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Effect of postinduction nutrient feed composition and use of lactose as inducer during production of thermostable xylanase in Escherichia coli glucose-limited fed-batch cultivations

Ramchuran, Santosh LU ; Holst, Olle LU and Nordberg Karlsson, Eva LU (2005) In Journal of Bioscience and Bioengineering 99(5). p.477-484
Abstract
Escherichia coli is a microorganism routinely used in the production of heterologous proteins. The overexpression of a xylanase (Xyn10A Delta NC), which originated from the thermophile Rhodo-thermus marinus cloned under the control of the strong T7/lac promoter in a defined medium (mAT) using a substrate-limited feed strategy, was however shown to impose a significant metabolic burden on host cells. This resulted in a decreased cell growth rate and ultimately also a decreased target protein production. The investigation hence centers on the effect of some selected nutrient feed additives (amino acid [Cys] or TCA-intermediates [citrate, succinate, malate]) used to relieve the metabolic burden imposed during the feeding and postinduction... (More)
Escherichia coli is a microorganism routinely used in the production of heterologous proteins. The overexpression of a xylanase (Xyn10A Delta NC), which originated from the thermophile Rhodo-thermus marinus cloned under the control of the strong T7/lac promoter in a defined medium (mAT) using a substrate-limited feed strategy, was however shown to impose a significant metabolic burden on host cells. This resulted in a decreased cell growth rate and ultimately also a decreased target protein production. The investigation hence centers on the effect of some selected nutrient feed additives (amino acid [Cys] or TCA-intermediates [citrate, succinate, malate]) used to relieve the metabolic burden imposed during the feeding and postinduction phases of these glucose-limited fed-batch cultivations. The use of either succinic acid or malic acid as feed-additives resulted in an increase in production of approximately 40% of the heterologous thermostable xylanase. Furthermore, use of lactose as an alternative inducer of the T7/lac promoter was also proven to be a suitable strategy that significantly prolonged the heterologous protein production phase as compared with induction using isopropyl P-D-thiogalactopyranoside (IPTG). (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Journal of Bioscience and Bioengineering
volume
99
issue
5
pages
477 - 484
publisher
Elsevier
external identifiers
  • pmid:16233820
  • wos:000230570700006
  • scopus:21044441279
ISSN
1347-4421
DOI
10.1263/jbb.99.477
language
English
LU publication?
yes
id
59f39520-c5ca-451a-b07a-57d0f6114f59 (old id 155104)
date added to LUP
2007-07-02 16:52:21
date last changed
2017-04-30 07:09:44
@article{59f39520-c5ca-451a-b07a-57d0f6114f59,
  abstract     = {Escherichia coli is a microorganism routinely used in the production of heterologous proteins. The overexpression of a xylanase (Xyn10A Delta NC), which originated from the thermophile Rhodo-thermus marinus cloned under the control of the strong T7/lac promoter in a defined medium (mAT) using a substrate-limited feed strategy, was however shown to impose a significant metabolic burden on host cells. This resulted in a decreased cell growth rate and ultimately also a decreased target protein production. The investigation hence centers on the effect of some selected nutrient feed additives (amino acid [Cys] or TCA-intermediates [citrate, succinate, malate]) used to relieve the metabolic burden imposed during the feeding and postinduction phases of these glucose-limited fed-batch cultivations. The use of either succinic acid or malic acid as feed-additives resulted in an increase in production of approximately 40% of the heterologous thermostable xylanase. Furthermore, use of lactose as an alternative inducer of the T7/lac promoter was also proven to be a suitable strategy that significantly prolonged the heterologous protein production phase as compared with induction using isopropyl P-D-thiogalactopyranoside (IPTG).},
  author       = {Ramchuran, Santosh and Holst, Olle and Nordberg Karlsson, Eva},
  issn         = {1347-4421},
  language     = {eng},
  number       = {5},
  pages        = {477--484},
  publisher    = {Elsevier},
  series       = {Journal of Bioscience and Bioengineering},
  title        = {Effect of postinduction nutrient feed composition and use of lactose as inducer during production of thermostable xylanase in Escherichia coli glucose-limited fed-batch cultivations},
  url          = {http://dx.doi.org/10.1263/jbb.99.477},
  volume       = {99},
  year         = {2005},
}