Exploring the secretome of Corynebacterium glutamicum ATCC 13032
The demand for alternative sources of food proteins is increasing due to the limitations and challenges associated with conventional food production. Advances in biotechnology have enabled the production of proteins using microorganisms, thus prompting the exploration of attractive microbial hosts c...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2024-02-01
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| Series: | Frontiers in Bioengineering and Biotechnology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fbioe.2024.1348184/full |
| _version_ | 1827352778776772608 |
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| author | Suvasini Balasubramanian Suvasini Balasubramanian Julie Bonne Køhler Carsten Jers Peter Ruhdal Jensen Ivan Mijakovic Ivan Mijakovic |
| author_facet | Suvasini Balasubramanian Suvasini Balasubramanian Julie Bonne Køhler Carsten Jers Peter Ruhdal Jensen Ivan Mijakovic Ivan Mijakovic |
| author_sort | Suvasini Balasubramanian |
| collection | DOAJ |
| description | The demand for alternative sources of food proteins is increasing due to the limitations and challenges associated with conventional food production. Advances in biotechnology have enabled the production of proteins using microorganisms, thus prompting the exploration of attractive microbial hosts capable of producing functional proteins in high titers. Corynebacterium glutamicum is widely used in industry for the production of amino acids and has many advantages as a host organism for recombinant protein production. However, its performance in this area is limited by low yields of target proteins and high levels of native protein secretion. Despite representing a challenge for heterologous protein production, the C. glutamicum secretome has not been fully characterized. In this study, state-of-the-art mass spectrometry-based proteomics was used to identify and analyze the proteins secreted by C. glutamicum. Both the wild-type strain and a strain that produced and secreted a recombinant β-lactoglobulin protein were analyzed. A total of 427 proteins were identified in the culture supernatants, with 148 predicted to possess a secretion signal peptide. MS-based proteomics on the secretome enabled a comprehensive characterization and quantification (based on abundance) of the secreted proteins through label-free quantification (LFQ). The top 12 most abundant proteins accounted for almost 80% of the secretome. These are uncharacterized proteins of unknown function, resuscitation promoting factors, protein PS1, Porin B, ABC-type transporter protein and hypothetical membrane protein. The data can be leveraged for protein production by, e.g., utilizing the signal peptides of the most abundant proteins to improve secretion of heterologous proteins. In addition, secretory stress can potentially be alleviated by inactivating non-essential secreted proteins. Here we provide targets by identifying the most abundant, secreted proteins of which majority are of unknown function. The data from this study can thus provide valuable insight for researchers looking to improve protein secretion and optimize C. glutamicum as a host for secretory protein production. |
| first_indexed | 2024-03-08T03:11:58Z |
| format | Article |
| id | doaj.art-a2f21daf022a4a03bdd6393918c09377 |
| institution | Directory Open Access Journal |
| issn | 2296-4185 |
| language | English |
| last_indexed | 2024-03-08T03:11:58Z |
| publishDate | 2024-02-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Bioengineering and Biotechnology |
| spelling | doaj.art-a2f21daf022a4a03bdd6393918c093772024-02-13T04:33:07ZengFrontiers Media S.A.Frontiers in Bioengineering and Biotechnology2296-41852024-02-011210.3389/fbioe.2024.13481841348184Exploring the secretome of Corynebacterium glutamicum ATCC 13032Suvasini Balasubramanian0Suvasini Balasubramanian1Julie Bonne Køhler2Carsten Jers3Peter Ruhdal Jensen4Ivan Mijakovic5Ivan Mijakovic6Microbial Biotechnology and Biorefining, National Food Institute, Technical University of Denmark, Kongens Lyngby, DenmarkNovo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kongens Lyngby, DenmarkNovo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kongens Lyngby, DenmarkNovo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kongens Lyngby, DenmarkMicrobial Biotechnology and Biorefining, National Food Institute, Technical University of Denmark, Kongens Lyngby, DenmarkNovo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kongens Lyngby, DenmarkSystems and Synthetic Biology Division, Department of Biology and Biological Engineering, Chalmers University of Technology, Gothenburg, SwedenThe demand for alternative sources of food proteins is increasing due to the limitations and challenges associated with conventional food production. Advances in biotechnology have enabled the production of proteins using microorganisms, thus prompting the exploration of attractive microbial hosts capable of producing functional proteins in high titers. Corynebacterium glutamicum is widely used in industry for the production of amino acids and has many advantages as a host organism for recombinant protein production. However, its performance in this area is limited by low yields of target proteins and high levels of native protein secretion. Despite representing a challenge for heterologous protein production, the C. glutamicum secretome has not been fully characterized. In this study, state-of-the-art mass spectrometry-based proteomics was used to identify and analyze the proteins secreted by C. glutamicum. Both the wild-type strain and a strain that produced and secreted a recombinant β-lactoglobulin protein were analyzed. A total of 427 proteins were identified in the culture supernatants, with 148 predicted to possess a secretion signal peptide. MS-based proteomics on the secretome enabled a comprehensive characterization and quantification (based on abundance) of the secreted proteins through label-free quantification (LFQ). The top 12 most abundant proteins accounted for almost 80% of the secretome. These are uncharacterized proteins of unknown function, resuscitation promoting factors, protein PS1, Porin B, ABC-type transporter protein and hypothetical membrane protein. The data can be leveraged for protein production by, e.g., utilizing the signal peptides of the most abundant proteins to improve secretion of heterologous proteins. In addition, secretory stress can potentially be alleviated by inactivating non-essential secreted proteins. Here we provide targets by identifying the most abundant, secreted proteins of which majority are of unknown function. The data from this study can thus provide valuable insight for researchers looking to improve protein secretion and optimize C. glutamicum as a host for secretory protein production.https://www.frontiersin.org/articles/10.3389/fbioe.2024.1348184/fullCorynebacterium glutamicumsecretome analysisrecombinant protein productionα-lactalbuminβ-lactoglobulin |
| spellingShingle | Suvasini Balasubramanian Suvasini Balasubramanian Julie Bonne Køhler Carsten Jers Peter Ruhdal Jensen Ivan Mijakovic Ivan Mijakovic Exploring the secretome of Corynebacterium glutamicum ATCC 13032 Frontiers in Bioengineering and Biotechnology Corynebacterium glutamicum secretome analysis recombinant protein production α-lactalbumin β-lactoglobulin |
| title | Exploring the secretome of Corynebacterium glutamicum ATCC 13032 |
| title_full | Exploring the secretome of Corynebacterium glutamicum ATCC 13032 |
| title_fullStr | Exploring the secretome of Corynebacterium glutamicum ATCC 13032 |
| title_full_unstemmed | Exploring the secretome of Corynebacterium glutamicum ATCC 13032 |
| title_short | Exploring the secretome of Corynebacterium glutamicum ATCC 13032 |
| title_sort | exploring the secretome of corynebacterium glutamicum atcc 13032 |
| topic | Corynebacterium glutamicum secretome analysis recombinant protein production α-lactalbumin β-lactoglobulin |
| url | https://www.frontiersin.org/articles/10.3389/fbioe.2024.1348184/full |
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