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| Status |
Public on Aug 04, 2020 |
| Title |
Physiological responses of Saccharopolyspora erythraea towards perturbation of intracellular NADH/NAD+ |
| Organism |
Saccharopolyspora erythraea |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
Knowledge about effects of cofactor perturbation on cellular metabolism is scarce with respect to Saccharopolyspora erythraea. The water-forming NADH oxidase (NOX) from Streptococcus pneumonia was expressed in S.erythraea E3, an important industrial strain for erythromycin production, at three different levels to investigate effects of intracellular redox status on secondary metabolism. NOX expression reduced the intracellular [NADH]/[NAD+] ratios significantly, although with a strong constitutive promoter NOX function was limited due to the shortage of oxygen. We demonstrated the negative correlation between [NADH]/[NAD+] ratios and biosynthesis of erythromycin in S.erythraea, but a positive correlation between the redox ratios and pigment production as well. We furthermore completed next-generation RNA sequencing of E3 and two NOX-expression strains. The transcription results showed that transfer processes of carbohydrates, DNA and chemical groups were altered resulting in metabolic shifts to supply more NADH for NOX fully functioning. Additionally, redox status affected transcription of several genes by allosteric effects on their transcription initiation. Specifically, transcriptional analysis along with enzymatic assay suggested that redox status influenced biosynthesis of erythromycin indirectly by allosteric effects on biosynthesis of the secondary messenger, c-di-GMP. The present work provides a basis for future cofactor manipulation in S.erythraea for further improvement of erythromycin production.
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| Overall design |
S.erythraea cultivated in the minimal liquid medium was harvested at early exponential phase (4 h). The total RNA were sequenced in duplicate for both the parental strain E3 as well as the engineered strain A2N and R3N.
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| Contributor(s) |
Li X |
| Citation(s) |
32766231 |
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https://doi.org/10.3389/fbioe.2020.00818
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| Submission date |
Jan 22, 2020 |
| Last update date |
Aug 24, 2020 |
| Contact name |
Xiaobo Li |
| Organization name |
Technical University of Denmark
|
| Street address |
Kemitorvet
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| City |
Kongens Lyngby |
| ZIP/Postal code |
2800 |
| Country |
Denmark |
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| Platforms (1) |
| GPL26951 |
BGISEQ-500 (Saccharopolyspora erythraea) |
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| Samples (6)
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| Relations |
| BioProject |
PRJNA602682 |
| SRA |
SRP243907 |
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