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Structure of the γ-ε complex of cyanobacterial F1-ATPase reveals a suppression mechanism of the γ subunit on ATP hydrolysis in phototrophs.
Murakami, S, Kondo, K, Katayama, S, Hara, S, Sunamura, EI, Yamashita, E, Groth, G, Hisabori, T
The Biochemical journal. 2018;(18):2925-2939
Abstract
F1-ATPase forms the membrane-associated segment of F0F1-ATP synthase - the fundamental enzyme complex in cellular bioenergetics for ATP hydrolysis and synthesis. Here, we report a crystal structure of the central F1 subcomplex, consisting of the rotary shaft γ subunit and the inhibitory ε subunit, from the photosynthetic cyanobacterium Thermosynechococcus elongatus BP-1, at 1.98 Å resolution. In contrast with their homologous bacterial and mitochondrial counterparts, the γ subunits of photosynthetic organisms harbour a unique insertion of 35-40 amino acids. Our structural data reveal that this region forms a β-hairpin structure along the central stalk. We identified numerous critical hydrogen bonds and electrostatic interactions between residues in the hairpin and the rest of the γ subunit. To elaborate the critical function of this β-hairpin in inhibiting ATP hydrolysis, the corresponding domain was deleted in the cyanobacterial F1 subcomplex. Biochemical analyses of the corresponding α3β3γ complex confirm that the clinch of the hairpin structure plays a critical role and accounts for a significant interaction in the α3β3 complex to induce ADP inhibition during ATP hydrolysis. In addition, we found that truncating the β-hairpin insertion structure resulted in a marked impairment of the interaction with the ε subunit, which binds to the opposite side of the γ subunit from the β-hairpin structure. Combined with structural analyses, our work provides experimental evidence supporting the molecular principle of how the insertion region of the γ subunit suppresses F1 rotation during ATP hydrolysis.
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2.
Cyanobacterial pigments: Perspectives and biotechnological approaches.
Saini, DK, Pabbi, S, Shukla, P
Food and chemical toxicology : an international journal published for the British Industrial Biological Research Association. 2018;:616-624
Abstract
Cyanobacteria are the oxygenic photosynthesis performing prokaryotes and show a connecting link between plastids of eukaryotic autotrophs and prokaryotes. A variety of pigments, like chlorophyll, carotenoids and phycobiliproteins which exhibit different colors are present in cyanobacteria. Increasing consciousness about the harmful effects of synthetic or chemical dyes encouraged people to give more preference towards the usage of natural products, such as plant or microbial-derived colors in food and cosmetics. That is why cyanobacteria are exploited as a source of natural colors and have high commercial value in many industries. This review mainly focuses on different cyanobacterial pigments, their applications and modern biotechnological approaches such as genetic engineering, systems biology to enhance the production of biopigments for their potential use in pharmaceuticals, food, research, and cosmetics industries.
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Production of polyhydroxybutyrates and carbohydrates in a mixed cyanobacterial culture: Effect of nutrients limitation and photoperiods.
Arias, DM, Uggetti, E, García-Galán, MJ, García, J
New biotechnology. 2018;:1-11
Abstract
In the present study, different photoperiods and nutritional conditions were applied to a mixed wastewater-borne cyanobacterial culture in order to enhance the intracellular accumulation of polyhydroxybutyrates (PHBs) and carbohydrates. Two different experimental set-ups were used. In the first, the culture was permanently exposed to illumination, while in the second it was submitted to light/dark alternation (12 h cycles). In both cases, two different nutritional regimes were also evaluated, N-limitation and P-limitation. Results showed that the highest PHB concentration (104 mg L-1) was achieved under P limited conditions and permanent illumination, whereas the highest carbohydrate concentration (838 mg L-1) was obtained under N limited condition and light/dark alternation. With regard to bioplastics and biofuel generation, this study demonstrates that the accumulation of PHBs (bioplastics) and carbohydrates (potential biofuel substrate) is favored in wastewater-borne cyanobacteria under conditions where nutrients are limited.
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Strains of the toxic and bloom-forming Nodularia spumigena (cyanobacteria) can degrade methylphosphonate and release methane.
Teikari, JE, Fewer, DP, Shrestha, R, Hou, S, Leikoski, N, Mäkelä, M, Simojoki, A, Hess, WR, Sivonen, K
The ISME journal. 2018;(6):1619-1630
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Abstract
Nodularia spumigena is a nitrogen-fixing cyanobacterium that forms toxic blooms in the Baltic Sea each summer and the availability of phosphorous is an important factor limiting the formation of these blooms. Bioinformatic analysis identified a phosphonate degrading (phn) gene cluster in the genome of N. spumigena suggesting that this bacterium may use phosphonates as a phosphorus source. Our results show that strains of N. spumigena could grow in medium containing methylphosphonic acid (MPn) as the sole source of phosphorous and released methane when growing in medium containing MPn. We analyzed the total transcriptomes of N. spumigena UHCC 0039 grown using MPn and compared them with cultures growing in Pi-replete medium. The phnJ, phosphonate lyase gene, was upregulated when MPn was the sole source of phosphorus, suggesting that the expression of this gene could be used to indicate the presence of bioavailable phosphonates. Otherwise, growth on MPn resulted in only a minor reconstruction of the transcriptome and enabled good growth. However, N. spumigena strains were not able to utilize any of the anthropogenic phosphonates tested. The phosphonate utilizing pathway may offer N. spumigena a competitive advantage in the Pi-limited cyanobacterial blooms of the Baltic Sea.
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Engineering Rubisco activase from thermophilic cyanobacteria into high-temperature sensitive plants.
Ogbaga, CC, Stepien, P, Athar, HU, Ashraf, M
Critical reviews in biotechnology. 2018;(4):559-572
Abstract
In the past decade, various strategies to improve photosynthesis and crop yield, such as leaf morphology, light interception and use efficiency, biochemistry of light reactions, stomatal conductance, carboxylation efficiency, and source to sink regulation, have been discussed at length. Leaf morphology and physiology are tightly coupled to light capturing efficiency, gas exchange capacity, and temperature regulation. However, apart from the photoprotective mechanism of photosystem-II (PSII), i.e. non-photochemical quenching, very low genetic variation in the components of light reactions has been observed in plants. In the last decade, biochemistry-based enhancement of carboxylation efficiency that improves photosynthesis in plants was one of the potential strategies for improving plant biomass production. Enhancement of activation of the ubiquitous enzyme ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco; EC 4.1.1.39) by Rubisco activase may be another potential strategy for improving a photosynthesis-driven increase in crop yield. Rubisco activase modifies the conformation of the active center in Rubisco by removing tightly bound inhibitors, thereby contributing to enzyme activation and rapid carboxylation. Thermophilic cyanobacteria are oxygenic photosynthetic bacteria that thrive in high-temperature environments. This critical review discusses the prospects for and the potential of engineering Rubisco activase from thermophilic cyanobacteria into temperature-sensitive plants, to increase the threshold temperature and survival of these plants in arid regions.
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The low spin - high spin equilibrium in the S2-state of the water oxidizing enzyme.
Boussac, A, Ugur, I, Marion, A, Sugiura, M, Kaila, VRI, Rutherford, AW
Biochimica et biophysica acta. Bioenergetics. 2018;(5):342-356
Abstract
In Photosystem II (PSII), the Mn4CaO5-cluster of the active site advances through five sequential oxidation states (S0 to S4) before water is oxidized and O2 is generated. Here, we have studied the transition between the low spin (LS) and high spin (HS) configurations of S2 using EPR spectroscopy, quantum chemical calculations using Density Functional Theory (DFT), and time-resolved UV-visible absorption spectroscopy. The EPR experiments show that the equilibrium between S2LS and S2HS is pH dependent, with a pKa ≈ 8.3 (n ≈ 4) for the native Mn4CaO5 and pKa ≈ 7.5 (n ≈ 1) for Mn4SrO5. The DFT results suggest that exchanging Ca with Sr modifies the electronic structure of several titratable groups within the active site, including groups that are not direct ligands to Ca/Sr, e.g., W1/W2, Asp61, His332 and His337. This is consistent with the complex modification of the pKa upon the Ca/Sr exchange. EPR also showed that NH3 addition reversed the effect of high pH, NH3-S2LS being present at all pH values studied. Absorption spectroscopy indicates that NH3 is no longer bound in the S3TyrZ state, consistent with EPR data showing minor or no NH3-induced modification of S3 and S0. In both Ca-PSII and Sr-PSII, S2HS was capable of advancing to S3 at low temperature (198 K). This is an experimental demonstration that the S2LS is formed first and advances to S3via the S2HS state without detectable intermediates. We discuss the nature of the changes occurring in the S2LS to S2HS transition which allow the S2HS to S3 transition to occur below 200 K. This work also provides a protocol for generating S3 in concentrated samples without the need for saturating flashes.
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Carbon/Nitrogen Metabolic Balance: Lessons from Cyanobacteria.
Zhang, CC, Zhou, CZ, Burnap, RL, Peng, L
Trends in plant science. 2018;(12):1116-1130
Abstract
Carbon and nitrogen are the two most abundant nutrient elements for all living organisms, and their metabolism is tightly coupled. What are the signaling mechanisms that cells use to sense and control the carbon/nitrogen (C/N) metabolic balance following environmental changes? Based on studies in cyanobacteria, it was found that 2-phosphoglycolate derived from the oxygenase activity of Rubisco (ribulose-1,5-bisphosphate carboxylase/oxygenase) and 2-oxoglutarate from the Krebs cycle act as the carbon- and nitrogen-starvation signals, respectively, and their concentration ratio likely reflects the status of the C/N metabolic balance. We will present and discuss the regulatory principles underlying the signaling mechanisms, which are likely to be conserved in other photosynthetic organisms. These concepts may also contribute to developments in the field of biofuel engineering or improvements in crop productivity.
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Developments of Cyanobacteria for Nano-Marine Drugs: Relevance of Nanoformulations in Cancer Therapies.
Bajpai, VK, Shukla, S, Kang, SM, Hwang, SK, Song, X, Huh, YS, Han, YK
Marine drugs. 2018;(6)
Abstract
Current trends in the application of nanomaterials are emerging in the nano-biotechnological sector for development of medicines. Cyanobacteria (blue-green algae) are photosynthetic prokaryotes that have applications to human health and numerous biological activities as dietary supplements. Cyanobacteria produce biologically active and chemically diverse compounds such as cyclic peptides, lipopeptides, fatty acid amides, alkaloids, and saccharides. More than 50% of marine cyanobacteria are potentially exploitable for the extraction of bioactive substances, which are effective in killing cancer cells by inducing apoptotic death. The current review emphasizes that not even 10% of microalgal bioactive components have reached commercialized platforms due to difficulties related to solubility. Considering these factors, they should be considered as a potential source of natural products for drug discovery and drug delivery approaches. Nanoformulations employing a wide variety of nanoparticles and their polymerized forms could be an emerging approach to the development of new cancer drugs. This review highlights recent research on microalgae-based medicines or compounds as well as their biomedical applications. This review further discusses the facts, limitations, and commercial market trends related to the use of microalgae for industrial and medicinal purposes.
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[Super Liquid Crystalline Polysaccharides Produced by Ultimately-ecological Microreactors].
Kaneko, T, Okajima, MK
Yakugaku zasshi : Journal of the Pharmaceutical Society of Japan. 2018;(4):489-496
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Abstract
Cyanobacteria fix carbon dioxide and nitrogen from the atmosphere using solar energy to produce various biomolecules, and thus are regarded as ultimately ecological microreactors. Sacran is a cyanobacterial polysaccharide with a very high molecular weight of 29 Mg/mol, which is extracted from Aphanothece sacrum cyanobacterium mass-cultivated in freshwater environments such as river or spring. Sacran is a water-soluble heteropolysaccharide comprising more than 6 kinds of sugar residues and contains 12% sulfate anionic groups and 27% carboxyls. Sacran has a super-absorbent function of water, which can retain 6000 mL for 1 g specimen, due to very long hydrating chains. The value is much higher than hyaluronic acid or conventional super-absorbent polymers. Sacran exhibits self-orienting behavior in dilute solution at a concentration range over 0.25 wt%, which is quite low when compared with conventional liquid crystalline polysaccharides. Mesogenic helical chains of sacrans have extremely high aspect ratios of 1600 for highly persistent lengths of 32 micrometer. Through the liquid crystallinity, sacran solution shows a shear-thinning behavior and the solution spread over a substrate such as biological skin very efficiently to create a thin layer. Applied on atopic dermatitis skin sacran solution exerts excellent moisturizing effect and anti-itching action.
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Isolation and identification of siderophores produced by cyanobacteria.
Řezanka, T, Palyzová, A, Sigler, K
Folia microbiologica. 2018;(5):569-579
Abstract
Cyanobacteria are one of the most successful and oldest forms of life that are present on Earth. They are prokaryotic photoautotrophic microorganisms that colonize so diverse environments as soil, seawater, and freshwater, but also stones, plants, or extreme habitats such as snow and ice as well as hot springs. This diversity in the type of environment they live in requires a successful adaptation to completely different conditions. For this reason, cyanobacteria form a wide range of different secondary metabolites. In particular, the cyanobacteria living in both freshwater and sea produce many metabolites that have biological activity. In this review, we focus on metabolites called siderophores, which are low molecular weight chemical compounds specifically binding iron ions. They have a relatively low molecular weight and are produced by bacteria and also by fungi. The main role of siderophores is to obtain iron from the environment and to create a soluble complex available to microbial cells. Siderophores play an important role in microbial ecology; for example, in agriculture they support the growth of many plants and increase their production by increasing the availability of Fe in plants. The aim of this review is to demonstrate the modern use of physico-chemical methods for the detection of siderophores in cyanobacteria and the use of these methods for the detection and characterization of the siderophore-producing microorganisms. Using high-performance liquid chromatography-mass spectrometry (LC-MS), it is possible not only to discover new chemical structures but also to identify potential interactions between microorganisms. Based on tandem mass spectrometry (MS/MS) analyses, previous siderophore knowledge can be used to interpret MS/MS data to examine both known and new siderophores.