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identification of the yqhe and yafb genes encoding two 2, 5-diketo-d-gluconate reductases in escherichia coli.the identification of a gene (yiae) encoding 2-ketoaldonate reductase (2kr) in our previous work led to the hypothesis that escherichia coli has other ketogluconate reductases including 2, 5-diketo-d-gluconate reductase (25dkgr) and to study of the related ketogluconate metabolism. by using the deduced amino acid sequences of 5-diketo-d-gluconate reductase (5kdgr) of gluconobacter oxydans and 25dkgr of corynebacterium sp., protein databases were screened to detect homologous proteins. among the ...199910427017
dna from uncultured organisms as a source of 2,5-diketo-d-gluconic acid reductases.total dna of a population of uncultured organisms was extracted from soil samples, and by using pcr methods, the genes encoding two different 2,5-diketo-d-gluconic acid reductases (dkgrs) were recovered. degenerate pcr primers based on published sequence information gave internal gene fragments homologous to known dkgrs. nested primers specific for the internal fragments were combined with random primers to amplify flanking gene fragments from the environmental dna, and two hypothetical full-len ...200111526025
conversion of glucose to 2-keto-l-gulonate, an intermediate in l-ascorbate synthesis, by a recombinant strain of erwinia citreus.a gene for 2,5-diketo-d-gluconate (25dkg) reductase, which encodes an enzyme composed of 277 amino acid residues catalyzing the reduction of 25dkg to 2-keto-l-gulonate (2klg), was cloned from corynebacterium sp. strain shs752001 and expressed in erwinia citreus shs2003, a strain which oxidizes glucose to 25dkg. the recombinant microorganism converted glucose to 2klg, a compound which can be readily converted to l-ascorbate (vitamin c). improvements in the yield of 2klg were obtained by changing ...198816347687
recombinant organisms for production of industrial products.a revolution in industrial microbiology was sparked by the discoveries of ther double-stranded structure of dna and the development of recombinant dna technology. traditional industrial microbiology was merged with molecular biology to yield improved recombinant processes for the industrial production of primary and secondary metabolites, protein biopharmaceuticals and industrial enzymes. novel genetic techniques such as metabolic engineering, combinatorial biosynthesis and molecular breeding te ...200921326937
recombinant organisms for production of industrial products.a revolution in industrial microbiology was sparked by the discoveries of ther double-stranded structure of dna and the development of recombinant dna technology. traditional industrial microbiology was merged with molecular biology to yield improved recombinant processes for the industrial production of primary and secondary metabolites, protein biopharmaceuticals and industrial enzymes. novel genetic techniques such as metabolic engineering, combinatorial biosynthesis and molecular breeding te ...200921326937
evaluation of the food grade expression systems nice and psip for the production of 2,5-diketo-d-gluconic acid reductase from corynebacterium glutamicum.2,5-diketo-d-gluconic acid reductase (2,5-dkg reductase) catalyses the reduction of 2,5-diketo-d-gluconic acid (2,5-dkg) to 2-keto-l-gulonic acid (2-klg), a direct precursor (lactone) of l-ascorbic acid (vitamin c). this reaction is an essential step in the biocatalytic production of the food supplement vitamin c from d-glucose or d-gluconic acid. as 2,5-dkg reductase is usually produced recombinantly, it is of interest to establish an efficient process for 2,5-dkg reductase production that also ...201323356419
genetic and biochemical characterization of the pathway in pantoea citrea leading to pink disease of pineapple.pink disease of pineapple, caused by pantoea citrea, is characterized by a dark coloration on fruit slices after autoclaving. this coloration is initiated by the oxidation of glucose to gluconate, which is followed by further oxidation of gluconate to as yet unknown chromogenic compounds. to elucidate the biochemical pathway leading to pink disease, we generated six coloration-defective mutants of p. citrea that were still able to oxidize glucose into gluconate. three mutants were found to be af ...200010735866
the yiae gene, located at 80.1 minutes on the escherichia coli chromosome, encodes a 2-ketoaldonate reductase.an open reading frame located in the bisc-cspa intergenic region, or at 80.1 min on the escherichia coli chromosome, encodes a hypothetical 2-hydroxyacid dehydrogenase, which was identified as a result of the e. coli genome sequencing project. we report here that the product of the gene (yiae) is a 2-ketoaldonate reductase (2kr). the gene was cloned and expressed with a c-terminal his tag in e. coli, and the protein was purified by metal-chelate affinity chromatography. the determination of the ...19989811658
purification and characterization of 5-ketofructose reductase from erwinia citreus.5-ketofructose reductase [d(-)fructose:(nadp+) 5-oxidoreductase] was purified to homogeneity from erwinia citreus and demonstrated to catalyse the reversible nadph-dependent reduction of 5-ketofructose (d-threo-2,5-hexodiulose) to d-fructose. the enzyme appeared as a single species upon analyses by sds/polyacrylamide-gel electrophoresis and isoelectric focusing with an apparent relative molecular mass of 40,000 and an isoelectric point of 4.4. the amino acid composition of the enzyme and the n-t ...19883178725
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