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

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 e...

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Main Authors: Morvarid, Akhavan Rezaei, Allaudin, Zeenathul Nazariah, Tam, Yew Joon, Hassan, Zuridah, Mohd Lila, Mohd Azmi, Othman, Azizon
Format: Article
Language:English
Published: Universiti Putra Malaysia Press 2012
Online Access:http://psasir.upm.edu.my/id/eprint/40425/
http://psasir.upm.edu.my/id/eprint/40425/1/3.%20Effect%20of%20glycerol%20feed%20in%20methanol%20induction%20phase%20for%20hepatitis%20B%20surface%20antigen%20expression%20in%20Pichia%20pastoris%20strain%20KM71.pdf
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author Morvarid, Akhavan Rezaei
Allaudin, Zeenathul Nazariah
Tam, Yew Joon
Hassan, Zuridah
Mohd Lila, Mohd Azmi
Othman, Azizon
author_facet Morvarid, Akhavan Rezaei
Allaudin, Zeenathul Nazariah
Tam, Yew Joon
Hassan, Zuridah
Mohd Lila, Mohd Azmi
Othman, Azizon
author_sort Morvarid, Akhavan Rezaei
building UPM Institutional Repository
collection Online Access
description 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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spelling upm-404252019-10-30T06:50:59Z http://psasir.upm.edu.my/id/eprint/40425/ Effect of glycerol feed in methanol induction phase for hepatitis B surface antigen expression in Pichia pastoris strain KM71 Morvarid, Akhavan Rezaei Allaudin, Zeenathul Nazariah Tam, Yew Joon Hassan, Zuridah Mohd Lila, Mohd Azmi Othman, Azizon 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. Universiti Putra Malaysia Press 2012 Article PeerReviewed application/pdf en http://psasir.upm.edu.my/id/eprint/40425/1/3.%20Effect%20of%20glycerol%20feed%20in%20methanol%20induction%20phase%20for%20hepatitis%20B%20surface%20antigen%20expression%20in%20Pichia%20pastoris%20strain%20KM71.pdf Morvarid, Akhavan Rezaei and Allaudin, Zeenathul Nazariah and Tam, Yew Joon and Hassan, Zuridah and Mohd Lila, Mohd Azmi and Othman, 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 http://www.pertanika.upm.edu.my/Pertanika%20PAPERS/JST%20Vol.%2020%20%281%29%20Jan.%202012/08%20Pg%2031-42.pdf
spellingShingle Morvarid, Akhavan Rezaei
Allaudin, Zeenathul Nazariah
Tam, Yew Joon
Hassan, Zuridah
Mohd Lila, Mohd Azmi
Othman, Azizon
Effect of glycerol feed in methanol induction phase for hepatitis B surface antigen expression in Pichia pastoris strain KM71
title Effect of glycerol feed in methanol induction phase for hepatitis B surface antigen expression in Pichia pastoris strain KM71
title_full Effect of glycerol feed in methanol induction phase for hepatitis B surface antigen expression in Pichia pastoris strain KM71
title_fullStr Effect of glycerol feed in methanol induction phase for hepatitis B surface antigen expression in Pichia pastoris strain KM71
title_full_unstemmed Effect of glycerol feed in methanol induction phase for hepatitis B surface antigen expression in Pichia pastoris strain KM71
title_short Effect of glycerol feed in methanol induction phase for hepatitis B surface antigen expression in Pichia pastoris strain KM71
title_sort effect of glycerol feed in methanol induction phase for hepatitis b surface antigen expression in pichia pastoris strain km71
url http://psasir.upm.edu.my/id/eprint/40425/
http://psasir.upm.edu.my/id/eprint/40425/
http://psasir.upm.edu.my/id/eprint/40425/1/3.%20Effect%20of%20glycerol%20feed%20in%20methanol%20induction%20phase%20for%20hepatitis%20B%20surface%20antigen%20expression%20in%20Pichia%20pastoris%20strain%20KM71.pdf