Diversity of Glutathione S-Transferases (GSTs) in Cyanobacteria with Reference to Their Structures, Substrate Recognition and Catalytic Functions.

Glutathione S-Transferases (GSTs) comprise a diverse group of protein superfamily involved in cellular detoxification of various harmful xenobiotics and endobiotics. Cyanobacteria, being the primordial photosynthetic prokaryotes, served as an origin for the evolution of GSTs with diversity in their structures, substrate recognition, and catalytic functions. This study analysed the diversity of GSTs in cyanobacteria for the first time. Based on the sequence alignment and phylogenetic tree analysis, 12 GST classes were identified, which are distributed variedly within cyanobacterial orders such as four in Pleurocapsales, eight in Chroococcales, seven in Oscillatoriales, five in Stigonematales, and nine in Nostocales. Detailed evolutionary analysis of cyanobacterial GSTs suggested that the order Pleurocapsales served as the ancestry for GST evolution. The analysis also identified a conserved motif S[GLNTARS][ADE]I[LAI] with signature residues, cysteine, serine, and tyrosine at the N-terminal end that serves as the initiating residue for detoxification. Alternatively, the grouping of cyanobacterial GSTs and their unique signature residues were located, which serve as a possible discriminating factor. The study also described the mode of glutathione binding between the identified cyanobacterial GST groups highlighting the differences among the GST classes. New GST sequence data may improve further our understanding on GST evolution and other possible divergences in cyanobacteria.

Analysis and elucidation of phosphoenolpyruvate carboxylase in cyanobacteria.

Phosphoenolpyruvate carboxylase (PEPC) a cytosolic enzyme of higher plants is also found in bacteria and cyanobacteria. Genetic and biochemical investigations have indicated that there are several isoforms of PEPC belonging to C3; C3/C4 and C4 groups but, the evolution of PEPC in cyanobacteria is not yet understood. The present study opens up an opportunity to understand the isoforms and functions of PEPC in cyanobacteria. The variations observed in PEPC among lower and higher orders of cyanobacteria, suggests convergent evolution of PEPC. There is a specific PEPC phosphorylation residue 'serine' at the N-terminus and PEPC determinant residue 'serine' at the C-terminal that facilitates high affinity for substrate binding. These residues were unique to higher orders of cyanobacteria, but, not in lower orders and other prokaryotes. The different PEPC forms of cyanobacteria were investigated for their kinetic properties with phosphoenolpyruvate as the substrate and the findings corroborated well with the in silico findings. In vitro enzymatic study of cyanobacteria belonging to three different orders demonstrated the role of aspartate as an allosteric effector, which inhibited PEPC by interacting with the highly conserved residues in the active site. The differences in mode of inhibition among the different order, thus, give a fair picture about the cyanobacterial PEPCs. The higher orders appear to possess the sequence coordinates and functionally conserved residues similar to isoforms of C4 type higher plants, whereas isoforms of PEPC of the lower orders did not resemble either that of C3 or C4 plants.

Siderophore mediated uranium sequestration by marine cyanobacterium Synechococcus elongatus BDU 130911.

Four different marine cyanobacterial morphotypes were tested for their efficacy to produce siderophores in Fe minus [Fe(-)], Fe minus Uranium dosed [Fe(-)U(+)], and Fe dosed Uranium dosed [Fe(-)U(+)] media. Of the four organisms tested, Synechococcus elongatus BDU 130911 produced the highest amount of siderophore of 58µgmg(-1) dryweight. The results clearly indicate that uranium induces siderophore production in marine cyanobacteria even in the presence of iron [Fe(-)U(+)] condition. The type of siderophore revealed by FeCl(3), Tetrazolium and Atkin's tests is a hydroxamate; and thin layer chromatogram also authenticates our finding. Uranium siderophore complexation was confirmed through modified Chrome Azurol S (CAS) assay as well as based on residual uranium presence. In silico docking studies further validate siderophore complexation with uranium.