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Expression and analysis of the glycosylation properties of recombinant human erythropoietin expressed in Pichia pastoris

The Pichia pastoris expression system was used to produce recombinant human erythropoietin, a protein synthesized by the adult kidney and responsible for the regulation of red blood cell production. The entire recombinant human erythropoietin (rhEPO) gene was constructed using the Splicing by Overla...

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Autores principales: Teh, Ser Huy, Fong, Mun Yik, Mohamed, Zulqarnain
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Sociedade Brasileira de Genética 2011
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3168189/
https://www.ncbi.nlm.nih.gov/pubmed/21931521
http://dx.doi.org/10.1590/S1415-47572011005000022
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author Teh, Ser Huy
Fong, Mun Yik
Mohamed, Zulqarnain
author_facet Teh, Ser Huy
Fong, Mun Yik
Mohamed, Zulqarnain
author_sort Teh, Ser Huy
collection PubMed
description The Pichia pastoris expression system was used to produce recombinant human erythropoietin, a protein synthesized by the adult kidney and responsible for the regulation of red blood cell production. The entire recombinant human erythropoietin (rhEPO) gene was constructed using the Splicing by Overlap Extension by PCR (SOE-PCR) technique, cloned and expressed through the secretory pathway of the Pichia expression system. Recombinant erythropoietin was successfully expressed in P. pastoris. The estimated molecular mass of the expressed protein ranged from 32 kDa to 75 kDa, with the variation in size being attributed to the presence of rhEPO glycosylation analogs. A crude functional analysis of the soluble proteins showed that all of the forms were active in vivo.
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spelling pubmed-31681892011-09-19 Expression and analysis of the glycosylation properties of recombinant human erythropoietin expressed in Pichia pastoris Teh, Ser Huy Fong, Mun Yik Mohamed, Zulqarnain Genet Mol Biol Genetics of Microorganisms The Pichia pastoris expression system was used to produce recombinant human erythropoietin, a protein synthesized by the adult kidney and responsible for the regulation of red blood cell production. The entire recombinant human erythropoietin (rhEPO) gene was constructed using the Splicing by Overlap Extension by PCR (SOE-PCR) technique, cloned and expressed through the secretory pathway of the Pichia expression system. Recombinant erythropoietin was successfully expressed in P. pastoris. The estimated molecular mass of the expressed protein ranged from 32 kDa to 75 kDa, with the variation in size being attributed to the presence of rhEPO glycosylation analogs. A crude functional analysis of the soluble proteins showed that all of the forms were active in vivo. Sociedade Brasileira de Genética 2011-07-01 2011 /pmc/articles/PMC3168189/ /pubmed/21931521 http://dx.doi.org/10.1590/S1415-47572011005000022 Text en Copyright © 2011, Sociedade Brasileira de Genética. Printed in Brazil License information: This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Genetics of Microorganisms
Teh, Ser Huy
Fong, Mun Yik
Mohamed, Zulqarnain
Expression and analysis of the glycosylation properties of recombinant human erythropoietin expressed in Pichia pastoris
title Expression and analysis of the glycosylation properties of recombinant human erythropoietin expressed in Pichia pastoris
title_full Expression and analysis of the glycosylation properties of recombinant human erythropoietin expressed in Pichia pastoris
title_fullStr Expression and analysis of the glycosylation properties of recombinant human erythropoietin expressed in Pichia pastoris
title_full_unstemmed Expression and analysis of the glycosylation properties of recombinant human erythropoietin expressed in Pichia pastoris
title_short Expression and analysis of the glycosylation properties of recombinant human erythropoietin expressed in Pichia pastoris
title_sort expression and analysis of the glycosylation properties of recombinant human erythropoietin expressed in pichia pastoris
topic Genetics of Microorganisms
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3168189/
https://www.ncbi.nlm.nih.gov/pubmed/21931521
http://dx.doi.org/10.1590/S1415-47572011005000022
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