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Construction of cellobiose-growing and fermenting Saccharomyces cerevisiae strains

van Rooyen, Ronél LU ; Hahn-Hägerdal, Bärbel LU ; La Grange, D C and van Zyl, W H (2005) In Journal of Biotechnology 120(3). p.284-295
Abstract
beta-Glucosidase genes of fungal origins were isolated and heterologously expressed in Saccharomyces cerevisiae to enable growth on the disaccharide, cellobiose. To promote secretion of the beta-glucosidases, the genes were fused to the secretion signal of the Trichoderma reesei xyn2 gene and constitutively expressed from a multi-copy yeast expression vector under transcriptional control of the S. cerevisiae PGK1 promoter and terminator. The resulting recombinant enzymes were characterized with respect to pH and temperature optimum, as well as kinetic properties. The two most promising enzymes, BGL1 from Saccharomycopsis fibuligera and Bg1A from Aspergillus kawachii, were anchored to the yeast cell surface by fusing the mature proteins to... (More)
beta-Glucosidase genes of fungal origins were isolated and heterologously expressed in Saccharomyces cerevisiae to enable growth on the disaccharide, cellobiose. To promote secretion of the beta-glucosidases, the genes were fused to the secretion signal of the Trichoderma reesei xyn2 gene and constitutively expressed from a multi-copy yeast expression vector under transcriptional control of the S. cerevisiae PGK1 promoter and terminator. The resulting recombinant enzymes were characterized with respect to pH and temperature optimum, as well as kinetic properties. The two most promising enzymes, BGL1 from Saccharomycopsis fibuligera and Bg1A from Aspergillus kawachii, were anchored to the yeast cell surface by fusing the mature proteins to the alpha-agglutinin (AG alpha 1) or cell wall protein 2 (Cwp2) peptides. The maximum specific growth rates (mu(max)) of the recombinant S. cerevisiae strains were determined in batch cultivation. S. cerevisiae secreting the recombinant S. fibuligera BGL1 enzyme sustained growth aerobically and anaerobically, in minimal medium containing 5 g L-1 cellobiose at 0.23 h(-1) (compared to 0.29 h(-1) on glucose) and 0.18 h(-1) (compared to 0.25 h(-1) on glucose), respectively. Substrate consumption and product formation were determined to evaluate product yields in glucose and cellobiose. (c) 2005 Elsevier B.V. All rights reserved. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Journal of Biotechnology
volume
120
issue
3
pages
284 - 295
publisher
Elsevier
external identifiers
  • pmid:16084620
  • wos:000233347900005
  • scopus:27544459042
ISSN
1873-4863
DOI
10.1016/j.jbiotec.2005.06.013
language
English
LU publication?
yes
id
4dc5474f-868a-4f6b-bf16-0a5b29236c78 (old id 151202)
date added to LUP
2007-06-28 15:36:51
date last changed
2017-10-29 03:41:37
@article{4dc5474f-868a-4f6b-bf16-0a5b29236c78,
  abstract     = {beta-Glucosidase genes of fungal origins were isolated and heterologously expressed in Saccharomyces cerevisiae to enable growth on the disaccharide, cellobiose. To promote secretion of the beta-glucosidases, the genes were fused to the secretion signal of the Trichoderma reesei xyn2 gene and constitutively expressed from a multi-copy yeast expression vector under transcriptional control of the S. cerevisiae PGK1 promoter and terminator. The resulting recombinant enzymes were characterized with respect to pH and temperature optimum, as well as kinetic properties. The two most promising enzymes, BGL1 from Saccharomycopsis fibuligera and Bg1A from Aspergillus kawachii, were anchored to the yeast cell surface by fusing the mature proteins to the alpha-agglutinin (AG alpha 1) or cell wall protein 2 (Cwp2) peptides. The maximum specific growth rates (mu(max)) of the recombinant S. cerevisiae strains were determined in batch cultivation. S. cerevisiae secreting the recombinant S. fibuligera BGL1 enzyme sustained growth aerobically and anaerobically, in minimal medium containing 5 g L-1 cellobiose at 0.23 h(-1) (compared to 0.29 h(-1) on glucose) and 0.18 h(-1) (compared to 0.25 h(-1) on glucose), respectively. Substrate consumption and product formation were determined to evaluate product yields in glucose and cellobiose. (c) 2005 Elsevier B.V. All rights reserved.},
  author       = {van Rooyen, Ronél and Hahn-Hägerdal, Bärbel and La Grange, D C and van Zyl, W H},
  issn         = {1873-4863},
  language     = {eng},
  number       = {3},
  pages        = {284--295},
  publisher    = {Elsevier},
  series       = {Journal of Biotechnology},
  title        = {Construction of cellobiose-growing and fermenting Saccharomyces cerevisiae strains},
  url          = {http://dx.doi.org/10.1016/j.jbiotec.2005.06.013},
  volume       = {120},
  year         = {2005},
}