High‐level recombinant protein production with Corynebacterium glutamicum using acetate as carbon source
Abstract In recent years, biotechnological conversion of the alternative carbon source acetate has attracted much attention. So far, acetate has been mainly used for microbial production of bioproducts with bulk applications. In this study, we aimed to investigate the potential of acetate as carbon...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Wiley
2022-11-01
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| Series: | Microbial Biotechnology |
| Online Access: | https://doi.org/10.1111/1751-7915.14138 |
| _version_ | 1797991827132907520 |
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| author | Dirk Kiefer Lea Rahel Tadele Lars Lilge Marius Henkel Rudolf Hausmann |
| author_facet | Dirk Kiefer Lea Rahel Tadele Lars Lilge Marius Henkel Rudolf Hausmann |
| author_sort | Dirk Kiefer |
| collection | DOAJ |
| description | Abstract In recent years, biotechnological conversion of the alternative carbon source acetate has attracted much attention. So far, acetate has been mainly used for microbial production of bioproducts with bulk applications. In this study, we aimed to investigate the potential of acetate as carbon source for heterologous protein production using the acetate‐utilizing platform organism Corynebacterium glutamicum. For this purpose, expression of model protein eYFP with the promoter systems T7lac and tac was characterized during growth of C. glutamicum on acetate as sole carbon source. The results indicated a 3.3‐fold higher fluorescence level for acetate‐based eYFP production with T7 expression strain MB001(DE3) pMKEx2‐eyfp compared to MB001 pEKEx2‐eyfp. Interestingly, comparative eyfp expression studies on acetate or glucose revealed an up to 83% higher biomass‐specific production for T7 RNAP‐dependent eYFP production using acetate as carbon source. Furthermore, high‐level protein accumulation on acetate was demonstrated for the first time in a high cell density cultivation process with pH‐coupled online feeding control, resulting in a final protein titer of 2.7 g/L and product yield of 4 g per 100 g cell dry weight. This study presents a first proof of concept for efficient microbial upgrading of potentially low‐cost acetate into high‐value bioproducts, such as recombinant proteins. |
| first_indexed | 2024-04-11T08:57:09Z |
| format | Article |
| id | doaj.art-59168b886912457c9926c75c4afe7b91 |
| institution | Directory Open Access Journal |
| issn | 1751-7915 |
| language | English |
| last_indexed | 2024-04-11T08:57:09Z |
| publishDate | 2022-11-01 |
| publisher | Wiley |
| record_format | Article |
| series | Microbial Biotechnology |
| spelling | doaj.art-59168b886912457c9926c75c4afe7b912022-12-22T04:33:09ZengWileyMicrobial Biotechnology1751-79152022-11-0115112744275710.1111/1751-7915.14138High‐level recombinant protein production with Corynebacterium glutamicum using acetate as carbon sourceDirk Kiefer0Lea Rahel Tadele1Lars Lilge2Marius Henkel3Rudolf Hausmann4Department of Bioprocess Engineering Institute of Food Science and Biotechnology, University of Hohenheim Stuttgart GermanyDepartment of Bioprocess Engineering Institute of Food Science and Biotechnology, University of Hohenheim Stuttgart GermanyDepartment of Bioprocess Engineering Institute of Food Science and Biotechnology, University of Hohenheim Stuttgart GermanyCellular Agriculture TUM School of Life Sciences, Technical University of Munich Freising GermanyDepartment of Bioprocess Engineering Institute of Food Science and Biotechnology, University of Hohenheim Stuttgart GermanyAbstract In recent years, biotechnological conversion of the alternative carbon source acetate has attracted much attention. So far, acetate has been mainly used for microbial production of bioproducts with bulk applications. In this study, we aimed to investigate the potential of acetate as carbon source for heterologous protein production using the acetate‐utilizing platform organism Corynebacterium glutamicum. For this purpose, expression of model protein eYFP with the promoter systems T7lac and tac was characterized during growth of C. glutamicum on acetate as sole carbon source. The results indicated a 3.3‐fold higher fluorescence level for acetate‐based eYFP production with T7 expression strain MB001(DE3) pMKEx2‐eyfp compared to MB001 pEKEx2‐eyfp. Interestingly, comparative eyfp expression studies on acetate or glucose revealed an up to 83% higher biomass‐specific production for T7 RNAP‐dependent eYFP production using acetate as carbon source. Furthermore, high‐level protein accumulation on acetate was demonstrated for the first time in a high cell density cultivation process with pH‐coupled online feeding control, resulting in a final protein titer of 2.7 g/L and product yield of 4 g per 100 g cell dry weight. This study presents a first proof of concept for efficient microbial upgrading of potentially low‐cost acetate into high‐value bioproducts, such as recombinant proteins.https://doi.org/10.1111/1751-7915.14138 |
| spellingShingle | Dirk Kiefer Lea Rahel Tadele Lars Lilge Marius Henkel Rudolf Hausmann High‐level recombinant protein production with Corynebacterium glutamicum using acetate as carbon source Microbial Biotechnology |
| title | High‐level recombinant protein production with Corynebacterium glutamicum using acetate as carbon source |
| title_full | High‐level recombinant protein production with Corynebacterium glutamicum using acetate as carbon source |
| title_fullStr | High‐level recombinant protein production with Corynebacterium glutamicum using acetate as carbon source |
| title_full_unstemmed | High‐level recombinant protein production with Corynebacterium glutamicum using acetate as carbon source |
| title_short | High‐level recombinant protein production with Corynebacterium glutamicum using acetate as carbon source |
| title_sort | high level recombinant protein production with corynebacterium glutamicum using acetate as carbon source |
| url | https://doi.org/10.1111/1751-7915.14138 |
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