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Cofactor Engineering Redirects Secondary Metabolism and Enhances Erythromycin Production in Saccharopolyspora erythraea .
- Source :
-
ACS synthetic biology [ACS Synth Biol] 2020 Mar 20; Vol. 9 (3), pp. 655-670. Date of Electronic Publication: 2020 Feb 28. - Publication Year :
- 2020
-
Abstract
- Saccharopolyspora erythraea is used for industrial erythromycin production. To explore the physiological role of intracellular energy state in metabolic regulation by S. erythraea , we initially overexpressed the F <subscript>1</subscript> part of the endogenous F <subscript>1</subscript> F <subscript>0</subscript> -ATPase in the high yielding erythromycin producing strain E3. The F <subscript>1</subscript> -ATPase expression resulted in lower [ATP]/[ADP] ratios, which was accompanied by a strong increase in the production of a reddish pigment and a decreased erythromycin production. Subsequent transcriptional analysis revealed that the lower intracellular [ATP]/[ADP] ratios exerted a pleotropic regulation on the metabolism of S. erythraea . The lower [ATP]/[ADP] ratios induced physiological changes to restore the energy balance, mainly via pathways that tend to produce ATP or regenerate NADH. The F <subscript>1</subscript> -ATPase overexpression strain exhibited a state of redox stress, which was correlated to an alteration of electron transport at the branch of the terminal oxidases, and S. erythraea channeled the enhanced glycolytic flux toward a reddish pigment in order to reduce NADH formation. The production of erythromycin was decreased, which is in accordance with the net ATP requirement and the excess NADH formed through this pathway. Partial growth inhibition by apramycin increased the intracellular [ATP]/[ADP] ratios and demonstrated a positive correlation between [ATP]/[ADP] ratios and erythromycin synthesis. Finally, overexpression of the entire F <subscript>1</subscript> F <subscript>0</subscript> -ATPase complex resulted in 28% enhanced erythromycin production and markedly reduced pigment synthesis in E3. The work illustrates a feasible strategy to optimize the distribution of fluxes in secondary metabolism.
- Subjects :
- Acetyl Coenzyme A metabolism
Adenosine Diphosphate metabolism
Adenosine Triphosphate metabolism
Coenzymes metabolism
Electron Transport
Gene Expression Regulation, Bacterial
Microorganisms, Genetically-Modified
NAD genetics
NAD metabolism
Pigments, Biological genetics
Pigments, Biological metabolism
Proton-Translocating ATPases genetics
Proton-Translocating ATPases metabolism
Secondary Metabolism
Coenzymes genetics
Erythromycin biosynthesis
Metabolic Engineering methods
Saccharopolyspora genetics
Saccharopolyspora metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 2161-5063
- Volume :
- 9
- Issue :
- 3
- Database :
- MEDLINE
- Journal :
- ACS synthetic biology
- Publication Type :
- Academic Journal
- Accession number :
- 32078772
- Full Text :
- https://doi.org/10.1021/acssynbio.9b00528