors.initials: C
authors.lastname: Bräsen
authors.affiliation: Molecular Enzyme Technology and Biochemistry, University of Duisburg-Essen, Universitätsstrasse 5, 45117 Essen, Germany. Electronic address: bettina.siebers@uni-due.de.
authors.forename: Bettina
authors.initials: B
authors.lastname: Siebers
authors.affiliation: Sigma-Aldrich, Industriestrasse 25, CH-9470 Buchs, Switzerland. Electronic address: roland.wohlgemuth@sial.com.
authors.forename: Roland
authors.initials: R
authors.lastname: Wohlgemuth
mesh_terms: D055162:Biocatalysis; D003681:Dehydration / Q000378:metabolism; D005942:Gluconates / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism*; D006836:Hydro-Lyases / Q000737:chemistry / Q000378:metabolism*; D008070:Lipopolysaccharides / Q000096:biosynthesis / Q000737:chemistry; D053858:Metabolic Networks and Pathways; D013400:Sugar Acids / Q000737:chemistry
chemical_list: D005942:Gluconates; D008070:Lipopolysaccharides; D013400:Sugar Acids; C002957:2-keto-3-deoxygluconate; D006836:Hydro-Lyases; C020576:gluconate dehydratase
keywords: 2-Keto-3-deoxy-d-gluconate; Biocatalytic dehydration; Gluconate dehydratase; LC–MS; Metabolite