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Incorporation of alternative amino acids into cyanophycin by different cyanophycin synthetases heterologously expressed in Corynebacterium glutamicum
Cyanophycin (multi-l-arginyl-poly-l-aspartic acid; also known as cyanophycin grana peptide [CGP]) is a biopolymer that could be used in various fields, for example, as a potential precursor for the synthesis of polyaspartic acid or for the production of CGP-derived dipeptides. To extend the applicat...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Springer Berlin Heidelberg
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8050183/ https://www.ncbi.nlm.nih.gov/pubmed/33856569 http://dx.doi.org/10.1186/s13568-021-01217-5 |
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author | Wördemann, Ramona Wiefel, Lars Wendisch, Volker F. Steinbüchel, Alexander |
author_facet | Wördemann, Ramona Wiefel, Lars Wendisch, Volker F. Steinbüchel, Alexander |
author_sort | Wördemann, Ramona |
collection | PubMed |
description | Cyanophycin (multi-l-arginyl-poly-l-aspartic acid; also known as cyanophycin grana peptide [CGP]) is a biopolymer that could be used in various fields, for example, as a potential precursor for the synthesis of polyaspartic acid or for the production of CGP-derived dipeptides. To extend the applications of this polymer, it is therefore of interest to synthesize CGP with different compositions. A recent re-evaluation of the CGP synthesis in C. glutamicum has shown that C. glutamicum is a potentially interesting microorganism for CGP synthesis with a high content of alternative amino acids. This study shows that the amount of alternative amino acids can be increased by using mutants of C. glutamicum with altered amino acid biosynthesis. With the DM1729 mutant, the lysine content in the polymer could be increased up to 33.5 mol%. Furthermore, an ornithine content of up to 12.6 mol% was achieved with ORN2(P(gdh4)). How much water-soluble or insoluble CGP is synthesized is strongly related to the used cyanophycin synthetase. CphA(Dh) synthesizes soluble CGP exclusively. However, soluble CGP could also be isolated from cells expressing CphA(6308)Δ1 or CphA(6308)Δ1_C595S in addition to insoluble CGP in all examined strains. The point mutation in CphA(6308)Δ1_C595S partially resulted in a higher lysine content. In addition, the CGP content could be increased to 36% of the cell dry weight under optimizing growth conditions in C. glutamicum ATCC13032. All known alternative major amino acids for CGP synthesis (lysine, ornithine, citrulline, and glutamic acid) could be incorporated into CGP in C. glutamicum. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13568-021-01217-5. |
format | Online Article Text |
id | pubmed-8050183 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Springer Berlin Heidelberg |
record_format | MEDLINE/PubMed |
spelling | pubmed-80501832021-04-30 Incorporation of alternative amino acids into cyanophycin by different cyanophycin synthetases heterologously expressed in Corynebacterium glutamicum Wördemann, Ramona Wiefel, Lars Wendisch, Volker F. Steinbüchel, Alexander AMB Express Original Article Cyanophycin (multi-l-arginyl-poly-l-aspartic acid; also known as cyanophycin grana peptide [CGP]) is a biopolymer that could be used in various fields, for example, as a potential precursor for the synthesis of polyaspartic acid or for the production of CGP-derived dipeptides. To extend the applications of this polymer, it is therefore of interest to synthesize CGP with different compositions. A recent re-evaluation of the CGP synthesis in C. glutamicum has shown that C. glutamicum is a potentially interesting microorganism for CGP synthesis with a high content of alternative amino acids. This study shows that the amount of alternative amino acids can be increased by using mutants of C. glutamicum with altered amino acid biosynthesis. With the DM1729 mutant, the lysine content in the polymer could be increased up to 33.5 mol%. Furthermore, an ornithine content of up to 12.6 mol% was achieved with ORN2(P(gdh4)). How much water-soluble or insoluble CGP is synthesized is strongly related to the used cyanophycin synthetase. CphA(Dh) synthesizes soluble CGP exclusively. However, soluble CGP could also be isolated from cells expressing CphA(6308)Δ1 or CphA(6308)Δ1_C595S in addition to insoluble CGP in all examined strains. The point mutation in CphA(6308)Δ1_C595S partially resulted in a higher lysine content. In addition, the CGP content could be increased to 36% of the cell dry weight under optimizing growth conditions in C. glutamicum ATCC13032. All known alternative major amino acids for CGP synthesis (lysine, ornithine, citrulline, and glutamic acid) could be incorporated into CGP in C. glutamicum. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13568-021-01217-5. Springer Berlin Heidelberg 2021-04-15 /pmc/articles/PMC8050183/ /pubmed/33856569 http://dx.doi.org/10.1186/s13568-021-01217-5 Text en © The Author(s) 2021 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Original Article Wördemann, Ramona Wiefel, Lars Wendisch, Volker F. Steinbüchel, Alexander Incorporation of alternative amino acids into cyanophycin by different cyanophycin synthetases heterologously expressed in Corynebacterium glutamicum |
title | Incorporation of alternative amino acids into cyanophycin by different cyanophycin synthetases heterologously expressed in Corynebacterium glutamicum |
title_full | Incorporation of alternative amino acids into cyanophycin by different cyanophycin synthetases heterologously expressed in Corynebacterium glutamicum |
title_fullStr | Incorporation of alternative amino acids into cyanophycin by different cyanophycin synthetases heterologously expressed in Corynebacterium glutamicum |
title_full_unstemmed | Incorporation of alternative amino acids into cyanophycin by different cyanophycin synthetases heterologously expressed in Corynebacterium glutamicum |
title_short | Incorporation of alternative amino acids into cyanophycin by different cyanophycin synthetases heterologously expressed in Corynebacterium glutamicum |
title_sort | incorporation of alternative amino acids into cyanophycin by different cyanophycin synthetases heterologously expressed in corynebacterium glutamicum |
topic | Original Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8050183/ https://www.ncbi.nlm.nih.gov/pubmed/33856569 http://dx.doi.org/10.1186/s13568-021-01217-5 |
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