Identification of the direct regulon of NtcA during early acclimation to nitrogen starvation in the cyanobacterium Synechocystis sp. PCC 6803

Polyhydroxybutyrate (PHB) is a polymer of great interest as a substitute for conventional plastics, which are becoming an enormous environmental problem. PHB can be produced directly from CO2 in photoautotrophic cyanobacteria. The model cyanobacterium Synechocystis sp. PCC 6803 produces PHB under conditions of nitrogen starvation. However, it is so far unclear which metabolic pathways provide the precursor molecules for PHB synthesis during nitrogen starvation. In this study, we investigated if PHB could be derived from the main intracellular carbon pool, glycogen. A mutant of the major glycogen phosphorylase, GlgP2 (slr1367 product), was almost completely impaired in PHB synthesis. Conversely, in the absence of glycogen synthase GlgA1 (sll0945 product), cells not only produced less PHB, but were also impaired in acclimation to nitrogen depletion. To analyze the role of the various carbon catabolic pathways (EMP, ED and OPP pathways) for PHB production, mutants of key enzymes of these pathways were analyzed, showing different impact on PHB synthesis. Together, this study clearly indicates that PHB in glycogen-producing Synechocystis sp. PCC 6803 cells is produced from this carbon-pool during nitrogen starvation periods. This knowledge can be used for metabolic engineering to get closer to the overall goal of a sustainable, carbon-neutral bioplastic production.

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On the Role and Production of Polyhydroxybutyrate (PHB) in the Cyanobacterium Synechocystis sp. PCC 6803

Life ◽

10.3390/life10040047 ◽

2020 ◽

Vol 10 (4) ◽

pp. 47 ◽

Cited By ~ 5

Author(s):

Moritz Koch ◽

Kenneth W. Berendzen ◽

Karl Forchhammer

Keyword(s):

Cell Sorting ◽

Nitrogen Starvation ◽

Single Cells ◽

Continuous Light ◽

Cell Level ◽

Physiological Conditions ◽

Fitness Advantage ◽

Physiological Relevance ◽

Synechocystis Sp ◽

Pcc 6803

The cyanobacterium Synechocystis sp. PCC 6803 is known for producing polyhydroxybutyrate (PHB) under unbalanced nutrient conditions. Although many cyanobacteria produce PHB, its physiological relevance remains unknown, since previous studies concluded that PHB is redundant. In this work, we try to better understand the physiological conditions that are important for PHB synthesis. The accumulation of intracellular PHB was higher when the cyanobacterial cells were grown under an alternating day–night rhythm as compared to continuous light. In contrast to previous reports, a reduction of PHB was observed when the cells were grown under conditions of limited gas exchange. Since previous data showed that PHB is not required for the resuscitation from nitrogen starvation, a series of different abiotic stresses were applied to test if PHB is beneficial for its fitness. However, under none of the tested conditions did cells containing PHB show a fitness advantage compared to a PHB-free-mutant (ΔphaEC). Additionally, the distribution of PHB in single cells of a population Synechocystis cells was analyzed via fluorescence-activated cell sorting (FACS). The results showed a considerable degree of phenotypic heterogeneity at the single cell level concerning the content of PHB, which was consistent over several generations. These results improve our understanding about how and why Synechocystis synthesizes PHB and gives suggestions how to further increase its production for a biotechnological process.

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