Format

Send to:

Choose Destination
See comment in PubMed Commons below
Appl Microbiol Biotechnol. 2009 Feb;82(2):249-59. doi: 10.1007/s00253-008-1756-z. Epub 2008 Nov 18.

Impact of oxygen supply on rtPA expression in Escherichia coli BL21 (DE3): ammonia effects.

Author information

  • 1Newworld Institute of Biotechnology, State Key Laboratory of Bioreactor Engineering, East China University of Science & Technology, P.O. Box 311, 130 Meilong Road, Shanghai 200237, People's Republic of China.

Abstract

In shake flasks, good oxygen supply tended to decrease rtPA expression in media containing only yeast extract and tryptone, while oxygen limitation would increase rtPA synthesis in the same medium. Our data showed that though the drop of rtPA expression in the 20-ml cultures of LBG or 2YTG was accompanied with a severe acetate accumulation, it was actually caused by low ammonia. The rtPA expression level could be significantly improved by increasing culture ammonium ion up to 500 mM. The effects of exogenous high ammonia on cell growth and rtPA expression were further examined in shake flasks and a 4-l fermentor. The high ammonia had no significant impact on cell growth and oxygen respiratory activity but significantly depressed the activities of glutamine synthetase/glutamate synthase and glutamate dehydrogenase, suggesting that ammonium ion as a nitrogen source improved the protein expression by mediating ammonia-assimilating enzymes. We thus propose in our work that E. coli cells, which were grown to a certain density to produce rtPA, would undergo nitrogen starvation under the low ammonia conditions even when the organic nitrogen sources remained abundant. The scale-up of rtPA production from shake flasks to fermentors could be readily achieved in the media containing rich ammonium ion.

PMID:
19015848
[PubMed - indexed for MEDLINE]
PubMed Commons home

PubMed Commons

0 comments
How to join PubMed Commons

    Supplemental Content

    Full text links

    Icon for Springer
    Loading ...
    Write to the Help Desk