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A Cost-Effective ELP-Intein Coupling System for Recombinant Protein Purification from Plant Production Platform
BACKGROUND: Plant bioreactor offers an efficient and economical system for large-scale production of recombinant proteins. However, high cost and difficulty in scaling-up of downstream purification of the target protein, particularly the common involvement of affinity chromatography and protease in...
Autores principales: | , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2011
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3168869/ https://www.ncbi.nlm.nih.gov/pubmed/21918684 http://dx.doi.org/10.1371/journal.pone.0024183 |
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author | Tian, Li Sun, Samuel S. M. |
author_facet | Tian, Li Sun, Samuel S. M. |
author_sort | Tian, Li |
collection | PubMed |
description | BACKGROUND: Plant bioreactor offers an efficient and economical system for large-scale production of recombinant proteins. However, high cost and difficulty in scaling-up of downstream purification of the target protein, particularly the common involvement of affinity chromatography and protease in the purification process, has hampered its industrial scale application, therefore a cost-effective and easily scale-up purification method is highly desirable for further development of plant bioreactor. METHODOLOGY/PRINCIPAL FINDINGS: To tackle this problem, we investigated the ELP-intein coupling system for purification of recombinant proteins expressed in transgenic plants using a plant lectin (PAL) with anti-tumor bioactivity as example target protein and rice seeds as production platform. Results showed that ELP-intein-PAL (EiP) fusion protein formed novel irregular ER-derived protein bodies in endosperm cells by retention of endogenous prolamins. The fusion protein was partially self-cleaved in vivo, but only self-cleaved PAL protein was detected in total seed protein sample and deposited in protein storage vacuoles (PSV). The in vivo uncleaved EiP protein was accumulated up to 2–4.2% of the total seed protein. The target PAL protein could be purified by the ELP-intein system efficiently without using complicated instruments and expensive chemicals, and the yield of pure PAL protein by the current method was up to 1.1 mg/g total seed protein. CONCLUSION/SIGNIFICANCE: This study successfully demonstrated the purification of an example recombinant protein from rice seeds by the ELP-intein system. The whole purification procedure can be easily scaled up for industrial production, providing the first evidence on applying the ELP-intein coupling system to achieve cost-effective purification of recombinant proteins expressed in plant bioreactors and its possible application in industry. |
format | Online Article Text |
id | pubmed-3168869 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2011 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-31688692011-09-14 A Cost-Effective ELP-Intein Coupling System for Recombinant Protein Purification from Plant Production Platform Tian, Li Sun, Samuel S. M. PLoS One Research Article BACKGROUND: Plant bioreactor offers an efficient and economical system for large-scale production of recombinant proteins. However, high cost and difficulty in scaling-up of downstream purification of the target protein, particularly the common involvement of affinity chromatography and protease in the purification process, has hampered its industrial scale application, therefore a cost-effective and easily scale-up purification method is highly desirable for further development of plant bioreactor. METHODOLOGY/PRINCIPAL FINDINGS: To tackle this problem, we investigated the ELP-intein coupling system for purification of recombinant proteins expressed in transgenic plants using a plant lectin (PAL) with anti-tumor bioactivity as example target protein and rice seeds as production platform. Results showed that ELP-intein-PAL (EiP) fusion protein formed novel irregular ER-derived protein bodies in endosperm cells by retention of endogenous prolamins. The fusion protein was partially self-cleaved in vivo, but only self-cleaved PAL protein was detected in total seed protein sample and deposited in protein storage vacuoles (PSV). The in vivo uncleaved EiP protein was accumulated up to 2–4.2% of the total seed protein. The target PAL protein could be purified by the ELP-intein system efficiently without using complicated instruments and expensive chemicals, and the yield of pure PAL protein by the current method was up to 1.1 mg/g total seed protein. CONCLUSION/SIGNIFICANCE: This study successfully demonstrated the purification of an example recombinant protein from rice seeds by the ELP-intein system. The whole purification procedure can be easily scaled up for industrial production, providing the first evidence on applying the ELP-intein coupling system to achieve cost-effective purification of recombinant proteins expressed in plant bioreactors and its possible application in industry. Public Library of Science 2011-08-30 /pmc/articles/PMC3168869/ /pubmed/21918684 http://dx.doi.org/10.1371/journal.pone.0024183 Text en Tian, Sun. http://creativecommons.org/licenses/by/4.0/ 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 author and source are properly credited. |
spellingShingle | Research Article Tian, Li Sun, Samuel S. M. A Cost-Effective ELP-Intein Coupling System for Recombinant Protein Purification from Plant Production Platform |
title | A Cost-Effective ELP-Intein Coupling System for Recombinant Protein Purification from Plant Production Platform |
title_full | A Cost-Effective ELP-Intein Coupling System for Recombinant Protein Purification from Plant Production Platform |
title_fullStr | A Cost-Effective ELP-Intein Coupling System for Recombinant Protein Purification from Plant Production Platform |
title_full_unstemmed | A Cost-Effective ELP-Intein Coupling System for Recombinant Protein Purification from Plant Production Platform |
title_short | A Cost-Effective ELP-Intein Coupling System for Recombinant Protein Purification from Plant Production Platform |
title_sort | cost-effective elp-intein coupling system for recombinant protein purification from plant production platform |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3168869/ https://www.ncbi.nlm.nih.gov/pubmed/21918684 http://dx.doi.org/10.1371/journal.pone.0024183 |
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