On the Role and Production of Polyhydroxybutyrate (PHB) in the Cyanobacterium Synechocystis sp. PCC 6803
Abstract
:1. Introduction
2. Materials and Methods
2.1. Cyanobacterial Cultivation Conditions
2.2. Physical Stress Conditions
2.3. Oxygen Measurements
2.4. Microscopy and Staining Procedures
2.5. Electron Microscopy
2.6. Spectral Analysis
2.7. PAM
2.8. PHB Quantification
2.9. FACS and Flow Cytometry
3. Results
3.1. The Influence of Environmental Conditions and Central Pathways on PHB Production
3.2. Recovery from Nitrogen Starvation
3.3. Heterogeneity of PHB Production
4. Discussion
4.1. Heterogeneity of PHB Content
4.2. Physiological Function of PHB
5. Conclusions
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Appendix A
Strain | Relevant Marker of Genotype | Reference |
---|---|---|
Synechocystis sp. PCC 6803 | - | Pasteur culture collection |
Δzwf | Slr1843::cmR | Chen et al. 2016 |
ΔpfkB1/2 | sll1196::kmR, sll0745::spR | Chen et al. 2016 |
ΔphaEC | slr1829,slr1830::kmR | Klotz et al. 2016 |
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Abiotic Stress | Treatment | Effect |
---|---|---|
(1) | ||
Cold | 4 °C over night 1 h at −20°C; 14 h at −20 °C | No difference |
Heat | 45 min at 40 °C; 45 min at 40°C and 20 min at 50°C | No difference All cells were dead |
Physical Force | Centrifugation for 30 min at 20,000 g; 1 h at 25,000 g 3 × 5 min at 4 m/s glass-bead milling in Ribolyser; 3 × 5 min at 7 m/s Ribolyser | No difference No difference All cells were dead |
Darkness | 1 day; 2 days, 5 days; 8 days; 10 days; 15 days | No difference |
Low Light | 14 days at 5 µE | No difference |
Alternating Light | 12/12 light/dark | No difference |
Drought | 30 min at 30 °C SpeedVac | All cells were dead |
Nitrogen Starvation | 3 weeks 10 weeks | No difference ∆phaEC showed weak growth advantage |
High Light | 1 day incubation at 500 µE | No difference |
(2) | ||
Buffered Medium | BG11 agar plates containing 300 µL, 1 mL, 3 mL TES buffer | No difference |
Salt | Recovery at BG11 agar plates with: 100, 150, 300 mM NaCl 2 × BG11 salts | No difference No difference |
Carbon Availability | 0, 10, 50, 150 mM bicarbonate (added to BG11 agar plates) | No difference |
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Koch, M.; Berendzen, K.W.; Forchhammer, K. On the Role and Production of Polyhydroxybutyrate (PHB) in the Cyanobacterium Synechocystis sp. PCC 6803. Life 2020, 10, 47. https://doi.org/10.3390/life10040047
Koch M, Berendzen KW, Forchhammer K. On the Role and Production of Polyhydroxybutyrate (PHB) in the Cyanobacterium Synechocystis sp. PCC 6803. Life. 2020; 10(4):47. https://doi.org/10.3390/life10040047
Chicago/Turabian StyleKoch, Moritz, Kenneth W. Berendzen, and Karl Forchhammer. 2020. "On the Role and Production of Polyhydroxybutyrate (PHB) in the Cyanobacterium Synechocystis sp. PCC 6803" Life 10, no. 4: 47. https://doi.org/10.3390/life10040047