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. 1988 Jul 15;253(2):511–516. doi: 10.1042/bj2530511

Purification and characterization of 5-ketofructose reductase from Erwinia citreus.

J L Schrimsher 1, P T Wingfield 1, A Bernard 1, R Mattaliano 1, M A Payton 1
PMCID: PMC1149327  PMID: 3178725

Abstract

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-terminal sequence of the first 39 residues are described. The steady-state kinetic mechanism was an ordered one with NADPH binding first to the enzyme and then to 5-ketofructose, and the order of product release was D-fructose followed by NADP+. The reversible nature of the reaction offers the possibility of using this enzyme for the determination of D-fructose.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Ameyama M., Shinagawa E., Matsushita K., Adachi O. D-fructose dehydrogenase of Gluconobacter industrius: purification, characterization, and application to enzymatic microdetermination of D-fructose. J Bacteriol. 1981 Feb;145(2):814–823. doi: 10.1128/jb.145.2.814-823.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Avigad G., Englard S., Pifko S. 5-Keto-D-fructose. IV. A specific reduced nicotinamide adenine dinucleotide phosphate-linked reductase from Gluconobacter cerinus. J Biol Chem. 1966 Jan 25;241(2):373–378. [PubMed] [Google Scholar]
  3. Cleland W. W. Statistical analysis of enzyme kinetic data. Methods Enzymol. 1979;63:103–138. doi: 10.1016/0076-6879(79)63008-2. [DOI] [PubMed] [Google Scholar]
  4. Edelhoch H. Spectroscopic determination of tryptophan and tyrosine in proteins. Biochemistry. 1967 Jul;6(7):1948–1954. doi: 10.1021/bi00859a010. [DOI] [PubMed] [Google Scholar]
  5. Huber R. E., Brockbank R. L. Strong inhibitory effect of furanoses and sugar lactones on beta-galactosidase Escherichia coli. Biochemistry. 1987 Mar 24;26(6):1526–1531. doi: 10.1021/bi00380a005. [DOI] [PubMed] [Google Scholar]
  6. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  7. Wingfield P., Payton M., Tavernier J., Barnes M., Shaw A., Rose K., Simona M. G., Demczuk S., Williamson K., Dayer J. M. Purification and characterization of human interleukin-1 beta expressed in recombinant Escherichia coli. Eur J Biochem. 1986 Nov 3;160(3):491–497. doi: 10.1111/j.1432-1033.1986.tb10066.x. [DOI] [PubMed] [Google Scholar]
  8. Yamada Y., Aida K., Uemura T. Enzymatic studies on the oxidation of sugar and sugar alcohol. II. Purification and properties of NADPH-linked 5-ketofructose reductase. J Biochem. 1967 Jun;61(6):803–811. doi: 10.1093/oxfordjournals.jbchem.a128616. [DOI] [PubMed] [Google Scholar]

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