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Optimization of conditions for secretion of dengue virus type 2 envelope domain III using Pichia pastoris

Paper ID Volume ID Publish Year Pages File Format Full-Text
21395 43220 2010 7 PDF Available
Title
Optimization of conditions for secretion of dengue virus type 2 envelope domain III using Pichia pastoris
Abstract

We have developed a recombinant clone of the methylotrophic yeast Pichia pastoris capable of secreting dengue virus type 2 envelope domain III (sEDIII-2). We explored various induction parameters including media composition, temperature, pH, and methanol concentration, to optimize conditions for sEDIII-2 expression in shake flask culture. Induction at 20 °C in the presence of 2% (v/v) methanol in a medium buffered to pH 5.8 resulted in highest secretion of sEDIII-2. This yield could be further enhanced up to 70% by repeated induction of the same initial biomass. Using a fed-batch cultivation strategy, we observed that shake-flask yields can be scaled up ∼ 8-fold in a bioreactor. We obtained ∼ 94% purity with > 70% recovery after purification. This study, which demonstrates for the first time the feasibility of secreting envelope domain III using the P. pastoris host, will be relevant to sub-unit approaches to dengue vaccine development.

Keywords
Dengue; Flavivirus; Envelope domain III; Pichia pastoris; Fed-batch cultivation; Vaccine
First Page Preview
Optimization of conditions for secretion of dengue virus type 2 envelope domain III using Pichia pastoris
Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Bioscience and Bioengineering - Volume 110, Issue 4, October 2010, Pages 408–414
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering