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Effect of glycerol feed in methanol induction phase for hepatitis B surface antigen expression in Pichia pastoris strain KM71


Citation

Morvarid, Akhavan Rezaei and Allaudin, Zeenathul Nazariah and Tam, Yew Joon and Hassan, Zuridah and Mohd Lila, Mohd Azmi and O., Azizon (2012) Effect of glycerol feed in methanol induction phase for hepatitis B surface antigen expression in Pichia pastoris strain KM71. Pertanika Journal of Science & Technology, 20 (1). pp. 31-42. ISSN 0128-7680; ESSN: 2231-8526

Abstract / Synopsis

This study describes expression of HBs Ag in methylotrophic yeast, Pichia Pastoris under alcohol oxidase promoter. A single copy number of HBs Ag gene was transformed into pichia strain of KM 71, a Mutˢ type, by using pA0815 pichia expression vector. The recombinant was cultivated in a shake flask either using methanol or a mixed feed of glycerol -methanol for induction. The HBs Ag gene integrity was justified using direct PCR method. The expressed products in the soluble cell extracts were analyzed by Western blot, SDS page, Bradford assay and ELISA tests. The recombinant HBs Ag was expressed successfully in Pichia pastoris strain KM71 at a high level of HBs Ag protein expression. Thus, an addition of glycerol in the ratio of glycerol per methanol 1/1 (g g-1) consistently produced 2-fold increment in both biomass accumulation and HBs Ag productivity.


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Additional Metadata

Item Type: Article
Divisions: Institute of Bioscience
Publisher: Universiti Putra Malaysia Press
Keywords: Biomass; Hepatitis B; KM71 strain; pAO815 expression vector; Pichia pastoris;Surface Ag
Depositing User: Najah Mohd Ali
Date Deposited: 04 Nov 2015 09:36
Last Modified: 04 Nov 2015 09:36
URI: http://psasir.upm.edu.my/id/eprint/40425
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