Vibrio natriegens as a pET-Compatible Expression Host Complementary to Escherichia coli
Efficient and novel recombinant protein expression systems can further reduce the production cost of enzymes. Vibrio natriegens is the fastest growing free-living bacterium with a doubling time of less than 10 min, which makes it highly attractive as a protein expression host. Here, 196 pET plasmids...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2021-02-01
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| Series: | Frontiers in Microbiology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fmicb.2021.627181/full |
| _version_ | 1818655893993029632 |
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| author | Jiaqi Xu Feng Dong Meixian Wu Rongsheng Tao Rongsheng Tao Junjie Yang Mianbin Wu Yu Jiang Yu Jiang Sheng Yang Sheng Yang Lirong Yang |
| author_facet | Jiaqi Xu Feng Dong Meixian Wu Rongsheng Tao Rongsheng Tao Junjie Yang Mianbin Wu Yu Jiang Yu Jiang Sheng Yang Sheng Yang Lirong Yang |
| author_sort | Jiaqi Xu |
| collection | DOAJ |
| description | Efficient and novel recombinant protein expression systems can further reduce the production cost of enzymes. Vibrio natriegens is the fastest growing free-living bacterium with a doubling time of less than 10 min, which makes it highly attractive as a protein expression host. Here, 196 pET plasmids with different genes of interest (GOIs) were electroporated into the V. natriegens strain VnDX, which carries an integrated T7 RNA polymerase expression cassette. As a result, 65 and 75% of the tested GOIs obtained soluble expression in V. natriegens and Escherichia coli, respectively, 20 GOIs of which showed better expression in the former. Furthermore, we have adapted a consensus “what to try first” protocol for V. natriegens based on Terrific Broth medium. Six sampled GOIs encoding biocatalysts enzymes thus achieved 50–128% higher catalytic efficiency under the optimized expression conditions. Our study demonstrated V. natriegens as a pET-compatible expression host with a spectrum of highly expressed GOIs distinct from E. coli and an easy-to-use consensus protocol, solving the problem that some GOIs cannot be expressed well in E. coli. |
| first_indexed | 2024-12-17T03:16:56Z |
| format | Article |
| id | doaj.art-f8d530dd3cb0478c9659b186d0ed645b |
| institution | Directory Open Access Journal |
| issn | 1664-302X |
| language | English |
| last_indexed | 2024-12-17T03:16:56Z |
| publishDate | 2021-02-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Microbiology |
| spelling | doaj.art-f8d530dd3cb0478c9659b186d0ed645b2022-12-21T22:05:38ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2021-02-011210.3389/fmicb.2021.627181627181Vibrio natriegens as a pET-Compatible Expression Host Complementary to Escherichia coliJiaqi Xu0Feng Dong1Meixian Wu2Rongsheng Tao3Rongsheng Tao4Junjie Yang5Mianbin Wu6Yu Jiang7Yu Jiang8Sheng Yang9Sheng Yang10Lirong Yang11Institute of Bioengineering, College of Chemical and Biological Engineering, Zhejiang University, Hangzhou, ChinaHuzhou Center of Industrial Biotechnology, Shanghai Institutes for Biological Sciences, Huzhou, ChinaKey Laboratory of Synthetic Biology, CAS Center for Excellence in Molecular Plant Sciences, Chinese Academy of Sciences, Shanghai, ChinaHuzhou Center of Industrial Biotechnology, Shanghai Institutes for Biological Sciences, Huzhou, ChinaHuzhou Yisheng Biotechnology Co., Ltd., Huzhou, ChinaKey Laboratory of Synthetic Biology, CAS Center for Excellence in Molecular Plant Sciences, Chinese Academy of Sciences, Shanghai, ChinaInstitute of Bioengineering, College of Chemical and Biological Engineering, Zhejiang University, Hangzhou, ChinaHuzhou Center of Industrial Biotechnology, Shanghai Institutes for Biological Sciences, Huzhou, ChinaShanghai Taoyusheng Biotechnology Co., Ltd., Shanghai, ChinaHuzhou Center of Industrial Biotechnology, Shanghai Institutes for Biological Sciences, Huzhou, ChinaKey Laboratory of Synthetic Biology, CAS Center for Excellence in Molecular Plant Sciences, Chinese Academy of Sciences, Shanghai, ChinaInstitute of Bioengineering, College of Chemical and Biological Engineering, Zhejiang University, Hangzhou, ChinaEfficient and novel recombinant protein expression systems can further reduce the production cost of enzymes. Vibrio natriegens is the fastest growing free-living bacterium with a doubling time of less than 10 min, which makes it highly attractive as a protein expression host. Here, 196 pET plasmids with different genes of interest (GOIs) were electroporated into the V. natriegens strain VnDX, which carries an integrated T7 RNA polymerase expression cassette. As a result, 65 and 75% of the tested GOIs obtained soluble expression in V. natriegens and Escherichia coli, respectively, 20 GOIs of which showed better expression in the former. Furthermore, we have adapted a consensus “what to try first” protocol for V. natriegens based on Terrific Broth medium. Six sampled GOIs encoding biocatalysts enzymes thus achieved 50–128% higher catalytic efficiency under the optimized expression conditions. Our study demonstrated V. natriegens as a pET-compatible expression host with a spectrum of highly expressed GOIs distinct from E. coli and an easy-to-use consensus protocol, solving the problem that some GOIs cannot be expressed well in E. coli.https://www.frontiersin.org/articles/10.3389/fmicb.2021.627181/fullVibrio natriegensrecombinant proteinpET expression systemfermentation optimizationsynthetic biology |
| spellingShingle | Jiaqi Xu Feng Dong Meixian Wu Rongsheng Tao Rongsheng Tao Junjie Yang Mianbin Wu Yu Jiang Yu Jiang Sheng Yang Sheng Yang Lirong Yang Vibrio natriegens as a pET-Compatible Expression Host Complementary to Escherichia coli Frontiers in Microbiology Vibrio natriegens recombinant protein pET expression system fermentation optimization synthetic biology |
| title | Vibrio natriegens as a pET-Compatible Expression Host Complementary to Escherichia coli |
| title_full | Vibrio natriegens as a pET-Compatible Expression Host Complementary to Escherichia coli |
| title_fullStr | Vibrio natriegens as a pET-Compatible Expression Host Complementary to Escherichia coli |
| title_full_unstemmed | Vibrio natriegens as a pET-Compatible Expression Host Complementary to Escherichia coli |
| title_short | Vibrio natriegens as a pET-Compatible Expression Host Complementary to Escherichia coli |
| title_sort | vibrio natriegens as a pet compatible expression host complementary to escherichia coli |
| topic | Vibrio natriegens recombinant protein pET expression system fermentation optimization synthetic biology |
| url | https://www.frontiersin.org/articles/10.3389/fmicb.2021.627181/full |
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