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Two-phase fed-batch modification for 48 hour peak expression of hepatitis B surface antigen in Pichia pastoris shake flask system


Citation

Tam, Yew Joon and Allaudin, Zeenathul Nazariah and Morvarid, Akhavan Rezaei and Mohd Lila, Mohd Azmi and Bahaman, Abdul Rani and Lo, Sewn Cen and Tan, Joo Shun (2014) Two-phase fed-batch modification for 48 hour peak expression of hepatitis B surface antigen in Pichia pastoris shake flask system. Central European Journal of Biology, 9 (8). pp. 749-760. ISSN 1895-104X; ESSN: 1644-3632

Abstract

A study of the Mut+ phenotype for the expression of recombinant hepatitis B surface antigen (HBsAg) in Pichia pastoris strain GS115 using the pPIC3.5K vector with a two-phase fed-batch protocol in a shake flask system is described. Expression levels of HBsAg protein of 6.74 g L−1 Dry Cell Weight (DCW) and 26.07 mg L−1 of HBsAg concentration were achieved 48 h from the induction point which added to a 120 h reduction in production period in comparison with MutS expression (168 h). The use of the pPIC3.5K-HBsAg plasmid in the Mut+ phenotype enhanced the expression of HBsAg by a nearly 13 times higher volumetric productivity in the first 24 h and 35 times higher at peak production concentration. Comparison of AOX expression cassettes relative to the HBsAg gene in the role of mRNA secondary structure during translation initiation revealed that HBsAg possesses lower folding stability with AOX1 Mut+ phenotype. The results from this study demonstrated that expression of HBsAg with Mut+ AOX1 promoter is adequate as an alternative for the production of HBsAg. In addition, the AOX promoter of the Mut+ phenotype was observed to be better suited for HBsAg expression, which correlates with the ease of translation initiation under shake flask conditions.


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Additional Metadata

Item Type: Article
Divisions: Faculty of Veterinary Medicine
Institute of Bioscience
DOI Number: https://doi.org/10.2478/s11535-014-0309-y
Publisher: Springer
Keywords: Hepatitis B surface antigen; Genetic cloning; Protein expression; Pichia pastoris; Recombinant protein; mRNA secondary structure
Depositing User: Nurul Ainie Mokhtar
Date Deposited: 11 Feb 2016 05:10
Last Modified: 30 Oct 2019 07:20
Altmetrics: http://www.altmetric.com/details.php?domain=psasir.upm.edu.my&doi=10.2478/s11535-014-0309-y
URI: http://psasir.upm.edu.my/id/eprint/35906
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