Biocatalytic production of the antibiotic aurachin D in Escherichia coli
Abstract Aurachin D is a potent inhibitor of cytochrome bd oxidases, which are potential targets in the treatment of infectious diseases. In this study, our aim was to improve the biocatalytic production of aurachin D from a quinolone precursor molecule with recombinant Escherichia coli cells expres...
| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
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SpringerOpen
2022-11-01
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| Series: | AMB Express |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s13568-022-01478-8 |
| _version_ | 1811328515074686976 |
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| author | Sebastian Kruth Lina Schibajew Markus Nett |
| author_facet | Sebastian Kruth Lina Schibajew Markus Nett |
| author_sort | Sebastian Kruth |
| collection | DOAJ |
| description | Abstract Aurachin D is a potent inhibitor of cytochrome bd oxidases, which are potential targets in the treatment of infectious diseases. In this study, our aim was to improve the biocatalytic production of aurachin D from a quinolone precursor molecule with recombinant Escherichia coli cells expressing the biosynthesis enzyme AuaA. In order to achieve a high-level production of this membrane-bound farnesyltransferase in E. coli, the expression of the auaA gene was translationally coupled to an upstream cistron in accordance with a bicistronic design (BCD) strategy. Screening of various BCD elements led to the identification of optimized auaA expression cassettes, which increased the aurachin D titer in E. coli up to 29-fold in comparison to T7-mediated expression. This titer could be further raised by codon optimization of auaA and by introducing the mevalonate pathway into the production strain. The latter measure was intended to improve the availability of farnesyl pyrophosphate, which is needed as a cosubstrate for the AuaA-catalyzed reaction. In sum, the described efforts resulted in a strain producing aurachin D with a titer that is 424 times higher than that obtained with the original, non-optimized expression host. Graphical Abstract |
| first_indexed | 2024-04-13T15:27:28Z |
| format | Article |
| id | doaj.art-3303a7dfad5c46f09e754074305b7c28 |
| institution | Directory Open Access Journal |
| issn | 2191-0855 |
| language | English |
| last_indexed | 2024-04-13T15:27:28Z |
| publishDate | 2022-11-01 |
| publisher | SpringerOpen |
| record_format | Article |
| series | AMB Express |
| spelling | doaj.art-3303a7dfad5c46f09e754074305b7c282022-12-22T02:41:28ZengSpringerOpenAMB Express2191-08552022-11-0112111010.1186/s13568-022-01478-8Biocatalytic production of the antibiotic aurachin D in Escherichia coliSebastian Kruth0Lina Schibajew1Markus Nett2Department of Biochemical and Chemical Engineering, TU Dortmund UniversityDepartment of Biochemical and Chemical Engineering, TU Dortmund UniversityDepartment of Biochemical and Chemical Engineering, TU Dortmund UniversityAbstract Aurachin D is a potent inhibitor of cytochrome bd oxidases, which are potential targets in the treatment of infectious diseases. In this study, our aim was to improve the biocatalytic production of aurachin D from a quinolone precursor molecule with recombinant Escherichia coli cells expressing the biosynthesis enzyme AuaA. In order to achieve a high-level production of this membrane-bound farnesyltransferase in E. coli, the expression of the auaA gene was translationally coupled to an upstream cistron in accordance with a bicistronic design (BCD) strategy. Screening of various BCD elements led to the identification of optimized auaA expression cassettes, which increased the aurachin D titer in E. coli up to 29-fold in comparison to T7-mediated expression. This titer could be further raised by codon optimization of auaA and by introducing the mevalonate pathway into the production strain. The latter measure was intended to improve the availability of farnesyl pyrophosphate, which is needed as a cosubstrate for the AuaA-catalyzed reaction. In sum, the described efforts resulted in a strain producing aurachin D with a titer that is 424 times higher than that obtained with the original, non-optimized expression host. Graphical Abstracthttps://doi.org/10.1186/s13568-022-01478-8AurachinBicistronic designEscherichia coliFarnesyltransferaseHeterologous expressionMembrane protein |
| spellingShingle | Sebastian Kruth Lina Schibajew Markus Nett Biocatalytic production of the antibiotic aurachin D in Escherichia coli AMB Express Aurachin Bicistronic design Escherichia coli Farnesyltransferase Heterologous expression Membrane protein |
| title | Biocatalytic production of the antibiotic aurachin D in Escherichia coli |
| title_full | Biocatalytic production of the antibiotic aurachin D in Escherichia coli |
| title_fullStr | Biocatalytic production of the antibiotic aurachin D in Escherichia coli |
| title_full_unstemmed | Biocatalytic production of the antibiotic aurachin D in Escherichia coli |
| title_short | Biocatalytic production of the antibiotic aurachin D in Escherichia coli |
| title_sort | biocatalytic production of the antibiotic aurachin d in escherichia coli |
| topic | Aurachin Bicistronic design Escherichia coli Farnesyltransferase Heterologous expression Membrane protein |
| url | https://doi.org/10.1186/s13568-022-01478-8 |
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