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Use of in vivo C-13 nuclear magnetic resonance Spectroscopy to elucidate L-arabinose metabolism in Yeasts

Fonseca, César; Neves, Ana Rute; Antunes, Alexandra M M; Noronha, João Paulo; Hahn-Hägerdal, Bärbel LU ; Santos, Helena and Spencer-Martins, Isabel (2008) In Applied and Environmental Microbiology 74(6). p.1845-1855
Abstract
Candida arabinofermentans PYCC 5603(T) and Pichia guilliermondii PYCC 3012 were shown to grow well on L-arabinose, albeit exhibiting distinct features that justify an in-depth comparative study of their respective pentose catabolism. Carbon-13 labeling experiments coupled with in vivo NMR were used to investigate L-arabinose metabolism in these yeasts, thereby complementing recently reported physiological and enzymatic data. The label supplied in L-[2-(13)C]arabinose to non-growing cells, under aerobic conditions, was found on C1 and C2 of arabitol and ribitol, C2 of xylitol, and on C1, C2 and C3 of trehalose. The detection of labeled arabitol and xylitol constitutes additional evidence for the operation in yeast of the redox catabolic... (More)
Candida arabinofermentans PYCC 5603(T) and Pichia guilliermondii PYCC 3012 were shown to grow well on L-arabinose, albeit exhibiting distinct features that justify an in-depth comparative study of their respective pentose catabolism. Carbon-13 labeling experiments coupled with in vivo NMR were used to investigate L-arabinose metabolism in these yeasts, thereby complementing recently reported physiological and enzymatic data. The label supplied in L-[2-(13)C]arabinose to non-growing cells, under aerobic conditions, was found on C1 and C2 of arabitol and ribitol, C2 of xylitol, and on C1, C2 and C3 of trehalose. The detection of labeled arabitol and xylitol constitutes additional evidence for the operation in yeast of the redox catabolic pathway widespread in filamentous fungi. Furthermore, labeling at positions C1 of trehalose and arabitol demonstrates that glucose-6-phosphate is recycled through the oxidative pentose phosphate pathway (PPP). This result was interpreted as a metabolic strategy to regenerate NADPH, the cofactor essential to sustain L-arabinose catabolism at the level of L-arabinose reductase and L-xylulose reductase. Moreover, the observed synthesis of D-arabitol and ribitol provides a route to supply NAD(+) under oxygen-limiting conditions. In P. guilliermondii PYCC 3012, the strong accumulation of L-arabitol (up to 0.4 M, intracellular concentration) during aerobic L-arabinose metabolism denotes the existence of a bottleneck at the level of L-arabitol 4-dehydrogenase. This report provides the first experimental evidence of a link between L-arabinose metabolism in fungi and the oxidative branch of PPP, and suggests rational guidelines for the design of strategies towards the production of new and efficient L-arabinose-fermenting yeasts. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Applied and Environmental Microbiology
volume
74
issue
6
pages
1845 - 1855
publisher
American Society for Microbiology
external identifiers
  • pmid:18245253
  • scopus:40849139270
  • wos:000254065600021
ISSN
0099-2240
DOI
10.1128/AEM.02453-07
language
English
LU publication?
yes
id
d4dd9e49-8211-4594-880c-cf290d72dd1b (old id 1042300)
date added to LUP
2008-03-25 14:39:19
date last changed
2017-01-01 04:50:25
@article{d4dd9e49-8211-4594-880c-cf290d72dd1b,
  abstract     = {Candida arabinofermentans PYCC 5603(T) and Pichia guilliermondii PYCC 3012 were shown to grow well on L-arabinose, albeit exhibiting distinct features that justify an in-depth comparative study of their respective pentose catabolism. Carbon-13 labeling experiments coupled with in vivo NMR were used to investigate L-arabinose metabolism in these yeasts, thereby complementing recently reported physiological and enzymatic data. The label supplied in L-[2-(13)C]arabinose to non-growing cells, under aerobic conditions, was found on C1 and C2 of arabitol and ribitol, C2 of xylitol, and on C1, C2 and C3 of trehalose. The detection of labeled arabitol and xylitol constitutes additional evidence for the operation in yeast of the redox catabolic pathway widespread in filamentous fungi. Furthermore, labeling at positions C1 of trehalose and arabitol demonstrates that glucose-6-phosphate is recycled through the oxidative pentose phosphate pathway (PPP). This result was interpreted as a metabolic strategy to regenerate NADPH, the cofactor essential to sustain L-arabinose catabolism at the level of L-arabinose reductase and L-xylulose reductase. Moreover, the observed synthesis of D-arabitol and ribitol provides a route to supply NAD(+) under oxygen-limiting conditions. In P. guilliermondii PYCC 3012, the strong accumulation of L-arabitol (up to 0.4 M, intracellular concentration) during aerobic L-arabinose metabolism denotes the existence of a bottleneck at the level of L-arabitol 4-dehydrogenase. This report provides the first experimental evidence of a link between L-arabinose metabolism in fungi and the oxidative branch of PPP, and suggests rational guidelines for the design of strategies towards the production of new and efficient L-arabinose-fermenting yeasts.},
  author       = {Fonseca, César and Neves, Ana Rute and Antunes, Alexandra M M and Noronha, João Paulo and Hahn-Hägerdal, Bärbel and Santos, Helena and Spencer-Martins, Isabel},
  issn         = {0099-2240},
  language     = {eng},
  number       = {6},
  pages        = {1845--1855},
  publisher    = {American Society for Microbiology},
  series       = {Applied and Environmental Microbiology},
  title        = {Use of in vivo C-13 nuclear magnetic resonance Spectroscopy to elucidate L-arabinose metabolism in Yeasts},
  url          = {http://dx.doi.org/10.1128/AEM.02453-07},
  volume       = {74},
  year         = {2008},
}