Overexpressing <i>CrePAPS</i> Polyadenylate Activity Enhances Protein Translation and Accumulation in <i>Chlamydomonas reinhardtii</i>
The alga <i>Chlamydomonas reinhardtii</i> is a potential platform for recombinant protein expression in the future due to various advantages. Dozens of <i>C. reinhardtii</i> strains producing genetically engineered recombinant therapeutic protein have been reported. However,...
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| Format: | Article |
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MDPI AG
2022-04-01
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| Series: | Marine Drugs |
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| Online Access: | https://www.mdpi.com/1660-3397/20/5/276 |
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| author | Quan Wang Jieyi Zhuang Shuai Ni Haolin Luo Kaijie Zheng Xinyi Li Chengxiang Lan Di Zhao Yongsheng Bai Bin Jia Zhangli Hu |
| author_facet | Quan Wang Jieyi Zhuang Shuai Ni Haolin Luo Kaijie Zheng Xinyi Li Chengxiang Lan Di Zhao Yongsheng Bai Bin Jia Zhangli Hu |
| author_sort | Quan Wang |
| collection | DOAJ |
| description | The alga <i>Chlamydomonas reinhardtii</i> is a potential platform for recombinant protein expression in the future due to various advantages. Dozens of <i>C. reinhardtii</i> strains producing genetically engineered recombinant therapeutic protein have been reported. However, owing to extremely low protein expression efficiency, none have been applied for industrial purposes. Improving protein expression efficiency at the molecular level is, therefore, a priority. The 3′-end poly(A) tail of mRNAs is strongly correlated with mRNA transcription and protein translation efficiency. In this study, we identified a canonical <i>C. reinhardtii</i> poly(A) polymerase (CrePAPS), verified its polyadenylate activity, generated a series of overexpressing transformants, and performed proteomic analysis. Proteomic results demonstrated that overexpressing CrePAPS promoted ribosomal assembly and enhanced protein accumulation. The accelerated translation was further verified by increased crude and dissolved protein content detected by Kjeldahl and bicinchoninic acid (BCA) assay approaches. The findings provide a novel direction in which to exploit photosynthetic green algae as a recombinant protein expression platform. |
| first_indexed | 2024-03-10T03:32:22Z |
| format | Article |
| id | doaj.art-f4c7501d2e5043939b678c28c49d160b |
| institution | Directory Open Access Journal |
| issn | 1660-3397 |
| language | English |
| last_indexed | 2024-03-10T03:32:22Z |
| publishDate | 2022-04-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Marine Drugs |
| spelling | doaj.art-f4c7501d2e5043939b678c28c49d160b2023-11-23T11:53:48ZengMDPI AGMarine Drugs1660-33972022-04-0120527610.3390/md20050276Overexpressing <i>CrePAPS</i> Polyadenylate Activity Enhances Protein Translation and Accumulation in <i>Chlamydomonas reinhardtii</i>Quan Wang0Jieyi Zhuang1Shuai Ni2Haolin Luo3Kaijie Zheng4Xinyi Li5Chengxiang Lan6Di Zhao7Yongsheng Bai8Bin Jia9Zhangli Hu10Guangdong Technology Research Center for Marine Algal Bioengineering, College of Life Sciences and Oceanography, Shenzhen University, Shenzhen 518055, ChinaGuangdong Technology Research Center for Marine Algal Bioengineering, College of Life Sciences and Oceanography, Shenzhen University, Shenzhen 518055, ChinaGuangdong Technology Research Center for Marine Algal Bioengineering, College of Life Sciences and Oceanography, Shenzhen University, Shenzhen 518055, ChinaGuangdong Technology Research Center for Marine Algal Bioengineering, College of Life Sciences and Oceanography, Shenzhen University, Shenzhen 518055, ChinaGuangdong Technology Research Center for Marine Algal Bioengineering, College of Life Sciences and Oceanography, Shenzhen University, Shenzhen 518055, ChinaGuangdong Technology Research Center for Marine Algal Bioengineering, College of Life Sciences and Oceanography, Shenzhen University, Shenzhen 518055, ChinaGuangdong Technology Research Center for Marine Algal Bioengineering, College of Life Sciences and Oceanography, Shenzhen University, Shenzhen 518055, ChinaGuangdong Technology Research Center for Marine Algal Bioengineering, College of Life Sciences and Oceanography, Shenzhen University, Shenzhen 518055, ChinaGuangdong Technology Research Center for Marine Algal Bioengineering, College of Life Sciences and Oceanography, Shenzhen University, Shenzhen 518055, ChinaGuangdong Technology Research Center for Marine Algal Bioengineering, College of Life Sciences and Oceanography, Shenzhen University, Shenzhen 518055, ChinaGuangdong Technology Research Center for Marine Algal Bioengineering, College of Life Sciences and Oceanography, Shenzhen University, Shenzhen 518055, ChinaThe alga <i>Chlamydomonas reinhardtii</i> is a potential platform for recombinant protein expression in the future due to various advantages. Dozens of <i>C. reinhardtii</i> strains producing genetically engineered recombinant therapeutic protein have been reported. However, owing to extremely low protein expression efficiency, none have been applied for industrial purposes. Improving protein expression efficiency at the molecular level is, therefore, a priority. The 3′-end poly(A) tail of mRNAs is strongly correlated with mRNA transcription and protein translation efficiency. In this study, we identified a canonical <i>C. reinhardtii</i> poly(A) polymerase (CrePAPS), verified its polyadenylate activity, generated a series of overexpressing transformants, and performed proteomic analysis. Proteomic results demonstrated that overexpressing CrePAPS promoted ribosomal assembly and enhanced protein accumulation. The accelerated translation was further verified by increased crude and dissolved protein content detected by Kjeldahl and bicinchoninic acid (BCA) assay approaches. The findings provide a novel direction in which to exploit photosynthetic green algae as a recombinant protein expression platform.https://www.mdpi.com/1660-3397/20/5/276<i>Chlamydomonas reinhardtii</i>protein productionexpression platformCrePAPSpolyadenylate activityprotein accumulation |
| spellingShingle | Quan Wang Jieyi Zhuang Shuai Ni Haolin Luo Kaijie Zheng Xinyi Li Chengxiang Lan Di Zhao Yongsheng Bai Bin Jia Zhangli Hu Overexpressing <i>CrePAPS</i> Polyadenylate Activity Enhances Protein Translation and Accumulation in <i>Chlamydomonas reinhardtii</i> Marine Drugs <i>Chlamydomonas reinhardtii</i> protein production expression platform CrePAPS polyadenylate activity protein accumulation |
| title | Overexpressing <i>CrePAPS</i> Polyadenylate Activity Enhances Protein Translation and Accumulation in <i>Chlamydomonas reinhardtii</i> |
| title_full | Overexpressing <i>CrePAPS</i> Polyadenylate Activity Enhances Protein Translation and Accumulation in <i>Chlamydomonas reinhardtii</i> |
| title_fullStr | Overexpressing <i>CrePAPS</i> Polyadenylate Activity Enhances Protein Translation and Accumulation in <i>Chlamydomonas reinhardtii</i> |
| title_full_unstemmed | Overexpressing <i>CrePAPS</i> Polyadenylate Activity Enhances Protein Translation and Accumulation in <i>Chlamydomonas reinhardtii</i> |
| title_short | Overexpressing <i>CrePAPS</i> Polyadenylate Activity Enhances Protein Translation and Accumulation in <i>Chlamydomonas reinhardtii</i> |
| title_sort | overexpressing i crepaps i polyadenylate activity enhances protein translation and accumulation in i chlamydomonas reinhardtii i |
| topic | <i>Chlamydomonas reinhardtii</i> protein production expression platform CrePAPS polyadenylate activity protein accumulation |
| url | https://www.mdpi.com/1660-3397/20/5/276 |
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