Cyanobacterial Research Research Articles

An Exopolysaccharide from the Cyanobacterium Arthrospira platensis May Utilize CH-π Bonding: A Review of the Isolation, Purification, and Chemical Structure of Calcium-Spirulan.

The CH-π bonding potential of a saccharide is determined primarily by the number of hydrogen atoms available for bonding and is reduced by side groups that interfere with the CH-π bond. Each hydrogen bond increases the total bond energy, while interfering hydroxyl groups and other side groups reduce the bond energy by repulsion. The disaccharide repeating units of Calcium-Spirulan (Ca-SP), a large exopolysaccharide sub fractionated from the supernatant of the cyanobacterium Arthrospira platensis, contain a unique monosaccharide that is completely devoid of hydroxyl groups and side groups on its entire beta surface, leaving five hydrogen atoms available for CH-π bonding in the planar conformation. While planar conformations of independent pyranose rings are rare-to-nonexistent, due to ring strain associated with that conformation, the binding site of a protein could provide the conformational energy needed to overcome that energy barrier. By enabling a planar conformation, a protein could also enable the sugar to form a novel 5-hydrogen CH-π bond configuration. One study of the anticoagulant property of Ca-SP shows that the molecule acts as an activator of Heparin Cofactor II (HC-II), boosting its anticoagulant kinetics by 104. In comparison, the longstanding anticoagulant drug Heparin boosts the HC-II kinetics by 103. The difference may be explained by this unique CH-π configuration. Here, we review current knowledge and experience on the isolation techniques, analytical methods, and chemical structures of Ca-SP. We emphasize a discussion of the CH-π bonding potential of this unique polysaccharide because it is a topic that has not yet been addressed. By introducing the topic of CH-π bonding to the cyanobacterial research community, this review may help to set the stage for further investigation of these unique molecules, their genetics, their biosynthetic pathways, their chemistry, and their biological functions.

What is holding back cyanobacterial research and applications? A survey of the cyanobacterial research community

Cyanobacteria are a diverse group of prokaryotic organisms that have been the subject of intense basic research, resulting in a wealth of knowledge about fundamental cellular processes such as photosynthesis. However, the translation of that research towards industry-relevant applications is still limited. To understand the reasons for this contradictory situation, we conducted a quantitative survey among researchers in the cyanobacterial community, a set of individual interviews with established researchers, and a literature analysis. Our results show that the community seems to be committed to embracing cyanobacterial diversity and promoting collaboration. Additionally, participants expressed a strong desire to develop standardized protocols for research and establish larger consortia to accelerate progress. The results of the survey highlight the need for a more integrated approach to cyanobacterial research that encompasses both basic and applied aspects. Based on the survey and interview results as well as our literature analysis, we highlight areas for potential improvement, strategies to enhance cyanobacterial research, and open questions that demand further exploration. Addressing these challenges should accelerate the development of industrial applications based on cyanobacterial research.

Cyanobacterial Genomes from a Brackish Coastal Lagoon Reveal Potential for Novel Biogeochemical Functions and Their Evolution.

Cyanobacteria are recognised for their pivotal roles in aquatic ecosystems, serving as primary producers and major agents in diazotrophic processes. Currently, the primary focus of cyanobacterial research lies in gaining a more detailed understanding of these well-established ecosystem functions. However, their involvement and impact on other crucial biogeochemical cycles remain understudied. This knowledge gap is partially attributed to the challenges associated with culturing cyanobacteria in controlled laboratory conditions and the limited understanding of their specific growth requirements. This can be circumvented partially by the culture-independent methods which can shed light on the genomic potential of cyanobacterial species and answer more profound questions about the evolution of other key biogeochemical functions. In this study, we assembled 83 cyanobacterial genomes from metagenomic data generated from environmental DNA extracted from a brackish water lagoon (Chilika Lake, India). We taxonomically classified these metagenome-assembled genomes (MAGs) and found that about 92.77% of them are novel genomes at the species level. We then annotated these cyanobacterial MAGs for all the encoded functions using KEGG Orthology. Interestingly, we found two previously unreported functions in Cyanobacteria, namely, DNRA (Dissimilatory Nitrate Reduction to Ammonium) and DMSP (Dimethylsulfoniopropionate) synthesis in multiple MAGs using nirBD and dsyB genes as markers. We validated their presence in several publicly available cyanobacterial isolate genomes. Further, we identified incongruities between the evolutionary patterns of species and the marker genes and elucidated the underlying reasons for these discrepancies. This study expands our overall comprehension of the contribution of cyanobacteria to the biogeochemical cycling in coastal brackish ecosystems.

CyanoCyc cyanobacterial web portal.

CyanoCyc is a web portal that integrates an exceptionally rich database collection of information about cyanobacterial genomes with an extensive suite of bioinformatics tools. It was developed to address the needs of the cyanobacterial research and biotechnology communities. The 277 annotated cyanobacterial genomes currently in CyanoCyc are supplemented with computational inferences including predicted metabolic pathways, operons, protein complexes, and orthologs; and with data imported from external databases, such as protein features and Gene Ontology (GO) terms imported from UniProt. Five of the genome databases have undergone manual curation with input from more than a dozen cyanobacteria experts to correct errors and integrate information from more than 1,765 published articles. CyanoCyc has bioinformatics tools that encompass genome, metabolic pathway and regulatory informatics; omics data analysis; and comparative analyses, including visualizations of multiple genomes aligned at orthologous genes, and comparisons of metabolic networks for multiple organisms. CyanoCyc is a high-quality, reliable knowledgebase that accelerates scientists' work by enabling users to quickly find accurate information using its powerful set of search tools, to understand gene function through expert mini-reviews with citations, to acquire information quickly using its interactive visualization tools, and to inform better decision-making for fundamental and applied research.

The quantitative proteome atlas of a model cyanobacterium

Cyanobacteria are a group of oxygenic photosynthetic bacteria with great potentials in biotechnological applications and advantages as models for photosynthesis research. The subcellular localizations of the majority of proteins in any cyanobacteria remain undetermined, representing a major challenge in using cyanobacteria for both basic and industrial researches. Here, using label-free quantitative proteomics, we map 2027 proteins of Synechocystis sp. PCC6803, a model cyanobacterium, to different subcellular compartments and generate a proteome atlas with such information. The atlas leads to numerous unexpected but important findings, including the predominant localization of the histidine kinases Hik33 and Hik27 on the thylakoid but not the plasma membrane. Such information completely changes the concept regarding how the two kinases are activated. Together, the atlas provides subcellular localization information for nearly 60% proteome of a model cyanobacterium, and will serve as an important resource for the cyanobacterial research community.

Mining of Cyanobacterial Genomes Indicates Natural Product Biosynthetic Gene Clusters Located in Conjugative Plasmids.

Microbial natural products are compounds with unique chemical structures and diverse biological activities. Cyanobacteria commonly possess a wide range of biosynthetic gene clusters (BGCs) to produce natural products. Although natural product BGCs have been found in almost all cyanobacterial genomes, little attention has been given in cyanobacterial research to the partitioning of these biosynthetic pathways in chromosomes and plasmids. Cyanobacterial plasmids are believed to disperse several natural product BGCs, such as toxins, by plasmids through horizontal gene transfer. Therefore, plasmids may confer the ability to produce toxins and may play a role in the evolution of diverse natural product BGCs from cyanobacteria. Here, we performed an analysis of the distribution of natural product BGCs in 185 genomes and mapped the presence of genes involved in the conjugation in plasmids. The 185 analyzed genomes revealed 1817 natural products BGCs. Individual genomes contained 1–42 biosynthetic pathways (mean 8), 95% of which were present in chromosomes and the remaining 5% in plasmids. Of the 424 analyzed cyanobacterial plasmids, 12% contained homologs of genes involved in conjugation and natural product biosynthetic pathways. Among the biosynthetic pathways in plasmids, manual curation identified those to produce aeruginosin, anabaenopeptin, ambiguine, cryptophycin, hassallidin, geosmin, and microcystin. These compounds are known toxins, protease inhibitors, odorous compounds, antimicrobials, and antitumorals. The present study provides in silico evidence using genome mining that plasmids may be involved in the distribution of natural product BGCs in cyanobacteria. Consequently, cyanobacterial plasmids have importance in the context of biotechnology, water management, and public health risk assessment. Future research should explore in vivo conjugation and the end products of natural product BGCs in plasmids via chemical analyses.

Progressive transitional studies of engineered Synechococcus from laboratory to outdoor pilot-scale cultivation for production of ethylene

Cyanobacterial research is impeded by the substantial discrepancies between laboratory studies and outdoor performances, despite successful demonstrations of genetically engineered strains for array of compounds. Therefore, evaluation of adaptive responses is necessary to achieve outdoor scale-up cultivation of cyanobacteria. Under current study, cyanobacterium Synechococcus elongatusPCC7942 engineered for ethylene biosynthesis, was gradually acclimatised, ensuring sustained and progressive transition from laboratory to outdoor conditions. Bubble size of 4.9 ± 0.2 mm and air-flow rate of 0.05 vvm in BG11 supplemented with 5 g/L bicarbonate giving mass transfer coefficient (KLa) of 10.48 h−1 yielded highest specific growth rate (0.24 h−1) with the transformants. At the 100 L photobioreactor scale, ethylene productivity of 1.5 mL.L-1.h−1 was achieved. A comprehensive investigation on photosynthetic responses of the transformants adapted to the outdoor conditions exhibited interesting photosynthetic electron transport regulations, involving antenna density modulation in response to diurnal and dynamic light transitions, indicating successful transition.

Response of Key Metabolites during a UV-A Exposure Time-Series in the Cyanobacterium Chlorogloeopsis fritschii PCC 6912.

Ultraviolet A (UV-A) is the major component of UV radiation reaching the Earth’s surface, causing indirect damage to photosynthetic organisms via the production of reactive oxygen species (ROS). In comparison, UV-B causes both direct damage to biomolecules and indirect damage. UV-B is well studied in cyanobacterial research due to their long evolutionary history and adaptation to high levels of UV, with less work on the effects of UV-A. In this study, the response of key metabolites in Chlorogloeopsis fritschii (C. fritschii) during 48 h of photosynthetically active radiation (PAR, 15 µmol·m−2·s−1) supplemented with UV-A (11 µmol·m−2·s−1) was investigated using gas chromatography- mass spectrometry (GC-MS). Results showed an overall significant increase in metabolite levels up to 24 h of UV-A exposure. Compared with previously reported UV-B (PAR + UV-B) and PAR only results, UV-A showed more similarity compared to PAR only exposure as opposed to supplemented UV-B. The amino acids glutamate, phenylalanine and leucine showed differences in levels between UV (both supplemented UV-A and supplemented UV-B) and PAR only (non-supplemented PAR), hinting to their relevance in UV stress response. The fatty acids, palmitic and stearic acid, showed positive log2 fold-change (FC) in supplemented UV-A and PAR only experiments but negative log2 FC in UV-B, indicating the more harmful effect of UV-B on primary metabolism. Less research has been conducted on UV-A exposure and cyanobacteria, a potential environmental stimuli for the optimisation of metabolites for industrial biotechnology. This study will add to the literature and knowledge on UV-A stress response at the metabolite level in cyanobacteria, especially within the less well-known species C. fritschii.

Optimization of a T7-RNA polymerase system in Synechococcus sp. PCC 7002 mirrors the protein overproduction phenotype from E. coli BL21(DE3)

With the goal of expanding the diversity of tools available for controlling gene expression in cyanobacteria, the T7-RNA polymerase gene expression system from E. coli BL21(DE3) was adapted and systematically engineered for robust function Synechococcus sp. PCC 7002, a fast-growing saltwater strain. Expression of T7-RNA polymerase was controlled via LacI regulation, while functionality was optimized by both further tuning its expression level along with optimizing the translation initiation region of the expressed gene, in this case an enhanced YFP reporter. Under high CO2 conditions, the resulting system displayed a 60-fold dynamic range in expression levels. Furthermore, when maximally induced, T7-RNA polymerase-dependent protein production constituted up to two-thirds of total cellular protein content in Synechococcus sp. PCC 7002. Ultimately, however, this came at the cost of 40% reductions in both biomass and pigmentation levels. Taken together, the developed T7-RNA polymerase gene expression system is effective for controlling and achieving high-level expression of heterologous genes in Synechococcus sp. PCC 7002, making it a valuable tool for cyanobacterial research. KEY POINTS: • Promoter driving T7-RNA polymerase was optimized. • Up to 60-fold dynamic range in expression, depending on CO2 conditions. • Two-thirds of total protein is T7-RNA polymerase dependent.

Blue-Green Algae in Surface Water: Problems and Opportunities

Cyanobacteria, commonly known as blue-green algae, are often seen as a problem. Their accumulation (bloom) in surface water can cause toxicity and aesthetic concerns. Efforts have been made in preventing and managing cyanobacterial blooms. By contrast, purposeful cultivation of cyanobacteria can create great opportunities in food, chemical and biofuel applications. This review summarises the current stage of research and the socio-economic impacts associated with both the problems and opportunities induced from the presence of cyanobacteria in surface water. Insightful knowledge of factors that trigger cyanobacterial blooms has allowed for the development of prevention and control strategies. Advanced technologies are utilised to detect, quantify and treat cyanobacterial biomass and cyanotoxins in a timely manner. Additionally, understanding of cyanobacterial biochemical properties enables their applications in food and health industry, agriculture and biofuel production. Researchers have been able to genetically modify several cyanobacterial strains to obtain a direct pathway for ethanol and hydrogen production. Cyanobacterial blooms have been effectively addressed with advances technologies and cyanobacterial research. However, this review identified a knowledge gap regarding cyanotoxin synthesis and the relevant environmental triggers. This information is essential for developing measures to prevent cyanobacterial blooms. Additionally, this review affirms the promising opportunities that cyanobacteria offer in the food, cosmetics, pigments and agriculture. Biofuel production from cyanobacterial biomass presents an immense potential but is currently constrained by the cultivation process. Thus, future research should strive to achieve effective mass harvesting of cyanobacterial biomass and obtain a profound understanding of cyanotoxin production.

Emerging Species and Genome Editing Tools: Future Prospects in Cyanobacterial Synthetic Biology.

Recent advances in synthetic biology and an emerging algal biotechnology market have spurred a prolific increase in the availability of molecular tools for cyanobacterial research. Nevertheless, work to date has focused primarily on only a small subset of model species, which arguably limits fundamental discovery and applied research towards wider commercialisation. Here, we review the requirements for uptake of new strains, including several recently characterised fast-growing species and promising non-model species. Furthermore, we discuss the potential applications of new techniques available for transformation, genetic engineering and regulation, including an up-to-date appraisal of current Clustered Regularly Interspaced Short Palindromic Repeats/CRISPR associated protein (CRISPR/Cas) and CRISPR interference (CRISPRi) research in cyanobacteria. We also provide an overview of several exciting molecular tools that could be ported to cyanobacteria for more advanced metabolic engineering approaches (e.g., genetic circuit design). Lastly, we introduce a forthcoming mutant library for the model species Synechocystis sp. PCC 6803 that promises to provide a further powerful resource for the cyanobacterial research community.

The Diversity of Cyanobacterial Toxins on Structural Characterization, Distribution and Identification: A Systematic Review.

The widespread distribution of cyanobacteria in the aquatic environment is increasing the risk of water pollution caused by cyanotoxins, which poses a serious threat to human health. However, the structural characterization, distribution and identification techniques of cyanotoxins have not been comprehensively reviewed in previous studies. This paper aims to elaborate the existing information systematically on the diversity of cyanotoxins to identify valuable research avenues. According to the chemical structure, cyanotoxins are mainly classified into cyclic peptides, alkaloids, lipopeptides, nonprotein amino acids and lipoglycans. In terms of global distribution, the amount of cyanotoxins are unbalanced in different areas. The diversity of cyanotoxins is more obviously found in many developed countries than that in undeveloped countries. Moreover, the threat of cyanotoxins has promoted the development of identification and detection technology. Many emerging methods have been developed to detect cyanotoxins in the environment. This communication provides a comprehensive review of the diversity of cyanotoxins, and the detection and identification technology was discussed. This detailed information will be a valuable resource for identifying the various types of cyanotoxins which threaten the environment of different areas. The ability to accurately identify specific cyanotoxins is an obvious and essential aspect of cyanobacterial research.

CyanoBase:a large-scale update on its 20th anniversary.

The first ever cyanobacterial genome sequence was determined two decades ago and CyanoBase (http://genome.microbedb.jp/cyanobase), the first database for cyanobacteria was simultaneously developed to allow this genomic information to be used more efficiently. Since then, CyanoBase has constantly been extended and has received several updates. Here, we describe a new large-scale update of the database, which coincides with its 20th anniversary. We have expanded the number of cyanobacterial genomic sequences from 39 to 376 species, which consists of 86 complete and 290 draft genomes. We have also optimized the user interface for large genomic data to include the use of semantic web technologies and JBrowse and have extended community-based reannotation resources through the re-annotation of Synechocystis sp. PCC 6803 by the cyanobacterial research community. These updates have markedly improved CyanoBase, providing cyanobacterial genome annotations as references for cyanobacterial research.

Cyanobacterial KnowledgeBase (CKB), a Compendium of Cyanobacterial Genomes and Proteomes.

Cyanobacterial KnowledgeBase (CKB) is a free access database that contains the genomic and proteomic information of 74 fully sequenced cyanobacterial genomes belonging to seven orders. The database also contains tools for sequence analysis. The Species report and the gene report provide details about each species and gene (including sequence features and gene ontology annotations) respectively. The database also includes cyanoBLAST, an advanced tool that facilitates comparative analysis, among cyanobacterial genomes and genomes of E. coli (prokaryote) and Arabidopsis (eukaryote). The database is developed and maintained by the Sub-Distributed Informatics Centre (sponsored by the Department of Biotechnology, Govt. of India) of the National Facility for Marine Cyanobacteria, a facility dedicated to marine cyanobacterial research. CKB is freely available at http://nfmc.res.in/ckb/index.html.

<p style="page-break-inside: avoid;">Nodularia (Cyanobacteria, Nostocaceae): a phylogenetically uniform genus with variable phenotypes

The taxonomy of cyanobacteria currently faces the challenge of overhauling the traditional system to better reflect the results of phylogenetic analyses. In the present study, we assessed the phylogenetic position, morphological variability, ability to produce the toxin nodularin, and source habitat of 17 benthic and soil isolates of Nodularia. A combined analysis of two loci (partial 16S rRNA gene and rbcLX region) confirmed the genus as a monophyletic unit and the close relationship of its members. However, the taxonomic resolution at the subgeneric level was extremely problematic. The phylogenetic clustering did not show any reasonable congruence with either morphological or ecological features commonly used to separate taxa in heterocytous cyanobacteria. Despite the near phylogenetic similarity of planktonic, benthic and soil Nodularia strains, we did not find any new nodularin-producing strains among the non-planktonic isolates. The relatively low variability in conserved molecular markers within the genus Nodularia exemplifies the limitations of the currently accepted taxonomic workflow and polyphasic approach. Elucidation of mechanisms that drive the phenotypic variability in such groups presents a major challenge in cyanobacterial research.

An overview of cyanobacterial research and management in South Africa post-2000

The quality of South Africa’s water resources is becoming increasingly impaired by eutrophication, to the extent that regional crises now prevail. Thirty-five percent of the total storage is eutrophic to hypertrophic – including grosslyenriched reservoirs that far exceed the globally-accepted definition of hypertrophy. Failing infrastructure contributes to the problem of polluted urban runoff which comprises a significant fraction of flows to inland reservoirs, particularly in the landlocked, economic heartland of the country. In the period post-2000, a number of developments have underpinned theprovision of a research and management infrastructure that ensures that South African water resource management structures are capable of dealing with cyanobacterial problems. This paper details the progress and achievements that have been made during this period.

Current trends in high throughput proteomics in cyanobacteria

Advancements in genome sequencing and high throughput proteomics of cyanobacterial strains led to 13 published reports, from a small number of laboratories. These successful studies focused on Synechocystis, Nostoc and Anabaena strains, prochlorococcus, and halotolerant Euhalothece. The implications of emerging quantitative aspects developed and applied in these large-scale studies are assessed in the wake of advanced cyanobacterial research. Furthermore, contributions from traditional and early high throughput analysis of cyanobacterial proteomics are compared and summarised. Finally, opinions are provided to link both the trends and the future challenges. This review aims to push the synergy between proteomics and cyanobacterial research to improve both the technical and biological significance.

Quantitative analysis of expression of two circadian clock-controlled gene clusters coding for the bidirectional hydrogenase in the cyanobacterium Synechococcus sp. PCC7942.

Hydrogen metabolism is of central interest in cyanobacterial research because of its potential applications. The gene expression and physiological role of the cyanobacterial bidirectional NAD(P)+-reducing hydrogenase are poorly understood. Transcription rates of hoxEF and hoxUYH encoding this enzyme have been studied in Synechococcus sp. PCC7942. PhoxU activity was about three times higher than that of PhoxE. Circadian phasing of both promoters was found to be synchronous and influenced expression levels by at least one order of magnitude. This is the first demonstration of circadian control of gene expression for any hydrogenase. For the majority of PhoxU-driven messages, transcription presumably terminates between hoxU and hoxH. Being part of a polycistronic hoxUYHW... operon, hoxW, encoding a protease involved in C-terminal processing of the hydrogenase large-subunit HoxH, is mainly expressed by its own promoter, PhoxW. The complex transcript formation may be a key feature for controlling bidirectional hydrogenase expression in vivo.

The First Maj and Tor Nessling Foundation Symposium on ‘Recent Developments in Cyanobacterial Research’

The First Maj and Tor Nessling Foundation Symposium on ‘Recent Developments in Cyanobacterial Research’

Cyanobacteria in New Zealand inland waters: Experimental studies

Abstract A small volume of experimental work with cyanobacteria in New Zealand has been produced through the last two decades. Except for some studies with geothermal organisms biased to New Zealand conditions, most work has followed the world‐wide trends in cyanobacterial research. There has been some emphasis on N2‐fixing species, but symbiotic relationships, metabolic kinetics, nutrient and other environmental factors promoting ecological dominance, and toxicity have been investigated.