Advanced

Control of xylose consumption by xylose transport in recombinant Saccharomyces cerevisiae

Gárdonyi, Márk LU ; Jeppsson, Marie LU ; Lidén, Gunnar LU ; Gorwa-Grauslund, Marie-Francoise LU and Hahn-Hägerdal, Bärbel LU (2003) In Biotechnology and Bioengineering 7(82). p.818-824
Abstract
Saccharomyces cerevisiae TMB3001 has previously been engineered to utilize xylose by integrating the genes coding for xylose reductase (XR) and xylitol dehydrogenase (XDH) and overexpressing the native xylulokinase (XK) gene. The resulting strain is able to metabolize xylose, but its xylose utilization rate is low compared to that of natural xylose utilizing yeasts, like Pichia stipitis or Candida shehatae. One difference between S. cerevisiae and the latter species is that these possess specific xylose transporters, while S. cerevisiae takes up xylose via the high-affinity hexose transporters. For this reason, in part, it has been suggested that xylose transport in S. cerevisiae may limit the xylose utilization.

We investigated... (More)
Saccharomyces cerevisiae TMB3001 has previously been engineered to utilize xylose by integrating the genes coding for xylose reductase (XR) and xylitol dehydrogenase (XDH) and overexpressing the native xylulokinase (XK) gene. The resulting strain is able to metabolize xylose, but its xylose utilization rate is low compared to that of natural xylose utilizing yeasts, like Pichia stipitis or Candida shehatae. One difference between S. cerevisiae and the latter species is that these possess specific xylose transporters, while S. cerevisiae takes up xylose via the high-affinity hexose transporters. For this reason, in part, it has been suggested that xylose transport in S. cerevisiae may limit the xylose utilization.

We investigated the control exercised by the transport over the specific xylose utilization rate in two recombinant S. cerevisiae strains, one with low XR activity, TMB3001, and one with high XR activity, TMB3260. The strains were grown in aerobic sugar-limited chemostat and the specific xylose uptake rate was modulated by changing the xylose concentration in the feed, which allowed determination of the flux response coefficients. Separate measurements of xylose transport kinetics allowed determination of the elasticity coefficients of transport with respect to extracellular xylose concentration. The flux control coefficient, C, for the xylose transport was calculated from the response and elasticity coefficients. The value of C for both strains was found to be < 0.1 at extracellular xylose concentrations > 7.5 g L-1. However, for strain TMB3260 the flux control coefficient was higher than 0.5 at xylose concentrations < 0.6 g L-1, while C stayed below 0.2 for strain TMB3001 irrespective of xylose concentration. © 2003 Wiley Periodicals, Inc. Biotechnol Bioeng 82: 818-824, 2003. (Less)
Please use this url to cite or link to this publication:
author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
xylose transport, Saccharomyces cerevisiae, metabolic control analysis, xylose reductase
in
Biotechnology and Bioengineering
volume
7
issue
82
pages
818 - 824
publisher
John Wiley & Sons
external identifiers
  • wos:000182913300008
  • pmid:12701148
  • scopus:0038363853
ISSN
1097-0290
DOI
10.1002/bit.10631
language
English
LU publication?
yes
id
c0447da4-3e63-43dd-9acb-4b69fbd782ee (old id 636593)
date added to LUP
2007-11-30 12:11:21
date last changed
2017-07-30 04:47:30
@article{c0447da4-3e63-43dd-9acb-4b69fbd782ee,
  abstract     = {Saccharomyces cerevisiae TMB3001 has previously been engineered to utilize xylose by integrating the genes coding for xylose reductase (XR) and xylitol dehydrogenase (XDH) and overexpressing the native xylulokinase (XK) gene. The resulting strain is able to metabolize xylose, but its xylose utilization rate is low compared to that of natural xylose utilizing yeasts, like Pichia stipitis or Candida shehatae. One difference between S. cerevisiae and the latter species is that these possess specific xylose transporters, while S. cerevisiae takes up xylose via the high-affinity hexose transporters. For this reason, in part, it has been suggested that xylose transport in S. cerevisiae may limit the xylose utilization. <br/><br>
We investigated the control exercised by the transport over the specific xylose utilization rate in two recombinant S. cerevisiae strains, one with low XR activity, TMB3001, and one with high XR activity, TMB3260. The strains were grown in aerobic sugar-limited chemostat and the specific xylose uptake rate was modulated by changing the xylose concentration in the feed, which allowed determination of the flux response coefficients. Separate measurements of xylose transport kinetics allowed determination of the elasticity coefficients of transport with respect to extracellular xylose concentration. The flux control coefficient, C, for the xylose transport was calculated from the response and elasticity coefficients. The value of C for both strains was found to be &lt; 0.1 at extracellular xylose concentrations &gt; 7.5 g L-1. However, for strain TMB3260 the flux control coefficient was higher than 0.5 at xylose concentrations &lt; 0.6 g L-1, while C stayed below 0.2 for strain TMB3001 irrespective of xylose concentration. © 2003 Wiley Periodicals, Inc. Biotechnol Bioeng 82: 818-824, 2003.},
  author       = {Gárdonyi, Márk and Jeppsson, Marie and Lidén, Gunnar and Gorwa-Grauslund, Marie-Francoise and Hahn-Hägerdal, Bärbel},
  issn         = {1097-0290},
  keyword      = {xylose transport,Saccharomyces cerevisiae,metabolic control analysis,xylose reductase},
  language     = {eng},
  number       = {82},
  pages        = {818--824},
  publisher    = {John Wiley & Sons},
  series       = {Biotechnology and Bioengineering},
  title        = {Control of xylose consumption by xylose transport in recombinant Saccharomyces cerevisiae},
  url          = {http://dx.doi.org/10.1002/bit.10631},
  volume       = {7},
  year         = {2003},
}