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Growth-uncoupled isoprenoid synthesis in Rhodobacter sphaeroides
- Source :
- Biotechnology for Biofuels, Vol 13, Iss 1, Pp 1-13 (2020), Biotechnology for Biofuels 13 (2020) 1, Biotechnology for biofuels, England, Biotechnology for Biofuels, 13(1)
- Publication Year :
- 2020
- Publisher :
- BMC, 2020.
-
Abstract
- Background Microbial cell factories are usually engineered and employed for cultivations that combine product synthesis with growth. Such a strategy inevitably invests part of the substrate pool towards the generation of biomass and cellular maintenance. Hence, engineering strains for the formation of a specific product under non-growth conditions would allow to reach higher product yields. In this respect, isoprenoid biosynthesis represents an extensively studied example of growth-coupled synthesis with rather unexplored potential for growth-independent production. Rhodobacter sphaeroides is a model bacterium for isoprenoid biosynthesis, either via the native 2-methyl-d-erythritol 4-phosphate (MEP) pathway or the heterologous mevalonate (MVA) pathway, and for poly-β-hydroxybutyrate (PHB) biosynthesis. Results This study investigates the use of this bacterium for growth-independent production of isoprenoids, with amorpha-4,11-diene as reporter molecule. For this purpose, we employed the recently developed Cas9-based genome editing tool for R. sphaeroides to rapidly construct single and double deletion mutant strains of the MEP and PHB pathways, and we subsequently transformed the strains with the amorphadiene producing plasmid. Furthermore, we employed 13C-metabolic flux ratio analysis to monitor the changes in the isoprenoid metabolic fluxes under different cultivation conditions. We demonstrated that active flux via both isoprenoid pathways while inactivating PHB synthesis maximizes growth-coupled isoprenoid synthesis. On the other hand, the strain that showed the highest growth-independent isoprenoid yield and productivity, combined the plasmid-based heterologous expression of the orthogonal MVA pathway with the inactivation of the native MEP and PHB production pathways. Conclusions Apart from proposing a microbial cell factory for growth-independent isoprenoid synthesis, this work provides novel insights about the interaction of MEP and MVA pathways under different growth conditions.
- Subjects :
- 0106 biological sciences
Bio Process Engineering
PHB
lcsh:Biotechnology
Heterologous
Rhodobacter sphaeroides
Management, Monitoring, Policy and Law
Isoprenoid biosynthesis
01 natural sciences
Applied Microbiology and Biotechnology
lcsh:Fuel
03 medical and health sciences
chemistry.chemical_compound
Plasmid
Biosynthesis
lcsh:TP315-360
010608 biotechnology
lcsh:TP248.13-248.65
VLAG
030304 developmental biology
2. Zero hunger
0303 health sciences
Strain (chemistry)
biology
Renewable Energy, Sustainability and the Environment
Chemistry
BacGen
MEP
MVA
biology.organism_classification
Terpenoid
General Energy
Biochemistry
BIOS Applied Metabolic Systems
lipids (amino acids, peptides, and proteins)
Heterologous expression
Growth-independent production
Physical Chemistry and Soft Matter
Bacteria
Biotechnology
Subjects
Details
- Language :
- English
- ISSN :
- 17546834
- Volume :
- 13
- Issue :
- 1
- Database :
- OpenAIRE
- Journal :
- Biotechnology for Biofuels
- Accession number :
- edsair.doi.dedup.....35f4ebbc44567fa1e0457fa46f683184