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Amino acid supplementation, controlled oxygen limitation and sequential double induction improves heterologous xylanase production by Pichia stipitis

Görgens, Johann F; Passoth, Volkmar; van Zyl, WH; Knoetze, JH and Hahn-Hägerdal, Bärbel LU (2005) In FEMS Yeast Research 5(6-7). p.677-683
Abstract
Heterologous endo-beta-1,4-xylanase was produced by Pichia stipitis under control of the hypoxia-inducible PsADH2-promoter in a high-cell-clensity culture. After promoter induction by a shift to oxygen limitation, different aeration rates (oxygen transfer rates) were applied while maintaining oxygen-limitation. Initially, enzyme production was higher in oxygen-limited cultures with high rates of oxygen transfer, although the maximum xylanase activity was not significantly influenced. Amino acid supplementation increased the production of the heterologous endo-beta-1,4-xylanase significantly in highly aerated oxygen-limited cultures, until glucose was depleted. A slight second induction of the promoter was observed in all cultures after the... (More)
Heterologous endo-beta-1,4-xylanase was produced by Pichia stipitis under control of the hypoxia-inducible PsADH2-promoter in a high-cell-clensity culture. After promoter induction by a shift to oxygen limitation, different aeration rates (oxygen transfer rates) were applied while maintaining oxygen-limitation. Initially, enzyme production was higher in oxygen-limited cultures with high rates of oxygen transfer, although the maximum xylanase activity was not significantly influenced. Amino acid supplementation increased the production of the heterologous endo-beta-1,4-xylanase significantly in highly aerated oxygen-limited cultures, until glucose was depleted. A slight second induction of the promoter was observed in all cultures after the glucose had been consumed. The second induction was most obvious in amino acid-supplemented cultures with higher oxygen transfer rates during oxygen limitation. When such oxygen-limited cultures were shifted back to fully aerobic conditions, a significant re-induction of heterologous endo-beta-1,4-xylanase production was observed. Re-induction was accompanied by ethanol consumption. A similar protein production pattern was observed when cultures were first grown on ethanol as sole carbon source and subsequently glucose and oxygen limitation were applied. Thus, we present the first expression system in yeast with a sequential double-inducible promoter. (c) 2004 Federation of European Microbiological Societies. Published by Elsevier B.V. All rights reserved. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
FEMS Yeast Research
volume
5
issue
6-7
pages
677 - 683
publisher
Elsevier
external identifiers
  • wos:000228153900019
  • pmid:15780668
  • scopus:15044351457
ISSN
1567-1364
DOI
10.1016/j.femsyr.2004.12.003
language
English
LU publication?
yes
id
dedbcfa6-3438-4a52-a471-c9e260835a75 (old id 151266)
date added to LUP
2007-06-28 11:48:51
date last changed
2017-01-01 05:08:54
@article{dedbcfa6-3438-4a52-a471-c9e260835a75,
  abstract     = {Heterologous endo-beta-1,4-xylanase was produced by Pichia stipitis under control of the hypoxia-inducible PsADH2-promoter in a high-cell-clensity culture. After promoter induction by a shift to oxygen limitation, different aeration rates (oxygen transfer rates) were applied while maintaining oxygen-limitation. Initially, enzyme production was higher in oxygen-limited cultures with high rates of oxygen transfer, although the maximum xylanase activity was not significantly influenced. Amino acid supplementation increased the production of the heterologous endo-beta-1,4-xylanase significantly in highly aerated oxygen-limited cultures, until glucose was depleted. A slight second induction of the promoter was observed in all cultures after the glucose had been consumed. The second induction was most obvious in amino acid-supplemented cultures with higher oxygen transfer rates during oxygen limitation. When such oxygen-limited cultures were shifted back to fully aerobic conditions, a significant re-induction of heterologous endo-beta-1,4-xylanase production was observed. Re-induction was accompanied by ethanol consumption. A similar protein production pattern was observed when cultures were first grown on ethanol as sole carbon source and subsequently glucose and oxygen limitation were applied. Thus, we present the first expression system in yeast with a sequential double-inducible promoter. (c) 2004 Federation of European Microbiological Societies. Published by Elsevier B.V. All rights reserved.},
  author       = {Görgens, Johann F and Passoth, Volkmar and van Zyl, WH and Knoetze, JH and Hahn-Hägerdal, Bärbel},
  issn         = {1567-1364},
  language     = {eng},
  number       = {6-7},
  pages        = {677--683},
  publisher    = {Elsevier},
  series       = {FEMS Yeast Research},
  title        = {Amino acid supplementation, controlled oxygen limitation and sequential double induction improves heterologous xylanase production by Pichia stipitis},
  url          = {http://dx.doi.org/10.1016/j.femsyr.2004.12.003},
  volume       = {5},
  year         = {2005},
}