GroEL/ES mediated the in vivo recovery of TRAIL inclusion bodies in Escherichia coli
Abstract Inclusion body (IB) formation generates substantial bio-waste in the pharmaceutical industry and remains a major challenge for heterologous protein expression. Although chaperones can be co-expressed to improve soluble protein yield, their contribution to IB processing in vivo has not been...
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Nature Portfolio
2018-10-01
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| Series: | Scientific Reports |
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| Online Access: | https://doi.org/10.1038/s41598-018-34090-7 |
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| author | Zhanqing Wang Min Zhang Xin Lv Jiying Fan Jian Zhang Jing Sun Yaling Shen |
| author_facet | Zhanqing Wang Min Zhang Xin Lv Jiying Fan Jian Zhang Jing Sun Yaling Shen |
| author_sort | Zhanqing Wang |
| collection | DOAJ |
| description | Abstract Inclusion body (IB) formation generates substantial bio-waste in the pharmaceutical industry and remains a major challenge for heterologous protein expression. Although chaperones can be co-expressed to improve soluble protein yield, their contribution to IB processing in vivo has not been thoroughly studied. Here, a GroEL-GroES co-expressing strain and a deficient strain were constructed to study the in vivo recovery of recombinant human tumor necrosis factor-related apoptosis-inducing ligand (TRAIL). The interaction between GroEL/ES and TRAIL was simulated by molecular docking and identified by co-immunoprecipitation. The in vitro cytotoxicity of TRAIL IBs before and after in vivo recovery was subsequently determined by MTT assay. Additionally, IB structures were measured by Fourier transform infrared (FT-IR) spectroscopy and fluorescence spectroscopy. The results showed that after in vivo refolding, IBs retained lower levels of anti-tumor activity and fewer native-like β-sheet structures. Fewer recoverable polypeptides were trapped in IBs after GroEL/ES co-expression and refolding in vivo. Therefore, GroEL/ES mediated the in vivo recovery of TRAIL IBs in Escherichia coli. These results may identify potential uses for IBs and provide additional insight into the detailed mechanisms of in vivo protein recovery. |
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| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-12-13T17:20:24Z |
| publishDate | 2018-10-01 |
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| spelling | doaj.art-9c001b387a9d4afc83e57e681db6a0bc2022-12-21T23:37:19ZengNature PortfolioScientific Reports2045-23222018-10-018111010.1038/s41598-018-34090-7GroEL/ES mediated the in vivo recovery of TRAIL inclusion bodies in Escherichia coliZhanqing Wang0Min Zhang1Xin Lv2Jiying Fan3Jian Zhang4Jing Sun5Yaling Shen6State Key Laboratory of Bioreactor Engineering, Shanghai Collaborative Innovation Center for Biomanufacturing Technology, East China University of Science and TechnologyState Key Laboratory of Bioreactor Engineering, Shanghai Collaborative Innovation Center for Biomanufacturing Technology, East China University of Science and TechnologyState Key Laboratory of Bioreactor Engineering, Shanghai Collaborative Innovation Center for Biomanufacturing Technology, East China University of Science and TechnologyState Key Laboratory of Bioreactor Engineering, Shanghai Collaborative Innovation Center for Biomanufacturing Technology, East China University of Science and TechnologyState Key Laboratory of Bioreactor Engineering, Shanghai Collaborative Innovation Center for Biomanufacturing Technology, East China University of Science and TechnologyState Key Laboratory of Bioreactor Engineering, Shanghai Collaborative Innovation Center for Biomanufacturing Technology, East China University of Science and TechnologyState Key Laboratory of Bioreactor Engineering, Shanghai Collaborative Innovation Center for Biomanufacturing Technology, East China University of Science and TechnologyAbstract Inclusion body (IB) formation generates substantial bio-waste in the pharmaceutical industry and remains a major challenge for heterologous protein expression. Although chaperones can be co-expressed to improve soluble protein yield, their contribution to IB processing in vivo has not been thoroughly studied. Here, a GroEL-GroES co-expressing strain and a deficient strain were constructed to study the in vivo recovery of recombinant human tumor necrosis factor-related apoptosis-inducing ligand (TRAIL). The interaction between GroEL/ES and TRAIL was simulated by molecular docking and identified by co-immunoprecipitation. The in vitro cytotoxicity of TRAIL IBs before and after in vivo recovery was subsequently determined by MTT assay. Additionally, IB structures were measured by Fourier transform infrared (FT-IR) spectroscopy and fluorescence spectroscopy. The results showed that after in vivo refolding, IBs retained lower levels of anti-tumor activity and fewer native-like β-sheet structures. Fewer recoverable polypeptides were trapped in IBs after GroEL/ES co-expression and refolding in vivo. Therefore, GroEL/ES mediated the in vivo recovery of TRAIL IBs in Escherichia coli. These results may identify potential uses for IBs and provide additional insight into the detailed mechanisms of in vivo protein recovery.https://doi.org/10.1038/s41598-018-34090-7Tumor Necrosis Factor-related Apoptosis-inducing Ligand (TRAIL)Soluble TRAILrhTRAILGroE ChaperoninColi Strain C600 |
| spellingShingle | Zhanqing Wang Min Zhang Xin Lv Jiying Fan Jian Zhang Jing Sun Yaling Shen GroEL/ES mediated the in vivo recovery of TRAIL inclusion bodies in Escherichia coli Scientific Reports Tumor Necrosis Factor-related Apoptosis-inducing Ligand (TRAIL) Soluble TRAIL rhTRAIL GroE Chaperonin Coli Strain C600 |
| title | GroEL/ES mediated the in vivo recovery of TRAIL inclusion bodies in Escherichia coli |
| title_full | GroEL/ES mediated the in vivo recovery of TRAIL inclusion bodies in Escherichia coli |
| title_fullStr | GroEL/ES mediated the in vivo recovery of TRAIL inclusion bodies in Escherichia coli |
| title_full_unstemmed | GroEL/ES mediated the in vivo recovery of TRAIL inclusion bodies in Escherichia coli |
| title_short | GroEL/ES mediated the in vivo recovery of TRAIL inclusion bodies in Escherichia coli |
| title_sort | groel es mediated the in vivo recovery of trail inclusion bodies in escherichia coli |
| topic | Tumor Necrosis Factor-related Apoptosis-inducing Ligand (TRAIL) Soluble TRAIL rhTRAIL GroE Chaperonin Coli Strain C600 |
| url | https://doi.org/10.1038/s41598-018-34090-7 |
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