J. Biochem, 1998, Vol. 123, No. 1 189-193
© 1998 Japanese Biochemical Society
research-article |
Purification and Some Properties of a Hepatic NADPH-Dependent Reductase That Specifically Acts on 1,5-Anhydro-D-Fructose
Department of Life Sciences (Chemistry), Graduate School of Arts and Sciences, The University of Tokyo Meguro-ku, Tokyo 153
2To whom correspondence should be addressed. Fax: +81-3-3485-2904, Tel: +81-3-5454-6584
Glycogen gives rise to 1,5-anhydro-D-fructose (AF), which is then reduced to 1,5-anhydro-D-glucitol (AG) in animal livers. An enzyme that catalyzes NADPH-dependent reduction of AF to AG was isolated and purified to homogeneity from porcine liver. Its apparent molecular mass was about 38 kDa on the basis of SDS-PAGE, and its monomeric dispersion in aqueous solution was indicated by gel filtration on a Superose 12 column. Amino acid sequences were determined for four peptides obtained from the purified enzyme. The resulting sequences covered about 50% of the whole sequence and indicated a remarkable similarity between the enzyme and aldose reductase. The purified enzyme showed molecular activity of 8.7 s–1 on the basis of a molecular mass of 38 kDa, and a Km value of 0.44 mM for AF at the optimum pH of 7.0. It reduced pyridine-3-aldehyde and 2, 3-butanedione effectively, acetaldehyde, glucosone, and glucuronic acid poorly and showed no detectable action on glucose, mannose and fructose. It was inactivated by p-chloromercuribenzoic acid to a considerable extent, and the inactivation was partially reversed by 2-mercaptoethanol treatment. It was also sparingly inhibited by relatively high concentrations of glucose, glucose-1(6)-phosphate and 1,5-anhydroglucitol. The reverse reaction, i.e., NADP+ -dependent AG oxidation, was not observed. The observed catalytic properties and partial amino acid sequences rule out the possibility that the isolated protein is identical with any known reductase.
1Present address: Biophysical Pharmaceutics Lab., Medicinal Research Laboratories, Taisho Pharmaceutical Co., Ltd., Yoshino, Ohmiya, Saitama 330.
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  A. Kuhn, S. Yu, and F. Giffhorn Catabolism of 1,5-Anhydro-D-Fructose in Sinorhizobium morelense S-30.7.5: Discovery, Characterization, and Overexpression of a New 1,5-Anhydro-D-Fructose Reductase and Its Application in Sugar Analysis and Rare Sugar Synthesis Appl. Envir. Microbiol., February 1, 2006; 72(2): 1248 - 1257. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K. Hirano, M. Ziak, K. Kamoshita, Y. Sukenaga, S. Kametani, Y. Shiga, J. Roth, and H. Akanuma N-linked oligosaccharide processing enzyme glucosidase II produces 1,5-anhydrofructose as a side product Glycobiology, December 1, 2000; 10(12): 1283 - 1289. [Abstract] [Full Text] [PDF]
|
|