Home News About Us Contact Contributors Disclaimer Privacy Policy Help FAQ

Home
Search
Quick Search
Advanced
Fulltext
Browse
Collections
Persons
My eDoc
Session History
Login
Name:
Password:
Documentation
Help
Support Wiki
Direct access to
document ID:


          Institute: MPI für Dynamik komplexer technischer Systeme     Collection: Bioprocess Engineering     Display Documents



  history
ID: 245291.0, MPI für Dynamik komplexer technischer Systeme / Bioprocess Engineering
Production of hantavirus Puumala nucleocapsid protein in Saccharomyces cerevisiae for vaccine and diagnostics
Authors:Antoniukas, L.; Grammel, H.; Reichl, U.
Name of Conference/Meeting:BioPerspectives 2005
Place of Conference/Meeting:Wiesbaden, Germany
(Start) Date of Event 
 (YYYY-MM-DD):
2005-05-10
End Date of Conference/Meeting 
 (YYYY-MM-DD):
2005-05-12
Audience:Experts Only
Abstract / Description:Some European hantaviruses can infect humans and cause a serious disease - HFRS. There are no commercially available vaccine and diagnostics against those hantaviruses. The recombinat nucleocapsid protein of hantavirus was demonstrated being a good vaccine candidate and suitable antigen for diagnostics. This study focused on the growth of the recombinant Saccharomyces cerevisiae FH4C strain and respective production of the hantavirus Puumala nucleocapsid protein (N). The recombinant sequence (which encodes N protein) was expressed intracellulary from a S. cerevisiae 2-micron plasmid vector under the control of fused, galactose inducible GAL10-PYK promoter. Different cultivation strategies and media were tested in shake flasks and a 5 L bioreactor, and the respective cell growth, metabolism and nucleocapsid protein production were analyzed. According to requirements for human vaccine production, the traditional medium for yeast cultivation (YEP) can not be used. Therefore the search and screening of an optimal animal origin free medium were performed. The commercial minimal medium for yeast cultivation (YNB) was tested first and it showed a very high specific yield of the recombinant N protein (18.9 mg/g DCW) in batch cultivation, but very low biomass yield (7.5 g/L), therefore the volumetric yield of the recombinant N protein was not very high (142.2 mg/L). When a concentrated minimal medium and fed-batch strategy were used for yeast cultivation, the biomass yield was significantly improved (50 g/L), but the recombinant protein yield decreased drastically. Therefore the commercial (YNB) medium was enriched with plant origin complex components: malt extract and soybean peptone. The best results were obtained from the batch cultivation with malt extract enriched YNB medium: the specific yield of the recombinant N protein was slightly higher (19.4 mg/g DCW) when compared to pure YNB and the biomass yield was higher (11 g/L), therefore the volumetric yield of the recombinant protein was significantly improved (213.6 mg/L). These findings provided the basement of the final and optimal strategy for the recombinant S. cerevisiae cultivation and N protein production.
Document Type:Poster
Communicated by:Udo Reichl
Affiliations:MPI für Dynamik komplexer technischer Systeme/Bioprocess Engineering
The scope and number of records on eDoc is subject to the collection policies defined by each institute - see "info" button in the collection browse view.