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Use of Ubp1 protease analog to produce recombinant human growth hormone in Escherichia coli

BACKGROUND: Numerous bacterial human growth hormone (hGH) expression methods under conventional fermentation and induction conditions have been described. Despite significant progress made in this area over the past several years, production of recombinant hGH by using cellular expression systems st...

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Autores principales: Wojtowicz-Krawiec, Anna, Sokolowska, Iwona, Smorawinska, Maria, Chojnacka-Puchta, Luiza, Mikiewicz, Diana, Lukasiewicz, Natalia, Marciniak-Rusek, Alina, Wolinowska, Renata, Bierczynska-Krzysik, Anna, Porebska, Anna Joanna, Kuthan-Styczen, Jolanta, Gurba, Lidia, Borowicz, Piotr, Mazurkiewicz, Anna, Plucienniczak, Grazyna, Plucienniczak, Andrzej
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4154517/
https://www.ncbi.nlm.nih.gov/pubmed/25158991
http://dx.doi.org/10.1186/s12934-014-0113-4
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author Wojtowicz-Krawiec, Anna
Sokolowska, Iwona
Smorawinska, Maria
Chojnacka-Puchta, Luiza
Mikiewicz, Diana
Lukasiewicz, Natalia
Marciniak-Rusek, Alina
Wolinowska, Renata
Bierczynska-Krzysik, Anna
Porebska, Anna Joanna
Kuthan-Styczen, Jolanta
Gurba, Lidia
Borowicz, Piotr
Mazurkiewicz, Anna
Plucienniczak, Grazyna
Plucienniczak, Andrzej
author_facet Wojtowicz-Krawiec, Anna
Sokolowska, Iwona
Smorawinska, Maria
Chojnacka-Puchta, Luiza
Mikiewicz, Diana
Lukasiewicz, Natalia
Marciniak-Rusek, Alina
Wolinowska, Renata
Bierczynska-Krzysik, Anna
Porebska, Anna Joanna
Kuthan-Styczen, Jolanta
Gurba, Lidia
Borowicz, Piotr
Mazurkiewicz, Anna
Plucienniczak, Grazyna
Plucienniczak, Andrzej
author_sort Wojtowicz-Krawiec, Anna
collection PubMed
description BACKGROUND: Numerous bacterial human growth hormone (hGH) expression methods under conventional fermentation and induction conditions have been described. Despite significant progress made in this area over the past several years, production of recombinant hGH by using cellular expression systems still requires further optimization. Fusion of the ubiquitin (Ub) tag to the hGH protein allowed to increase of the overall efficiency of the biosynthesis and improve the protein stability. Ub is a protein composed of 76 amino acid residues with a molecular mass of 8.6 kDa, expressed in all eukaryotes. This protein is an element of the universal protein modification system, which does not occur in bacteria, and is a useful carrier for heterologous proteins obtained through expression in Escherichia coli. Purification of Ub-fusion proteins is easier than that of unconjugated recombinant proteins, and Ub can be removed by deubiquitinating proteases (DUBs or UBPs). RESULTS AND CONCLUSION: In the present study the UBPD2C protease, a stable UBP1 analog, was produced as a recombinant protein in E. coli and used for production of recombinant human growth hormone (rhGH). hGH was expressed as a fusion protein with Ub as a tag. Our findings show that the UBPD2C protease is very effective in removing the Ub moiety from recombinant Ub-fused hGH. The described approach enables obtaining a considerable yield of rhGH in a purity required for pharmaceutical products.
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spelling pubmed-41545172014-09-05 Use of Ubp1 protease analog to produce recombinant human growth hormone in Escherichia coli Wojtowicz-Krawiec, Anna Sokolowska, Iwona Smorawinska, Maria Chojnacka-Puchta, Luiza Mikiewicz, Diana Lukasiewicz, Natalia Marciniak-Rusek, Alina Wolinowska, Renata Bierczynska-Krzysik, Anna Porebska, Anna Joanna Kuthan-Styczen, Jolanta Gurba, Lidia Borowicz, Piotr Mazurkiewicz, Anna Plucienniczak, Grazyna Plucienniczak, Andrzej Microb Cell Fact Research BACKGROUND: Numerous bacterial human growth hormone (hGH) expression methods under conventional fermentation and induction conditions have been described. Despite significant progress made in this area over the past several years, production of recombinant hGH by using cellular expression systems still requires further optimization. Fusion of the ubiquitin (Ub) tag to the hGH protein allowed to increase of the overall efficiency of the biosynthesis and improve the protein stability. Ub is a protein composed of 76 amino acid residues with a molecular mass of 8.6 kDa, expressed in all eukaryotes. This protein is an element of the universal protein modification system, which does not occur in bacteria, and is a useful carrier for heterologous proteins obtained through expression in Escherichia coli. Purification of Ub-fusion proteins is easier than that of unconjugated recombinant proteins, and Ub can be removed by deubiquitinating proteases (DUBs or UBPs). RESULTS AND CONCLUSION: In the present study the UBPD2C protease, a stable UBP1 analog, was produced as a recombinant protein in E. coli and used for production of recombinant human growth hormone (rhGH). hGH was expressed as a fusion protein with Ub as a tag. Our findings show that the UBPD2C protease is very effective in removing the Ub moiety from recombinant Ub-fused hGH. The described approach enables obtaining a considerable yield of rhGH in a purity required for pharmaceutical products. BioMed Central 2014-08-27 /pmc/articles/PMC4154517/ /pubmed/25158991 http://dx.doi.org/10.1186/s12934-014-0113-4 Text en © Wojtowicz-Krawiec et al.; licensee BioMed Central Ltd. 2014 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research
Wojtowicz-Krawiec, Anna
Sokolowska, Iwona
Smorawinska, Maria
Chojnacka-Puchta, Luiza
Mikiewicz, Diana
Lukasiewicz, Natalia
Marciniak-Rusek, Alina
Wolinowska, Renata
Bierczynska-Krzysik, Anna
Porebska, Anna Joanna
Kuthan-Styczen, Jolanta
Gurba, Lidia
Borowicz, Piotr
Mazurkiewicz, Anna
Plucienniczak, Grazyna
Plucienniczak, Andrzej
Use of Ubp1 protease analog to produce recombinant human growth hormone in Escherichia coli
title Use of Ubp1 protease analog to produce recombinant human growth hormone in Escherichia coli
title_full Use of Ubp1 protease analog to produce recombinant human growth hormone in Escherichia coli
title_fullStr Use of Ubp1 protease analog to produce recombinant human growth hormone in Escherichia coli
title_full_unstemmed Use of Ubp1 protease analog to produce recombinant human growth hormone in Escherichia coli
title_short Use of Ubp1 protease analog to produce recombinant human growth hormone in Escherichia coli
title_sort use of ubp1 protease analog to produce recombinant human growth hormone in escherichia coli
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4154517/
https://www.ncbi.nlm.nih.gov/pubmed/25158991
http://dx.doi.org/10.1186/s12934-014-0113-4
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