Metabolomics-based Analysis of the Importance of Slr0643 Protein in Regulating Synechocystis sp. PCC6803 to Acclimate to Glucose Mixotrophic Conditions
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Abstract:
In this study, a wild-type strain of Synechocystis sp. PCC6803 and a Slr0643 knockout mutant (Δslr0643) strain were used. The mechanism underlying the regulatory role of Slr0643 protein in the acclimation of Synechocystis sp. PCC6803 to glucose mixotrophic conditions was investigated through physiological experiments and gas chromatography-mass spectrometry (GC-MS)-based plant metabolomic analysis. Under mixotrophic conditions with 2.5 mM glucose, the growth rate of the wild-type was significantly higher than that under autotrophic condition, while the growth rate of Δslr0643 strain was significantly lower than that under autotrophic condition. After four days of mixotrophic growth with glucose, the OD730 value of Δslr0643 mutant was only 47.25% of that for the wild type, and the glucose utilization rate of Δslr0643 mutant was consistently lower than that the wild-type strain. Through the identification of the differential metabolites of the Δslr0643 mutant strain and wild-type strain over the mixotrophic cultivation with 2.5 mM glucose, 11 significantly different metabolites were found, with the contents of 9 differential metabolites in the wild-type strain increasing significantly during mixotrophic cultivation. Through the enrichment analysis of the metabolic pathways involved in the different metabolites of the mutant strain relative to the wild -type strain at each culture time point, it was found that the differential metabolites were mainly enriched in 8 metabolic pathways, the tricarboxylic acid cycle, pyruvate metabolism, glutamate metabolism, suggesting that Slr0643 protein was involved in regulating these metabolic pathways to make Synechocystis sp. PCC6803 adapt to glucose mixotrophic conditions. This study showed that the deletion of slr0643 damaged multiple metabolic pathways in cells, thereby making the cells unable to utilize glucose normally, and Slr0643 plays an important role in the acclimation of Synechocystis sp. PCC6803 to glucose mixotrophic conditions.
