Induction of a metabolic switch in insect cells by substrate-limited fed batch cultures

Applied Microbiology and Biotechnology
L OhmanL Häggström

Abstract

Insect cell metabolism was studied in substrate-limited fed batch cultures of Spodoptera frugiperda (Sf-9) cells. Results from a glucose-limited culture, a glutamine-limited culture, a culture limited in both glucose and glutamine, and a batch culture were compared. A stringent relation between glucose excess and alanine formation was found. In contrast, glucose limitation induced ammonium formation, while, at the same time, alanine formation was completely suppressed. Simultaneous glucose and glucosamine limitation suppressed both alanine and ammonium formation. Although the metabolism was influenced by substrate limitation, the specific growth rate was similar in all cultures. Alanine formation must involve incorporation of free ammonium, if ammonium formation is mediated by glutaminase and glutamate dehydrogenase, as our data suggest. On the basis of the results, two possible pathways for the formation of alanine in the intermediary metabolism are suggested. The cellular yield on glucose was increased 6.6 times during glucose limitation, independently of the cellular yield on glutamine, which was increased 50-100 times during glutamine limitation. The results indicate that alanine overflow metabolism is energetically wastefu...Continue Reading

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Citations

Oct 17, 2008·Applied Microbiology and Biotechnology·Nuno CarinhasPaula M Alves
Feb 22, 2008·In Vitro Cellular & Developmental Biology. Animal·Leila MatindoostFateme Rahbarizade
Sep 29, 1995·Journal of Biotechnology·J Ljunggren, L Häggström
Mar 6, 1998·Journal of Biotechnology·M DoverskogL Häggström
Sep 11, 2003·Biotechnology and Bioengineering·Luis MarangaManuel J T Carrondo
Oct 11, 2011·Biotechnology Advances·Jean-Christophe DrugmandSpiros N Agathos
Mar 9, 2010·Journal of Insect Physiology·Patricia Y ScaraffiaRoger L Miesfeld
May 22, 2009·Biotechnology and Bioengineering·Vicente BernalPaula M Alves
Dec 3, 2017·In Vitro Cellular & Developmental Biology. Animal·Charlotte PushparajanGabriel Visnovsky

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