Physiological responses of the cyanobacterium Synechocystis sp. PCC 6803 under rhythmic light variations.

Cyanobacteria are challenged by daily fluctuations of light intensities and photoperiod in their natural habitats, which affect the physiology and fitness of cyanobacteria. Circadian rhythms (CRs), an important endogenous process found in all organisms including cyanobacteria, control their physiological activities and helps in coping with 24-h light/dark (LD) cycle. In cyanobacteria, physiological responses under rhythmic ultraviolet radiation (UVR) are poorly studied. Therefore, we studied the changes in photosynthetic pigments, and physiological parameters of Synechocystis sp. PCC 6803 under UVR and photosynthetically active radiation (PAR) of light/dark (LD) oscillations having the combinations of 0, 4:20, 8:16, 12:12, 16:8, 20:4, and 24:24 h. The LD 16:8 enhanced the growth, pigments, proteins, photosynthetic efficiency, and physiology of Synechocystis sp. PCC6803. Continuous light (LL 24) of UVR and PAR exerted negative impact on the photosynthetic pigments, and chlorophyll fluorescence. Significant increase in reactive oxygen species (ROS) resulted in loss of plasma membrane integrity followed by decreased viability of cells. The dark phase played a significant role in Synechocystis to withstand the LL 24 under PAR and UVR. This study offers detailed understanding of the physiological responses of the cyanobacterium to changing light environment.

Phylogenetic distribution, structural analysis and interaction of nucleotide excision repair proteins in cyanobacteria.

Cyanobacteria are photosynthetic Gram-negative, oxygen evolving prokaryotes with cosmopolitan distribution. Ultraviolet radiation (UVR) and other abiotic stresses result in DNA lesions in cyanobacteria. Nucleotide excision repair (NER) pathway removes the DNA lesions produced by UVR to normal DNA sequence. In cyanobacteria, detailed knowledge about NER proteins is poorly studied. Therefore, we have studied the NER proteins in cyanobacteria. Analyses of 289 amino acids sequence from 77 cyanobacterial species have revealed the presence of a minimum of one copy of NER protein in their genome. Phylogenetic analysis of NER protein shows that UvrD has maximal rate of amino acid substitutions which resulted in increased branch length. The motif analysis shows that UvrABC proteins is more conserved than UvrD, Further, UvrA with UvrB protein interacts with each other and form stable complex which have DNA binding domain on the surface of the complex. UvrB also have DNA binding domain. Positive electrostatic potential was found in the DNA binding region, which is followed by negative and neutral electrostatic potential. Additionally, the surface accessibility values at the DNA strands of T5-T6 dimer binding site were maximal. Protein nucleotide interaction shows the strong binding of T5-T6 dimer with NER proteins of Synechocystis sp. PCC 6803. This process repairs the UV-induced DNA lesions in dark when photoreactivation is inactive. Regulation of NER proteins protect cyanobacterial genome and maintain the fitness of organism under different abiotic stresses.

Anticancer Compounds from Cyanobacteria and their Implications in Apoptosis.

Cyanobacteria have been recognized as a rich source of bioactive metabolites with potential biotechnological applications in the pharmacological industry. The chemically diverse natural compounds or their analogues cause cytotoxicity. They may kill various cancer cells by inducing apoptosis or changing the activation of cell signaling, particularly involving the protein kinase-C family of enzymes, mitochondrial dysfunctions, and oxidative damage. B cell lymphoma 2 (Bcl-2) is an essential component of apoptosis and is an antiapoptotic molecule. The key apoptotic regulators associated with cancer are members of the Bcl-2 protein family, the key member of which is Bcl-2. The Bcl-2 protein is a promising target for the emergence of new anti-tumor therapies because of its critical role in controlling apoptosis. This review explores the significance of Bcl-2 in the onset of cancer; it may be used as a target for developing high-quality drug therapies to treat various tumors. In addition, a number of computational techniques were used to identify novel hit compounds that may act as inhibitors of the apoptotic protein Bcl-2, including virtual screening, toxicity prediction, and drug-likeness analysis. Twenty-three compounds were assessed as potential hits against Bcl-2, and these compounds were subjected to ADMET property prediction. Dendroamide A and Welwitindolinone A appear to be the most stable and effective drugs against Bcl-2 out of all those evaluated. This article gives an overview of the bioactive compounds produced by cyanobacteria that have anticancer properties and may be exploited to create novel anticancer medications in the future.

Computational studies on photolyase (Phr) proteins of cyanobacteria.

Photolyases (Phrs) are enzymes that utilize the blue/ultraviolet (UV-A) region of light for repairing UV-induced cyclopyramidine dimers. We studied Phr groups by bioinformatic analyses as well as active-site and structural modeling. Analysis of 238 amino acid sequences from 85 completely sequenced cyanobacterial genomes revealed five classes of Phrs, CPD Gr I, 6-4 Phrs/cryptochrome, Cry-DASH, Fe-S bacteria Phrs, and a group with fewer amino acids (276-385) in length. The distribution of Phr groups in cyanobacteria belonging to the order Synechococcales was found to be influenced by the habitats of the organisms. Class V Phrs are exclusively present in cyanobacteria. Unique motifs and binding sites were reported in groups II and III. The Fe-S protein binding site was only present in group V and the active site residues and putative CPD/6-4PP binding residues are charged amino acids present on the surface of the proteins. The majority of hydrophilic amino acid residues were present on the surface of the Phrs. Sequence analysis confirmed the diverse nature of Phrs, although sequence diversity did not affect the overall three-dimensional structure. Protein-ligand interaction analysis identified novel CPD/6-4PP binding sites on Phrs. This structural information of Phrs can be used for the preparation of efficient Phr-based formulations.

Responses of a hot spring cyanobacterium under ultraviolet and photosynthetically active radiation: photosynthetic performance, antioxidative enzymes, mycosporine-like amino acid profiling and its antioxidative potentials.

This study summarizes the response of a hot spring cyanobacterium Fischerella sp. strain HKAR-14, under simulated light conditions of ultraviolet radiation (UVR), photosynthetically active radiation (PAR), PAR + UV-A (PA) and PAR + UV-A + UV-B (PAB). Exposure to UVR caused a decline in growth and Chl a while total carotene content increased under PA and PAB. Maximum photochemical efficiency of photosystem II (F v /F m) and relative electron transport rate decreased significantly in PA and PAB exposure. Higher non-photochemical quenching and lower photochemical quenching values were observed in UVR-exposed samples as compared to the control. Levels of intracellular reactive oxygen species (ROS) increased significantly in PAB and PA. Fluorescence microscopic images showed an increase in green fluorescence, indicating the generation of ROS in UVR. The antioxidant machinery including superoxide dismutase, catalase and peroxidase showed an increase of 1.76-fold and 2.5-fold superoxide dismutase, 2.4-fold and 3.7-fold catalase, 1.83-fold and 2.5-fold peroxidase activities under PA and PAB, respectively. High-performance liquid chromatography equipped with photodiode array detector, electrospray ionization mass spectrometry, Fourier-transform infrared spectroscopy and nuclear magnetic resonance spectroscopy analyses reveal the occurrence of a single mycosporine-like amino acid, shinorine (λ max 332.3 ± 2 nm, m/z 333.1), with a retention time of 1.157 min. The electrochemical characterization of shinorine was determined by cyclic voltammetry. The shinorine molecule possesses electrochemical activity and represents diffusion-controlled process in 0.1 M (pH 7.0) phosphate buffer. An antioxidant assay of shinorine showed its efficient activity as antioxidant which increased in a dose-dependent manner.

Composition and functional property of photosynthetic pigments under circadian rhythm in the cyanobacterium Spirulina platensis.

Circadian rhythm is an important endogenous biological signal for sustainable growth and development of cyanobacteria in natural ecosystems. Circadian effects of photosynthetically active radiation (PAR), ultraviolet-A (UV-A) and ultraviolet-B (UV-B) radiations on pigment composition have been studied in the cyanobacterium Spirulina platensis under light (L)/dark (D) oscillation with a combination of 4/20, 8/16, 12/12, 16/8, 20/4 and 24/24 h time duration. Circadian exposure of PAR + UV-A (PA) and PAR + UV-A + UV-B (PAB) showed more than twofold decline in Chl a, total protein and phycocyanin (PC) in light phase and significant recovery was achieved in dark phase. The fluorescence emission wavelength of PC was shifted towards lower wavelengths in the light phase of PAB in comparison to P and PA whereas the same wavelength was retrieved in the dark phase. The production of free radicals was accelerated twofold in the light phase (24 h L) whereas the same was retrieved to the level of control during the dark phase. Oxidatively induced damage was alleviated by antioxidative enzymes such as catalase (CAT), peroxidase (POD), superoxide dismutase (SOD) and ascorbate peroxidase (APX) in the light phase (0-24-h L) whereas the dark phase showed significant inhibition of the same enzymes. Similar characteristic inhibition of free radicals and recovery of PC was observed inside cellular filament after circadian rhythm of 24/24 h (L/D). Circadian exposure of P, PA and PAB significantly altered the synthesis and recovery of pigments that could be crucial for optimization and sustainable production of photosynthetic products for human welfare.

Codon usage analysis of photolyase encoding genes of cyanobacteria inhabiting diverse habitats.

Nucleotide and amino acid compositions were studied to determine the genomic and structural relationship of photolyase gene in freshwater, marine and hot spring cyanobacteria. Among three habitats, photolyase encoding genes from hot spring cyanobacteria were found to have highest GC content. The genomic GC content was found to influence the codon usage and amino acid variability in photolyases. The third position of codon was found to have more effect on amino acid variability in photolyases than the first and second positions of codon. The variation of amino acids Ala, Asp, Glu, Gly, His, Leu, Pro, Gln, Arg and Val in photolyases of three different habitats was found to be controlled by first position of codon (G1C1). However, second position (G2C2) of codon regulates variation of Ala, Cys, Gly, Pro, Arg, Ser, Thr and Tyr contents in photolyases. Third position (G3C3) of codon controls incorporation of amino acids such as Ala, Phe, Gly, Leu, Gln, Pro, Arg, Ser, Thr and Tyr in photolyases from three habitats. Photolyase encoding genes of hot spring cyanobacteria have 85% codons with G or C at third position, whereas marine and freshwater cyanobacteria showed 82 and 60% codons, respectively, with G or C at third position. Principal component analysis (PCA) showed that GC content has a profound effect in separating the genes along the first major axis according to their RSCU (relative synonymous codon usage) values, and neutrality analysis indicated that mutational pressure has resulted in codon bias in photolyase genes of cyanobacteria.

Detection of Reactive Oxygen Species (ROS) in Cyanobacteria Using the Oxidant-sensing Probe 2',7'-Dichlorodihydrofluorescein Diacetate (DCFH-DA).

Reactive oxygen species (ROS) are cell signaling molecules synthesized inside the cells as a response to routine metabolic processes. In stress conditions such as ultraviolet radiation (UVR), ROS concentration increases several folds in the cells that become toxic for the cell survival. Here we present the method for in vivo detection of ROS by using an oxidant-sensing probe 2',7'-dichlorodihydrofluorescein diacetate (DCFH-DA) in cyanobacteria. This method provides reliable, simple, rapid and cost effective means for detection of ROS in cyanobacteria.