One-step synthesis of 2-keto-3-deoxy-d-gluconate by biocatalytic dehydration of d-gluconate.

  title={One-step synthesis of 2-keto-3-deoxy-d-gluconate by biocatalytic dehydration of d-gluconate.},
  author={K. Matsubara and R. K{\"o}hling and B. Sch{\"o}nenberger and T. Kouril and D. Esser and C. Br{\"a}sen and B. Siebers and Roland Wohlgemuth},
  journal={Journal of biotechnology},
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… Expand
Enzymatic Synthesis of 2-Keto-3-Deoxy-6-Phosphogluconate by the 6-Phosphogluconate-Dehydratase From Caulobacter crescentus
The efficient biocatalytic KDPG production through enzymatic dehydration of 6-phosphogluconate (6PG) up to gram scale is reported using the 6PG dehydratase/Entner-Doudoroff dehydrat enzyme from Caulobacter crescentus (CcEDD). Expand
Key Enzymes of the Semiphosphorylative Entner-Doudoroff Pathway in the Haloarchaeon Haloferax volcanii: Characterization of Glucose Dehydrogenase, Gluconate Dehydratase, and 2-Keto-3-Deoxy-6-Phosphogluconate Aldolase
The identification and characterization of the key enzymes glucose dehydrogenase, gluconate dehydratase, and 2-keto-3-deoxy-6-phosphogluconate aldolase and their encoding genes of the proposed semiphosphorylative Entner-Doudoroff pathway in the haloarchaeon Haloferax volcanii are described and the functional involvement of the three enzymes was proven by analyses of the corresponding knockout mutants. Expand
Characterization of recombinantly expressed dihydroxy-acid dehydratase from Sulfobus solfataricus-A key enzyme for the conversion of carbohydrates into chemicals.
The recombinant production of DHAD from Sulfolobus solfataricus (SsDHAD) in E. coli is reported on with special focus on activity toward non-natural substrates, thermo-stability, therMO-inactivation kinetics and activation capabilities and its application within multi-step cascades for chemicals production. Expand
Reaction Design for the Compartmented Combination of Heterogeneous and Enzyme Catalysis
The combination of a heterogeneously catalyzed reaction with a biotransformation as a one-pot cascade process is an important strategy to reduce costs, time, and labor efforts in the production ofExpand
A continuous assay for l-talarate/galactarate dehydratase using circular dichroism.
The continuous CD-based assay offers a practical and efficient alternative method to the coupled assay that requires access to 5-KDG aldolase, and to the labor-intensive, fixed-time assays. Expand
Enzymology of Alternative Carbohydrate Catabolic Pathways
The Embden–Meyerhof–Parnas (EMP) and Entner–Doudoroff (ED) pathways are considered the most abundant catabolic pathways found in microorganisms, and ED enzymes have been shown to also be widespreadExpand
Biocatalysis - key enabling tools from biocatalytic one-step and multi-step reactions to biocatalytic total synthesis.
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Horizons of Systems Biocatalysis and Renaissance of Metabolite Synthesis
A Systems Biocatalysis approach to the synthesis of metabolites aims at biocatalytic route designs with high molecular economy and provides valuable clues for resource‐ and energy‐efficient synthetic routes to natural metabolites. Expand
A combined experimental and modelling approach for the Weimberg pathway optimisation
An iterative approach is reported to construct and validate a quantitative model for the pentose degradation Weimberg pathway in Caulobacter crescentus and its biotechnological applications in cell-free system and standard metabolic engineering. Expand
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Directed evolution of D-2-keto-3-deoxy-6-phosphogluconate aldolase to new variants for the efficient synthesis of D- and L-sugars.
The effectiveness of using in vitro evolution to rapidly alter the properties of an aldolase to improve its utility in asymmetric synthesis is demonstrated and the principles and strategies described in this study should be applicable to other a Aldolases to further expand the scope of their synthetic utility. Expand
Enzymatic synthesis of 2-keto-3-deoxy-d-glucose from D-gluconate.
KDG 1 was prepared from d -gluconate with a cell-free extract of Clostridium pasteurianum containing gluconate dehydratase and because of the heat tolerance of the dehydrat enzyme KDG synthesis could be carried out at 50°C. Expand
Dioxalanones as synthetic intermediates. Part 6. Synthesis of 3-deoxy-D-manno-2-octulosonic acid (KDO), 3-deoxy-D-arabino-2-heptulosonic acid (DAH) and 2-keto-3-deoxy-D-gluconic acid (KDG)
Abstract Three biosynthetically significant α-keto acids KDO ( 7 ), DAH ( 8 ) and KDG ( 9 ) have been synthesised via 5-ylidene-1,3-dioxalan-4-one intermediates formed by Wittig reactions ofExpand
The Structural Basis for Substrate Promiscuity in 2-Keto-3-deoxygluconate Aldolase from the Entner-Doudoroff Pathway in Sulfolobus solfataricus*
The structure of KDGA is reported, determined by multiwavelength anomalous diffraction phasing, and it is confirmed that it is a member of the tetrameric N-acetylneuraminate lyase superfamily of Schiff base-forming aldolases, which have allowed rationalization of the structural basis of promiscuous substrate recognition and catalysis. Expand
Nouvelle Synthese de L'Acide 3-Desoxy-D-Erythro-2-Hexulosonique (KDG). A Partir De La D-Glucono-1,5-Lactone Synthese Et Etude De RMN De Derives O-Methyles Du KDG
ABSTRACT 3-Deoxy-D-erythro-2-hexulosonic acid (KDG), an important metabolite of bacterial polysaccharide degradation, was prepared from D-glucono-1,5-lactone through a six-step sequence, with a 45%Expand
Identification and characterization of Sulfolobus solfataricus D-gluconate dehydratase: a key enzyme in the non-phosphorylated Entner-Doudoroff pathway.
The purification and characterization of D-gluconate dehydratase from S. solfataricus found that it is a phosphoprotein and that its catalytic activity is regulated by a phosphorylation-dephosphorylation mechanism, the first report on biochemical and genetic characterization of this enzyme involved in the non-phosphorylated Entner-Doudoroff pathway. Expand
2-keto-3-deoxyl-L-arabonate aldolase and its role in a new pathway of L-arabinose degradation.
  • A. Dahms, R. Anderson
  • Chemistry, Medicine
  • Biochemical and biophysical research communications
  • 1969
Abstract A new aldolase which catalyzes the cleavage of 2-keto-3-deoxy-L-arabonate to pyruvate and glycolaldehyde was demonstrated in extracts of a pseudomonad (strain MSU-1). Extracts also catalyzedExpand
The occurrence of a modified Entner-Doudoroff pathway in Clostridium aceticum
The three enzymes involved in gluconate breakdown have been characterized with respect to their requirements for reducing agents and metal ions, and Gluconate dehydrase requires a sulfhydryl compound and ferrous ions for activity, KDG kinase a divalent metal ion for activity. Expand
Crystal structure and stereochemical studies of KD(P)G aldolase from Thermoproteus tenax
This work has shown that the 2‐keto‐3‐deoxy‐(6‐phospho)‐gluconate (KD(P)G) aldolase is a trade‐off between required catabolic flexibility needed for the conversion of phosphorylated and nonphosphorylated substrates and required stereo control of cellular/physiological enzymatic reactions. Expand
Synthesis and properties of 3,6-dideoxyhexulosonic acids and related compounds
Synthesis and characterizatio of the epimeric 3,6-dideoxy- d (and l )-hexulosonic acids are described and it is shown that each compound exists in solution mainly as the 2,5-hemiacetal. Expand