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GDP-mannose 3',5'-epimerase forms GDP-L-gulose, a putative intermediate for the de novo biosynthesis of vitamin C in plants

Beata Wolucka UGent and Marc Van Montagu UGent (2003) JOURNAL OF BIOLOGICAL CHEMISTRY. 278(48). p.47483-47490
abstract
Despite its importance for agriculture, bioindustry, and nutrition, the fundamental process of L-ascorbic acid ( vitamin C) biosynthesis in plants is not completely elucidated, and little is known about its regulation. The recently identified GDP-Man 3', 5'-epimerase catalyzes a reversible epimerization of GDP-D-mannose that precedes the committed step in the biosynthesis of vitamin C, resulting in the hydrolysis of the highly energetic glycosyl-pyrophosphoryl linkage. Here, we characterize the native and recombinant GDP-Man 3', 5'-epimerase of Arabidopsis thaliana. GDP and GDP-D-glucose are potent competitive inhibitors of the enzyme, whereas GDP-L-fucose gives a complex type of inhibition. The epimerase contains a modified version of the NAD binding motif and is inhibited by NAD(P) H and stimulated by NAD(P)(+). A feedback inhibition of vitamin C biosynthesis is observed apparently at the level of GDP-Man 3', 5'-epimerase. The epimerase catalyzes at least two distinct epimerization reactions and releases, besides the well known GDP-L-galactose, a novel intermediate: GDP- L-gulose. The yield of the epimerization varies and seems to depend on the molecular form of the enzyme. Both recombinant and native enzymes co-purified with a Hsp70 heat-shock protein ( Escherichia coli DnaK and A. thaliana Hsc70.3, respectively). We speculate, therefore, that the Hsp70 molecular chaperones might be involved in folding and/or regulation of the epimerase. In summary, the plant epimerase undergoes a complex regulation and could control the carbon flux into the vitamin C pathway in response to the redox state of the cell, stress conditions, and GDP- sugar demand for the cell wall/glycoprotein biosynthesis. Exogenous L-gulose and L-gulono-1,4-lactone serve as direct precursors of L-ascorbic acid in plant cells. We propose an L-gulose pathway for the de novo biosynthesis of vitamin C in plants.
Please use this url to cite or link to this publication:
author
organization
year
type
journalArticle (original)
publication status
published
subject
keyword
CATALYTIC MECHANISM, ESCHERICHIA-COLI, D-GLUCOSE, L-GALACTOSE, ARABIDOPSIS-THALIANA, SHOCK COGNATE PROTEIN, 5'-DIPHOSPHATE D-MANNOSE, PERFORMANCE LIQUID-CHROMATOGRAPHY, GAMMA-LACTONE DEHYDROGENASE, L-ASCORBIC-ACID
journal title
JOURNAL OF BIOLOGICAL CHEMISTRY
J. Biol. Chem.
volume
278
issue
48
pages
47483 - 47490
Web of Science type
Article
Web of Science id
000186731400019
JCR category
BIOCHEMISTRY & MOLECULAR BIOLOGY
JCR impact factor
6.482 (2003)
JCR rank
31/261 (2003)
JCR quartile
1 (2003)
ISSN
0021-9258
DOI
10.1074/jbc.M309135200
language
English
UGent publication?
yes
classification
A1
copyright statement
I have transferred the copyright for this publication to the publisher
id
215361
handle
http://hdl.handle.net/1854/LU-215361
date created
2004-05-10 18:40:00
date last changed
2016-12-19 15:40:22
@article{215361,
  abstract     = {Despite its importance for agriculture, bioindustry, and nutrition, the fundamental process of L-ascorbic acid ( vitamin C) biosynthesis in plants is not completely elucidated, and little is known about its regulation. The recently identified GDP-Man 3', 5'-epimerase catalyzes a reversible epimerization of GDP-D-mannose that precedes the committed step in the biosynthesis of vitamin C, resulting in the hydrolysis of the highly energetic glycosyl-pyrophosphoryl linkage. Here, we characterize the native and recombinant GDP-Man 3', 5'-epimerase of Arabidopsis thaliana. GDP and GDP-D-glucose are potent competitive inhibitors of the enzyme, whereas GDP-L-fucose gives a complex type of inhibition. The epimerase contains a modified version of the NAD binding motif and is inhibited by NAD(P) H and stimulated by NAD(P)(+). A feedback inhibition of vitamin C biosynthesis is observed apparently at the level of GDP-Man 3', 5'-epimerase. The epimerase catalyzes at least two distinct epimerization reactions and releases, besides the well known GDP-L-galactose, a novel intermediate: GDP- L-gulose. The yield of the epimerization varies and seems to depend on the molecular form of the enzyme. Both recombinant and native enzymes co-purified with a Hsp70 heat-shock protein ( Escherichia coli DnaK and A. thaliana Hsc70.3, respectively). We speculate, therefore, that the Hsp70 molecular chaperones might be involved in folding and/or regulation of the epimerase. In summary, the plant epimerase undergoes a complex regulation and could control the carbon flux into the vitamin C pathway in response to the redox state of the cell, stress conditions, and GDP- sugar demand for the cell wall/glycoprotein biosynthesis. Exogenous L-gulose and L-gulono-1,4-lactone serve as direct precursors of L-ascorbic acid in plant cells. We propose an L-gulose pathway for the de novo biosynthesis of vitamin C in plants.},
  author       = {Wolucka, Beata and Van Montagu, Marc},
  issn         = {0021-9258},
  journal      = {JOURNAL OF BIOLOGICAL CHEMISTRY},
  keyword      = {CATALYTIC MECHANISM,ESCHERICHIA-COLI,D-GLUCOSE,L-GALACTOSE,ARABIDOPSIS-THALIANA,SHOCK COGNATE PROTEIN,5'-DIPHOSPHATE D-MANNOSE,PERFORMANCE LIQUID-CHROMATOGRAPHY,GAMMA-LACTONE DEHYDROGENASE,L-ASCORBIC-ACID},
  language     = {eng},
  number       = {48},
  pages        = {47483--47490},
  title        = {GDP-mannose 3',5'-epimerase forms GDP-L-gulose, a putative intermediate for the de novo biosynthesis of vitamin C in plants},
  url          = {http://dx.doi.org/10.1074/jbc.M309135200},
  volume       = {278},
  year         = {2003},
}

Chicago
Wolucka, Beata, and Marc Van Montagu. 2003. “GDP-mannose 3’,5'-epimerase Forms GDP-L-gulose, a Putative Intermediate for the De Novo Biosynthesis of Vitamin C in Plants.” Journal of Biological Chemistry 278 (48): 47483–47490.
APA
Wolucka, B., & Van Montagu, M. (2003). GDP-mannose 3’,5'-epimerase forms GDP-L-gulose, a putative intermediate for the de novo biosynthesis of vitamin C in plants. JOURNAL OF BIOLOGICAL CHEMISTRY, 278(48), 47483–47490.
Vancouver
1.
Wolucka B, Van Montagu M. GDP-mannose 3’,5'-epimerase forms GDP-L-gulose, a putative intermediate for the de novo biosynthesis of vitamin C in plants. JOURNAL OF BIOLOGICAL CHEMISTRY. 2003;278(48):47483–90.
MLA
Wolucka, Beata, and Marc Van Montagu. “GDP-mannose 3’,5'-epimerase Forms GDP-L-gulose, a Putative Intermediate for the De Novo Biosynthesis of Vitamin C in Plants.” JOURNAL OF BIOLOGICAL CHEMISTRY 278.48 (2003): 47483–47490. Print.