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
(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)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/155104
- author
- Ramchuran, Santosh LU ; Holst, Olle LU and Nordberg Karlsson, Eva LU
- organization
- publishing date
- 2005
- 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
- 2022-03-20 18:22:41
@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}}, }