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Production of Recombinant Horseradish Peroxidase in an Engineered Cell-free Protein Synthesis System
One of the main advantages of a cell-free synthesis system is that the synthetic machinery of cells can be modularized and re-assembled for desired purposes. In this study, we attempted to combine the translational activity of Escherichia coli extract with a heme synthesis pathway for the functional...
Autores principales: | , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Frontiers Media S.A.
2021
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8579056/ https://www.ncbi.nlm.nih.gov/pubmed/34778239 http://dx.doi.org/10.3389/fbioe.2021.778496 |
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author | Park, Yu-Jin Kim, Dong-Myung |
author_facet | Park, Yu-Jin Kim, Dong-Myung |
author_sort | Park, Yu-Jin |
collection | PubMed |
description | One of the main advantages of a cell-free synthesis system is that the synthetic machinery of cells can be modularized and re-assembled for desired purposes. In this study, we attempted to combine the translational activity of Escherichia coli extract with a heme synthesis pathway for the functional production of horseradish peroxidase (HRP). We first optimized the reaction conditions and the sequence of template DNA to enhance protein expression and folding. The reaction mixture was then supplemented with 5-aminolevulinic acid synthase to facilitate co-synthesis of the heme prosthetic group from glucose. Combining the different synthetic modules required for protein synthesis and cofactor generation led to successful production of functional HRP in a cell-free synthesis system. |
format | Online Article Text |
id | pubmed-8579056 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-85790562021-11-11 Production of Recombinant Horseradish Peroxidase in an Engineered Cell-free Protein Synthesis System Park, Yu-Jin Kim, Dong-Myung Front Bioeng Biotechnol Bioengineering and Biotechnology One of the main advantages of a cell-free synthesis system is that the synthetic machinery of cells can be modularized and re-assembled for desired purposes. In this study, we attempted to combine the translational activity of Escherichia coli extract with a heme synthesis pathway for the functional production of horseradish peroxidase (HRP). We first optimized the reaction conditions and the sequence of template DNA to enhance protein expression and folding. The reaction mixture was then supplemented with 5-aminolevulinic acid synthase to facilitate co-synthesis of the heme prosthetic group from glucose. Combining the different synthetic modules required for protein synthesis and cofactor generation led to successful production of functional HRP in a cell-free synthesis system. Frontiers Media S.A. 2021-10-27 /pmc/articles/PMC8579056/ /pubmed/34778239 http://dx.doi.org/10.3389/fbioe.2021.778496 Text en Copyright © 2021 Park and Kim. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Bioengineering and Biotechnology Park, Yu-Jin Kim, Dong-Myung Production of Recombinant Horseradish Peroxidase in an Engineered Cell-free Protein Synthesis System |
title | Production of Recombinant Horseradish Peroxidase in an Engineered Cell-free Protein Synthesis System |
title_full | Production of Recombinant Horseradish Peroxidase in an Engineered Cell-free Protein Synthesis System |
title_fullStr | Production of Recombinant Horseradish Peroxidase in an Engineered Cell-free Protein Synthesis System |
title_full_unstemmed | Production of Recombinant Horseradish Peroxidase in an Engineered Cell-free Protein Synthesis System |
title_short | Production of Recombinant Horseradish Peroxidase in an Engineered Cell-free Protein Synthesis System |
title_sort | production of recombinant horseradish peroxidase in an engineered cell-free protein synthesis system |
topic | Bioengineering and Biotechnology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8579056/ https://www.ncbi.nlm.nih.gov/pubmed/34778239 http://dx.doi.org/10.3389/fbioe.2021.778496 |
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