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...
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2011-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC3168869?pdf=render |
| _version_ | 1828377620674248704 |
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| author | Li Tian Samuel S M Sun |
| author_facet | Li Tian Samuel S M Sun |
| author_sort | Li Tian |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-04-14T08:14:34Z |
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| id | doaj.art-bd4e7fbe8b4b4b3a8f432aa976dae243 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-04-14T08:14:34Z |
| publishDate | 2011-01-01 |
| publisher | Public Library of Science (PLoS) |
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| series | PLoS ONE |
| spelling | doaj.art-bd4e7fbe8b4b4b3a8f432aa976dae2432022-12-22T02:04:27ZengPublic Library of Science (PLoS)PLoS ONE1932-62032011-01-0168e2418310.1371/journal.pone.0024183A cost-effective ELP-intein coupling system for recombinant protein purification from plant production platform.Li TianSamuel S M SunBACKGROUND: 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.http://europepmc.org/articles/PMC3168869?pdf=render |
| spellingShingle | Li Tian Samuel S M Sun A cost-effective ELP-intein coupling system for recombinant protein purification from plant production platform. PLoS ONE |
| 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 |
| url | http://europepmc.org/articles/PMC3168869?pdf=render |
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