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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 orcid (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
; and
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
2016-04-01 11:44:53
date last changed
2025-10-14 10:41:20
@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}},
  doi          = {{10.1263/jbb.99.477}},
  volume       = {{99}},
  year         = {{2005}},
}