Investigating the effect of carbon source on rabies virus glycoprotein production in Pichia pastoris by a transcriptomic approach

MicrobiologyOpen
Safa Ben Azoun, Héla Kallel

Abstract

Several factors affect protein expression in Pichia pastoris, one among them is the carbon source. In this work, we studied the effect of this factor on the expression level of rabies virus glycoprotein (RABV-G) in two recombinant clones harboring seven copies of the gene of interest. The expression was driven either by the constitutive glyceraldehyde-3-phosphate dehydrogenase (GAP) promoter or the inducible alcohol oxidase1 (AOX1) promoter. Clones were compared in terms of cell physiology and carbon source metabolism. The transcription levels of 16 key genes involved in the central metabolic pathway, the methanol catabolism, and the oxidative stress were investigated in both clones. Cell size, as a parameter reflecting cell physiological changes, was also monitored. Our results showed that when glucose was used as the sole carbon source, large cells were obtained. Transcript levels of the genes of the central metabolic pathway were also upregulated, whereas antioxidative gene transcript levels were low. By contrast, the use of methanol as a carbon source generated small cells and a shift in carbon metabolism toward the dissimilatory pathway by the upregulation of formaldehyde dehydrogenase gene and the downregulation of those ...Continue Reading

References

Oct 23, 2001·Biotechnology and Bioengineering·S Frykman, F Srienc
Feb 3, 2004·Biotechnology and Bioengineering·Hubertus HohenblumDiethard Mattanovich
Jul 31, 2004·BioTechniques·Keertan DhedaAlimuddin Zumla
Oct 20, 2005·Journal of Bioscience and Bioengineering·M Inan, M M Meagher
Jul 12, 2008·The FEBS Journal·Rogerio MargisMarcia Margis-Pinheiro
Feb 17, 2009·Journal of Proteome Research·Martin DragositsDiethard Mattanovich
Oct 26, 2010·BMC Systems Biology·Kristin BaumannPau Ferrer
Nov 6, 2010·Applied Microbiology and Biotechnology·Ildar NisamedtinovToomas Paalme
Feb 26, 2011·Biotechnology and Bioengineering·Jan HeylandAndreas Schmid
May 11, 2011·Biotechnology and Bioengineering·Markus BucheticsDiethard Mattanovich
May 12, 2012·Current Biology : CB·Jonathan J TurnerJan M Skotheim
Feb 5, 2013·Future Microbiology·Brigitte GasserDiethard Mattanovich
Apr 20, 2014·Applied Microbiology and Biotechnology·Mudassar AhmadHelmut Schwab
May 24, 2014·Metabolic Engineering·Justyna NoconDiethard Mattanovich
Feb 28, 2015·Applied Microbiology and Biotechnology·Verena PuxbaumBrigitte Gasser
Feb 18, 2016·Microbial Biotechnology·Safa Ben AzounHéla Kallel
Mar 30, 2016·Applied Microbiology and Biotechnology·Justyna NoconDiethard Mattanovich

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Datasets Mentioned

BETA
KT878717

Methods Mentioned

BETA
protein folding
PCR
electrophoresis

Software Mentioned

Image
Primer

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