Production of hantavirus Puumala nucleocapsid protein in Saccharomyces cerevisiae for vaccine and diagnostics

Journal of Biotechnology
L AntoniukasU Reichl

Abstract

The production of hantavirus Puumala nucleocapsid (N) protein for potential applications as a vaccine and for diagnostic purposes was investigated with Saccharomyces cerevisiae as a recombinant host. The N protein gene and the hexahistidine tagged N (h-N) protein gene were expressed intracellular from a 2-microm plasmid vectors under the control of a fused galactose inducible GAL10-PYK promoter. For monitoring the recombinant gene expression, a h-N and a GFP fusion protein was used. Different cultivation strategies and growth media compositions were tested in shake flasks and a 5 l bioreactor. When using defined YNB growth medium, we found the biomass yield to be unsatisfactorily low. Higher concentrated YNB medium, promoted cell growth but showed a pronounced inhibitory effect on heterologous gene expression. This phenomenon could not be attributed to plasmid losses, as we could demonstrate high stability of the vector under the applied cultivation conditions. Supplementation of YNB medium with extracts of plant origin resulted in increased biomass yields with concomitant high expression levels of the recombinant gene. The modified medium was used for fed-batch cultivations where basic metabolic features as well as growth para...Continue Reading

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Citations

Dec 26, 2006·Applied and Environmental Microbiology·Alane E Wentz, Eric V Shusta
Feb 2, 2008·Biotechnology Letters·Douglas MayaSebastián Chávez
Jan 6, 2015·The Journal of Pharmacy and Pharmacology·Roslyn M Bill
Apr 9, 2008·FEMS Microbiology Reviews·Ai-Sheng XiongQuan-Hong Yao
Dec 20, 2008·Expert Review of Vaccines·Piet MaesMarc Van Ranst
Feb 23, 2020·Frontiers in Microbiology·Rongrong LiuXingan Wu
May 2, 2020·Expert Review of Clinical Pharmacology·Kamila Botelho Sampaio de OliveiraSimoni Campos Dias

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