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1

Establishment of the reporter system for a thylakoid-lacking cyanobacterium, Gloeobacter violaceus PCC 7421  

PubMed Central

Gloeobacter violaceus PCC 7421 is considered, by molecular phylogenetic analyses, to be an early-branching cyanobacterium within the cyanobacterial clade. G. violaceus is the only known oxygenic photosynthetic organism that lacks thylakoid membranes. There is only one report on the development of a transformation system for G. violaceus [H. Guo, X. Xu, Prog. Nat. Sci. 14 (2004) 31–35] and further studies using the system have not been reported. In the present study, we succeeded in introducing an expression vector (pKUT1121) derived from a broad-host-range plasmid, RSF1010, into G. violaceus by conjugation. The frequency of transformation of our system is significantly higher than that described in the previous report. In addition, luciferase heterologously expressed in G. violaceus functioned as a reporter. The established system will promote the molecular genetic studies on G. violaceus.

Araki, Mie; Shimada, Yuichiro; Mimuro, Mamoru; Tsuchiya, Tohru

2012-01-01

2

The plasma membrane of the cyanobacterium Gloeobacter violaceus contains segregated bioenergetic domains.  

PubMed

The light reactions of oxygenic photosynthesis almost invariably take place in the thylakoid membranes, a highly specialized internal membrane system located in the stroma of chloroplasts and the cytoplasm of cyanobacteria. The only known exception is the primordial cyanobacterium Gloeobacter violaceus, which evolved before the appearance of thylakoids and harbors the photosynthetic complexes in the plasma membrane. Thus, studies on G. violaceus not only shed light on the evolutionary origin and the functional advantages of thylakoid membranes but also might include insights regarding thylakoid formation during chloroplast differentiation. Based on biochemical isolation and direct in vivo characterization, we report here structural and functional domains in the cytoplasmic membrane of a cyanobacterium. Although G. violaceus has no internal membranes, it does have localized domains with apparently specialized functions in its plasma membrane, in which both the photosynthetic and the respiratory complexes are concentrated. These bioenergetic domains can be visualized by confocal microscopy, and they can be isolated by a simple procedure. Proteomic analysis of these domains indicates their physiological function and suggests a protein sorting mechanism via interaction with membrane-intrinsic terpenoids. Based on these results, we propose specialized domains in the plasma membrane as evolutionary precursors of thylakoids. PMID:21642550

Rexroth, Sascha; Mullineaux, Conrad W; Ellinger, Dorothea; Sendtko, Esther; Rögner, Matthias; Koenig, Friederike

2011-06-03

3

The Plasma Membrane of the Cyanobacterium Gloeobacter violaceus Contains Segregated Bioenergetic Domains[C][W  

PubMed Central

The light reactions of oxygenic photosynthesis almost invariably take place in the thylakoid membranes, a highly specialized internal membrane system located in the stroma of chloroplasts and the cytoplasm of cyanobacteria. The only known exception is the primordial cyanobacterium Gloeobacter violaceus, which evolved before the appearance of thylakoids and harbors the photosynthetic complexes in the plasma membrane. Thus, studies on G. violaceus not only shed light on the evolutionary origin and the functional advantages of thylakoid membranes but also might include insights regarding thylakoid formation during chloroplast differentiation. Based on biochemical isolation and direct in vivo characterization, we report here structural and functional domains in the cytoplasmic membrane of a cyanobacterium. Although G. violaceus has no internal membranes, it does have localized domains with apparently specialized functions in its plasma membrane, in which both the photosynthetic and the respiratory complexes are concentrated. These bioenergetic domains can be visualized by confocal microscopy, and they can be isolated by a simple procedure. Proteomic analysis of these domains indicates their physiological function and suggests a protein sorting mechanism via interaction with membrane-intrinsic terpenoids. Based on these results, we propose specialized domains in the plasma membrane as evolutionary precursors of thylakoids.

Rexroth, Sascha; Mullineaux, Conrad W.; Ellinger, Dorothea; Sendtko, Esther; Rogner, Matthias; Koenig, Friederike

2011-01-01

4

The phycocyanin-associated rod linker proteins of the phycobilisome of Gloeobacter violaceus PCC 7421 contain unusually located rod-capping domains  

Microsoft Academic Search

Gloeobacter violaceus PCC 7421 is a unique cyanobacterium that has no thylakoids and whose genome has been sequenced [Y. Nakamura, T. Kaneko, S. Sato, M. Mimuro, H. Miyashita, T. Tsuchiya, S. Sasamoto, A. Watanabe, K. Kawashima, Y. Kishida, C. Kiyokawa, M. Kohara, M. Matsumoto, A. Matsuno, N. Nakazaki, S. Shimpo, C. Takeuchi, M. Yamada, S. Tabata, Complete Genome Structure of

Emma Berta Gutiérrez-Cirlos; Bertha Pérez-Gómez; David W. Krogmann; Carlos Gómez-Lojero

2006-01-01

5

Conformational Transitions Underlying Pore Opening and Desensitization in Membrane-embedded Gloeobacter violaceus Ligand-gated Ion Channel (GLIC)  

PubMed Central

Direct structural insight into the mechanisms underlying activation and desensitization remain unavailable for the pentameric ligand-gated channel family. Here, we report the structural rearrangements underlying gating transitions in membrane-embedded GLIC, a prokaryotic homologue, using site-directed spin labeling and electron paramagnetic resonance (EPR) spectroscopy. We particularly probed the conformation of pore-lining second transmembrane segment (M2) under conditions that favor the closed and the ligand-bound desensitized states. The spin label mobility, intersubunit spin-spin proximity, and the solvent-accessibility parameters in the two states clearly delineate the underlying protein motions within M2. Our results show that during activation the extracellular hydrophobic region undergoes major changes involving an outward translational movement, away from the pore axis, leading to an increase in the pore diameter, whereas the lower end of M2 remains relatively immobile. Most notably, during desensitization, the intervening polar residues in the middle of M2 move closer to form a solvent-occluded barrier and thereby reveal the location of a distinct desensitization gate. In comparison with the crystal structure of GLIC, the structural dynamics of the channel in a membrane environment suggest a more loosely packed conformation with water-accessible intrasubunit vestibules penetrating from the extracellular end all the way to the middle of M2 in the closed state. These regions have been implicated to play a major role in alcohol and drug modulation. Overall, these findings represent a key step toward understanding the fundamentals of gating mechanisms in this class of channels.

Velisetty, Phanindra; Chalamalasetti, Sreevatsa V.; Chakrapani, Sudha

2012-01-01

6

Assimilation efficiency of a temperate-zone intertidal fish ( Cebidichthys violaceus ) fed diets of macroalgae  

Microsoft Academic Search

We studied assimilation efficiencies of the temperate-zone intertidal fish Cebidichthys violaceus (Girard, 1854) fed in the laboratory on each of the following species of macroalgae: Spongomorpha coalita (Chlorophyta), Ulva lobata (Chlorophyta), Iridaea flaccida (Rhodophyta) and Porphyra perforata (Rhodophyta). Together, these 4 algae make up over 75% of the natural summer diet of C. violaceus. Assimilation efficiency was calculated by proximate

T. W. Edwards; M. H. Horn

1982-01-01

7

Salt bridge in the conserved His-Asp cluster in Gloeobacter rhodopsin contributes to trimer formation.  

PubMed

Gloeobacter rhodopsin (GR) is a eubacterial proton pump having a highly conserved histidine near the retinal Schiff base counter-ion, aspartate. Various interactions between His and Asp of the eubacterial proton pump have been reported. Here, we showed the pH-dependent trimer/monomer transition of GR in the presence of dodecyl-?-D-maltoside by size-exclusion chromatography. The pH dependence was closely related to the protonation state of the counter-ion, Asp121. For the H87M mutant, pH dependence disappeared and a monomer became dominant. We concluded that the formation or breaking of the salt bridge between His87 and Asp121 inside the protein changes the quaternary structure. PMID:23313943

Tsukamoto, Takashi; Kikukawa, Takashi; Kurata, Takuro; Jung, Kwang-Hwan; Kamo, Naoki; Demura, Makoto

2013-01-09

8

Parental Genome Separation and Elimination of Cells and Chromosomes Revealed by AFLP and GISH analyses in a Brassica carinata · Orychophragmus violaceus Cross  

Microsoft Academic Search

? Background and Aims The phenomenon of parental genome separation during the mitotic divisions of hybrid cells was proposed to occur under genetic control in intergeneric hybrids between cultivated Brassica species and Orychophragmus violaceus (2n = 24). To elucidate further the cytological and molecular mechanisms behind parental genome separation, Brassica carinata (2n = 34) · O. violaceus hybrids were resynthesized

YU-WEI H UA; AI-YUN L I

9

Parental Genome Separation and Elimination of Cells and Chromosomes Revealed by AFLP and GISH analyses in a Brassica carinata x Orychophragmus violaceus Cross  

Microsoft Academic Search

? Background and Aims The phenomenon of parental genome separation during the mitotic divisions of hybrid cells was proposed to occur under genetic control in intergeneric hybrids between cultivated Brassica species and Orychophragmus violaceus (2n = 24). To elucidate further the cytological and molecular mechanisms behind parental genome separation, Brassica carinata (2n = 34) · O. violaceus hybrids were resynthesized

YU-WEI HUA; MIN LIU; ZAI-YUN LI

2006-01-01

10

Two copepod species largely confused: Asterocheres echinicola (Norman, 1868) and A. violaceus (Claus, 1889). Taxonomical implications  

NASA Astrophysics Data System (ADS)

Due to its extremely brief description, Asterocheres echinicola (Norman, 1868) has been confused with some Asterocheres species such as Asterocheres suberitis Giesbrecht, 1897, Asterocheres parvus Giesbrecht, 1897 and Asterocheres latus (Brady, 1872). Furthermore, this species has been considered conspecific with Cyclopicera lata (Brady, 1872) and Asterocheres kervillei Canu, 1898. The objective of this paper is to study the syntypes of Asterocheres echinicola deposited in the Museum of Natural History of London together with abundant material from this and other institutions. Re-examination of these syntypes revealed that Asterocheres echinicola was conspecific with the currently known Asterocheres species, A. violaceus. Therefore, this latter species should be considered as a junior synonym of the former. The specimens described by Brady as Cyclopicera lata represent distinctively Asterocheres echinicola (= Asterocheres violaceus) and are identical to Sars’s Ascomyzom parvum and to Giesbrecht’s Asterocheres echinicola. We propose to rename Cyclopicera lata as Asterocheres latus (Brady, 1872), and raise Sars’ Ascomyzon latus, a species which is different from Asterocheres echinicola (= Asterocheres violaceus) and from Asterocheres latus (= Cyclopicera lata), as a new species. In this paper, we not only redescribe both species A. echinicola and A. latus, but also compare them with their previous descriptions, with the new material available and with their congeners. The redescription of Asterocheres latus revealed new specific differences between this species and Asterocheres kervillei, a species considered as synonymous of Asterocheres latus for almost 40 years. We strongly recommend that these differences are sufficient to consider these two species different. Finally, we analyzed the implications of all these taxonomical changes with respect to the diversity of the hosts utilized by these copepods and their geographical distribution.

Bandera, M. Eugenia; Conradi, Mercedes

2009-12-01

11

Growth, consumption, assimilation and excretion in the marine herbivorous fish Cebidichthys violaceus (Girard) fed natural and high protein diets  

Microsoft Academic Search

Laboratory feeding trials were conducted to determine the growth and food processing rates and gut retention times of a wild, cool-temperate zone fish fed its simulated summer diet of seaweeds and two higher protein diets more typical of those of carnivorous fishes. When this stichaeid fish, Cebidichthys violaceus (Girard), was fed for 12 wk on a pelleted natural summer diet

Michael H. Horn; Kristin F. Mailhiot; Michael B. Fris; Lon L. McClanahan

1995-01-01

12

Assimilatory sulfur metabolism in marine microorganisms: sulfur metabolism, protein synthesis, and growth of Alteromonas luteo - violaceus and Pseudomonas halodurans during perturbed batch growth  

Microsoft Academic Search

The antibiotic protein synthesis inhibitor chloramphenicol specifically blocked the incorporation of (35 S) sulfate into the residue protein of two marine bacteria, Pseudomonas halodurans and Alteromonas luteo-violaceus. Simultaneous inhibition of total protein synthesis occurred, but incorporation of 35 S into low-molecular-weight organic compounds continued. A. luteo-violaceus rapidly autolyzed, with similar reduction in cell counts, total culture protein and cellular sulfur,

R. L. Cuhel; C. D. Taylor; H. W. Jannasch

1982-01-01

13

Parental Genome Separation and Elimination of Cells and Chromosomes Revealed by AFLP and GISH analyses in a Brassica carinata x Orychophragmus violaceus Cross  

PubMed Central

• Background and Aims The phenomenon of parental genome separation during the mitotic divisions of hybrid cells was proposed to occur under genetic control in intergeneric hybrids between cultivated Brassica species and Orychophragmus violaceus (2n = 24). To elucidate further the cytological and molecular mechanisms behind parental genome separation, Brassica carinata (2n = 34) × O. violaceus hybrids were resynthesized and their chromosome/genomic complements analysed. • Methods F1 hybrids of the cross were obtained following embryo rescue, and were investigated for their cytological behaviour and subjected to genomic in situ hybridization (GISH) and amplified fragment length polymorphism (AFLP) to determine the contribution of parental genomes. • Key Results All the F1 plants with high fertility closely resembled B. carinata in morphological attributes. These were mixoploids with 2n chromosome numbers ranging from 17 to 35; however, 34, the same number as in B. carinata, was the most frequent number of chromosomes in ovary and pollen mother cells (PMCs). GISH clearly identified 16 chromosomes of B. nigra in ovary cells and PMCs with 2n = 34 and 35. However, no O. violaceus chromosome was detected, indicating the presence of the intact B. carinata genome and elimination of the entire O. violaceus genome. However, some AFLP bands specific for O. violaceus and novel for the two parents were detected in the leaves. Cells with fewer than 34 chromosomes had lost some B. oleracea chromosomes. F2 plants were predominantly like B. carinata, but some contained O. violaceus characters. • Conclusions The cytological mechanism for the results involves complete and partial genome separation at mitosis in embryos of F1 plants followed by chromosome doubling, elimination of cells with O. violaceus chromosomes and some introgression of O. violaceus genetic information.

HUA, YU-WEI; LIU, MIN; LI, ZAI-YUN

2006-01-01

14

Two new bioactive triterpene glycosides from the sea cucumber Pseudocolochirus violaceus.  

PubMed

By activity-guided fractionation, two new triterpene glycosides, violaceusides A (1) and B (2), were isolated from the sea cucumber Pseudocolochirus violaceus as active compounds causing morphological abnormality of Pyricularia oryzae mycelia. By extensive 2D NMR techniques and chemical evidence, the structures of the two new glycosides were established as 16beta-acetoxy-3-O-[3-O-methyl-beta-D-glucopyranosyl-(1 --> 3)-beta-D-xylopyranosyl-(1 --> 4)-beta-D-quinovopyranosyl-(1 --> 2)-4-O-sodiumsulphate-beta-D-xylopyranosyl]-holosta-7,24-diene-3beta-ol (1) and 16beta-acetoxy-3-O-[3-O-methyl-beta-D-glucopyranosyl-(1 --> 3)-beta-D-xylopyranosyl-(1 --> 4)-beta-D-glucopyranosyl-(1 --> 2)-4-O-sodiumsulphate-beta-D-xylopyranosyl]-holosta-7,24-diene-3beta-ol (2), respectively. The two glycosides also exhibited significant cytotoxicity against HL-60 and BEL-7402 cancer cell lines. PMID:16753775

Zhang, Shu-Yu; Yi, Yang-Hua; Tang, Hai-Feng; Li, Ling; Sun, Peng; Wu, Jun

15

Genetic variation within and among populations of Orychophragmus violaceus (Cruciferae) in China as detected by ISSR analysis  

Microsoft Academic Search

Orychophragmus violaceus, a ground covering plant that is widely distributed in China. It has both high economical value in food, forage, health care\\u000a and ornamental value in gardening. In this study, the genetic diversity of 245 individuals from nine populations in China\\u000a were investigated using the inter-simple sequence repeat markers. Of the 100 primers screened, eight were highly polymorphic.\\u000a Using

Li-Jun Zhang; Si-Lan Dai

2010-01-01

16

Assimilatory sulfur metabolism in marine microorganisms: Sulfur metabolism, growth, and protein synthesis of Pseudomonas halodurans and Alteromonas luteo-violaceus during sulfate limitation  

Microsoft Academic Search

Sulfate concentration in the growth medium exerted a strong influence on the sulfur content of protein in two marine bacteria, Pseudomonas halodurans and Alteromonasluteo-violaceus, but the distribution of sulfur in major biochemical fractions was not affected. 90% of the total cellular sulfur was contained in low molecular weight organic compounds and protein; inorganic sulfate was not an important component. The

Russell L. Cuhel; Craig D. Taylor; Holger W. Jannasch

1981-01-01

17

Growth and reproductive cycles of the marine fouling ascidians Ciona intestinalis, Styela plicata, Botrylloides violaceus , and Leptoclinum mitsukurii at Aburatsubo-Moroiso Inlet (central Japan)  

Microsoft Academic Search

Experimental cultivations of post-metamorphosis juveniles were repeated in different seasons for Ciona intestinalis (L.) and Styela plicata (Lesueur) (simple ascidians) as well as for Leptolinum mitsukurii (Oka) and Botrylloides violaceus Oka (compound ascidians) at Aburatsubo-Moroiso Inlet near Misaki Biological Station (Japan). Growth in body length of simple ascidians was exponential during juvenile development up to near sexual maturity. C. intestinalis

M. Yamaguchi

1975-01-01

18

Assimilatory sulfur metabolism in marine microorganisms: Sulfur metabolism, protein synthesis, and growth of Pseudomonas halodurans and Alteromonas luteo-violaceus during unperturbed batch growth  

Microsoft Academic Search

Analysis of the distribution of 35S-sulfate and 14C-glutamate in major biochemical components of the two marine bacteria, Pseudomonas halodurans and Alteromonas luteo-violaceus, was compared with cell density and total cellular protein during exponential growth in batch culture. For both organisms, the sulfur distribution was restricted principally to the low molecular weight organic and protein fractions, which together accounted for over

Russell L. Cuhel; Craig D. Taylor; Holger W. Jannasch

1981-01-01

19

Assimilatory sulfur metabolism in marine microorganisms: sulfur metabolism, protein synthesis, and growth of Alteromonas luteo - violaceus and Pseudomonas halodurans during perturbed batch growth  

SciTech Connect

The antibiotic protein synthesis inhibitor chloramphenicol specifically blocked the incorporation of (35 S) sulfate into the residue protein of two marine bacteria, Pseudomonas halodurans and Alteromonas luteo-violaceus. Simultaneous inhibition of total protein synthesis occurred, but incorporation of 35 S into low-molecular-weight organic compounds continued. A. luteo-violaceus rapidly autolyzed, with similar reduction in cell counts, total culture protein and cellular sulfur, whereas P. halodurans remained viable. Treatment with chloramphenicol, growth during nitrogen and carbon limitation, and the carbon and energy sources used for growth did not alter the sulfur content of P. halodurans protein. The mean value (1.09%, by weight), representing a wide variety of environmentally relevant growth conditions, was in agreement with model protein composition. The variability of cellular composition of P. halodurnas and A. luteo-violaceus is discussed with respect to the measurement of bacterial growth in natural environments. Total carbon and nitrogen per cell varied greatly (coefficient of variation, ca. 100%) depending on growth conditions. Variation in total sulfur and protein per cell was much less (coefficient of variation, less than 50%), but the least variation was found for sulfate incorporation into residue protein (coefficient of variation, ca. 15%). Thus, sulfate incorporation into residue protein can be used as an accurate measurement of de novo protein synthesis in these bacteria. (Refs. 26).

Cuhel, R.L.; Taylor, C.D.; Jannasch, H.W.

1982-01-01

20

The effect of temperature on the germination of Melocactus violaceus Pfeiff. (Cactaceae), a threatened species in restinga sandy coastal plain of Brazil.  

PubMed

Melocactus violaceus is an endangered species due to habitat destruction and the overcollection of this species for ornamental use. The aim of this study was to test the effect of different temperatures on the germination of M. violaceus. Three treatments were conducted: a constant temperature of 25ºC, a 20-35ºC alternating temperature, both inside germination chamber, and an alternating temperature under room temperature (mean temperature ranged from 25-37ºC). The final seed germination rates at the alternating temperature treatments were not significantly different (65% in the seed germinator and 62.5% at room condition). However, both treatments with alternating temperatures had significantly higher germination rates compared to the treatment kept at the constant temperature (8%). Our study showed that alternating temperatures between 20 and 37ºC provides satisfactory conditions to induce a high percentage of seed germination of M. violaceus, without the passage of seeds through the digestive tract of its natural disperser, the lizard Tropidurus torquatus. This condition contributes to efficiently producing seedlings that can be reintroduced into conservation areas or used as ornamentals that may help reduce the overcollection of the remaining native populations. PMID:23828368

Zamith, Luiz R; Cruz, Denise D; Richers, Bárbara T T

21

Glyoxylate metabolism in the cyanobacterium Coccochloris peniocystis  

SciTech Connect

The possible metabolism of glyoxylate via an incomplete glycolate pathway and a glyoxylate cycle in the cyanobacterium C. peniocystis was investigated. Levels of enzyme activities determined from partially purified preparations were: glycolate dehydrogenase, 1.18; isocitrate lyase, 6.67; malate synthetase, 1.03. Metabolism of {sup 14}C glycolate by intact cells was inhibited by the glycolate dehydrogenase inhibitor {alpha}-hydroxypyridyl methane sulfonate. Metabolism of {sup 14}C-glyoxylate was not inhibited by the amino-transferase inhibitor, aminooxyacetate, suggesting that formation of glycine is not an absolute requirement for glyoxylate metabolism. The lack of formation of labelled serine from {sup 14}C glyoxylate or {sup 14}C-glycine indicates that the glycolate pathway is incomplete. These results suggest that a glycolate pathway, such as found in higher plants, is absent in cyanobacteria and that some alternate pathway, possible a glyoxylate cycle, operates in the metabolism of glycolate pathway intermediates.

Norman, E.G.; Colman, B. (York Univ., Toronto, Ontario (Canada))

1989-04-01

22

Phylogeography of the Thermophilic Cyanobacterium Mastigocladus laminosus?  

PubMed Central

We have taken a phylogeographic approach to investigate the demographic and evolutionary processes that have shaped the geographic patterns of genetic diversity for a sample of isolates of the cosmopolitan thermophilic cyanobacterial Mastigocladus laminosus morphotype collected from throughout most of its range. Although M. laminosus is found in thermal areas throughout the world, our observation that populations are typically genetically differentiated on local geographic scales suggests the existence of dispersal barriers, a conclusion corroborated by evidence for genetic isolation by distance. Genealogies inferred using nitrogen metabolism gene sequence data suggest that a significant amount of the extant global diversity of M. laminosus can be traced back to a common ancestor associated with the western North American hot spot currently located below Yellowstone National Park. Estimated intragenic recombination rates are comparable to those of pathogenic bacteria known for their capacity to exchange DNA, indicating that genetic exchange has played an important role in generating novel variation during M. laminosus diversification. Selection has constrained protein changes at loci involved in the assimilation of both dinitrogen and nitrate, suggesting the historic use of both nitrogen sources in this heterocystous cyanobacterium. Lineage-specific differences in thermal performance were also observed.

Miller, Scott R.; Castenholz, Richard W.; Pedersen, Deana

2007-01-01

23

Chlorpyrifos degradation by the cyanobacterium Synechocystis sp. strain PUPCCC 64  

Microsoft Academic Search

Background, aim, and scope  Indiscriminate use of insecticides leads to environmental problems and poses a great threat to beneficial microorganisms.\\u000a The aim of the present work was to study chlorpyrifos degradation by a rice field cyanobacterium Synechocystis sp. strain PUPCCC 64 so that the organism is able to reduce insecticide pollution in situ.\\u000a \\u000a \\u000a \\u000a \\u000a Material and methods  The unicellular cyanobacterium isolated and purified

D. P. Singh; J. I. S. Khattar; J. Nadda; Y. Singh; A. Garg; N. Kaur; A. Gulati

24

Two tuf genes in the cyanobacterium Spirulina platensis.  

PubMed Central

Probes derived from the tufA gene of Escherichia coli have been utilized to detect homologous sequences on Spirulina platensis DNA. A 6-kilobase-pair fragment of S. platensis DNA appears to contain two sequences homologous to the E. coli gene. Thus, as reported for gram-negative bacteria, the cyanobacterium presumably contains two tuf genes. Images

Tiboni, O; Di Pasquale, G; Ciferri, O

1984-01-01

25

Aerobic nitrogen fixation by Lyngbya sp., a marine tropical cyanobacterium  

Microsoft Academic Search

Lyngbya majuscula, a frond-forming, non-heterocystous cyanobacterium found in the sublittoral zone off Oahu, Hawaii, is capable of high rates of aerobic nitrogen fixation in the light but incapable of nitrogen fixation in the dark. L. majuscula differs therefore from heterocystous Calothrix spp., which are found in the same habitat and are capable of N2 fixation in the dark.

Keith Jones

1990-01-01

26

Fermentative metabolism to produce hydrogen gas and organic compounds in a cyanobacterium, Spirulina platensis  

Microsoft Academic Search

The non nitrogen-fixing and filamentous cyanobacterium Spirulina platensis NIES-46 produced hydrogen gas, ethanol, and low molecular organic acids auto-fermentatively under dark and anaerobic conditions. The fermentative productivity was enhanced by incubating the cyanobacterium under nitrogen-starved conditions. Cell-free extracts of the cyanobacterium catalyzed hydrogen production by the addition of acetyl-coenzyme A and pyruvate. Pyruvate-degrading and acetaldehyde dehydrogenase activities were observed in

Katsuhiro Aoyama; Ieaki Uemura; Jun Miyake; Yasuo Asada

1997-01-01

27

A primitive cyanobacterium as pioneer microorganism for terraforming Mars.  

PubMed

The primitive characteristics of the cyanobacterium Chroococcidiopsis suggest that it represents a very ancient type of the group. Its morphology is simple but shows a wide range of variability, and it resembles certain Proterozoic microfossils. Chroococcidiopsis is probably the most desiccation-resistant cyanobacterium, the sole photosynthetic organism in extreme arid habitats. It is also present in a wide range of other extreme environments, from Antarctic rocks to thermal springs and hypersaline habitats, but it is unable to compete with more specialized organisms. Genetic evidence suggests that all forms belong to a single species. Its remarkable tolerance of environmental extremes makes Chroococcidiopsis a prime candidate for use as a pioneer photosynthetic microorganism for terraforming of Mars. The hypolithic microbial growth form (which lives under stones of a desert pavement) could be used as a model for development of technologies for large-scale Martian farming. PMID:11539232

Friedmann, E I; Ocampo-Friedmann, R

1995-03-01

28

Unicellular cyanobacterium symbiotic with a single-celled eukaryotic alga.  

PubMed

Symbioses between nitrogen (N)(2)-fixing prokaryotes and photosynthetic eukaryotes are important for nitrogen acquisition in N-limited environments. Recently, a widely distributed planktonic uncultured nitrogen-fixing cyanobacterium (UCYN-A) was found to have unprecedented genome reduction, including the lack of oxygen-evolving photosystem II and the tricarboxylic acid cycle, which suggested partnership in a symbiosis. We showed that UCYN-A has a symbiotic association with a unicellular prymnesiophyte, closely related to calcifying taxa present in the fossil record. The partnership is mutualistic, because the prymnesiophyte receives fixed N in exchange for transferring fixed carbon to UCYN-A. This unusual partnership between a cyanobacterium and a unicellular alga is a model for symbiosis and is analogous to plastid and organismal evolution, and if calcifying, may have important implications for past and present oceanic N(2) fixation. PMID:22997339

Thompson, Anne W; Foster, Rachel A; Krupke, Andreas; Carter, Brandon J; Musat, Niculina; Vaulot, Daniel; Kuypers, Marcel M M; Zehr, Jonathan P

2012-09-21

29

Fermentation in the unicellular cyanobacterium Microcystis PCC7806  

Microsoft Academic Search

The cyanobacterium Microcystis PCC7806 fermented endogenously stored glycogen to ethanol, acetate, CO2, and H2 when incubated anaerobically in the dark. The switch from photoautotrophic to fermentative metabolism did not require de novo protein synthesis, and fermentation started immediately after cells had been transferred to dark anoxic conditions. From the molar ratios of the products and from enzyme activities in cell-free

Roy Moezelaar; Lucas J. Stal

1994-01-01

30

Expression of mouse metallothionein in the cyanobacterium Synechococcus PCC7942  

Microsoft Academic Search

A cDNA encoding mouse metallothionein was cloned into the shuttle vector pUC303, creating a translational fusion with the bacterial chloramphenicol acetyltransferase gene. The resulting fusion protein has been expressed in the cyanobacteriumSynechococcus PCC7942. Cyanobacterial transformants expressed mouse metallothionein-specific mRNA species as detected by RNA slot blots. In addition, the transformants expressed a unique cadmium ionbinding protein corresponding to the predicted

J L Erbe; K B Taylor; L M Hall

1996-01-01

31

Photoinhibition of photosynthesis in the cyanobacterium Microcystis aeruginosa  

Microsoft Academic Search

We have examined characteristics of the photoinhibition of photosynthesis which occur in the unicellular cyanobacterium Microcystis aeruginosa, following exposure to photon fluence rates in excess of those required for growth. Photoinhibition occurs following exposure of cells to a photon fluence rate of 1,000 µmol m-2 s-1, which is manifested as a decrease in either light-limited CO2 fixation or light-saturated CO2-dependent

G. C. Whitelam; G. A. Cold

1983-01-01

32

Modelling of growth conditions for cyanobacterium Spirulina platensis in microcosms  

Microsoft Academic Search

The influence of cultivation conditions on the growth of the cyanobacterium Spirulina platensis was investigated by using two types of photobioreactors. In a rotative photobioreactor the doubling time (td) was 3.54 days. The better value found for td in an aerated photobioreactor by changing the initial nitrogen concentration (NaNO3) at 0.003, 0.015, 0.030 and 0.060?M was 2.5 days. A factorial

Jorge Alberto Vieira Costa; Giani Andrea Linde; Daniel Ibraim Pires Atala; Guilherme Martinez Mibielli; Roselini Trapp Krüger

2000-01-01

33

Removal of lead (Pb 2+) by the Cyanobacterium Gloeocapsa sp  

Microsoft Academic Search

Pb2+ removal ability of the viable-freshwater cyanobacterium Gloeocapsa sp. was studied in batch experiments. Gloeocapsa sp. was cultured in the Medium 18 with pH adjusted to 3, 4, 5, 6 and 7. Growth was subsequently determined based on the increase of chlorophyll-a content. Gloeocapsa sp. was able to grow at all pH levels tested, except at pH 3. Removal of

Suneerat Raungsomboon; Amnat Chidthaisong; Boosya Bunnag; Duangrat Inthorn; Narumon W. Harvey

2008-01-01

34

Introduction of a nitrogen-fixing cyanobacterium into tobacco shoot regenerates  

Microsoft Academic Search

Tobacco (Nicotiana tabacum L.) shoots associated with the nitrogen-fixing cyanobacterium Anabaena variabilis Kütz. (ATCC 29413) were regenerated in mixed cultures of tobacco callus and the cyanobacterium. The cyanobacteria were localized inside the tissues as well as on the surface of regenerated shoots, formed heterocysts, and were capable of acetylene reduction.

M. V. Gusev; T. G. Korzhenevskaya; L. V. Pyvovarova; O. I. Baulina; R. G. Butenko

1986-01-01

35

Regulated nitrate transport in the cyanobacterium Anacystis nidulans.  

PubMed Central

Intracellular accumulation of nitrate, indicative of the operation of an active nitrate transport system, has been measured in intact cells of the cyanobacterium Anacystis nidulans. The ability of the cells to accumulate nitrate was effectively hindered by either ammonium addition or selective inhibition of CO2 fixation by DL-glyceraldehyde, with the effect of either compound being prevented by previously blocking ammonium assimilation. The results support the contention that nitrate utilization in cyanobacteria is regulated at the level of nitrate transport through the concerted action of ammonium assimilation and CO2 fixation.

Lara, C; Romero, J M; Guerrero, M G

1987-01-01

36

Effect of 1.7 MHz ultrasound on a gas-vacuolate cyanobacterium and a gas-vacuole negative cyanobacterium.  

PubMed

Ultrasonic signals propagated through medium were directly applied to unicellular cyanobacterium cell surfaces to investigate the biological effects induced by ultrasound. The gas-vacuolate cyanobacterium Microcystis aeruginosa and the gas-vacuole negative cyanobacterium Synechococcus PCC 7942 responded differently to ultrasound. When M. aeruginosa was irradiated by 1.7 MHz ultrasound at 0.6 W cm(-2) every day, it showed a decrease of nearly 65% in biomass increment, and this group's generation time increased twice as much as the control. While Synechococcus culture irradiated every day still grew as fast as the control, and its final biomass was as much as the control. The value of the electric conductivity change (Deltasigma) sharply increased in Microcystis suspension during the exposure process, which revealed more ultrasonic cavitation yield in liquid related to the gas-vacuolate cyanobacteria. The relative malondialdehyde (MDA) content, a quantitative indicator of lipid peroxidation, increased by 65% in Microcystis cells and 9% in Synechoccus cells after ultrasonic irradiation. Moreover, the membrane permeability, quantified by measuring the relative amount of electrolyte leaking out of cells, increased to more than 60% in the Microcystis cells. The results indicated that Microcystis cells were susceptible to ultrasonic stress. According to Rayleigh-Plesset's bubble activation theory, 1.7 MHz ultrasound approached the eigenfrequency of gas-vacuolate cells. The present investigation suggested the importance of the cavitational effect relative to intracellular gas-vacuoles in the loss of cell viability. In summary, 1.7 MHz ultrasonic irradiation was effective in preventing water-bloom forming cyanobacteria from growing rapidly due to changes in the functioning and integrity of cellular and subcellular structures. PMID:15261016

Tang, Jiao Wen; Wu, Qing Yu; Hao, Hong Wei; Chen, Yifang; Wu, Minsheng

2004-07-15

37

The photocycle and proton translocation pathway in a cyanobacterial ion-pumping rhodopsin.  

PubMed

The genome of thylakoidless cyanobacterium Gloeobacter violaceus encodes a fast-cycling rhodopsin capable of light-driven proton transport. We characterize the dark state, the photocycle, and the proton translocation pathway of GR spectroscopically. The dark state of GR contains predominantly all-trans-retinal and, similar to proteorhodopsin, does not show the light/dark adaptation. We found an unusually strong coupling between the conformation of the retinal and the site of Glu132, the homolog of Asp96 of BR. Although the photocycle of GR is similar to that of proteorhodopsin in general, it differs in accumulating two intermediates typical for BR, the L-like and the N-like states. The latter state has a deprotonated cytoplasmic proton donor and is spectrally distinct from the strongly red-shifted N intermediate known for proteorhodopsin. The proton uptake precedes the release and occurs during the transition to the O intermediate. The proton translocation pathway of GR is similar to those of other proton-pumping rhodopsins, involving homologs of BR Schiff base proton acceptor and donor Asp85 and Asp96 (Asp121 and Glu132). We assigned a pair of FTIR bands (positive at 1749 cm(-1) and negative at 1734 cm(-1)) to the protonation and deprotonation, respectively, of these carboxylic acids. PMID:19217863

Miranda, Mylene R M; Choi, Ah Rheum; Shi, Lichi; Bezerra, Arandi G; Jung, Kwang-Hwan; Brown, Leonid S

2009-02-18

38

Identification and functional analysis of a phytoene desaturase gene from the extremely radioresistant bacterium Deinococcus radiodurans.  

PubMed

The phytoene-related desaturases are the key enzymes in the carotenoid biosynthetic pathway. The gene encoding phytoene desaturase in the deinoxanthin synthesis pathway of Deinococcus radiodurans was identified and characterized. Two putative phytoene desaturase homologues (DR0861 and DR0810) were identified by analysis of conserved amino acid regions, and the former displayed the highest identity (68 %) with phytoene desaturase of the cyanobacterium Gloeobacter violaceus. DR0861 gene knockout and dinucleotide-binding motif deletion resulted in the arrest of lycopene synthesis and the accumulation of phytoene. The colourless DR0861 knockout mutant became more sensitive to acute ionizing radiation and oxygen stress. Complementation of the mutant with a heterologous or homologous gene restored its pigment and resistance. The desaturase activity of DR0861 (crtI) was further confirmed by the assay of enzyme activity in vitro and heterologous expression in Escherichia coli containing crtE and crtB genes (responsible for phytoene synthesis) from Erwinia uredovora. In addition, the amount of lycopene synthesis in E. coli resulting from the expression of crtI from D. radiodurans was determined, and this had significant dose-dependent effects on the survival rate of E. coli exposed to hydrogen peroxide and ionizing radiation. PMID:17464079

Xu, Zhenjian; Tian, Bing; Sun, Zongtao; Lin, Jun; Hua, Yuejin

2007-05-01

39

Mössbauer study of cobalt and iron in the cyanobacterium (blue green alga)  

NASA Astrophysics Data System (ADS)

Mössbauer emission and absorption studies have been performed on cobalt and iron in the cyanobacterium (blue-green alga). The Mössbauer spectrum of the cyanobacterium cultivated with57Co is decomposed into two doublets. The parameters of the major doublet are in good agreement with those of cyanocobalamin (vitamin B12) labeled with57Co. The other minor doublet has parameters close to those of Fe(II) coordinated with six nitrogen atoms. These suggest that cobalt is used for the biosynthesis of vitamin B12 or its analogs in the cyanobacterium. The spectra of the cyanobacterium grown with57Fe show that iron is in the high-spin trivalent state and possibly in the form of ferritin, iron storage protein.

Ambe, Shizuko

1990-07-01

40

Daily rhythm of O 2 -evolution in the cyanobacterium Trichodesmium thiebautii under natural and constant conditions  

Microsoft Academic Search

The rate of oxygen evolution by the tropical marine cyanobacterium Trichodesmium thiebautii was recorded at different times during the day in samples collected in 1992 from the Bahama Islands and the NE Caribbean Sea. This cyanobacterium is unique in that it is the only non-heterocystous diazotroph capable of N2-fixation in daylight. Oxygen evolution was measured under conditions of natural day\\/night

T. Roenneberg; E. J. Carpenter

1993-01-01

41

Photoinhibition and reactivation of photosynthesis in the cyanobacterium Anacystis nidulans  

SciTech Connect

The susceptibility of photosynthesis to photoinhibition and its recovery were studied on cultures of the cyanobacterium Anacystis nidulans. Oxygen evolution and low temperature fluorescence kinetics were measured. Upon exposure to high light A. nidulans showed a rapid decrease in oxygen evolution followed by a quasi steady state rate of photosynthesis. This quasi steady state rate decreased with increasing photon flux density of the photoinhibitory light. Reactivation of photosynthesis in dim light after the photoinhibitory treatment was rapid: 85 to 95% recovery occurred within 2 hours. In the presence of the translation inhibitor, streptomycin (250 micrograms per milliliter), no reactivation occurred. We also found that the damage increased dramatically if the high light treatment was done with streptomycin added. A transcription inhibitor, rifampicin, did not inhibit the reactivation process. Based on these data we conclude that the photoinhibitory damage observed is the net result of a balance between the photoinhibitory process and the operation of the repairing mechanism(s).

Samuelsson, G.; Loenneborg, A.; Rosenqvist, E.; Gustafsson, P.; Oequist, G.

1985-12-01

42

Phosphate transport and arsenate resistance in the cyanobacterium Anabaena variabilis  

SciTech Connect

Cells of the cyanobacterium Anabaena variabilis starved for phosphate for 3 days took up phosphate at about 100 times the rate of unstarved cells.Kinetic data suggested that a new transport system had been induced by starvation for phosphate. The inducible phosphate transport system was quickly repressed by addition of P/sub i/. Phosphate-starved cells were more sensitive to the toxic effects of arsenate than were unstarved cells, but phosphate could alleviate some of the toxicity. Arsenate was a noncompetitive inhibitor of phosphate transport; however, the apparent K/sub i/ values were high, particularly for phosphate-replete cells. Preincubation of phosphate-starved cells with arsenate caused subsequent inhibition of phosphate transport, suggesting that intracellular arsenate inhibited phosphate transport. This effect was not seen in phosphate-replete cells.

Thiel, T.

1988-03-01

43

Induction of anaerobic, photoautotrophic growth in the cyanobacterium Oscillatoria limnetica.  

PubMed Central

Anaerobic photoautotrophic growth of the cyanobacterium Oscillatoria limnetica was demonstrated under nitrogen in the presence of 3-(3,4-dichlorophenyl)-1,1-dimethylurea (5micron), a constant concentration of Na2S (2.5 mM), and constant pH (7.3). The photoanaerobic growth rate (2 days doubling time) was similar to that obtained under oxygenic photoautotrophic growth conditions. The potential of oxygenic photosynthesis is constitutive in the cells; that of anoxygenic photosynthesis is rapidly (2 h) induced in the presence of Na2S in the light in a process requiring protein synthesis. The facultative anaerobic phototrophic growth physiology exhibited by O. limnetica would seem to represent an intermediate physiological pattern between the obligate anaerobic one of photosynthetic bacteria and the oxygenic one of eucaryotic algae.

Oren, A; Padan, E

1978-01-01

44

Phosphate transport and arsenate resistance in the cyanobacterium Anabaena variabilis.  

PubMed Central

Cells of the cyanobacterium Anabaena variabilis starved for phosphate for 3 days took up phosphate at about 100 times the rate of unstarved cells. Kinetic data suggested that a new transport system had been induced by starvation for phosphate. The inducible phosphate transport system was quickly repressed by addition of Pi. Phosphate-starved cells were more sensitive to the toxic effects of arsenate than were unstarved cells, but phosphate could alleviate some of the toxicity. Arsenate was a noncompetitive inhibitor of phosphate transport; however, the apparent Ki values were high, particularly for phosphate-replete cells. Preincubation of phosphate-starved cells with arsenate caused subsequent inhibition of phosphate transport, suggesting that intracellular arsenate inhibited phosphate transport. This effect was not seen in phosphate-replete cells.

Thiel, T

1988-01-01

45

[2 site-specific endonucleases of the cyanobacterium Nostoc linckia].  

PubMed

Two restrictases Nli387/7 I and Nli387/7 II have been isolated from cyanobacterium Nostoc linckia using chromatography on phosphocellulose, "Mono Q" column, and heparin sepharose 4B. The preparations are described by the method of electrophoresis in polyacrylamide gel under denaturing conditions. Catalytic properties of restrictases are determined: optimal pH of the action--9.0--9.5, optimal concentration of Na+--5 mM, that of Mg2+--6 mM, optimal temperature--37 degrees C. The isolated enzymes are isoschizomers of restrictases avaI and AvaII. The point of cutting is determined for enzyme Nli387/7 I. It is shown that restrictase Nli387/7 I is a false isoschizomer Ava I. PMID:1650423

Mel'nik, A I; Rebentish, B A; Bolotin, A V; Mendzhul, M I

46

Production of the Neurotoxin BMAA by a Marine Cyanobacterium  

PubMed Central

Diverse species of cyanobacteria have recently been discovered to produce the neurotoxic non-protein amino acid ?-methylamino-L-alanine (BMAA). In Guam, BMAA has been studied as a possible environmental toxin in the diets of indigenous Chamorro people known to have high levels of Amyotrophic Lateral Sclerosis/ Parkinsonism Dementia Complex (ALS/PDC). BMAA has been found to accumulate in brain tissues of patients with progressive neurodegenerative illness in North America. In Guam, BMAA was found to be produced by endosymbiotic cyanobacteria of the genus Nostoc which live in specialized cycad roots. We here report detection of BMAA in laboratory cultures of a free-living marine species of Nostoc. We successfully detected BMAA in this marine species of Nostoc with five different methods: HPLC-FD, UPLC-UV, Amino Acid Analyzer, LC/MS, and Triple Quadrupole LC/MS/MS. This consensus of five different analytical methods unequivocally demonstrates the presence of BMAA in this marine cyanobacterium. Since protein-associated BMAA can accumulate in increasing levels within food chains, it is possible that biomagnification of BMAA could occur in marine ecosystems similar to the biomagnification of BMAA in terrestrial ecosystems. Production of BMAA by marine cyanobacteria may represent another route of human exposure to BMAA. Since BMAA at low concentrations causes the death of motor neurons, low levels of BMAA exposure may trigger motor neuron disease in genetically vulnerable individuals.

Banack, Sandra Anne; Johnson, Holly E.; Cheng, Ran; Cox, Paul Alan

2007-01-01

47

Nitrile-Containing Fischerindoles from the Cultured Cyanobacterium Fischerella sp  

PubMed Central

Chemical investigation of the cultured cyanobacterium Fischerella sp. (SAG strain number 46.79) led to the isolation of four nitrile-containing indole alkaloids, namely 12-epi-fischerindole I nitrile (1), deschloro 12-epi-fischerindole I nitrile (2), 12-epi-fischerindole W nitrile (3), and deschloro 12-epi-fischerindole W nitrile (4) along with a known metabolite hapalosin. The structures were determined by detailed spectroscopic analyses on the basis of 1D and 2D NMR and HRESIMS data. All isolates were evaluated for cytotoxicity against human cancer cells and for 20S proteasome inhibition. Deschloro 12-epi-fischerindole I nitrile (2) was found to be weakly cytotoxic against HT-29 cells with an ED50 value of 23 ?M. Hapalosin showed weak cytotoxicity against HT-29 and MCF-7 cells with ED50 values of 22 and 27 ?M, respectively, as well as moderate 20S proteasome inhibition with an IC50 value of 12 ?M. Compounds 1-4 all contain a nitrile moiety instead of the isonitrile found in all fischerindoles reported to date. Compounds 3 and 4 also display a new carbon skeleton, in which a six-membered ring replaces the five-membered ring normally found in fischerindole-type alkaloids.

Kim, Hyunjung; Krunic, Aleksej; Lantvit, Daniel; Shen, Qi; Kroll, David J.; Swanson, Steven M.

2012-01-01

48

Oxidative inactivation of glutamine synthetase from the cyanobacterium Anabaena variabilis.  

PubMed Central

In crude extracts of the cyanobacterium Anabaena variabilis, glutamine synthetase (GS) could be effectively inactivated by the addition of NADH. GS inactivation was completed within 30 min. Both the inactivated GS and the active enzyme were isolated. No difference between the two enzyme forms was seen in sodium dodecyl sulfate-gels, and only minor differences were detectable by UV spectra, which excludes modification by a nucleotide. Mass spectrometry revealed that the molecular masses of active and inactive GS are equal. While the Km values of the substrates were unchanged, the Vmax values of the inactive GS were lower, reflecting the inactivation factor in the crude extract. This result indicates that the active site was affected. From the crude extract, a fraction mediating GS inactivation could be enriched by ammonium sulfate precipitation and gel filtration. GS inactivation by this fraction required the presence of NAD(P)H, Fe3+, and oxygen. In the absence of the GS-inactivating fraction, GS could be inactivated by Fe2+ and H2O2. The GS-inactivating fraction produced Fe2+ and H2O2, using NADPH, Fe3+, and oxygen. Accordingly, the inactivating fraction was inhibited by catalase and EDTA. This GS-inactivating system of Anabaena is similar to that described for oxidative GS inactivation in Escherichia coli. We conclude that GS inactivation by NAD(P)H is caused by irreversible oxidative damage and is not due to a regulatory mechanism of nitrogen assimilation.

Martin, G; Haehnel, W; Boger, P

1997-01-01

49

Ribulose diphosphate carboxylase of the cyanobacterium Spirulina platensis  

SciTech Connect

The ribulose diphosphate (RDP) carboxylase activity of the cyanobacterium Spirulina platensis is represented by two peaks when a cell homogenate is centrifuged in a sucrose density gradient. In the case of differential centrifugation (40,000 g, 1 h), the activity of the enzyme was distributed between the supernatant liquid (soluble form) and the precipitate (carboxysomal form). From the soluble fraction, in which 80-95% of the total activity of the enzyme is concentrated, electrophoretically homogeneous RDP carboxylase was isolated by precipitation with ammonium sulfate and centrifugation in a sucrose density gradient. The purified enzyme possessed greater electrophoretic mobility in comparison with the RDP carboxylase of beans Vicia faba. The molecular weight of the enzyme, determined by gel filtration, was 450,000. The enzyme consists of monotypic subunits with a molecular weight of 53,000. The small subunits were not detected in electrophoresis in polyacrylamide gel in the presence of SDS after fixation and staining of the gels by various methods.

Terekhova, I.V.; Chernyad'ev, I.I.; Doman, N.G.

1986-11-20

50

Production of the neurotoxin BMAA by a marine cyanobacterium.  

PubMed

Diverse species of cyanobacteria have recently been discovered to produce the neurotoxic non-protein amino acid beta-methylamino-L-alanine (BMAA). In Guam, BMAA has been studied as a possible environmental toxin in the diets of indigenous Chamorro people known to have high levels of Amyotrophic Lateral Sclerosis/ Parkinsonism Dementia Complex (ALS/PDC). BMAA has been found to accumulate in brain tissues of patients with progressive neurodegenerative illness in North America. In Guam, BMAA was found to be produced by endosymbiotic cyanobacteria of the genus Nostoc which live in specialized cycad roots. We here report detection of BMAA in laboratory cultures of a free-living marine species of Nostoc. We successfully detected BMAA in this marine species of Nostoc with five different methods: HPLC-FD, UPLC-UV, Amino Acid Analyzer, LC/MS, and Triple Quadrupole LC/MS/MS. This consensus of five different analytical methods unequivocally demonstrates the presence of BMAA in this marine cyanobacterium. Since protein-associated BMAA can accumulate in increasing levels within food chains, it is possible that biomagnification of BMAA could occur in marine ecosystems similar to the biomagnification of BMAA in terrestrial ecosystems. Production of BMAA by marine cyanobacteria may represent another route of human exposure to BMAA. Since BMAA at low concentrations causes the death of motor neurons, low levels of BMAA exposure may trigger motor neuron disease in genetically vulnerable individuals. PMID:18463731

Banack, Sandra Anne; Johnson, Holly E; Cheng, Ran; Cox, Paul Alan

2007-12-06

51

Time-course analysis of cyanobacterium transcriptome: detecting oscillatory genes.  

PubMed

The microarray technique allows the simultaneous measurements of the expression levels of thousands of mRNAs. By mining these data one can identify the dynamics of the gene expression time series. The detection of genes that are periodically expressed is an important step that allows us to study the regulatory mechanisms associated with the circadian cycle. The problem of finding periodicity in biological time series poses many challenges. Such challenge occurs due to the fact that the observed time series usually exhibit non-idealities, such as noise, short length, outliers and unevenly sampled time points. Consequently, the method for finding periodicity should preferably be robust against such anomalies in the data. In this paper, we propose a general and robust procedure for identifying genes with a periodic signature at a given significance level. This identification method is based on autoregressive models and the information theory. By using simulated data we show that the suggested method is capable of identifying rhythmic profiles even in the presence of noise and when the number of data points is small. By recourse of our analysis, we uncover the circadian rhythmic patterns underlying the gene expression profiles from Cyanobacterium Synechocystis. PMID:22028849

Layana, Carla; Diambra, Luis

2011-10-18

52

Export of extracellular polysaccharides modulates adherence of the cyanobacterium synechocystis.  

PubMed

The field of cyanobacterial biofuel production is advancing rapidly, yet we know little of the basic biology of these organisms outside of their photosynthetic pathways. We aimed to gain a greater understanding of how the cyanobacterium Synechocystis PCC 6803 (Synechocystis, hereafter) modulates its cell surface. Such understanding will allow for the creation of mutants that autoflocculate in a regulated way, thus avoiding energy intensive centrifugation in the creation of biofuels. We constructed mutant strains lacking genes predicted to function in carbohydrate transport or synthesis. Strains with gene deletions of slr0977 (predicted to encode a permease component of an ABC transporter), slr0982 (predicted to encode an ATP binding component of an ABC transporter) and slr1610 (predicted to encode a methyltransferase) demonstrated flocculent phenotypes and increased adherence to glass. Upon bioinformatic inspection, the gene products of slr0977, slr0982, and slr1610 appear to function in O-antigen (OAg) transport and synthesis. However, the analysis provided here demonstrated no differences between OAg purified from wild-type and mutants. However, exopolysaccharides (EPS) purified from mutants were altered in composition when compared to wild-type. Our data suggest that there are multiple means to modulate the cell surface of Synechocystis by disrupting different combinations of ABC transporters and/or glycosyl transferases. Further understanding of these mechanisms may allow for the development of industrially and ecologically useful strains of cyanobacteria. Additionally, these data imply that many cyanobacterial gene products may possess as-yet undiscovered functions, and are meritorious of further study. PMID:24040267

Fisher, Michael L; Allen, Rebecca; Luo, Yingqin; Curtiss, Roy

2013-09-10

53

Antagonistic interactions between filamentous heterotrophs and the cyanobacterium Nostoc muscorum  

PubMed Central

Background Little is known about interactions between filamentous heterotrophs and filamentous cyanobacteria. Here, interactions between the filamentous heterotrophic bacteria Fibrella aestuarina (strain BUZ 2) and Fibrisoma limi (BUZ 3) with an axenic strain of the autotrophic filamentous cyanobacterium Nostoc muscorum (SAG 25.82) were studied in mixed cultures under nutrient rich (carbon source present in medium) and poor (carbon source absent in medium) conditions. Findings F. aestuarina BUZ 2 significantly reduced the cyanobacterial population whereas F. limi BUZ 3 did not. Physical contact between heterotrophs and autotroph was observed and the cyanobacterial cells showed some level of damage and lysis. Therefore, either contact lysis or entrapment with production of extracellular compounds in close vicinity of host cells could be considered as potential modes of action. The supernatants from pure heterotrophic cultures did not have an effect on Nostoc cultures. However, supernatant from mixed cultures of BUZ 2 and Nostoc had a negative effect on cyanobacterial growth, indicating that the lytic compounds were only produced in the presence of Nostoc. The growth and survival of tested heterotrophs was enhanced by the presence of Nostoc or its metabolites, suggesting that the heterotrophs could utilize the autotrophs and its products as a nutrient source. However, the autotroph could withstand and out-compete the heterotrophs under nutrient poor conditions. Conclusions Our results suggest that the nutrients in cultivation media, which boost or reduce the number of heterotrophs, were the important factor influencing the outcome of the interplay between filamentous heterotrophs and autotrophs. For better understanding of these interactions, additional research is needed. In particular, it is necessary to elucidate the mode of action for lysis by heterotrophs, and the possible defense mechanisms of the autotrophs.

2011-01-01

54

Characterization of transposon Tn5469 from the cyanobacterium Fremyella diplosiphon.  

PubMed Central

A transposon, designated Tn5469, was isolated from mutant strain FdR1 of the filamentous cyanobacterium Fremyella diplosiphon following its insertion into the rcaC gene. Tn5469 is a 4,904-bp noncomposite transposon with 25-bp near-perfect terminal inverted repeats and has three tandemly arranged, slightly overlapping potential open reading frames (ORFs) encoding proteins of 104.6 kDa (909 residues), 42.5 kDa (375 residues), and 31.9 kDa (272 residues). Insertion of Tn5469 into the rcaC gene in strain FdR1 generated a duplicate 5-bp target sequence. On the basis of amino acid sequence identifies, the largest ORF, designated tnpA, is predicted to encode a composite transposase protein. A 230-residue domain near the amino terminus of the TnpA protein has 15.4% amino acid sequence identity with a corresponding domain for the putative transposase encoded by Lactococcus lactis insertion sequence S1 (ISS1). In addition, the sequence for the carboxyl-terminal 600 residues of the TnpA protein is 20.0% identical to that for the TniA transposase encoded by Tn5090 on Klebsiella aerogenes plasmid R751. The TnpA and TniA proteins contain the D,D(35)E motif characteristic of a recently defined superfamily consisting of bacterial transposases and integrase proteins of eukaryotic retroelements and retrotransposons. The two remaining ORFs on Tn5469 encode proteins of unknown function. Southern blot analysis showed that wild-type F. diplosiphon harbors five genomic copies of Tn5469. In comparison, mutant strain FdR1 harbors an extra genomic copy of Tn5469 which was localized to the inactivated rcaC gene. Among five morphologically distinct cyanobacterial strains examined, none was found to contain genomic sequences homologous to Tn5469.

Kahn, K; Schaefer, M R

1995-01-01

55

The gene complement for proteolysis in the cyanobacterium Synechocystis sp. PCC 6803 and Arabidopsis thaliana chloroplasts  

Microsoft Academic Search

A set of 62 genes that encode the entire peptidase complement of Synechocystis sp. PCC 6803 has been identified in the genome database of that cyanobacterium. Sequence comparisons with the Arabidopsis genome uncovered the presumably homologous chloroplast components inherited from their cyanobacterial ancestor. A systematic gene disruption approach was chosen to individually inactivate, by customary transformation strategies, the majority of

Anna Sokolenko; Elena Pojidaeva; Vladislav Zinchenko; Vladimir Panichkin; Vadim M. Glaser; Reinhold G. Herrmann; Sergey V. Shestakov

2002-01-01

56

HETEROCYST DIFFERENTIATION AND CELL DIVISION IN THE CYANOBACTERIUM ANABAENA CYLINDRICA: EFFECT OF HIGH LIGHT INTENSITY  

Microsoft Academic Search

SUMMARY Heterocyst differentiation in the cyanobacterium Anabaena cylindrica is initiated by the removal of fixed nitrogen from the medium. These specialized cells occur singly at regular intervals within filaments of vegetative cells. Incubation of cultures for periods of up to 12 h immediately prior to or following removal of fixed nitrogen, at a light intensity (500 \\/iEinsteins cm~2 s\\

DAVID G. ADAMS; NOEL G. CARR

57

Ecophysiology of the Cyanobacterium Dactylococcopsis salina: Effect of Light Intensity, Sulphide and Temperature  

Microsoft Academic Search

~~~ Dactylococcopsis salina is a planktonic gas-vacuolated cyanobacterium that forms a distinct bacterial plate at the metalimnion of Solar Lake, Sinai. Temperature, light intensity and sulphide concentration were examined as possible limiting factors determining the distribution of D. salina during the annual limnological cycle of Solar Lake. Both laboratory cultures and in situ samples were examined for their photosynthetic activity

J. Van Run; YEHUDA COHEN

1983-01-01

58

Benthic-pelagic coupling in the population dynamics of the harmful cyanobacterium Microcystis  

Microsoft Academic Search

SUMMARY 1. In eutrophic lakes, large amounts of the cyanobacterium Microcystis may overwinter in the sediment and re-inoculate the water column in spring. 2. We monitored changes in pelagic and benthic populations of Microcystis in Lake Volkerak, The Netherlands. In addition, sedimentation rates and the rate of recruitment from the sediment were measured using traps. These data were used to

JOLANDA M. H. V ERSPAGEN; KLAUS D. J OHNK; W. I BELINGS; LUUC R. M UR; J EF H UISMAN

59

Evidence for Paralytic Shellfish Poisons in the Freshwater Cyanobacterium Lyngbya wollei (Farlow ex Gomont) comb. nov  

Microsoft Academic Search

Lyngbya wollei (Farlow ex Gomont) comb. nov., a perennial mat-forming filamentous cyanobacterium prev- alent in lakes and reservoirs of the southeastern United States, was found to produce a potent, acutely lethal neurotoxin when tested in the mouse bioassay. Signs of poisoning were similar to those of paralytic shellfish poisoning. As part of the Tennessee Valley Authority master plan for Guntersville

W. W. CARMICHAEL; W. R. EVANS; Q. Q. YIN; P. BELL; E. MOCZYDLOWSKI

1997-01-01

60

Selective inhibitory potential of silver nanoparticles on the harmful cyanobacterium Microcystis aeruginosa  

Microsoft Academic Search

Silver nanoparticles (SNPs) at 1 mg\\/l inhibited the growth of the toxic cyanobacterium, Microcystis aeruginosa, by 87%. Similar results were obtained in field experiments. M. aeruginosa was more sensitive to SNPs than were green algae. SNPs may be a useful selective biocidal agent for the control of M. aeruginosa.

Myung-Hwan Park; Keun-Hee Kim; Huk-Hee Lee; Jin-Seog Kim; Soon-Jin Hwang

2010-01-01

61

Transient state characteristics of changes in light conditions for the cyanobacterium Oscillatoria agardhii  

Microsoft Academic Search

The cyanobacterium Oscillatoria agardhii was subjected to changes in irradiance and to changes in light period. During transient states parameters as growth rate, pigment contents, photosynthetic activities and pool sizes of carbohydrate and proteins were followed. The changes in pigments and photosynthesis were similar for irradiance transitions and transitions in light period length. Carbohydrates served for the supply of carbon

A. F. Post

1987-01-01

62

Draft Genome Sequence of the Brazilian Toxic Bloom-Forming Cyanobacterium Microcystis aeruginosa Strain SPC777.  

PubMed

Microcystis aeruginosa strain SPC777 is an important toxin-producing cyanobacterium, isolated from a water bloom of the Billings reservoir (São Paulo State, Brazil). Here, we report the draft genome sequence and initial findings from a preliminary analysis of strain SPC777, including several gene clusters involved in nonribosomal and ribosomal synthesis of secondary metabolites. PMID:23908289

Fiore, Marli F; Alvarenga, Danillo O; Varani, Alessandro M; Hoff-Risseti, Caroline; Crespim, Elaine; Ramos, Rommel T J; Silva, Artur; Schaker, Patricia D C; Heck, Karina; Rigonato, Janaina; Schneider, Maria Paula C

2013-08-01

63

Expression of Escherichia coli Phosphoenolpyruvate Carboxylase in a Cyanobacterium' Functional Complementation of Synechococcus PCC 7942 ppc  

Microsoft Academic Search

The gene (ppc) coding for phosphoenolpyruvate carboxylase (PEPCase) in the cyanobacterium Synechococcus PCC 7942 has been inactivated via insertional mutagenesis while being function- ally complemented by Escherichia coli ppc. cyanobacterial cells functionally complemented by E. coli ppc showed decreased PEP- Case activity in crude cell lysates and detergent-permeabilized whole cell assays. Decreased rates of growth, reduced levels of chlorophyll a,

John R. Coleman

64

Evidence for a trimeric organization of the photosystem I complex from the thermophilic cyanobacterium Synechococcus sp  

Microsoft Academic Search

A photosystem I (PS I) reaction center complex was isolated and purified from the cyanobacterium Synechococcus sp. The complex has a molecular mass of about 600 kDa and contains 120 Chl a molecules per photoactive Chl a1 (P-700). Electron micrographs show that the PS I complex has the shape of a disk with a diameter of about 19 nm and

W. Saenger; M. Rögner; M. G. van Heel; J. P. Dekker; E. J. Boekema; I. Witt; H. T. Witt

1987-01-01

65

Reductive transformation of methyl parathion by the cyanobacterium Anabaena sp. strain PCC7120  

Microsoft Academic Search

Organophosphorus compounds are toxic chemicals that are applied worldwide as household pesticides and for crop protection, and they are stockpiled for chemical warfare. As a result, they are routinely detected in air and water. Methods and routes of biodegradation of these compounds are being sought. We report that under aerobic, photosynthetic conditions, the cyanobacterium Anabaena sp. transformed methyl parathion first

J. W. Barton; T. Kuritz; L. E. O’Connor; C. Y. Ma; M. P. Maskarinec; B. H. Davison

2004-01-01

66

Causes for the Large Genome Size in a Cyanobacterium Anabaena sp. PCC7120  

Microsoft Academic Search

Three possible causes responsible for the large genome size of a cyanobacterium Anabaena sp. PCC7120 are investigated: 1) sequential tandem duplications of gene segments, genes or genomic segments, 2) horizontal gene transfers from other organisms, and 3) whole-genome duplication. We evaluated the frequency distribution of angles between paralog locations for the possibility 1), the fraction of genes deviated in GC

Nobuyoshi Sugaya; Makihiko Sato; Hiroo Murakami; Akira Imaizumi; Sachiyo Aburatani; Katsuhisa Horimoto

67

Toxin release in response to oxidative stress and programmed cell death in the cyanobacterium Microcystis aeruginosa  

Microsoft Academic Search

An unprecedented bloom of the cyanobacterium Microcystis aeruginosa Kütz. occurred in the St. Lucie Estuary, FL in the summer of 2005. Samples were analyzed for toxicity by ELISA and by use of the polymerase chain reaction (PCR) with specific oligonucleotide primers for the mcyB gene that has previously been correlated with the biosynthesis of toxic microcystins. Despite the fact that

Cliff Ross; Lory Santiago-Vázquez; Valerie Paul

2006-01-01

68

Aluminum effects on uptake and metabolism of phosphorus by the Cyanobacterium Anabaena cylindrica  

Microsoft Academic Search

Aluminum severely affects the growth of the cyanobacterium Anabaena cylindrica and induces symptoms indicating phosphorus starvation. Pre- or post-treating the cells with high (90 micromolar) phosphorus reduces the toxicity of aluminum compared to cells receiving a lower orthophosphate concentration. In this study aluminum (ranging from 9 to 36 micromolar) and phosphorus concentrations were chosen so that the precipitation of insoluble

A. Pettersson; Haellbom; B. L. Bergman

1988-01-01

69

Molybdate transport and its effect on nitrogen utilization in the cyanobacterium Anabaena variabilis ATCC 29413  

Microsoft Academic Search

Summary Molybdenum is an essential component of the cofactors of many metalloenzymes including nitrate reductase and Mo-nitrogenase. The cyanobacterium Anabaena variabilis ATCC 29413 uses nitrate and atmospheric N 2 as sources of nitrogen for growth. Two of the three nitrogenases in this strain are Mo-depen- dent enzymes, as is nitrate reductase; thus, transport of molybdate is important for growth of

Marta Zahalak; Brenda Pratte; Kelly J. Werth; Teresa Thiel

70

The role of extracellular polysaccharides produced by the terrestrial cyanobacterium Nostoc sp. strain HK-01 in NaCl tolerance  

Microsoft Academic Search

The terrestrial cyanobacterium Nostoc sp. HK-01 was more tolerant to NaCl stress than the aquatic cyanobacterium Anabaena sp. PCC 7120 (also called Nostoc sp. PCC 7120) which is similar to Nostoc sp. HK-01 in phylogeny. We determined the amount of extracellular polysaccharides (capsular and released polysaccharides)\\u000a from the cells of both strains cultured with or without 200 mM NaCl. The amount

Hidehisa Yoshimura; Toshihisa Kotake; Tsutomu Aohara; Yoichi Tsumuraya; Masahiko Ikeuchi; Masayuki Ohmori

71

Interaction of uranium with a filamentous, heterocystous, nitrogen-fixing cyanobacterium, Anabaena torulosa.  

PubMed

The filamentous, heterocystous, diazotrophic cyanobacterium, Anabaena torulosa was found to bind uranium from aqueous solutions containing 100 ?M uranyl carbonate at pH 7.8. The uranyl sequestration kinetics exhibited (a) an initial rapid phase, binding 48% uranium within 30 min resulting in a loading of 56 mg U g(-1) of dry wt, followed by (b) a slower phase, binding 65% uranium with resultant loading of 77.35 mg U g(-1) in 24h. Energy Dispersive X-ray fluorescence spectroscopy of uranium loaded biomass revealed all components of UL X-rays (UL(l), UL(?), UL(?1) and UL(?2)). Heat killed cells or extracellular polysaccharides derived from live cells exhibited limited uranyl binding (~26%) highlighting the importance of cell viability for optimum uranyl binding. The present study revealed the involvement of acid soluble polyphosphates in uranium accumulation by this brackish water cyanobacterium. PMID:22522016

Acharya, C; Chandwadkar, P; Apte, S K

2012-03-29

72

Fermentation and sulfur reduction in the mat-building cyanobacterium Microcoleus; chthonoplastes  

Microsoft Academic Search

The mat-building cyanobacterium Microcoleus chthonoplastes carried out a mixed-acid fermentation when incubated under anoxic conditions in the dark. Endogenous storage carbohydrate was fermented to acetate, ethanol, formate, lactate, H2, and CO2. Cells with a low glycogen content (about 0.3 mmol of glucose per mg of protein) produced acetate and ethanol in equimolar amounts. In addition to glycogen, part of the

ROY MOEZELAAR; SASKIA M. BIJVANK; L. J. Stal

1996-01-01

73

A Novel Nitrate\\/Nitrite Permease in the Marine Cyanobacterium Synechococcus sp. Strain PCC 7002  

Microsoft Academic Search

The nrtP and narB genes, encoding nitrate\\/nitrite permease and nitrate reductase, respectively, were isolated from the marine cyanobacterium Synechococcus sp. strain PCC 7002 and characterized. NrtP is a member of the major facilitator superfamily and is unrelated to the ATP-binding cassette-type nitrate transporters that previously have been described for freshwater strains of cyanobacteria. However, NrtP is similar to the NRT2-

TOSHIO SAKAMOTO; KAORI INOUE-SAKAMOTO; DONALD A. BRYANT

1999-01-01

74

Cytochrome c-553 Is not Required for Photosynthetic Activity in the Cyanobacterium Synechococcus  

Microsoft Academic Search

In cyanobacteria, the water-soluble cytochrome c-553 functions as a mobile carrier of electrons between the membrane-bound cytochrome bs-f complex and P-700 reaction centers of Photosystem 1. The structural gene for cytochrome c-553 (designated cytA) of the cyanobacterium Synechococcus sp. PCC 7942 was cloned, and the deduced amino acid sequence was shown to be similar to known cyanobacterial cytochrome c-553 proteins.

David E. Laudenbach; Stephen K. Herbert; David C. Fork; Arthur R. Grossman; Neil A. Strausb

1990-01-01

75

Flavonoid-Induced Expression of a Symbiosis-Related Gene in the Cyanobacterium Nostoc punctiforme  

PubMed Central

The flavonoid naringin was found to induce the expression of hrmA, a gene with a symbiotic phenotype in the cyanobacterium Nostoc punctiforme. A comparative analysis of several flavonoids revealed the 7-O-neohesperidoside, 4?-OH, and C-2 000000000000 000000000000 000000000000 000000000000 111111111111 000000000000 000000000000 000000000000 000000000000 C-3 double bond in naringin as structural determinants of its hrmA-inducing activity.

Cohen, Michael F.; Yamasaki, Hideo

2000-01-01

76

Phenotype and temperature affect the affinity for dissolved inorganic carbon in a cyanobacterium Microcystis  

Microsoft Academic Search

The cyanobacterium Microcystis is the most common bloom-forming species in eutrophicated water bodies. Known eco-physiological advantages of this organism\\u000a help it to compete effectively with other algae and cyanobacteria; however, little is known about the physiological characteristics\\u000a competence of colonial Microcystis. In the present study, carbonic anhydrase (CA) activity, the affinity for dissolved inorganic carbon (DIC), and the transcription\\u000a of

Xinghua Wu; Zhongxing Wu; Lirong Song

77

Purification, Characterization, and Gene Expression of All Sigma Factors of RNA Polymerase in a Cyanobacterium  

Microsoft Academic Search

The expression of RNA polymerase (RNAP) sigma factor genes and proteins was characterized as a first step toward understanding their functions in a unicellular cyanobacterium Synechocystis sp. PCC 6803, which can perform photosynthesis. All nine sigma factors (group 1, SigA; group 2, SigB to SigE; and group 3, SigF to SigI) and each RNAP core subunit (RpoA, RpoB, RpoC1 and

Sousuke Imamura; Satoshi Yoshihara; Serina Nakano; Noriko Shiozaki; Akiko Yamada; Kan Tanaka; Hideo Takahashi; Munehiko Asayama; Makoto Shirai

2003-01-01

78

Sensitivity of Two Disjunct Bacterioplankton Communities to Exudates from the Cyanobacterium Microcystis aeruginosa Kützing  

Microsoft Academic Search

Microcystis aeruginosa Kützing releases a variety of bioactive compounds during growth. This study determined whether bacteria from communities\\u000a co-occurring (M+) or not (M-) with this cosmopolitan cyanobacterium respond similarly to its products. Fifty M+ bacteria from\\u000a a M. aeruginosa bloom site (Western Basin of Lake Erie) and 50 M- bacteria from a Microcystis-free site (East Twin Lake, Portage Co., OH)

D. A. Casamatta; C. E. Wickstrom

2000-01-01

79

Optimal conditions for genetic transformations of the cyanobacterium Anacystis nidulans R2  

Microsoft Academic Search

Under optimal conditions, the cyanobacterium Anacystis nidulans R2 was transformed to ampicillin resistance at frequencies of >10⁷ transformants per ..mu..g of plasmid (pCH1) donor DNA. No stringent period of competency was detected, and high frequencies of transformation were achieved with cultures at various growth stages. Transformation increased with time after addition of donor DNA up to 15 to 18 h.

S. S. Golden; L. A. Sherman

1984-01-01

80

Cell differentiation and colony alteration of an edible terrestrial cyanobacterium Nostoc flagelliforme , in liquid suspension cultures  

Microsoft Academic Search

Morphological characteristics of an edible terrestrial cyanobacteriumNostoc flagelliforme in liquid suspension cultures under photoautotrophic conditions are presented. Different cell forms alternated in a regular\\u000a manner during the experimentation period (30 d).N. flagelliforme exhibited a very complex life cycle in terms of colony morphology, including mainly 4 different colony morphological forms,viz. hormogonia, filaments, seriate colonies and aseriate colonies. Under laboratory conditions

X.-J. Liu; F. Chen

2003-01-01

81

Action of selected heavy metal ions on the photosystem 2 activity of the cyanobacterium Spirulina platensis  

Microsoft Academic Search

Addition of different concentrations of heavy metal ions (Hg2+, Cu2+, Ni2+ and Pb2+) inhibited the photosystem 2 catalyzed electron transport activity (H2O?p-benzo-quinone) of the cyanobacteriumSpirulina platensis. Hg2+ caused the inhibition in electron transport activity in very low concentrations compared to the other metal ions. Hg2+ at this low concentration specifically altered the spectral properties of phycocyanin of the phycobilisomes in

S. D. S. Murthy; P. Mohanty

1995-01-01

82

First report of microcystins from a Brazilian isolate of the cyanobacterium Microcystis aeruginosa  

Microsoft Academic Search

This is the first report on microcystins, cyclic heptapeptide hepatotoxins, from Brazilian water supplies. A colony isolate (NPJB-1) of the colonial cyanobacteriumMicrocystis aeruginosa from Lagoa das Garças, São Paulo, was cultured under non-axenic conditions. Exponential phase cells were harvested, concentrated and lyophilized for mouse bioassays and toxin extraction. The LD100 of lyophilized cell suspensions was approximately 31 mg kg?1 (dry

Sandra M. F. O. Azevedo; William R. Evans; Wayne W. Carmichael; Michio Namikoshi

1994-01-01

83

Purification and Characterization of a Homodimeric Catalase-Peroxidase from the Cyanobacterium Anacystis nidulans  

Microsoft Academic Search

Cytosolic extracts of the cyanobacteriumAnacystis nidulansexhibit both catalase and o-dianisidine peroxidase activity. Native polyacrylamide gel electrophoresis demonstrates one distinct enzyme, which has been purified to essential homogeneity and found to be composed of two identical subunits of equal size (80.5 kDa). The isoelectric point is at pH 4.7. It is a very efficient catalase with a broad pH optimum between

C. Obinger; G. Regelsberger; G. Strasser; U. Burner; G. A. Peschek

1997-01-01

84

Extract of Microcystis water bloom affects cellular differentiation in filamentous cyanobacterium Trichormus variabilis (Nostocales, Cyanobacteria)  

Microsoft Academic Search

Cyanobacteria produce a wide spectrum of biologically active compounds such as microcystins, whose effects on photoautotrophic\\u000a organisms and role in the aquatic ecosystems have been most intensively discussed and studied. In our study, we examined effects\\u000a of semipurified Microcystis extract containing microcystins (0.2–20 nM corresponding to 0.2–20 ?g L?1) on age-induced cell differentiation of filamentous cyanobacterium Trichormus variabilis. The heterocyst and akinete formation

Kate?ina Bártová; Klára Hilscherová; Pavel Babica; Blahoslav Maršálek

85

Localization and distribution of hopanoids in membrane systems of the cyanobacterium Synechocystis PCC 6714.  

PubMed

Intracellular localization of triterpenic membrane stabilizers of the hopane series is described for the first time for a cyanobacterium. In Synechocystis PCC 6714, a bacteriohopanetetrol derivative (main compound) and diplopterol were detected in cell wall (CW) and thylakoid membrane (TM). Both hopanoids were enriched 4.5-fold and 9.0-fold in CW and outer membrane (OM) fractions, respectively, compared to TMs. PMID:1624128

Jürgens, U J; Simonin, P; Rohmer, M

1992-05-01

86

MERCURY INDUCED ALTERATION IN THE PHYCOBILIPROTEINS OF CYANOBACTERIUM, CYLINDROSPERMUM STAGNALE (kütz.) ADAPTED UNDER DIFFERENT LIGHT  

Microsoft Academic Search

Phycobilisomes extracted from the cyanobacterium, Cylindrospermum stagnale grown under different light conditions (white, red, blue and green) were subjected to graded concentration of mercuric chloride 6 µM, 9 µM and 12 µM. At all concentrations of Hgcl2, partial inhibition in the absorption of the phycobiliproteins was observed, contrarily in 6 µM concentration of red-light, no influence in phycocyanin absorbance was

Velu Vijaya; Venkatesan Padmapriya; Narayanaswamy Anand; Balan Karunai Sevi; J. A. John Paul

87

Photosynthetic performance of a helical tubular photobioreactor incorporating the cyanobacterium Spirulina platensis  

Microsoft Academic Search

The photosynthetic performance of a helical tubular photobioreactor (``Biocoil``), incorporating the filamentous cyanobacterium Spirulina platensis, was investigated. The photobioreactor was constructed in a cylindrical shape with a 0.25-m² basal area and a photostage comprising 60 m of transparent PVC tubing of 1.6-cm inner diameter. The inner surface of the cylinder was illuminated with cool white fluorescent lamps; the energy input

Yoshitomo Watanabe; D. O. Hall; J. De La Nouee

1995-01-01

88

The response of the filamentous cyanobacterium Spirulina platensis to salt stress  

Microsoft Academic Search

The responses of the filamentous cyanobacterium Spirulina platensis to increased NaCl concentrations (0.25–1.0 M) in addition to the concentration of sodium in the growth medium were studied. A two stage response to the salt stress was observed. This consisted of a relatively short shock stage, followed by adaptation process. It was shown that upon exposure to high salt concentrations of

Avigad Vonshak; Rachel Guy; Micha Guy

1988-01-01

89

Nitrogen stress induced changes in the marine cyanobacterium Oscillatoria willei BDU 130511  

Microsoft Academic Search

Exclusion of combined nitrogen (NaNO3) from the growth medium caused certain changes in metabolic processes leading to cessation in growth of the non-heterocystous, non nitrogen-fixing marine cyanobacterium Oscillatoria willei BDU 130511. But antioxidative enzymes, namely superoxide dismutase and peroxidase, helped the organism to survive the nitrogen stress. Prominent effects observed during nitrogen starvation\\/limitation were: (i) reduction of major and accessory

Sushanta Kumar Saha; Lakshmanan Uma; Gopalakrishnan Subramanian

2003-01-01

90

2-O-methyl-d-xylose containing sheath in the cyanobacterium Gloeothece sp. PCC 6501  

Microsoft Academic Search

The sheath of the unicellular cyanobacterium Gloeothece sp. PCC 6501 was isolated from cell homogenates by differential and sucrose gradient centrifugation, followed by lysozyme treatment and hot sodium dodecyl sulfate extraction. The sheath contains a major fraction of carbohydrate consisting of galactose, glucose, mannose, rhamnose, 2-O-methyl-d-xylose, xylose, glucuronic and galacturonic acids, but only traces of fatty acids and phosphate. A

J. Weckesser; C. Broll; S. P. Adhikary; U. J. Jürgens

1987-01-01

91

Fatty acids with antibacterial activity from the cyanobacterium Oscillatoria redekei HUB 051  

Microsoft Academic Search

Bioassay-guided fractionation of the n-hexane extract prepared from the biomass of the cyanobacterium Oscillatoria redekei syn. Limnothrix redekei HUB 051 by silica gel and RP-18 column chromatography followed by HPLC resulted in the isolation of a mixture of two unsaturated hydroxy fatty acids. Their further separation using normal phase HPLC resulted in the identification of a-dimorphecolic acid, a 9-hydroxy-10E, 12Z-octadecadienoic

Sabine Mundt; Susann Kreitlow; Rolf Jansen

2003-01-01

92

Eucapsitrione, an Anti-Mycobacterium tuberculosis Anthraquinone Derivative from the Cultured Freshwater Cyanobacterium Eucapsis sp.  

PubMed Central

Eucapsitrione (1), an anthraquinone derivative with an indeno-anthracene-trione skeleton, was isolated from the cyanobacterium Eucapsis sp. (UTEX 1519) by bioassay-guided fractionation. The chemical structure was determined by analyzing MS and 1D and 2D NMR spectroscopic data. Eucapsitrione (1) showed anti-Mycobacterium tuberculosis activity in the microplate Alamar blue assay and low-oxygen-recovery assay with MIC values of 3.1 and 6.4 µM, respectively.

Sturdy, Megan; Krunic, Aleksej; Cho, Sanghyun; Franzblau, Scott; Orjala, Jimmy

2010-01-01

93

Spectroscopic properties of PSI–IsiA supercomplexes from the cyanobacterium Synechococcus PCC 7942  

Microsoft Academic Search

The cyanobacterium Synechococcus PCC 7942 grown under iron starvation assembles a supercomplex consisting of a trimeric Photosystem I (PSI) complex encircled by a ring of 18 CP43? or IsiA light-harvesting complexes [Nature 412 (2001) 745]. Here we present a spectroscopic characterization by temperature-dependent absorption and fluorescence spectroscopy, site-selective fluorescence spectroscopy at 5 K, and circular dichroism of isolated PSI–IsiA, PSI

Elena G. Andrizhiyevskaya; Tatjana M. E. Schwabe; Marta Germano; Sandrine D'Haene; Jochen Kruip; Rienk van Grondelle; Jan P. Dekker

2002-01-01

94

Response of the estuarine cyanobacterium Lyngbya aestuarii to UV-B radiation  

Microsoft Academic Search

Growth response and changes in the spectral properties of methanolic extract of an estuarine cyanobacterium, Lyngbya aestuarii Agardh, to UV-B radiation were studied. Increase in growth accompanied by increase in chlorophyll a, protein and carbohydrate content was observed up to 12 h of UV-B irradiation followed by a decline with further increase\\u000a in the duration of UV exposure. Carotenoid content progressively

Jnanendra Rath; Siba P. Adhikary

2007-01-01

95

Genome Erosion in a Nitrogen-Fixing Vertically Transmitted Endosymbiotic Multicellular Cyanobacterium  

Microsoft Academic Search

BackgroundAn ancient cyanobacterial incorporation into a eukaryotic organism led to the evolution of plastids (chloroplasts) and subsequently to the origin of the plant kingdom. The underlying mechanism and the identities of the partners in this monophyletic event remain elusive.Methodology\\/Principal FindingsTo shed light on this evolutionary process, we sequenced the genome of a cyanobacterium residing extracellularly in an endosymbiosis with a

Liang Ran; John Larsson; Theoden Vigil-Stenman; Johan A. A. Nylander; Karolina Ininbergs; Wei-Wen Zheng; Alla Lapidus; Stephen Lowry; Robert Haselkorn; Birgitta Bergman; Niyaz Ahmed

2010-01-01

96

Influence of Leaching Parameters on the Biological Removal of Uranium from Coal by a Filamentous Cyanobacterium  

PubMed Central

Axenic cultures of the filamentous cyanobacterium LPP OL3 were incubated with samples of uraniumbearing coal from a German mining area. The influence of leaching parameters such as coal concentration (pulp density), initial biomass, particle size, temperature, and composition of the growth medium on the leaching of uranium from the ore by the cyanobacterial strain was studied. When low pulp densities were applied, the yield of biologically extracted uranium was optimal (reaching 96% at 1% [wt/vol] coal) and all released uranium was found in the culture liquid. Above 10% (wt/vol) coal in the medium, the amount of cell-bound uranium increased. Initial biomass concentration (protein content of the cultures) and particle size were not critical parameters of leaching by LPP OL3. However, temperature and composition of the growth medium profoundly influenced the leaching of uranium and growth of the cyanobacterium. The yield of leached uranium (at 10% [wt/vol] coal) could not be raised with a tank leaching apparatus. Also, coal ashes were not suitable substrates for the leaching of uranium by LPP OL3. In conclusion, the reactions of the cyanobacterium to variations in leaching parameters were different from reactions of acidic leaching organisms. Images

Lorenz, Michael G.; Krumbein, Wolfgang E.

1985-01-01

97

The extracellular-matrix-retaining cyanobacterium Nostoc verrucosum accumulates trehalose, but is sensitive to desiccation.  

PubMed

The aquatic cyanobacterium Nostoc verrucosum forms macroscopic colonies, which consist of both cellular filaments and massive extracellular matrix material. In this study, the physiological features of N. verrucosum were investigated and compared with those of the anhydrobiotic cyanobacterium Nostoc commune. Nostoc verrucosum cells were sensitive to desiccation, but tolerant of freeze-thawing treatment in terms of both cell viability and photosynthetic O(2) evolution. Natural colonies of these cyanobacteria contained similar levels of chlorophyll a, carotenoids, the UV-absorbing pigments scytonemin and mycosporine-like amino acids, and uronic acid [a component of extracellular polysaccharides (EPS)]. EPS from both N. verrucosum and N. commune indicated low acidity and a high affinity for divalent cations, although their sugar compositions differed. The WspA protein, known to be a major component of the extracellular matrix of N. commune, was detected in N. verrucosum. Desiccation caused similarly high levels of trehalose accumulation in both cyanobacteria. Although previously considered relevant to anhydrobiosis in the terrestrial cyanobacterium N. commune, the data presented here suggest that extracellular matrix production and trehalose accumulation are not enough for standing extreme desiccation in N. verrucosum. PMID:21507024

Sakamoto, Toshio; Kumihashi, Keisuke; Kunita, Shinpei; Masaura, Takuya; Inoue-Sakamoto, Kaori; Yamaguchi, Masaaki

2011-05-11

98

Autoradiographic studies of (methyl-/sup 3/H)thymidine incorporation in a cyanobacterium (Microcystis wesenbergii)-bacterium association and in selected algae and bacteria  

SciTech Connect

The present investigation showed by means of autoradiography that the cyanobacterium Microcystis wesenbergii did not incorporate (/sup 3/H)thymidine at nanomolar concentrations, whereas its associated heterotrophic bacteria appearing in the gelatinous cover of the cyanobacterium became labeled. Several other tested cyanobacteria and algae did not incorporate (/sup 3/H)thymidine.

Bern, L.

1985-01-01

99

Draft genome database construction from four strains (NIES-298, FCY- 26, -27, and -28) of the cyanobacterium Microcystis aeruginosa.  

PubMed

Microcystis aeruginosa is a cyanobacterium that can form harmful algal blooms (HABs) producing toxic secondary metabolites. We provide here draft genome information of four strains of this freshwater cyanobacterium that was obtained by the Next Generation Sequencing approach to provide a better understanding of molecular mechanisms at the physiological and ecological levels. After gene assembly, genes of each strain were identified and annotated, and a genome database and G-browser of M. aeruginosa were subsequently constructed. Such genome information resources will enable us to obtain useful information for molecular ecological studies with a better understanding of modulating mechanisms of environmental factors associated with blooming. PMID:22814493

Rhee, Jae-Sung; Choi, Beom-Soon; Han, Jeonghoon; Hwang, Soon-Jin; Choi, Ik-Young; Lee, Jae-Seong

2012-09-01

100

Extraction and purification of an unusual phycoerythrin in a terrestrial desiccation tolerant cyanobacterium Lyngbya arboricola  

Microsoft Academic Search

Presence and stability of an unusual phycoerythrin (PE) characteristically similar to R-PE are described in a terrestrial,\\u000a desiccation-tolerant cyanobacterium, Lyngbya arboricola. Extraction and purification of the PE by using acetone precipitation, gel filtration and ion-exchange chromatography resulted\\u000a in achieving a purity index (A560\\/A280) of up to 5.2. SDS-PAGE of the PE showed presence of 18 kDa, 20 kDa and 32 kDa bands corresponding

S. N. Tripathi; Shivali Kapoor; Alpana Shrivastava

2007-01-01

101

Cytotoxic and non-cytotoxic exometabolites of the cyanobacterium Nostoc insulare  

Microsoft Academic Search

The isolation, identification and quantification of exometabolites from culture media of the cyanobacterium Nostoc insulare are described. Besides the known exometabolite 4,4?-dihydroxybiphenyl (I), two more compounds, the ?-carboline 9H-pyrido(3,4-b)indole\\u000a (norharmane, II) and N,N?-(4,5-dimethyl-1,2-phenylene)bis-acetamide (III), were discovered. Concentrations of all three compounds\\u000a in media and biomass of five 250 L cultures of N. insulare were determined. Culture medium values for I ranged

Rainer-B. Volk; Sabine Mundt

2007-01-01

102

Short RNA half-lives in the slow-growing marine cyanobacterium Prochlorococcus  

Microsoft Academic Search

Background  RNA turnover plays an important role in the gene regulation of microorganisms and influences their speed of acclimation to\\u000a environmental changes. We investigated whole-genome RNA stability of Prochlorococcus, a relatively slow-growing marine cyanobacterium doubling approximately once a day, which is extremely abundant in the oceans.\\u000a \\u000a \\u000a \\u000a \\u000a Results  Using a combination of microarrays, quantitative RT-PCR and a new fitting method for determining RNA

Claudia Steglich; Debbie Lindell; Matthias Futschik; Trent Rector; Robert Steen; Sallie W Chisholm

2010-01-01

103

Photosynthetic production of the filamentous cyanobacterium Spirulina platensis in a cone-shaped helical tubular photobioreactor  

Microsoft Academic Search

The photosynthetic productivity of the filamentous cyanobacterium Spirulina platensis was investigated in a cone-shaped helical tubular photobioreactor. A laboratory-scale photobioreactor was constructed with a 0.255-m2 basal area and a conical shape (0.64rm highǴ.57rm top diameter). The photostage comprised transparent reinforced polyvinyl chloride (PVC) tubing with spirally wound, metal-wire reinforcing in the tubing wall (31rm in length and 1.6rcm internal diameter

Y. Watanabe; D. O. Hall

1996-01-01

104

New anabaenopeptins, potent carboxypeptidase-A inhibitors from the cyanobacterium Aphanizomenon flos-aquae.  

PubMed

Anabaenopeptins I (1) and J (2), two new ureido bond-containing cyclic peptides, were isolated from the cultured cyanobacterium Aphanizomenon flos-aquae (NIES-81) as potent carboxypeptidase-A (CPA) inhibitors. The gross structures of 1 and 2 were established by spectroscopic analysis, including the 2D NMR techniques. The absolute configurations of 1 and 2 were determined by spectral and chemical methods. Anabaenopeptins I and J inhibited CPA with IC(50) values of 5.2 and 7.6 ng/mL, respectively. PMID:11000037

Murakami, M; Suzuki, S; Itou, Y; Kodani, S; Ishida, K

2000-09-01

105

Effects of three pesticides on MSX-induced ammonia photoproduction by the cyanobacterium Nostoc linckia.  

PubMed

Three pesticides (2,4-D, basalin, aldrin) inhibited L-methionine-DL-sulfoximine (MSX)-induced photoproduction of ammonia by the nitrogen-fixing cyanobacterium Nostoc linckia. Combinations of pesticides and MSX were inhibitory except at low concentrations (100 and 500 micrograms/ml) of 2,4-D which stimulated NH+4 production. Similar results were obtained when pesticides were added 6 hr after the addition of MSX, but the inhibition was weaker. When MSX was added to the culture 6 hr after the addition of pesticides, the pesticides stimulated NH+4 photoproduction. Similar results were obtained on nitrogenase activity of the organism. PMID:2509191

Mishra, A K; Pandey, A B; Kumar, H D

1989-10-01

106

Planktocyclin, a cyclooctapeptide protease inhibitor produced by the freshwater cyanobacterium Planktothrix rubescens.  

PubMed

The freshwater cyanobacterium Planktothrix rubescens produces the cyclooctapeptide cyclo(Pro-Gly-Leu-Val-Met-Phe-Gly-Val). The chemical structure is new. This homodetic cyclic octapeptide was named planktocyclin ( 1). It consists solely of proteinogenic l-amino acids and is a strong inhibitor of mammalian trypsin and alpha-chymotrypsin and a moderately active inhibitor of human recombinant caspase-8. Mass spectrometric and 2D-NMR spectroscopic data allowed the determination of its structure. Synthetic planktocyclin was identical to the natural product. PMID:17935298

Baumann, Heike I; Keller, Simone; Wolter, Falko E; Nicholson, Graeme J; Jung, Günther; Süssmuth, Roderich D; Jüttner, Friedrich

2007-10-13

107

The blooms of a cyanobacterium, Microcystis cf. aeruginosa in a severely polluted estuary, the Golden Horn, Turkey  

Microsoft Academic Search

The distribution of toxic cyanobacterium Microcystis cf. aeruginosa in the severely polluted Golden Horn Estuary was studied from 1998 to 2000. Microcystis persisted at the upper estuary where the water circulation was poor and values ranged between 2.9 × 104 and 2.7 × 106 cells ml-1 throughout the study. Simultaneously measured physical (salinity, temperature, rainfall and secchi disc) and chemical

Seyfettin Tas; Erdogan Okus; Asli Aslan-Yilmaz

2006-01-01

108

Effects of the cyanobacterium Cylindrospermopsis raciborskii on feeding and life-history characteristics of the grazer Daphnia magna  

Microsoft Academic Search

Laboratory experiments were used to test the hypothesis that feeding and growth of the zooplankton grazer Daphnia magna will decrease with increasing proportions of the cyanobacterium Cylindrospermopsis raciborskii in the diet (mixed feeds with the green alga Scenedesmus obliquus). A strain of C. raciborskii, which does not produce cylindrospermopsin but contains saxitoxins and gonyautoxins, was not acutely toxic to Daphnia,

Maria Carolina S. Soares; M. F. L. L. W. Lürling; Renata Panosso; Vera Huszar

2009-01-01

109

Dominance of the noxious cyanobacterium Microcystis aeruginosa in low-nutrient lakes is associated with exotic zebra mussels  

Microsoft Academic Search

To examine the hypothesis that invasion by zebra mussels (Dreissena polymorpha) promotes phytoplankton dominance by the noxious cyanobacterium Microcystis aeruginosa, 61 Michigan lakes of varying nutrient levels that contain or lack zebra mussels were surveyed during late summer. After accounting for variation in total phos- phorus (TP) concentrations, lakes with Dreissena had lower total phytoplankton biomass, as measured by chloro-

David F. Raikow; Orlando Sarnelle; Alan E. Wilson; Stephen K. Hamilton

2004-01-01

110

Identification of a benthic microcystin-producing filamentous cyanobacterium (Oscillatoriales) associated with a dog poisoning in New Zealand  

Microsoft Academic Search

In November 2008 a dog died soon after ingesting benthic “algal” mat material from the Waitaki River, New Zealand. Based on a morphological examination of environmental material, the causative organism was putatively identified as the filamentous cyanobacterium Phormidium sp. Two strains (VUW25 and CYN61) were isolated and cultured to enable further taxonomic and cyanotoxin characterisation. Phylogenetic analyses based on a

Susanna A. Wood; Mark W. Heath; Patrick T. Holland; Rex Munday; Glenn B. McGregor; Ken G. Ryan

2010-01-01

111

Strategies for the co-existence of zooplankton with the toxic cyanobacterium Planktothrix rubescens in Lake Zurich  

Microsoft Academic Search

Since the cyanobacterium Planktothrix rubescens, which dominates the phytoplankton community in Lake Zürich, is generally considered toxic to zooplankton, we addressed the question whether co-occurring zooplankton species have developed adaptive responses. Artificially shortened filaments (<30 µm in length) of P.rubescens significantly reduced survival of Thamnocephalus platyurus (Crustacea, Branchiopoda, Anostraca) naturally occurring in temporary ponds. In contrast to Thamnocephalus, the survival

Rainer Kurmayer; Friedrich Jüttner

1999-01-01

112

Acute toxicity of excess mercury on the photosynthetic performance of cyanobacterium, S. platensis – assessment by chlorophyll fluorescence analysis  

Microsoft Academic Search

Measurement of chlorophyll fluorescence has been shown to be a rapid, non-invasive, and reliable method to assess photosynthetic performance in a changing environment. In this study, acute toxicity of excess Hg on the photosynthetic performance of the cyanobacterium S. platensis, was investigated by use of chlorophyll fluorescence analysis after cells were exposed to excess Hg (up to 20 ?M) for

C. M Lu; C. W Chau; J. H Zhang

2000-01-01

113

Com putational inference and experimental validation of the nitrogen assimilation regulatory network in cyanobacterium Synechococcus sp. WH 8102  

Microsoft Academic Search

Deciphering the regulatory networks encoded in the genome of an organism represents one of the most interesting and challenging tasks in the post- genome sequencing era. As an example of this problem, we have predicted a detailed model for the nitrogen assimilation network in cyanobacterium Synechococcus sp. WH 8102 (WH8102) using a com- putational protocol based on comparative genomics analysis

Zhengchang Su; Fenglou Mao; Phuongan Dam; Hongwei Wu; Victor Olman; Ian T. Paulsen; Brian Palenik; Ying Xu

114

Methods for prevention of mass development of the cyanobacterium Microcystis aeruginosa Kutz emend. Elenk. in aquatic systems  

Microsoft Academic Search

Methods for prevention of mass development of the cyanobacterium Microcystis aeruginosa Kutz emend. Elenk. in continental water bodies and industrial water supply systems are reviewed. The physicochemical, chemical,\\u000a and biological methods for prevention of M. aeruginosa development in water bodies and water supply systems are considered; examples of successful inhibition of M. aeruginosa growth in laboratory experiments are demonstrated. The

V. I. Kolmakov

2006-01-01

115

Precision of different methods used for estimating the abundance of the nitrogen-fixing marine cyanobacterium, Trichodesmium Ehrenberg  

Microsoft Academic Search

Variances involved in estimating the abundance of the nitrogen-fixing marine cyanobacterium, Trichodesmium Ehrenberg, were evaluated by repeated sampling using bottle casts and plankton net tows at two stations in the southern East China Sea. The variance among individual samples and the variance arising from subsampling processes were separated by the method of analysis of variance, and the coefficient of variation

Jeng Chang

2000-01-01

116

Viridamides A and B, lipodepsipeptides with antiprotozoal activity from the marine cyanobacterium Oscillatoria nigro-viridis.  

PubMed

Parallel chemical and phylogenetic investigation of a marine cyanobacterium from Panama led to the isolation of two new PKS-NRPS-derived compounds, viridamides A and B. Their structures were determined by NMR and mass spectroscopic methods, and the absolute configurations assigned by Marfey's method and chiral HPLC analysis. In addition to six standard, N-methylated amino and hydroxy acids, these metabolites contained the structurally novel 5-methoxydec-9-ynoic acid moiety and an unusual proline methyl ester terminus. Morphologically, this cyanobacterium was identified as Oscillatoria nigro-viridis, and its 16S rDNA sequence is reported here for the first time. Phylogenetic analysis of these sequence data has identified O. nigro-viridis strain OSC3L to be closely related to two other marine cyanobacterial genera, Trichodesmium and Blennothrix. Viridamide A showed antitrypanosomal activity with an IC50 of 1.1 microM and antileishmanial activity with an IC50 of 1.5 microM. PMID:18715036

Simmons, T Luke; Engene, Niclas; Ureña, Luis David; Romero, Luz I; Ortega-Barría, Eduardo; Gerwick, Lena; Gerwick, William H

2008-08-21

117

Diurnal rhythms result in significant changes in the cellular protein complement in the cyanobacterium Cyanothece 51142.  

PubMed

Cyanothece sp. ATCC 51142 is a diazotrophic cyanobacterium notable for its ability to perform oxygenic photosynthesis and dinitrogen fixation in the same single cell. Previous transcriptional analysis revealed that the existence of these incompatible cellular processes largely depends on tightly synchronized expression programs involving ?30% of genes in the genome. To expand upon current knowledge, we have utilized sensitive proteomic approaches to examine the impact of diurnal rhythms on the protein complement in Cyanothece 51142. We found that 250 proteins accounting for ?5% of the predicted ORFs from the Cyanothece 51142 genome and 20% of proteins detected under alternating light/dark conditions exhibited periodic oscillations in their abundances. Our results suggest that altered enzyme activities at different phases during the diurnal cycle can be attributed to changes in the abundance of related proteins and key compounds. The integration of global proteomics and transcriptomic data further revealed that post-transcriptional events are important for temporal regulation of processes such as photosynthesis in Cyanothece 51142. This analysis is the first comprehensive report on global quantitative proteomics in a unicellular diazotrophic cyanobacterium and uncovers novel findings about diurnal rhythms. PMID:21364985

Stöckel, Jana; Jacobs, Jon M; Elvitigala, Thanura R; Liberton, Michelle; Welsh, Eric A; Polpitiya, Ashoka D; Gritsenko, Marina A; Nicora, Carrie D; Koppenaal, David W; Smith, Richard D; Pakrasi, Himadri B

2011-02-22

118

Unique thylakoid membrane architecture of a unicellular N2-fixing cyanobacterium revealed by electron tomography.  

PubMed

Cyanobacteria, descendants of the endosymbiont that gave rise to modern-day chloroplasts, are vital contributors to global biological energy conversion processes. A thorough understanding of the physiology of cyanobacteria requires detailed knowledge of these organisms at the level of cellular architecture and organization. In these prokaryotes, the large membrane protein complexes of the photosynthetic and respiratory electron transport chains function in the intracellular thylakoid membranes. Like plants, the architecture of the thylakoid membranes in cyanobacteria has direct impact on cellular bioenergetics, protein transport, and molecular trafficking. However, whole-cell thylakoid organization in cyanobacteria is not well understood. Here we present, by using electron tomography, an in-depth analysis of the architecture of the thylakoid membranes in a unicellular cyanobacterium, Cyanothece sp. ATCC 51142. Based on the results of three-dimensional tomographic reconstructions of near-entire cells, we determined that the thylakoids in Cyanothece 51142 form a dense and complex network that extends throughout the entire cell. This thylakoid membrane network is formed from the branching and splitting of membranes and encloses a single lumenal space. The entire thylakoid network spirals as a peripheral ring of membranes around the cell, an organization that has not previously been described in a cyanobacterium. Within the thylakoid membrane network are areas of quasi-helical arrangement with similarities to the thylakoid membrane system in chloroplasts. This cyanobacterial thylakoid arrangement is an efficient means of packing a large volume of membranes in the cell while optimizing intracellular transport and trafficking. PMID:21173021

Liberton, Michelle; Austin, Jotham R; Berg, R Howard; Pakrasi, Himadri B

2010-12-20

119

Crucial Role of Extracellular Polysaccharides in Desiccation and Freezing Tolerance in the Terrestrial Cyanobacterium Nostoc commune  

PubMed Central

The cyanobacterium Nostoc commune is adapted to the terrestrial environment and has a cosmopolitan distribution. In this study, the role of extracellular polysaccharides (EPS) in the desiccation tolerance of photosynthesis in N. commune was examined. Although photosynthetic O2 evolution was not detected in desiccated colonies, the ability of the cells to evolve O2 rapidly recovered after rehydration. The air-dried colonies contained approximately 10% (wt/wt) water, and field-isolated, natural colonies with EPS were highly water absorbent and were rapidly hydrated by atmospheric moisture. The cells embedded in EPS in Nostoc colonies were highly desiccation tolerant, and O2 evolution was not damaged by air drying. Although N. commune was determined to be a mesophilic cyanobacterium, the cells with EPS were heat tolerant in a desiccated state. EPS could be removed from cells by homogenizing colonies with a blender and filtering with coarse filter paper. This treatment to remove EPS did not damage Nostoc cells or their ability to evolve O2, but O2 evolution was significantly damaged by desiccation treatment of the EPS-depleted cells. Similar to the EPS-depleted cells, the laboratory culture strain KU002 had only small amount of EPS and was highly sensitive to desiccation. In the EPS-depleted cells, O2 evolution was also sensitive to freeze-thaw treatment. These results strongly suggest that EPS of N. commune is crucial for the stress tolerance of photosynthesis during desiccation and during freezing and thawing.

Tamaru, Yoshiyuki; Takani, Yayoi; Yoshida, Takayuki; Sakamoto, Toshio

2005-01-01

120

Aluminum effects on uptake and metabolism of phosphorus by the Cyanobacterium Anabaena cylindrica  

SciTech Connect

Aluminum severely affects the growth of the cyanobacterium Anabaena cylindrica and induces symptoms indicating phosphorus starvation. Pre- or post-treating the cells with high (90 micromolar) phosphorus reduces the toxicity of aluminum compared to cells receiving a lower orthophosphate concentration. In this study aluminum (ranging from 9 to 36 micromolar) and phosphorus concentrations were chosen so that the precipitation of insoluble AlPO/sub 4/ never exceeded 10% of the total phosphate concentration. The uptake of /sup 32/P-phosphorus is not disturbed by aluminium either at high (100 micromolar) or low (10 micromolar) concentrations of phosphate. Also, the rapid accumulation of polyphosphate granules in cells exposed to aluminum indicates that the incorporation of phosphate is not disturbed. However, a significant decrease in the mobilization of the polyphosphates is observed, as is a lowered activity of the enzyme acid phosphatase, in aluminum treated cells. We conclude that aluminum acts on the intracellular metabolism of phosphate, which eventually leads to phosphorus starvation rather than on its uptake in the cyanobacterium A. cylindrica.

Pettersson, A.; Haellbom, L. Bergman, B.

1988-01-01

121

Antiherpetic efficacy of aqueous extracts of the cyanobacterium Arthrospira fusiformis from Chad.  

PubMed

Natural substances offer interesting pharmacological perspectives for antiviral drug development with regard to broad spectrum antiviral properties and novel modes of action. Drugs currently used to treat cutaneous or genital herpetic infections are effective in limiting disease, but the emergence of drug-resistant viruses in immunocompromised individuals can be problematic. A nontoxic cyanobacterium Arthrospira strain from Chad has been characterized by sequence analysis of the intergenic spacer region of the phycocyanin gene. This cyanobacterium was identified as Arthrospira fusiformis by phylogenetic tree analysis. The antiherpetic activity of crude aqueous extracts from the Chad A. fusiformis isolate was determined. Antiviral efficacy against herpes simplex virus of cold water extract, hot water extract and phosphate buffer extract was assessed in plaque reduction assays and their mode of antiherpetic action was analysed. In virus suspension assays, cold water extract, hot water extract and phosphate buffer extract inhibited virus infectivity by 54.9%, 64.6%, and 99.8%, respectively, in a dose-dependent manner. The mode of antiviral action was determined by addition of cyanobacterial extracts separately at different time periods during the viral infection cycle. Extracts of A. fusiformis strain clearly inhibited herpesvirus multiplication before and during virus infection of host cells. The phosphate buffer extract of the A. fusiformis strain affected free herpes simplex virus prior to infection of host cells and inhibited intracellular viral replication. It is concluded, that Arthrospira compounds warrant further investigation to examine their potential role in the treatment of herpetic infections. PMID:23802437

Sharaf, M; Amara, A; Aboul-Enein, A; Helmi, S; Ballot, A; Schnitzler, P

2013-05-01

122

Spectroscopic properties of PSI-IsiA supercomplexes from the cyanobacterium Synechococcus PCC 7942.  

PubMed

The cyanobacterium Synechococcus PCC 7942 grown under iron starvation assembles a supercomplex consisting of a trimeric Photosystem I (PSI) complex encircled by a ring of 18 CP43' or IsiA light-harvesting complexes [Nature 412 (2001) 745]. Here we present a spectroscopic characterization by temperature-dependent absorption and fluorescence spectroscopy, site-selective fluorescence spectroscopy at 5 K, and circular dichroism of isolated PSI-IsiA, PSI and IsiA complexes from this cyanobacterium grown under iron starvation. The results suggest that the IsiA ring increases the absorption cross-section of PSI by about 100%. Each IsiA subunit binds about 16-17 chlorophyll a (Chl a) molecules and serves as an efficient antenna for PSI. Each of the monomers of the trimeric PSI complex contains two red chlorophylls, which presumably give rise to one exciton-coupled dimer and at 5 K absorb and fluoresce at 703 and 713 nm, respectively. The spectral properties of these C-703 chlorophylls are not affected by the presence of the IsiA antenna ring. The spectroscopic properties of the purified IsiA complexes are similar to those of the related CP43 complex from plants, except that the characteristic narrow absorption band of CP43 at 682.5 nm is missing in IsiA. PMID:12460685

Andrizhiyevskaya, Elena G; Schwabe, Tatjana M E; Germano, Marta; D'Haene, Sandrine; Kruip, Jochen; van Grondelle, Rienk; Dekker, Jan P

2002-12-01

123

Allelopathic growth inhibition by the toxic, bloom-forming cyanobacterium Planktothrix rubescens.  

PubMed

Blooms of freshwater cyanobacteria are typically accompanied by an important decrease in phytoplankton biodiversity in the water bodies where they occur. This study examines the potential production of growth-inhibiting substances by the toxic, bloom-forming cyanobacterium Planktothrix rubescens, following the observation of physical segregation between this and another cyanobacterium during previously performed mixed-culture competition experiments. Inhibition assays examining the growth of target strains exposed to donor culture filtrates showed that the growth of Planktothrix agardhii TCC 83-2, P. agardhii PMC 75.02 and Mougeotia gracillima TCC 50-2 was significantly inhibited in the presence of culture filtrate from P. rubescens TCC 29-1, isolated from Lake Bourget, France. Filtrates from P. rubescens TCC 69-6 and P. rubescens TCC 69-7, isolated from Lakes Nantua and Paladru (France), respectively, did not, however, inhibit the growth of P. agardhii TCC 83-2. This brief exploration of the allelopathic activity of P. rubescens suggests that it may potentially inhibit coexisting competitors as well as phytoplankton isolated from other freshwater ecosystems, and that this capacity may vary among different strains of Planktothrix. The potential importance of this phenomenon in pelagic competition dynamics is discussed. PMID:18752621

Oberhaus, Laura; Briand, Jean-François; Humbert, Jean-François

2008-08-20

124

Evidence for a stable association of Psb30 (Ycf12) with photosystem II core complex in the cyanobacterium Synechocystis sp. PCC 6803  

Microsoft Academic Search

Ycf12 (Psb30) is a small hydrophobic subunit of photosystem II (PS II) complexes found in the cyanobacterium, Thermosynechococcus elongatus. However, earlier intense proteomic analysis on the PS II complexes from the cyanobacterium, Synechocystis 6803, could not detect Psb30. In this work, we generated a mutant of Synechocystis 6803 in which a hexa-histidine tag was fused to the C-terminus of Synechocystis

Natsuko Inoue-Kashino; Takeshi Takahashi; Akiko Ban; Miwa Sugiura; Yuichiro Takahashi; Kazuhiko Satoh; Yasuhiro Kashino

2008-01-01

125

Detection of bioactive exometabolites produced by the filamentous marine cyanobacterium Geitlerinema sp.  

PubMed

Marine cyanobacteria are noted for their ability to excrete metabolites with biotic properties. This paper focuses on such exometabolites obtained from the culture of the marine filamentous cyanobacterium Geitlerinema sp. strain, their purification and subsequent analyses. By this means the recoveries of the active compounds, a prerequisite for properly determining their concentration, are quantified here for the first time. We demonstrate a new procedure using Amberlite XAD-1180 resin in combination with the eluent isopropanol for extraction of the culture media and gas chromatography as simplified chemical analysis. This procedure reduced necessary bacteria cultivation time (from 150 to 21 days) at low volumes of culture media (300 mL) required for identification of two selected bioactive compounds: 4,4'-dihydroxybiphenyl and harmane. PMID:22160344

Caicedo, Nelson H; Kumirska, Jolanta; Neumann, Jennifer; Stolte, Stefan; Thöming, Jorg

2011-12-13

126

Changes in non-core regions stabilise plastocyanin from the thermophilic cyanobacterium Phormidium laminosum.  

PubMed

We report a theoretical investigation on the different stabilities of two plastocyanins. The first one belongs to the thermophilic cyanobacterium Phormidium laminosum and the second one belongs to its mesophilic relative Synechocystis sp. These proteins share the same topology and secondary-structure elements; however, the melting temperatures of their oxidised species differ by approximately 15 K. Long-time-scale molecular dynamics simulations, performed at different temperatures, show that the thermophilic protein optimises a set of intramolecular interactions (interstrand hydrogen bonding, salt bridging and hydrophobic clustering) within the region that comprises the strands beta 5 and beta 6, loop L5 and the helix. This region exhibits most of the differences in the primary sequence between the two proteins and, in addition, it is involved in the interaction with known physiological partners. Further work is in progress to unveil the specific structural features responsible for the different thermal stability of the two proteins. PMID:19915878

Muñoz-López, Francisco J; Raugei, Simone; De la Rosa, Miguel A; Díaz-Quintana, Antonio J; Carloni, Paolo

2010-03-01

127

Venturamides A and B: antimalarial constituents of the panamanian marine Cyanobacterium Oscillatoria sp.  

PubMed

Two new modified cyclic hexapeptides, venturamides A (1) and B (2), were isolated from the marine cyanobacterium Oscillatoria sp. by antimalarial bioassay-guided fractionation. The isolation of 1 and 2 represents the first example of the identification of cyanobacterial peptides with selective antimalarial activity. The planar structures of 1 and 2 were determined by 1D and 2D NMR analyses and, in the case of venturamide A (1), comparison with the literature data for a previously reported synthetic compound. The absolute configuration of the amino acid residues was determined by selective hydrolysis in conjunction with Marfey's analysis. Compounds 1 and 2 were tested for biological activity against a range of tropical parasites. PMID:17328572

Linington, Roger G; Gonzalez, José; Ureña, Luis-David; Romero, Luz I; Ortega-Barría, Eduardo; Gerwick, William H

2007-03-01

128

Unique modification of adenine in genomic DNA of the marine cyanobacterium Trichodesmium sp. strain NIBB 1067.  

PubMed Central

The genomic DNA of the marine nonheterocystous nitrogen-fixing cyanobacterium Trichodesmium sp. strain NIBB 1067 was found to be highly resistant to DNA restriction endonucleases. The DNA was digested extensively by the restriction enzyme DpnI, which requires adenine methylation for activity. The DNA composition, determined by high-performance liquid chromatography (HPLC), was found to be 69% AT. Surprisingly, it was found that a modified adenine which was not methylated at the usual N6 position was present and made up 4.7 mol% of the nucleosides in Trichodesmium DNA (15 mol% of deoxyadenosine). In order for adenine residues to be modified at this many positions, there must be many modifying enzymes or at least one of the modifying enzymes must have a degenerate recognition site. The reason(s) for this extensive methylation has not yet been determined but may have implications for the ecological success of this microorganism in nature. Images FIG. 1 FIG. 2

Zehr, J P; Ohki, K; Fujita, Y; Landry, D

1991-01-01

129

Natural Products Chemistry and Taxonomy of the Marine Cyanobacterium Blennothrix cantharidosmum  

PubMed Central

A Papua New Guinea field collection of the marine cyanobacterium Blennothrix cantharidosmum was investigated for its cytotoxic constituents. Bioassay-guided isolation defined the cytotoxic components as the known compounds lyngbyastatins 1 and 3. However, six new acyl proline derivatives, tumonoic acids D-I, plus the known tumonoic acid A, were also isolated. Their planar structures were defined from NMR and MS data while their stereostructures followed from a series of chiral chromatographies, degradation sequences and synthetic approaches. The new compounds were tested in an array of assays, but showed only modest antimalarial and inhibition of quorum sensing activities. Nevertheless, these are the first natural products to be reported from this genus, and this inspired a detailed morphologic and 16S rDNA-based phylogenetic analysis of the producing organism.

Clark, Benjamin R.; Engene, Niclas; Teasdale, Margaret E.; Rowley, David C.; Matainaho, Teatulohi; Valeriote, Frederick A.; Gerwick, William H.

2009-01-01

130

Coibamide A, a Potent Antiproliferative Cyclic Depsipeptide from the Panamanian Marine Cyanobacterium Leptolyngbya sp.  

PubMed Central

Coibamide A (1) is a new, potent antiproliferative depsipeptide which was isolated from a marine Leptolyngbya cyanobacterium collected from the Coiba National Park, Panama. The planar structure of 1 was elucidated by a combination of NMR spectroscopy and mass spectrometry. Exhaustive 1D and 2D NMR spectroscopy included natural abundance 15N and variable temperature experiments; mass spectrometry included TOF-ESI-MSn and FT-MSn experiments. Chemical degradation followed by chiral HPLC- and GC-MS analyses was used to assign the absolute configuration of 1. This highly methylated cyclized depsipeptide exhibited an unprecedented selectivity profile in the NCI 60 cancer cell line panel and appears to act via a novel mechanism.

Medina, Rebecca A.; Goeger, Douglas E.; Hills, Patrice; Mooberry, Susan L.; Huang, Nelson; Romero, Luz I.; Ortega-Barria, Eduardo; Gerwick, William H.; McPhail, Kerry L.

2008-01-01

131

Physiological effects of nickel chloride on the freshwater cyanobacterium Synechococcus sp. IU 625  

PubMed Central

Harmful algal blooms (HABs) are a serious environmental problem globally. The ability of cyanobacteria, one of the major causative agents of HABs, to grow in heavy metal polluted areas is proving a challenge to environmental restoration initiatives. Some cyanobacteria secrete toxins, such as microcystin, that are potentially dangerous to animals and humans. In this study, the physiology of a cyanobacterium was assessed to nickel chloride exposure. Cell growths were monitored throughout the study with various nickel chloride concentrations (0, 10, 25 or 50 mg/L). Morphological abnormalities were observed with microscopic image analyses. Inductively coupled plasma mass spectrometry (ICP-MS) was carried out to trace the distribution of nickel during the growth period. This study provides insight on potential nickel response mechanisms in freshwater cyanobacteria, which may lead to effective HAB prevention strategy development.

Nohomovich, Brian; Nguyen, Bao T.; Quintanilla, Michael; Lee, Lee H.; Murray, Sean R.; Chu, Tin-Chun

2013-01-01

132

Genotype x genotype interactions between the toxic cyanobacterium Microcystis and its grazer, the waterflea Daphnia  

PubMed Central

Toxic algal blooms are an important problem worldwide. The literature on toxic cyanobacteria blooms in inland waters reports widely divergent results on whether zooplankton can control cyanobacteria blooms or cyanobacteria suppress zooplankton by their toxins. Here we test whether this may be due to genotype × genotype interactions, in which interactions between the large-bodied and efficient grazer Daphnia and the widespread cyanobacterium Microcystis are not only dependent on Microcystis strain or Daphnia genotype but are specific to genotype × genotype combinations. We show that genotype × genotype interactions are important in explaining mortality in short-time exposures of Daphnia to Microcystis. These genotype × genotype interactions may result in local coadaptation and a geographic mosaic of coevolution. Genotype × genotype interactions can explain why the literature on zooplankton–cyanobacteria interactions is seemingly inconsistent, and provide hope that zooplankton can contribute to the suppression of cyanobacteria blooms in restoration projects.

Lemaire, Veerle; Brusciotti, Silvia; van Gremberghe, Ineke; Vyverman, Wim; Vanoverbeke, Joost; De Meester, Luc

2012-01-01

133

Growth and biopigment accumulation of cyanobacterium Spirulina platensis at different light intensities and temperature  

PubMed Central

In order to find out optimum culture condition for algal growth, the effect of light irradiance and temperature on growth rate, biomass composition and pigment production of Spirulina platensis were studied in axenic batch cultures. Growth kinetics of cultures showed a wide range of temperature tolerance from 20 °C to 40 °C. Maximum growth rate, cell production with maximum accumulation of chlorophyll and phycobilliproteins were found at temperature 35 °C and 2,000 lux light intensity. But with further increase in temperature and light intensity, reduction in growth rate was observed. Carotenoid content was found maximum at 3,500 lux. Improvement in the carotenoid content with increase in light intensity is an adaptive mechanism of cyanobacterium S.platensis for photoprotection, could be a good basis for the exploitation of microalgae as a source of biopigments.

Kumar, Manoj; Kulshreshtha, Jyoti; Singh, Gajendra Pal

2011-01-01

134

High rates of photobiological H2 production by a cyanobacterium under aerobic conditions.  

PubMed

Among the emerging renewable and green energy sources, biohydrogen stands out as an appealing choice. Hydrogen can be produced by certain groups of microorganisms that possess functional nitrogenase and/or bidirectional hydrogenases. In particular, the potential of photobiological hydrogen production by oxygenic photosynthetic microbes has attracted significant interest. However, nitrogenase and hydrogenase are generally oxygen sensitive, and require protective mechanisms to function in an aerobic extracellular environment. Here, we describe Cyanothece sp. ATCC 51142, a unicellular, diazotrophic cyanobacterium with the capacity to generate high levels of hydrogen under aerobic conditions. Wild-type Cyanothece 51142 can produce hydrogen at rates as high as 465 ?mol per mg of chlorophyll per hour in the presence of glycerol. Hydrogen production in this strain is mediated by an efficient nitrogenase system, which can be manipulated to convert solar energy into hydrogen at rates that are several fold higher, compared with any previously described wild-type hydrogen-producing photosynthetic microbe. PMID:21266989

Bandyopadhyay, Anindita; Stöckel, Jana; Min, Hongtao; Sherman, Louis A; Pakrasi, Himadri B

2010-01-01

135

Ribulose-1,5-bisphosphate carboxylase/oxygenase from thermophilic cyanobacterium Thermosynechococcus elongatus.  

PubMed

Ribulose-1,5-bisphosphate carboxylase/oxygenase (rubisco) can be divided into two branches: the "red-like type" of marine algae and the "green-like type" of cyanobacteria, green algae, and higher plants. We found that the "green-like type" rubisco from the thermophilic cyanobacterium Thermosynechococcus elongatus has an almost 2-fold higher specificity factor compared with rubiscos of mesophilic cyanobacteria, reaching the values of higher plants, and simultaneously revealing an improvement in enzyme thermostability. The difference in the activation energies at the transition stages between the oxygenase and carboxylase reactions for Thermosynechococcus elongatus rubisco is very close to that of Galdieria partita and significantly higher than that of spinach. This is the first characterization of a "green-like type" rubisco from thermophilic organism. PMID:17922215

Gubernator, Beata; Bartoszewski, Rafal; Kroliczewski, Jaroslaw; Wildner, Guenter; Szczepaniak, Andrzej

2007-10-06

136

Instability and variable toxicity of HBP-Tx, a toxin in the cyanobacterium Microcystis aeruginosa.  

PubMed

It was found that autoxidative degradation is responsible for the inactivation of the unstable Microcystis toxin HBP-Tx. The purified toxin was similar in its properties to the "fast-death-factor" in Microcystis, described as a cyclic peptide in the literature. The apparent presence of an entirely different toxin was simulated by the partially inactivated HBP-Tx. A number of associated fluorescent compounds were identified as the non-toxic degradation products of the toxin. As a consequence, as established method for the detection of other algal toxins was applied. This chemical assay, which uses fluorescent measurement of the oxidized toxin in the cyanobacterium Aphanizomenon flos-aquae, was applicable for HBP-Tx after the removal of interfering degradation products of the toxin. The results obtained with Microcystis toxin HBP-Tx do not confirm suggestions concerning the structure of the "fast-death-factor". PMID:6426092

Amann, M J

1984-01-01

137

Causes for the large genome size in a cyanobacterium Anabaena sp. PCC7120.  

PubMed

Three possible causes responsible for the large genome size of a cyanobacterium Anabaena sp. PCC7120 are investigated: 1) sequential tandem duplications of gene segments, genes or genomic segments, 2) horizontal gene transfers from other organisms, and 3) whole-genome duplication. We evaluated the frequency distribution of angles between paralog locations for the possibility 1), the fraction of genes deviated in GC content, GC skew, AT skew and codon adaptation index for the 2) and the gene-configuration comparison of paralogs for the 3). As a result, the possibility 3), the whole-genome duplication, was more reasonable as a molecular cause than the other causes for the large genome size in Anabaena sp. PCC7120. In addition, the whole-genome duplication was supported by the analysis of distribution pattern of protein genes with respect to functional categories. PMID:15712125

Sugaya, Nobuyoshi; Sato, Makihiko; Murakami, Hiroo; Imaizumi, Akira; Aburatani, Sachiyo; Horimoto, Katsuhisa

2004-01-01

138

Glycinebetaine as an osmoregulant and compatible solute in the marine cyanobacterium Spirulina subsalsa.  

PubMed

Glycinebetaine was found to be the major organic substrate accumulating under hypersaline growth conditions in the halotolerant cyanobacterium Spirulina subsalsa. In addition to its proposed role as osmolite, glycinebetaine is shown to specifically protect enzymatic activity. Glucose-6-phosphate dehydrogenase from S. subsalsa retained full activity in the presence of NaCl at concentrations as high as 1.5 M, provided that comparable concentrations of glycinebetaine were also present in the reaction mixture. A kinetic analysis indicated that glycinebetaine protected the enzyme against both NaCl-induced decrease in Vmax and reduction in affinity to glucose 6-phosphate. The alternative osmolites, glycerol and proline, protected the enzyme against the reduction in Vmax but not against the reduction in affinity to glucose 6-phosphate. PMID:3134857

Gabbay-Azaria, R; Tel-Or, E; Schönfeld, M

1988-07-01

139

Sorption and desorption studies of chromium(VI) from nonviable cyanobacterium Nostoc muscorum biomass.  

PubMed

This communication presents results pertaining to the sorptive and desorptive studies carried out on chromium(VI) removal onto nonviable freshwater cyanobacterium (Nostoc muscorum) biomass. Influence of varying the conditions for removal of chromium(VI), such as the pH of aqueous solution, the dosage of biosorbent, the contact time with the biosorbent, the temperature for the removal of chromium, the effect of light metal ions and the adsorption-desorption studies were investigated. Sorption interaction of chromium on to cyanobacterial species obeyed both the first and the second-order rate equation and the experimental data showed good fit with both the Langmuir and freundlich adsorption isotherm models. The maximum adsorption capacity was 22.92 mg/g at 25 degrees C and pH 3.0. The adsorption process was endothermic and the values of thermodynamic parameters of the process were calculated. Various properties of the cyanobacterium, as adsorbent, explored in the characterization part were chemical composition of the adsorbent, surface area calculation by BET method and surface functionality by FTIR. Sorption-desorption of chromium into inorganic solutions and distilled water were observed and this indicated the biosorbent could be regenerated using 0.1 M HNO3 and EDTA with upto 80% recovery. The biosorbents were reused in five biosorption-desorption cycles without a significant loss in biosorption capacity. Thus, this study demonstrated that the cyanobacterial biomass N. muscorum could be used as an efficient biosorbent for the treatment of chromium(VI) bearing wastewater. PMID:18053641

Gupta, V K; Rastogi, A

2007-10-13

140

Genome Evolution of the Cyanobacterium Nostoc linckia under Sharp Microclimatic Divergence at "Evolution Canyon," Israel.  

PubMed

We describe the genomic DNA diversity and divergence of the cyanobacterium Nostoc linckia from "Evolution Canyon," a microsite consisting of ecologically contrasting slopes, south-facing slope (SFS) and north-facing slope (NFS), at lower Nahal Oren, Mt. Carmel, Israel. The opposing slopes share their limestone lithology but vary greatly in their ecology, primarily because of different levels of solar radiation (which is six times higher on the SFS than on the NFS). The warm and xeric SFS displays a tropical African savanna, whereas the cool and mesic NFS displays a temperate South European Mediterranean live-oak maquis shrub forest. The cyanobacterium Nostoc linckia tested here is a sessile microorganism, growing as a carpet on rock surfaces and constantly exposed to environmental fluctuations of solar radiation, temperature, and desiccation. We demonstrate remarkable interslope and intraslope genetic divergence of the genome (including both coding and noncoding regions) of Nostoc linckia, by using 211 AFLP (amplified fragment length polymorphism) DNA molecular marker loci. Genetic polymorphism of N. linckia subpopulations on the ecologically harsher SFS was significantly (p <0.05) higher (p = 99.53%) than was that of the subpopulations on the climatically milder nfs (p = 85.78%). genetic polymorphism (p) and gene diversity (he) were significantly correlated with variables influencing aridity stress: solar radiation (sr) (rp = 0.956; p = 0.046), temperature (tm) (rp = 0.993; p = 0.0068), and day-night temperature difference (tdd) (rp = 0.975; p = 0.025). as in other tested organisms from "evolution canyon", but even more exceptionally because of its completely sedentary nature, we suggest that the climatically stressed sfs environment is responsible for this marked increase of genetic polymorphism, which is maintained by the combined evolutionary forces of diversifying and balancing selection. This could highlight the importance of ecological stress and selection in evolution and its remarkable effect on the genetic system across the prokaryotic genome. PMID:12024256

Satish, N.; Krugman, T.; Vinogradova, O.N.; Nevo, E.; Kashi, Y.

2001-10-01

141

Sucrose accumulation in salt-stressed cells of agp gene deletion-mutant in cyanobacterium Synechocystis sp PCC 6803.  

PubMed

The agp gene encoding the ADP-glucose pyrophosphorylase is involved in cyanobacterial glycogen synthesis and glucosylglycerol formation. By in vitro DNA recombination technology, a mutant with partial deletion of agp gene in the cyanobacterium Synechocystis sp. PCC 6803 was constructed. This mutant could not synthesize glycogen or the osmoprotective substance glucosylglycerol. In the mutant cells grown in the medium containing 0.9 M NaCl for 96 h, no glucosylglycerol was detected and the total amount of sucrose was 29 times of that of in wild-type cells. Furthermore, the agp deletion mutant could tolerate up to 0.9 M salt concentration. Our results suggest that sucrose might act as a similar potent osmoprotectant as glucosylglycerol in cyanobacterium Synechocystis sp. PCC 6803. PMID:12583900

Miao, Xiaoling; Wu, Qingyu; Wu, Guifang; Zhao, Nanming

2003-01-21

142

An iron stress operon involved in photosynthetic electron transport in the marine cyanobacterium Synechococcus sp. PCC 7002.  

PubMed

The iron-stress-induced genes isiA and isiB have been cloned and sequenced from the marine unicellular cyanobacterium Synechococcus sp. PCC 7002. These genes code for a photosystem II chlorophyll-binding protein and flavodoxin respectively. The genes form a dicistronic operon that is transcriptionally activated under iron-stress conditions to produce an abundant monocistronic message containing isiA and a much less abundant dicistronic message that also contains isiB. The arrangement of these genes, their transcriptional control and the relative abundance of the monocistronic and dicistronic messages produced under iron stress parallels the pattern shown by the freshwater cyanobacterium Synechococcus sp. PCC 7942. The genes for the corresponding proteins found under iron-replete conditions, CP-43 and ferredoxin, have also been cloned and sequenced. Northern blot analysis indicates that both of these genes are constitutively expressed under both iron-stress and iron-replete conditions. PMID:1527503

Leonhardt, K; Straus, N A

1992-08-01

143

A comparison of fermentation in the cyanobacterium Microcystis PCC7806 grown under a light\\/dark cycle and continuous light  

Microsoft Academic Search

The cyanobacterium Microcystis PCC7806, grown under continuous light, fermented endogenously stored glycogen to equimolar amounts of acetate and ethanol when incubated anaerobically in the dark. In addition, H-2, CO2 and some L-lactate were produced. This fermentation pattern differed from that displayed by cells which had been grown under an alternating light\\/dark (16\\/8 h) cycle. Such cells produced much more ethanol

ROY MOEZELAAR; LUCAS J. STAL

1997-01-01

144

Use of a transposon with luciferase as a reporter to identify environmentally responsive genes in a cyanobacterium  

Microsoft Academic Search

Anabaena, a filamentous cyanobacterium, is of developmental interest because, when deprived of fixed nitrogen, it shows patterned differentiation of Nâ-fixing cells called heterocysts. To help elucidate its early responses to a decrease in nitrogen, the authors used a derivative of transposon Tn5 to generate transcriptional fusions of promoterless bacterial luciferase genes, luxAB, to the Anabaena genome. Genes that responded to

C. P. Wolk; Yuping Cai; J. M. Panoff

1991-01-01

145

Functional reconstitution of photosystem I reaction center from cyanobacterium Synechocystis sp PCC6803 into liposomes using a new reconstitution procedure  

Microsoft Academic Search

Photosystem I reaction center from the cyanobacteriumSynechocystis sp PCC6803 was reconstituted into phosphatidylcholine\\/phosphatidic acid liposomes. Liposomes prepared by reversephase evaporation were treated with various amounts of different detergents and protein incorporation was analyzed at each step of the solubilization process. After detergent removal the activities of the resulting proteoliposomes were measured. The most efficient reconstitution was obtained by insertion of

Josep Cladera; Jean-Louis Rigaud; Hervé Bottin; Mireia Duñach

1996-01-01

146

A TRAP Transporter for Pyruvate and Other Monocarboxylate 2-Oxoacids in the Cyanobacterium Anabaena sp. Strain PCC 7120?  

PubMed Central

In the cyanobacterium Anabaena sp. strain PCC 7120, open reading frames (ORFs) alr3026, alr3027, and all3028 encode a tripartite ATP-independent periplasmic transporter (TRAP-T). Wild-type filaments showed significant uptake of [14C]pyruvate, which was impaired in the alr3027 and all3028 mutants and was inhibited by several monocarboxylate 2-oxoacids, identifying this TRAP-T system as a pyruvate/monocarboxylate 2-oxoacid transporter.

Pernil, Rafael; Herrero, Antonia; Flores, Enrique

2010-01-01

147

A TRAP transporter for pyruvate and other monocarboxylate 2-oxoacids in the cyanobacterium Anabaena sp. strain PCC 7120.  

PubMed

In the cyanobacterium Anabaena sp. strain PCC 7120, open reading frames (ORFs) alr3026, alr3027, and all3028 encode a tripartite ATP-independent periplasmic transporter (TRAP-T). Wild-type filaments showed significant uptake of [(14)C]pyruvate, which was impaired in the alr3027 and all3028 mutants and was inhibited by several monocarboxylate 2-oxoacids, identifying this TRAP-T system as a pyruvate/monocarboxylate 2-oxoacid transporter. PMID:20851902

Pernil, Rafael; Herrero, Antonia; Flores, Enrique

2010-09-17

148

Polyphasic characterization of a thermo-tolerant filamentous cyanobacterium isolated from the Euganean thermal muds (Padua, Italy)  

Microsoft Academic Search

In this paper we report a morphological, ultrastructural, biochemical and molecular (16S rRNA, 16S–23S ITS, rbcL and rpoC1 gene sequencing) survey on a very thin, non-heterocystous, filamentous cyanobacterium, isolated from mats covering several mud maturation tanks of the Euganean Thermal District, at temperatures ranging from 26 to 59°C. Denaturing gradient gel electrophoresis results, obtained using cyanobacterial primers targeting the 16S

Isabella Moro; Nicoletta Rascio; Nicoletta La Rocca; Katia Sciuto; Patrizia Albertano; Laura Bruno; Carlo Andreoli

2010-01-01

149

Complete nucleotide sequence of the freshwater unicellular cyanobacterium Synechococcus elongatus PCC 6301 chromosome: gene content and organization  

Microsoft Academic Search

The entire genome of the unicellular cyanobacterium Synechococcus elongatus PCC 6301 (formerly Anacystis nidulans Berkeley strain 6301) was sequenced. The genome consisted of a circular chromosome 2,696,255 bp long. A total of 2,525 potential\\u000a protein-coding genes, two sets of rRNA genes, 45 tRNA genes representing 42 tRNA species, and several genes for small stable\\u000a RNAs were assigned to the chromosome by

Chieko Sugita; Koretsugu Ogata; Masamitsu Shikata; Hiroyuki Jikuya; Jun Takano; Miho Furumichi; Minoru Kanehisa; Tatsuo Omata; Masahiro Sugiura; Mamoru Sugita

2007-01-01

150

The blooms of a cyanobacterium, Microcystis cf. aeruginosa in a severely polluted estuary, the Golden Horn, Turkey  

Microsoft Academic Search

The distribution of toxic cyanobacterium Microcystis cf. aeruginosa in the severely polluted Golden Horn Estuary was studied from 1998 to 2000. Microcystis persisted at the upper estuary where the water circulation was poor and values ranged between 2.9×104 and 2.7×106cellsml?1 throughout the study. Simultaneously measured physical (salinity, temperature, rainfall and secchi disc) and chemical parameters (nutrients and dissolved oxygen) were

Seyfettin Ta?; Erdo?an Oku?; Asl? Aslan-Y?lmaz

2006-01-01

151

Application of real-time PCR in the assessment of the toxic cyanobacterium Cylindrospermopsis raciborskii abundance and toxicological potential  

Microsoft Academic Search

Cyanobacteria are prokaryotic photosynthetic microorganisms that pose a serious threat to aquatic environments because they\\u000a are able to form blooms under eutrophic conditions and produce toxins. Cylindrospermopsis raciborskii is a planktonic heterocystous filamentous cyanobacterium initially assigned to the tropics but currently being found in more\\u000a temperate regions such as Portugal, the southernmost record for this species in Europe. Cylindrospermopsin originally

Cristiana Moreira; António Martins; Joana Azevedo; Marisa Freitas; Ana Regueiras; Micaela Vale; Agostinho Antunes; Vitor Vasconcelos

152

Enzymes of the Calvin Cycle and Intermediary Metabolism in the Cyanobacterium Anacystis nidulans Grown in Chemostat Culture  

Microsoft Academic Search

The cyanobacterium Anacystis nidulans grown in light-limited and C0,-limited chemostat cultures showed varying rates of CO, fixation with peaks at dilution rates of 0.10 to 0.12 h-l. The specific activities of a number of enzymes of the reductive and oxidative pentose phosphate and glycolytic pathways and the tricarboxylic acid and glyoxylate cycles varied significantly as a function of the growth

AMALIA D. KARAGOUNI; J. HOWARD SLATER

1979-01-01

153

Integrity and Activity of Photosystem 2 Complexes Isolated from the Thermophilic Cyanobacterium Synechococcus Elongatus Using Various Detergents  

Microsoft Academic Search

The efficiency in selective extraction of photosystem (PS) 2 oxygen evolving complexes was compared among seven detergents.\\u000a These were applied to thylakoid membranes of the thermophilic cyanobacterium Synechococcus elongatus. Used were five non-ionic detergents with one ionic and one zwitterionic for comparison. To compare the suitability and efficiency\\u000a of the detergents the following properties of the extracts were examined: maximum

E. Šetlíková; D. Sofrová; O. Prášil; P. Budá?; M. Koblížek; I. Šetlík

1999-01-01

154

Effects of ultraviolet radiation on productivity and nitrogen fixation in the Cyanobacterium, Anabaena sp. (Newton’s strain)  

Microsoft Academic Search

The biological effects of ultraviolet radiation (UVR; 290–400 nm), especially the UV-B (320–400 nm) component of the spectrum,\\u000a include both direct and indirect effects on many cellular processes. In cyanobacteria both photosynthesis and nitrogen fixation\\u000a can be affected directly by UVR, and indirectly by UVR through the production of reactive oxygen species (ROS). For the heterocystous\\u000a cyanobacterium, Anabaena sp. (Newton’s strain), exposure

Michael P. Lesser

2008-01-01

155

Role of calcium in the inhibition of nitrogenase activity by Methylparathion and Benthiocarb in the cyanobacterium Nostoc muscorum  

Microsoft Academic Search

Methylparathion and Benthiocarb inhibition of N2 fixation in the cyanobacterium Nostoc muscorum was reversed by Ca2+ at 1 mm but not at 0.1 mm. The concentration of intracellular Ca2+ was relatively high in the presence of these pesticides when 1 mm Ca2+ was also present, indicating that intracellular Ca2+ may participate in protecting nitrogenase activity against Methylparathion and Benthiocarb.

A. K. Bhunia; N. K. Basu; D. Roy; S. K. Banerjee

1994-01-01

156

Regulation of S-adenosylhomocysteine hydrolase in the halophilic cyanobacterium Aphanothece halophytica : a possible role in glycinebetaine biosynthesis  

Microsoft Academic Search

Aphanothece halophytica, a halophilic cyanobacterium capable of growing in saturated NaCl, accumulates high intracellular concentrations of glycinebetaine in response to increasing environmental NaCl. In this organism, intracellular levels of K+ rise dramatically with increasing external NaCl before an increase in glycinebetaine can be detected. Glycinebetaine synthesis requires three S-adenosylmethionine (AdoMet)-mediated transmethylations; each transmethylation reaction generates one molecule of the transmethylation

M. H. Sibley; J. H. Yopp

1987-01-01

157

Differential response of NaCl-Resistant mutants of the cyanobacterium Nostoc muscorum to salinity and osmotic stresses  

Microsoft Academic Search

Summary  In the parent strain of Nostoc muscorum, the percentage survival, nitrogenase activity and oxygenic photosynthesis were severely impaired by NaCl (ionic) and sucrose (non-ionic) stresses. Spontaneously occurring NaCl-Resistant mutant clones of the cyanobacterium N. muscorum were found to exhibit differential responses under ionic and non-ionic stresses. One of the mutants (NaCl-R) was found to show resistance in terms of percentage

Santosh Bhargava; Kiran Singh

2006-01-01

158

Biosorption of toxic metal ions by alkali-extracted biomass of a marine cyanobacterium, Phormidium valderianum BDU 30501  

Microsoft Academic Search

Alkali-extracted biomass of Phormidium valderianum BDU 30501, a marine filamentous, non-heterocystous cyanobacterium adsorbed more than 90% of cadmium ions from solutions containing 0.1–40?mM. Cadmium binding accounted up to 18% of biomass weight (w\\/w). The algal biosorbent was also efficient is sequestering metal ions (Cd2+, Co2+, Cu2+, Ni2+) from a mixture. Biosorbent placed in dialysis tubing could concentrate Cd2+ (50–65%) from

RamaRao Karna; L. Uma; G. Subramanian; P. Maruthi Mohan

1999-01-01

159

Uptake and use of the osmoprotective compounds trehalose, glucosylglycerol, and sucrose by the cyanobacterium Synechocystis sp. PCC6803  

Microsoft Academic Search

Accumulation of exogenously supplied osmoprotective compounds was analyzed in the cyanobacterium Synechocystis sp. PCC6803, which synthesizes glucosylglycerol as the principal osmoprotective compound. Glucosylglycerol and trehalose\\u000a were accumulated to high levels and protected cells of a mutant unable to synthesize glucosylglycerol against the deleterious\\u000a effects of salt stress. In the wild-type, uptake of trehalose repressed the synthesis of glucosylglycerol and caused

Stefan Mikkat; Uta Effmert; Martin Hagemann

1997-01-01

160

Inhibition of hydrogen uptake in Escherichia coli by expressing the hydrogenase from the cyanobacterium Synechocystis sp. PCC 6803  

Microsoft Academic Search

BACKGROUND: Molecular hydrogen is an environmentally-clean fuel and the reversible (bi-directional) hydrogenase of the cyanobacterium Synechocystis sp. PCC 6803 as well as the native Escherichia coli hydrogenase 3 hold great promise for hydrogen generation. These enzymes perform the simple reaction 2H+ + 2e- ? H2 (g). RESULTS: Hydrogen yields were enhanced up to 41-fold by cloning the bidirectional hydrogenase (encoded

Toshinari Maeda; Gönül Vardar; William T Self; Thomas K Wood

2007-01-01

161

Characterization of native and histidine-tagged deoxyxylulose 5-phosphate reductoisomerase from the cyanobacterium Synechocystis sp. PCC6803  

Microsoft Academic Search

The dxr gene encoding the 1-deoxy-d-xylulose 5-phosphate reductoisomerase (DXR) from the cyanobacterium Synechocystis sp. PCC6803 was expressed in Escherichia coli to produce both the native and N-terminal histidine-tagged forms of DXR. The enzymes were purified from the cell extracts using either anion exchange chromatography or metal affinity chromatography and gel filtration. The purified recombinant native and histidine-tagged enzymes each displayed

Xihou Yin; Philip J Proteau

2003-01-01

162

An Mrp-Like Cluster in the Halotolerant Cyanobacterium Aphanothece halophytica Functions as a Na+/H+ Antiporter?  

PubMed Central

The mrp homolog gene cluster mrpCD1D2EFGAB (Ap-mrp) was found in a halotolerant cyanobacterium, Aphanothece halophytica, amplified, and expressed in Escherichia coli mutant TO114. Ap-mrp complemented the salt-sensitive phenotype of TO114 and exhibited Na+/H+ and Li+/H+ exchange activities, indicating that Ap-Mrp functions as a Na+/H+ antiporter.

Fukaya, Fuminori; Promden, Worrawat; Hibino, Takashi; Tanaka, Yoshito; Nakamura, Tatsunosuke; Takabe, Teruhiro

2009-01-01

163

Genetic analysis of two new mutations resulting in herbicide resistance in the cyanobacterium Synechcoccus sp. PCC 7002  

Microsoft Academic Search

Two herbicide-resistant strains of the cyanobacterium Synechococcus sp. PCC 7002 are compared to the wild-type with respect to the DNA changes which result in herbicide resstance. The mutations have previously been mapped to a region of the cyanobacterial genome which encodes oneof three copies of psbA, the gene which encodes the 32 kDa Qb-binding protein also known as D1 (Buzby

Jeffrey C. Gingrich; Jeffrey S. Buzby; Veronica L. Stirewalt; Donald A. Bryant

1988-01-01

164

Optical characterization of the oceanic unicellular cyanobacterium Synechococcus grown under a day-night cycle in natural irradiance  

Microsoft Academic Search

The optical properties of the oceanic cyanobacterium Synechococcus (clone WH8103) were examined in a nutrient-replete laboratory culture grown under a day-night cycle in natural irradiance. Measurements of the spectral absorption and beam attenuation coefficients, the size distribution of cells in suspension, and microscopic analysis of samples were made at intervals of 2-4 hours for 2 days. These measurements were used

Dariusz Stramski; Alexi Shalapyonok; Rick A. Reynolds

1995-01-01

165

Photodynamic biocidal action of methylene blue and hydrogen peroxide on the cyanobacterium Synechococcus leopoliensis under visible light irradiation  

Microsoft Academic Search

Biofilm growth on stone surfaces is a significant contributing factor to stone biodeterioration. Current market based biocides are hazardous to the environment and to public health. We have investigated the photo-dynamic effect of methylene blue (MB) in the presence of hydrogen peroxide (H2O2) on the destruction of the cyanobacterium Synechococcus leopoliensis (S. leopoliensis) under irradiation with visible light. Data presented

Cathy McCullagh; Peter K. J. Robertson

2006-01-01

166

Efficient grazing and utilization of the marine cyanobacterium Synechococcus sp. by larvae of the bivalve Mercenaria mercenaria  

Microsoft Academic Search

Efficient grazing by marine bivalve larvae has been thought to be limited to particles larger than 4 µm in diameter, thereby eliminating photosynthetic and non-photosynthetic picoplankton as contributers to larval diets. Documentation of ingestion, carbon retention and growth of laboratory-reared larvae of the bivalve Mercenaria mercenaria L. on Synechococcus sp. (WH7803), a small unicellular cyanobacterium 1 µm in diameter, was

S. M. Gallager; J. B. Waterbury; D. K. Stoecker

1994-01-01

167

Low cellular P-quota and poor metabolic adaptations of the freshwater cyanobacterium Anabaena fertilissima Rao during Pi-limitation.  

PubMed

Anabaena fertilissima is a filamentous freshwater N(2)-fixing cyanobacterium, isolated from a paddy field. Growth of the cyanobacterium was limited by the non-availability of inorganic phosphate (Pi) in the growth medium and was found to be directly related to the cellular P quota, which declined rapidly in Pi-deficient cells. To overcome Pi-deficiency, cells induced both cell-bound and cell-free alkaline phosphatase activities (APase). The activity of cell-bound APase was rapid and 5-6 times higher than that of the cell-free APase activity. Native gel electrophoresis revealed the presence of two APase activity bands for both the cell bound and cell-free APase (Mr ?42 and 34 kDa). For Pi-deficient cells, APase activity was inversely related to cellular P-quota. In A. fertilissima phosphate uptake was facilitated by single high-affinity phosphate transporter (K ( s ), 4.54 ?M; V(max), 4.84 ?mol mg protein(-1) min(-1)). Pi-deficiency severely reduced the photosynthetic rate, respiration rate and nitrate uptake, as well as the activities of nitrate reductase, nitrite reductase and nitrogenase enzymes. In photosynthesis, PSII activity was maximally inhibited, followed by PSI and whole chain activities. Transcript levels of five key glycolytic enzymes showed the poor adaptability of the cyanobacterium to switch its metabolic activity to PPi-dependent enzyme variants, which has rather constant cellular concentrations. PMID:22968428

Tripathi, Keshawanand; Sharma, Naveen K; Rai, Vandna; Rai, Ashwani K

2012-09-12

168

Purification and properties of glutamine synthetase from the non-N2-fixing cyanobacterium Phormidium laminosum.  

PubMed Central

Soluble glutamine synthetase activity (L-glutamate:ammonia ligase, ADP forming, EC 6.3.1.2) was purified to electrophoretic homogeneity from the filamentous non-N2-fixing cyanobacterium Phormidium laminosum (OH-1-p.Cl1) by using conventional purification procedures in the absence of stabilizing ligands. The pure enzyme showed a specific activity of 152 mumol of gamma-glutamylhydroxamate formed.min-1 (transferase activity), which corresponded to 4.4 mumol of Pi released.min-1 (biosynthetic activity). The relative molecular mass of the native enzyme was 602 kilodaltons and was composed of 12 identically sized subunits of 52 kilodaltons. Biosynthetic activity required the presence of Mg2+ as an essential activator, although Co2+ and Zn2+ were partially effective. The kinetics of activation by Mg2+, Co2+, and Zn2+ were sigmoidal, and concentrations required for half-maximal activity were 18 mM (h = 2.2), 6.3 mM (h = 5.6), and 6.3 mM (h = 2.45), respectively. However, transferase activity required Mn2+ (Ka = 3.5 microM), Cu2+, Co2+, or Mg2+ being less effective. The substrate affinities calculated for L-Glu, ammonium, ATP, L-Gln, and hydroxylamine were 15, 0.4, 1.9 (h = 0.75), 14, and 4.1 mM, respectively. Optimal pH and temperature were 7.2 and 55 degrees C for biosynthetic activity and 7.5 and 45 degrees C for transferase activity. The biosynthetic reaction mechanism proceeded according to an ordered three-reactant system, the binding order being ammonium, L-Glu, and ATP. The presence of Mn2+ or Mg2+ drastically affected the thermostability of transferase and biosynthetic activities. Heat inactivation of biosynthetic activity in the presence of Mn2+ obeyed first-order kinetics, with an Ea of 76.8 kcal (ca. 321 kJ) mol-1. Gly, L-Asp, L-Ala, L-Ser and, with lower efficiency, L-Lys and L-Met, L-Lys, and L-Glu inhibited only transferase activity. No cumulative inhibition was observed when mixtures of amino acids were used. Biosynthetic activity was inhibited by AMP (Ki= 7 mM), ADP (Ki= 2.3 mM), p-hydroxymercuribenzoate (Ki= 25 microM), and L-methionine-D, L-sulfoximine (Ki= 2 microM). The enzyme was not activated in vitro by chemically reduced Anabaena thioredoxin. This is the first report of glutamine synthetase activity purified from a filamentous non-N2-fixing cyanobacterium.

Blanco, F; Alana, A; Llama, M J; Serra, J L

1989-01-01

169

Purification and properties of glutathione reductase from the cyanobacterium Anabaena sp. strain 7119.  

PubMed Central

An NADPH-glutathione reductase (EC 1.6.4.2) has been purified 6,000-fold to electrophoretic homogeneity from the filamentous cyanobacterium Anabaena sp. strain 7119. The purified enzyme exhibits a specific activity of 249 U/mg and is characterized by being a dimeric flavin adenine dinucleotide-containing protein with a ratio of absorbance at 280 nm to absorbance at 462 nm of 5.8, a native molecular weight of 104,000, a Stokes radius of 4.13 nm, and a pI of 4.02. The enzyme activity is inhibited by sulfhydryl reagents and heavy-metal ions, especially in the presence of NADPH, with oxidized glutathione behaving as a protective agent. As is the case with the same enzyme from other sources, the kinetic data are consistent with a branched mechanism. Nevertheless, the cyanobacterial enzyme presents three distinctive features with respect to that isolated from non-photosynthetic organisms: (i) absolute specificity for NADPH, (ii) an alkaline optimum pH value of ca. 9.0, and (iii) strong acidic character of the protein, as estimated by column chromatofocusing. The kinetic parameters are very similar to those found for the chloroplast enzyme, but the molecular weight is lower, being comparable to that of non-photosynthetic microorganisms. A protective function, analogous to that assigned to the chloroplast enzyme, is suggested. Images

Serrano, A; Rivas, J; Losada, M

1984-01-01

170

Biochemical effect of carbaryl on oxidative stress, antioxidant enzymes and osmolytes of cyanobacterium Calothrix brevissima.  

PubMed

Carbaryl is used in Indian agriculture for control of rice field pests and it is next to Benzene hexachloride in pesticide consumption. In present study, carbaryl (0, 10, 20, 30 and 40 mg/L) induced toxic effects were observed after 21 days exposure on a non target rice field biofertilizer Calothrix brevissima with special reference to oxidative stress, antioxidant enzymes and osmolytes. At 40 mg/L carbaryl the decrease in carotenoid, chlorophyll, phycobilin and protein were 63%, 43%, 40% and 40% respectively in comparison to control. Total carbohydrate, malondialdehyde, superoxide dismutase, ascorbate peroxidase, catalase and osmolytes showed enhancement at all the treated concentration. Increased amount of MDA (46% at 40 mg/L) indicated free radical mediated deleterious effect of carbaryl. Enhancement of SOD, APX, CAT and osmolytes in presence of carbaryl indicated their involvement in free radical scavenging. SOD, CAT and APX showed maximum activities (79%, 64% and 39% respectively) at 40 mg/L carbaryl. The order of enhancement in osmolytes was glycine-betaine (66%) > proline (54%) > sucrose (50%) at 40 mg/L which might be another adaptive defense strategy of the cyanobacterium against the pesticide. PMID:21979138

Habib, Khalid; Kumar, Satyendra; Manikar, Ningthoujam; Zutshi, Sunaina; Fatma, Tasneem

2011-10-07

171

[Comparison of daily alkaline phosphatase activity of a cyanobacterium (Microcystis aeruginosa) and a diatom (Synedra capitata)].  

PubMed

Alkaline phosphatase activity (ALP) (EC: 3.1.3.1) presents a nycthemeral variation in both Microcystis aeruginosa (cyanobacterium) and Synedra capitata (diatom) species. Nevertheless, a comparative study reveals differences between the enzymatic behaviour of these two species. ALP is 33 times higher in cyanobacteria than in diatoms under similar experimental conditions. Microcystis aeruginosa presents therefore a larger capacity for mineralizing organic phosphorus per unit of biomass. Under LD (16:8) conditions, diatoms show a higher enzymatic activity during the day time (around 0.12 mumol pNPP/mn/mg); on the contrary, cyanobacterial enzymatic activity is rather low during the day time and rises at the beginning of night time (around 3.5 mumol pNPP/mn/mg). Finally, the mean of ALP of Synedra capitata is maximal (around 0.12 mumol pNPP/mn/mg) under total darkness (DD) while the mean of enzymatic activity is maximal (around 3.58 mumol pNPP/mn/mg) under permanent light (LL) for the cyanobacteria. These observed differences in the alkaline phosphatase activity between Microcystis aeruginosa and Synedra capitata might, to some extent, explain the observed alternances within the planktonic settlements between algae and cyanobacteria in hypereutrophic lakes such as the Grangent reservoir (Loire). PMID:9247024

Giraudet, H; Berthon, J L; Buisson, B

1997-06-01

172

Molecular population genetics and phenotypic diversification of two populations of the thermophilic cyanobacterium Mastigocladus laminosus.  

PubMed

We investigated the distributions of genetic and phenotypic variation for two Yellowstone National Park populations of the heterocyst-forming cyanobacterium Mastigocladus (Fischerella) laminosus that exhibit dramatic phenotypic differences as a result of environmental differences in nitrogen availability. One population develops heterocysts and fixes nitrogen in situ in response to a deficiency of combined nitrogen in its environment, whereas the other population does neither due to the availability of a preferred nitrogen source. Slowly evolving molecular markers, including the 16S rRNA gene and the downstream internal transcribed spacer, are identical among all laboratory isolates from both populations but belie considerable genetic and phenotypic diversity. The total nucleotide diversity at six nitrogen metabolism loci was roughly three times greater than that observed for the human global population. The two populations are genetically differentiated, although variation in performance on different nitrogen sources among genotypes could not be explained by local adaptation to available nitrogen in the respective environments. Population genetic models suggest that local adaptation is mutation limited but also that the populations are expected to continue to diverge due to low migratory gene flow. PMID:16597984

Miller, Scott R; Purugganan, Michael D; Curtis, Stephanie E

2006-04-01

173

Genomic structure of an economically important cyanobacterium, Arthrospira (Spirulina) platensis NIES-39.  

PubMed

A filamentous non-N(2)-fixing cyanobacterium, Arthrospira (Spirulina) platensis, is an important organism for industrial applications and as a food supply. Almost the complete genome of A. platensis NIES-39 was determined in this study. The genome structure of A. platensis is estimated to be a single, circular chromosome of 6.8 Mb, based on optical mapping. Annotation of this 6.7 Mb sequence yielded 6630 protein-coding genes as well as two sets of rRNA genes and 40 tRNA genes. Of the protein-coding genes, 78% are similar to those of other organisms; the remaining 22% are currently unknown. A total 612 kb of the genome comprise group II introns, insertion sequences and some repetitive elements. Group I introns are located in a protein-coding region. Abundant restriction-modification systems were determined. Unique features in the gene composition were noted, particularly in a large number of genes for adenylate cyclase and haemolysin-like Ca(2+)-binding proteins and in chemotaxis proteins. Filament-specific genes were highlighted by comparative genomic analysis. PMID:20203057

Fujisawa, Takatomo; Narikawa, Rei; Okamoto, Shinobu; Ehira, Shigeki; Yoshimura, Hidehisa; Suzuki, Iwane; Masuda, Tatsuru; Mochimaru, Mari; Takaichi, Shinichi; Awai, Koichiro; Sekine, Mitsuo; Horikawa, Hiroshi; Yashiro, Isao; Omata, Seiha; Takarada, Hiromi; Katano, Yoko; Kosugi, Hiroki; Tanikawa, Satoshi; Ohmori, Kazuko; Sato, Naoki; Ikeuchi, Masahiko; Fujita, Nobuyuki; Ohmori, Masayuki

2010-03-04

174

Biochemical Studies of the Lagunamides, Potent Cytotoxic Cyclic Depsipeptides from the Marine Cyanobacterium Lyngbya majuscula  

PubMed Central

Lagunamides A (1) and B (2) are potent cytotoxic cyclic depsipeptides isolated from the filamentous marine cyanobacterium, Lyngbya majuscula, from Pulau Hantu, Singapore. These compounds are structurally related to the aurilide-class of molecules, which have been reported to possess exquisite antiproliferative activities against cancer cells. The present study presents preliminary findings on the selectivity of lagunamides against various cancer cell lines as well as their mechanism of action by studying their effects on programmed cell death or apoptosis. Lagunamide A exhibited a selective growth inhibitory activity against a panel of cancer cell lines, including P388, A549, PC3, HCT8, and SK-OV3 cells, with IC50 values ranging from 1.6 nM to 6.4 nM. Morphological studies showed blebbing at the surface of cancer cells as well as cell shrinkage accompanied by loss of contact with the substratum and neighboring cells. Biochemical studies using HCT8 and MCF7 cancer cells suggested that the cytotoxic effect of 1 and 2 might act via induction of mitochondrial mediated apoptosis. Data presented in this study warrants further investigation on the mode of action and underscores the importance of the lagunamides as potential anticancer agents.

Tripathi, Ashootosh; Fang, Wanru; Leong, David Tai; Tan, Lik Tong

2012-01-01

175

Membrane development in the cyanobacterium, Anacystis nidulans, during recovery from iron starvation  

SciTech Connect

Deprivation of iron from the growth medium results in physiological as well as structural changes in the unicellular cyanobacterium Anacystis nidulans R2. Important among these changes are alterations in the composition and function of the photosynthetic membranes. Room-temperature absorption spectra of iron-starved cyanobacterial cells show a chlorophyll absorption peak at 672 nanometers, 7 nanometers blue-shifted from its normal position at 679 nanometers. Iron-starved cells have decreased amounts of chlorophyll and phycobilins. Their fluorescence spectra (77K) have one prominent chlorophyll emission peak at 684 nanometers as compared to three peaks at 687, 696, and 717 nanometers from normal cells. Chlorophyll-protein analysis of iron-deprived cells indicated the absence of high molecular weight bands. Addition of iron to iron-starved cells induced a restoration process in which new components were initially synthesized and integrated into preexisting membranes; at later times, new membranes were assembled and cell division commenced. Synthesis of chlorophyll and phycocyanins started almost immediately after the addition of iron. The origin of the fluorescence emission at 687 and 696 nanometers is discussed in relation to the specific chlorophyll-protein complexes formed during iron reconstitution. 26 references, 2 figures, 1 table.

Pakrasi, H.B.; Goldenberg, A.; Sherman, L.A.

1985-09-01

176

Dinitrogen Fixation Is Restricted to the Terminal Heterocysts in the Invasive Cyanobacterium Cylindrospermopsis raciborskii CS-505  

PubMed Central

The toxin producing nitrogen-fixing heterocystous freshwater cyanobacterium Cylindrospermopsis raciborskii recently radiated from its endemic tropical environment into sub-tropical and temperate regions, a radiation likely to be favored by its ability to fix dinitrogen (diazotrophy). Although most heterocystous cyanobacteria differentiate regularly spaced intercalary heterocysts along their trichomes when combined nitrogen sources are depleted, C. raciborskii differentiates only two terminal heterocysts (one at each trichome end) that can reach >100 vegetative cells each. Here we investigated whether these terminal heterocysts are the exclusive sites for dinitrogen fixation in C. raciborskii. The highest nitrogenase activity and NifH biosynthesis (western-blot) were restricted to the light phase of a 12/12 light/dark cycle. Separation of heterocysts and vegetative cells (sonication and two-phase aqueous polymer partitioning) demonstrated that the terminal heterocysts are the sole sites for nifH expression (RT-PCR) and NifH biosynthesis. The latter finding was verified by the exclusive localization of nitrogenase in the terminal heterocysts of intact trichomes (immunogold-transmission electron microscopy and in situ immunofluorescence-light microscopy). These results suggest that the terminal heterocysts provide the combined nitrogen required by the often long trichomes (>100 vegetative cells). Our data also suggests that the terminal-heterocyst phenotype in C. raciborskii may be explained by the lack of a patL ortholog. These data help identify mechanisms by which C. raciborskii and other terminal heterocyst-forming cyanobacteria successfully inhabit environments depleted in combined nitrogen.

Plominsky, Alvaro M.; Larsson, John; Bergman, Birgitta; Delherbe, Nathalie; Osses, Igor; Vasquez, Monica

2013-01-01

177

Differential Expression of Photosynthesis and Nitrogen Fixation Genes in the Cyanobacterium Plectonema boryanum  

PubMed Central

The filamentous non-heterocystous cyanobacterium Plectonema boryanum fixes dinitrogen at a high rate during microaerobic growth in continuous illumination by temporal separation of oxygen-evolving photosynthesis and oxygen-sensitive dinitrogen fixation. The onset of nitrogen fixation is preceded by a depression in photosynthesis that establishes a sufficiently low level of dissolved oxygen in the growth medium. A several-fold reduction in the level of transcripts coding for phycocyanin (cpcBA) and the chlorophyll a binding protein of photosystem II (psbC) and psbA accompanied the depression in photosynthetic oxygen evolution. Unlike most of the other organisms examined to date, in P. boryanum, psbC and psbD do not appear to be co-transcribed. The psbC transcripts were down-regulated several fold, while the psbD transcript declined marginally during the nitrogen fixation phase. A decrease in dissolved oxygen and a dramatic increase in the level of nifH transcripts and the enzyme activity of nitrogenase were characteristic of the nitrogen fixation phase. The level of transcript for glnA, which encodes glutamine synthetase, was not altered. Reciprocal regulation of gene expression was well orchestrated with the alternating cycles of photosynthesis and nitrogen fixation in P. boryanum.

Misra, Hari S.; Tuli, Rakesh

2000-01-01

178

Inhibitory effects of sanguinarine against the cyanobacterium Microcystis aeruginosa NIES-843 and possible mechanisms of action.  

PubMed

Sanguinarine showed strong inhibitory effect against Microcystis aeruginosa, a typical water bloom-forming and microcystins-producing cyanobacterium. The EC50 of sanguinarine against the growth of M. aeruginosa NIES-843 was 34.54±1.17?g/L. Results of chlorophyll fluorescence transient analysis indicated that all the electron donating side, accepting side, and the reaction center of the Photosystem II (PS II) were the targets of sanguinarine against M. aeruginosa NIES-843. The elevation of reactive oxygen species (ROS) level in the cells of M. aeruginosa NIES-843 upon exposure indicated that sanguinarine induced oxidative stress in the active growing cells of M. aeruginosa NIES-843. Further results of gene expression analysis indicated that DNA damage and cell division inhibition were also involved in the inhibitory action mechanism of sanguinarine against M. aeruginosa NIES-843. The inhibitory characteristics of sanguinarine against M. aeruginosa suggest that the ecological- and public health-risks need to be evaluated before its application in cyanobacterial bloom control to avoid devastating events irreversibly. PMID:24060579

Shao, Jihai; Liu, Deming; Gong, Daoxin; Zeng, Qingru; Yan, Zhiyong; Gu, Ji-Dong

2013-09-08

179

Cyclic Depsipeptides, Grassypeptolides D, E and Ibu epidemethoxylyngbyastatin 3, from a Red Sea Leptolyngbya Cyanobacterium  

PubMed Central

Two new grassypeptolides and a lyngbyastatin analogue, together with the known dolastatin 12, have been isolated from field collections and laboratory cultures of the marine cyanobacterium Leptolyngbya sp. collected from the SS Thistlegorm shipwreck in the Red Sea. The overall stereostructures of grassypeptolides D (1) and E (2) and Ibu-epidemethoxylyngbyastatin 3 (3) were determined by a combination of 1D and 2D NMR experiments, MS analysis, Marfey's methodology, and HPLC-MS. Compounds 1 and 2 contain 2-methyl-3-aminobutyric acid (Maba) and 2-aminobutyric acid (Aba), while biosynthetically distinct 3 contains 3-amino-2-methylhexanoic acid (Amha) and the ?-keto amino acid 4-amino-2,-2-dimethyl-3-oxopentanoic acid (Ibu). Grassypeptolides D (1) and E (2) showed significant cytotoxicity to HeLa (IC50 = 335 and 192 nM, respectively) and mouse neuro-2a blastoma cells (IC50 = 599 and 407 nM, respectively), in contrast to Ibu-epidemethoxylyngbyastatin 3 (neuro-2a cells, IC50 > 10 ?M) and dolastatin 12 (neuro-2a cells, IC50 > 1 ?M).

Thornburg, Christopher C.; Thimmaiah, Muralidhara; Shaala, Lamiaa A.; Hau, Andrew M.; Malmo, Jay M.; Ishmael, Jane E.; Youssef, Diaa T.A.; McPhail, Kerry L.

2011-01-01

180

Self-suppression of biofilm formation in the cyanobacterium Synechococcus elongatus.  

PubMed

Biofilms are consortia of bacteria that are held together by an extracellular matrix. Cyanobacterial biofilms, which are highly ubiquitous and inhabit diverse niches, are often associated with biological fouling and cause severe economic loss. Information on the molecular mechanisms underlying biofilm formation in cyanobacteria is scarce. We identified a mutant of the cyanobacterium Synechococcus elongatus, which unlike the wild type, developed biofilms. This biofilm-forming phenotype is caused by inactivation of homologues of type II secretion /type IV pilus assembly systems and is associated with impairment of protein secretion. The conditioned medium from a wild-type culture represses biofilm formation by the secretion-mutants. This suggested that the planktonic nature of the wild-type strain is a result of a self-suppression mechanism, which depends on the deposition of a factor to the extracellular milieu. We also identified two genes that are essential for biofilm formation. Transcript levels of these genes are elevated in the mutant compared with the wild type, and are initially decreased in mutant cells cultured in conditioned medium of wild-type cells. The particular niche conditions will determine whether the inhibitor will accumulate to effective levels and thus the described mechanism allows switching to a sessile mode of existence. PMID:23298171

Schatz, Daniella; Nagar, Elad; Sendersky, Eleonora; Parnasa, Rami; Zilberman, Shaul; Carmeli, Shmuel; Mastai, Yitzhak; Shimoni, Eyal; Klein, Eugenia; Yeger, Orna; Reich, Ziv; Schwarz, Rakefet

2013-01-09

181

On the dynamics and constraints of batch culture growth of the cyanobacterium Cyanothece sp. ATCC 51142.  

PubMed

The unicellular, nitrogen fixing cyanobacterium Cyanothece sp. ATCC 51142 is of a remarkable potential for production of third-generation biofuels. As the biotechnological potential of Cyanothece 51142 varies with the time of the day, we argue that it will, similarly, depend on the phase of the culture growth. Here, we study the batch culture dynamics to discover the dominant constraints in the individual growth phases and identify potential for inducing or delaying transitions between culture growth phases in Cyanothece 51142. We found that specific growth rate in the exponential phase of the culture is much less dependent on incident irradiance than the photosynthetic activity. We propose that surplus electrons that are released by water splitting are used in futile processes providing photoprotection additional to non-photochemical quenching. We confirm that the transition from exponential to linear phase is caused by a light limitation and the transition from linear to stationary phase by nitrogen limitation. We observe spontaneous diurnal metabolic oscillations in stationary phase culture that are synchronized over the entire culture without an external clue. We tentatively propose that the self-synchronization of the metabolic oscillations is due to a cell-to-cell communication of the cyanobacteria that is necessary for nitrogenase activity in nitrate depleted medium. PMID:22575787

Sinetova, Maria A; Cervený, Jan; Zav?el, Tomáš; Nedbal, Ladislav

2012-04-28

182

A model of cyclic transcriptomic behavior in the cyanobacterium Cyanothece sp. ATCC 51142.  

PubMed

Systems biology attempts to reconcile large amounts of disparate data with existing knowledge to provide models of functioning biological systems. The cyanobacterium Cyanothece sp. ATCC 51142 is an excellent candidate for such systems biology studies because: (i) it displays tight functional regulation between photosynthesis and nitrogen fixation; (ii) it has robust cyclic patterns at the genetic, protein and metabolomic levels; and (iii) it has potential applications for bioenergy production and carbon sequestration. We have represented the transcriptomic data from Cyanothece 51142 under diurnal light/dark cycles as a high-level functional abstraction and describe development of a predictive in silico model of diurnal and circadian behavior in terms of regulatory and metabolic processes in this organism. We show that incorporating network topology into the model improves performance in terms of our ability to explain the behavior of the system under new conditions. The model presented robustly describes transcriptomic behavior of Cyanothece 51142 under different cyclic and non-cyclic growth conditions, and represents a significant advance in the understanding of gene regulation in this important organism. PMID:21698331

McDermott, Jason E; Oehmen, Christopher S; McCue, Lee Ann; Hill, Eric; Choi, Daniel M; Stöckel, Jana; Liberton, Michelle; Pakrasi, Himadri B; Sherman, Louis A

2011-06-23

183

Molecular Population Genetics and Phenotypic Diversification of Two Populations of the Thermophilic Cyanobacterium Mastigocladus laminosus  

PubMed Central

We investigated the distributions of genetic and phenotypic variation for two Yellowstone National Park populations of the heterocyst-forming cyanobacterium Mastigocladus (Fischerella) laminosus that exhibit dramatic phenotypic differences as a result of environmental differences in nitrogen availability. One population develops heterocysts and fixes nitrogen in situ in response to a deficiency of combined nitrogen in its environment, whereas the other population does neither due to the availability of a preferred nitrogen source. Slowly evolving molecular markers, including the 16S rRNA gene and the downstream internal transcribed spacer, are identical among all laboratory isolates from both populations but belie considerable genetic and phenotypic diversity. The total nucleotide diversity at six nitrogen metabolism loci was roughly three times greater than that observed for the human global population. The two populations are genetically differentiated, although variation in performance on different nitrogen sources among genotypes could not be explained by local adaptation to available nitrogen in the respective environments. Population genetic models suggest that local adaptation is mutation limited but also that the populations are expected to continue to diverge due to low migratory gene flow.

Miller, Scott R.; Purugganan, Michael D.; Curtis, Stephanie E.

2006-01-01

184

Fluorapatite as Inorganic Phosphate Source for the Cyanobacterium Anabaena PCC 7120  

NASA Astrophysics Data System (ADS)

We investigated the hypothesis that the cyanobacterium Anabaena PCC 7120 is able to use fluorapatite (FAP) as sole phosphate source for growth. In the experimental setup the dissolution of FAP was tested in a phosphate free growth medium in the presence and absence of the Anabaena, as well as the cell free supernatant of an Anabaena culture. The results were compared with that of an Anabaena culture grown without fluorapatite. Parameters measured were pH, dissolved P and Ca, as well as cell density. The FAP grains were analyzed using SEM and XPS. Additionally, the differential expression of secreted proteins in cultures with and without dissolved phosphate was examined. P-limited Anabaena cultures tend to aggregate and in the presence of FAP the cells attached themselves to the mineral grains. The cultures benefit from the presence of FAP. The cells have a very effective P-uptake system that is able to take up dissolved phosphate very efficiently and draw the concentrations down to very low levels. Furthermore, the SEM analysis of FAP showed an etching of the mineral grains in the samples from the Anabaena cultures. The mechanism of apatite dissolution with and without Anabaena will be discussed in terms of these experimental observations.

Schaperdoth, I.; Brantley, S.

2003-12-01

185

Solution structure of cytochrome c6 from the thermophilic cyanobacterium Synechococcus elongatus.  

PubMed Central

Cytochrome c6 is a small, soluble electron carrier between the two membrane-bound complexes cytochrome b6f and photosystem I (PSI) in oxygenic photosynthesis. We determined the solution structure of cytochrome c6 from the thermophilic cyanobacterium Synechococcus elongatus by NMR spectroscopy and molecular dynamics calculations based on 1586 interresidual distance and 28 dihedral angle restraints. The overall fold exhibits four alpha-helices and a small antiparallel beta-sheet in the vicinity of Met58, one of the axial heme ligands. The flat hydrophobic area in this cytochrome c6 is conserved in other c6 cytochromes and even in plastocyanin of higher plants. This docking region includes the site of electron transfer to PSI and possibly to the cytochrome b6f complex. The binding of cytochrome c6 to PSI in green algae involves interaction of a negative patch with a positive domain of PSI. This positive domain has not been inserted at the evolutionary level of cyanobacteria, but the negatively charged surface region is already present in S. elongatus cytochrome c6 and may thus have been optimized during evolution to improve the interaction with the positively charged cytochrome f. As the structure of PSI is known in S.elongatus, the reported cytochrome c6 structure can provide a basis for mutagenesis studies to delineate the mechanism of electron transfer between both.

Beissinger, M; Sticht, H; Sutter, M; Ejchart, A; Haehnel, W; Rosch, P

1998-01-01

186

Identification, cloning, and characterization of a novel ketoreductase from the cyanobacterium Synechococcus sp. strain PCC 7942.  

PubMed

A new ketoreductase useful for asymmetric synthesis of chiral alcohols was identified in the cyanobacterium Synechococcus sp. strain PCC 7942. Mass spectrometry of trypsin-digested peptides identified the protein as 3-ketoacyl-[acyl-carrier-protein] reductase (KR) (EC 1.1.1.100). The gene, referred to as fabG, was cloned, functionally expressed in Escherichia coli, and subsequently purified to homogeneity. The enzyme displayed a temperature optimum at 44 degrees C and a broad pH optimum between pH 7 and pH 9. The NADPH-dependent KR was able to asymmetrically reduce a variety of prochiral ketones with good to excellent enantioselectivities (>99.8%). The KR showed particular high specific activity for asymmetric reduction of ethyl 4-chloroacetoacetate (38.29 +/- 2.15 U mg(-1)) and 2',3',4',5',6'-pentafluoroacetophenone (8.57 +/- 0.49 U mg(-1)) to the corresponding (S)-alcohols. In comparison with an established industrial enzyme like the alcohol dehydrogenase from Lactobacillus brevis, the KR showed seven-times-higher activity toward 2',3',4',5',6'-pentafluoroacetophenone, with a remarkably higher enantiomeric excess (>99.8% [S] versus 43.3% [S]). PMID:18791006

Hölsch, Kathrin; Havel, Jan; Haslbeck, Martin; Weuster-Botz, Dirk

2008-09-12

187

Optimization of photobioreactor growth conditions for a cyanobacterium expressing mosquitocidal Bacillus thuringiensis Cry proteins.  

PubMed

An Anabaena strain (PCC 7120#11) that was genetically engineered to express Bacillus thuringiensis subsp. israelensis cry genes has shown good larvicidal activity against Anopheles arabiensis, a major vector of malaria in Africa. Response surface methodology was used to evaluate the relationship between key growth factors and the volumetric productivity of PCC 7120#11 in an indoor, flat-plate photobioreactor. The interaction of input CO? concentration and airflow rate had a statistically significant effect on the volumetric productivity of PCC 7120#11, as did the interaction of airflow rate and photosynthetic photon flux density. Model-based numerical optimization indicated that the optimal factor level combination for maximizing PCC 7120#11 volumetric productivity was a photosynthetic photon flux density of 154 ?mol m?² s?¹ and air enriched with 3.18% (v/v) CO? supplied at a flow rate of 1.02 vessel volumes per minute. At the levels evaluated in the study, none of the growth factors had a significant effect on the median lethal concentration of PCC 7120#11 against An. arabiensis larvae. This finding is important because loss of mosquitocidal activity under growth conditions that maximize volumetric productivity would impact on the feasibility of using PCC 7120#11 in malaria vector control programs. The study showed the usefulness of response surface methodology for determination of the optimal growth conditions for a cyanobacterium that is genetically engineered to have larvicidal activity against malaria vectors. PMID:23732832

Ketseoglou, Irene; Bouwer, Gustav

2013-05-31

188

Glycosylated Porphyra-334 and Palythine-Threonine from the Terrestrial Cyanobacterium Nostoc commune.  

PubMed

Mycosporine-like amino acids (MAAs) are water-soluble UV-absorbing pigments, and structurally different MAAs have been identified in eukaryotic algae and cyanobacteria. In this study novel glycosylated MAAs were found in the terrestrial cyanobacterium Nostoc commune (N. commune). An MAA with an absorption maximum at 334 nm was identified as a hexose-bound porphyra-334 derivative with a molecular mass of 508 Da. Another MAA with an absorption maximum at 322 nm was identified as a two hexose-bound palythine-threonine derivative with a molecular mass of 612 Da. These purified MAAs have radical scavenging activities in vitro, which suggests multifunctional roles as sunscreens and antioxidants. The 612-Da MAA accounted for approximately 60% of the total MAAs and contributed approximately 20% of the total radical scavenging activities in a water extract, indicating that it is the major water-soluble UV-protectant and radical scavenger component. The hexose-bound porphyra-334 derivative and the glycosylated palythine-threonine derivatives were found in a specific genotype of N. commune, suggesting that glycosylated MAA patterns could be a chemotaxonomic marker for the characterization of the morphologically indistinguishable N. commune. The glycosylation of porphyra-334 and palythine-threonine in N. commune suggests a unique adaptation for terrestrial environments that are drastically fluctuating in comparison to stable aquatic environments. PMID:24065157

Nazifi, Ehsan; Wada, Naoki; Yamaba, Minami; Asano, Tomoya; Nishiuchi, Takumi; Matsugo, Seiichi; Sakamoto, Toshio

2013-08-26

189

Polysaccharide covalently linked to the peptidoglycan of the cyanobacterium Synechocystis sp. strain PCC6714.  

PubMed Central

A polysaccharide was found to be covalently linked to the peptidoglycan of the unicellular cyanobacterium Synechocystis sp. strain PCC6714 via phosphodiester bonds. It could be cleaved from the peptidoglycan-polysaccharide (PG-PS) complex by hydrofluoric acid (HF) treatment in the cold (48% HF, 0 degrees C, 48 h) yielding a pure, HF-insoluble peptidoglycan fraction and an HF-soluble polysaccharide fraction. The PG-PS complex was isolated from the Triton X-100-insoluble cell wall fraction by hot sodium dodecyl sulfate treatment and digestion with proteases. Digestion of the complex with N-acetylmuramidase released the glycopeptide-linked polysaccharide, which was further purified by dialysis and gel filtration on Sephadex G-50 and G-200. The polysaccharide consisted of glucosamine, mannosamine, galactosamine, mannose, and glucose and had a molecular weight of 25,000 to 30,000. Muramic acid-6-phosphate was identified as the binding site of the covalently linked, nonphosphorylated polysaccharide as revealed by chemical analysis of linkage fragments of the PG-PS complex. Images

Jurgens, U J; Weckesser, J

1986-01-01

190

Cytoplasmic Membrane Changes during Adaptation of the Fresh Water Cyanobacterium Synechococcus 6311 to Salinity 1  

PubMed Central

In this investigation, changes were characterized in cell structure and cytoplasmic membrane organization that occur when the freshwater cyanobacterium Synechococcus 6311 is transferred from `low salt' (0.03 molar NaCl) to `high salt' (0.5 molar NaCl) media (i.e. sea water concentration). Cells were examined at several time points after the imposition of the salt stress and compared to control cells, in thin sections and freeze fracture electron microscopy, and by flow cytometry. One minute after exposure to high salt, i.e. `salt shock,' virtually all intracellular granules disappeared, the density of the cytoplasm decreased, and the appearance of DNA material was changed. Glycogen and other granules, however, reappeared by 4 hours after salt exposure. The organization of the cytoplasmic membrane undergoes major reorganization following salt shock. Freeze-fracture electron microscopy showed that small intramembrane particles (diameters 7.5 and 8.5 nanometers) are reduced in number by two- to fivefold, whereas large particles, (diameters 14.5 and 17.5 nanometers) increase two- to fourfold in frequency, compared to control cells grown in low salt medium. The changes in particle size distribution suggest synthesis of new membrane proteins, in agreement with the known increases in respiration, cytochrome oxidase, and sodium proton exchange activity of the cytoplasmic membrane. Images Fig. 2 Fig. 1 Fig. 5

Lefort-Tran, Marcelle; Pouphile, Monique; Spath, Susan; Packer, Lester

1988-01-01

191

Identification, Cloning, and Characterization of a Novel Ketoreductase from the Cyanobacterium Synechococcus sp. Strain PCC 7942?  

PubMed Central

A new ketoreductase useful for asymmetric synthesis of chiral alcohols was identified in the cyanobacterium Synechococcus sp. strain PCC 7942. Mass spectrometry of trypsin-digested peptides identified the protein as 3-ketoacyl-[acyl-carrier-protein] reductase (KR) (EC 1.1.1.100). The gene, referred to as fabG, was cloned, functionally expressed in Escherichia coli, and subsequently purified to homogeneity. The enzyme displayed a temperature optimum at 44°C and a broad pH optimum between pH 7 and pH 9. The NADPH-dependent KR was able to asymmetrically reduce a variety of prochiral ketones with good to excellent enantioselectivities (>99.8%). The KR showed particular high specific activity for asymmetric reduction of ethyl 4-chloroacetoacetate (38.29 ± 2.15 U mg?1) and 2?,3?,4?,5?,6?-pentafluoroacetophenone (8.57 ± 0.49 U mg?1) to the corresponding (S)-alcohols. In comparison with an established industrial enzyme like the alcohol dehydrogenase from Lactobacillus brevis, the KR showed seven-times-higher activity toward 2?,3?,4?,5?,6?-pentafluoroacetophenone, with a remarkably higher enantiomeric excess (>99.8% [S] versus 43.3% [S]).

Holsch, Kathrin; Havel, Jan; Haslbeck, Martin; Weuster-Botz, Dirk

2008-01-01

192

[Mutational analysis of the genes encoding the photosystem II proteins in Cyanobacterium synechocystis sp. 6803].  

PubMed

Chlorophyll--binding protein CP43 and cytochrome b559, encoded by psbC and psbE/F genes, are the components of photosystem II (PS II). Three psbC- and four psbE/F- mutants were isolated from the collection of PS II-deficient mutants of the cyanobacterium Synechocystis sp. 6803. Restoration of photosynthetic activity was achieved by transformation of psbE/F- mutants with cloned psbE/F gene cluster from wild type cells and each of psbC- mutants--with specific part of wild type psbC gene. DNA fragments carrying the mutations were isolated from mutant cells and sequenced. The mutations which affect PS II activity were identified in psbC gene as "frameshift" mutation, stop-codon formation, or as deletion of three nucleotides resulting in loss of one of three Phe residues in position 422-424 of CP43. Sequence of mutant psbE/F genes revealed single mutations resulting in deletion of Phe-36 or substitution of Pro-63 for Leu in alpha-subunit and Val-29 for Phe in beta-subunit of cytochrome b559. PMID:8307353

Elanskaia, I V; Broun, M N; Gadzhiev, A G; Shestakov, S V

1993-10-01

193

Amino acid transport systems required for diazotrophic growth in the cyanobacterium Anabaena sp. strain PCC 7120.  

PubMed Central

Uptake of 16 amino acids by the filamentous, heterocyst-forming cyanobacterium Anabaena sp. strain PCC 7120 was characterized with regard to kinetic parameters of transport, intracellular accumulation of the transported amino acids, and sensitivity of the transport process to energy metabolism inhibitors. Mutants resistant to certain toxic analogs of some amino acids were isolated that were impaired in amino acid transport. Results obtained in this study, together with those reported previously (A. Herrero and E. Flores, J. Biol. Chem. 265:3931-3935, 1990), suggest that there are at least five amino acid transport systems in strain PCC 7120: one high-affinity, active system for basic amino acids; one low-affinity, passive system for basic amino acids; two high-affinity, active systems with overlapping, but not identical, specificities for neutral amino acids; and one putative system for acidic amino acids. Some of the amino acid transport mutants were impaired in diazotrophic growth. These mutants were unable to develop a normal percentage of heterocysts and normal nitrogenase activity in response to nitrogen stepdown. Putative roles for the amino acid transport systems in uptake of extracellular amino acids, recapture of amino acids that have leaked from the cells, and intercellular transfer of amino acids in the filaments of Anabaena sp. strain PCC 7120 are discussed.

Montesinos, M L; Herrero, A; Flores, E

1995-01-01

194

Glutaraldehyde treatment of proteinaceous gas vesicles from cyanobacterium Anabaena flos-aquae.  

PubMed

As potential gas microcarriers, gas vesicles (GVs) were isolated from cultures of the filamentous cyanobacterium Anabaena flos-aquae and treated with glutaraldehyde. The effects of glutaraldehyde treatment on the stability of GVs, against elevated temperatures (40-121 degrees C) and protein-stripping agents such as urea and sodium dodecyl sulfate (SDS), were then examined with the pressure collapse curves generated using pressure nephelometry. The treatment was very beneficial to GVs against the exposure to SDS and urea; however, it did not make the evolution-optimized vesicle structure stronger or more temperature-resistant. In the presence of these protein-stripping agents, the treated vesicles had higher median (50%) collapse pressures (by > or =1 atm) than the untreated ones, at both room temperature and 40 degrees C. This increase has been presumably attributed to the cross-linking of the large GvpC protein to the ribbed GvpA shell, thereby resisting the stripping of GvpC that provides the primary mechanical strength to the vesicle wall. The glutaraldehyde treatment also restored the strength of GVs weakened by a 5-week storage in a refrigerator and, therefore, is expected to improve the stability of GVs for long-term storage. GVs could not be autoclaved. If necessary for the intended applications, glutaraldehyde treatment may also serve to chemically sterilize the vesicles, with the glutaraldehyde subsequently removed by dialysis. PMID:11101344

Sundararajan, A; Ju, L K

195

The Psb32 protein aids in repairing photodamaged photosystem II in the cyanobacterium Synechocystis 6803.  

PubMed

Photosystem II (PSII), a membrane protein complex, catalyzes the photochemical oxidation of water to molecular oxygen. This enzyme complex consists of approximately 20 stoichiometric protein components. However, due to the highly energetic reactions it catalyzes as part of its normal activity, PSII is continuously damaged and repaired. With advances in protein detection technologies, an increasing number of sub-stoichiometric PSII proteins have been identified, many of which aid in the biogenesis and assembly of this protein complex. Psb32 (Sll1390) has previously been identified as a protein associated with highly active purified PSII preparations from the cyanobacterium Synechocystis sp. PCC 6803. To investigate its function, the subcellular localization of Psb32 and the impact of deletion of the psb32 gene on PSII were analyzed. Here, we show that Psb32 is an integral membrane protein, primarily located in the thylakoid membranes. Although not required for cell viability, Psb32 protects cells from oxidative stress and additionally confers a selective fitness advantage in mixed culture experiments. Specifically, Psb32 protects PSII from photodamage and accelerates its repair. Thus, the data suggest that Psb32 plays an important role in minimizing the effect of photoinhibition on PSII. PMID:21653280

Wegener, Kimberly M; Bennewitz, Stefan; Oelmüller, Ralf; Pakrasi, Himadri B

2011-06-07

196

Apratoxin H and Apratoxin A Sulfoxide from the Red Sea Cyanobacterium Moorea producens.  

PubMed

Cultivation of the marine cyanobacterium Moorea producens, collected from the Nabq Mangroves in the Gulf of Aqaba (Red Sea), led to the isolation of new apratoxin analogues apratoxin H (1) and apratoxin A sulfoxide (2), together with the known apratoxins A-C, lyngbyabellin B, and hectochlorin. The absolute configuration of these new potent cytotoxins was determined by chemical degradation, MS, NMR, and CD spectroscopy. Apratoxin H (1) contains pipecolic acid in place of the proline residue present in apratoxin A, expanding the known suite of naturally occurring analogues that display amino acid substitutions within the final module of the apratoxin biosynthetic pathway. The oxidation site of apratoxin A sulfoxide (2) was deduced from MS fragmentation patterns and IR data, and 2 could not be generated experimentally by oxidation of apratoxin A. The cytotoxicity of 1 and 2 to human NCI-H460 lung cancer cells (IC50 = 3.4 and 89.9 nM, respectively) provides further insight into the structure-activity relationships in the apratoxin series. Phylogenetic analysis of the apratoxin-producing cyanobacterial strains belonging to the genus Moorea, coupled with the recently annotated apratoxin biosynthetic pathway, supports the notion that apratoxin production and structural diversity may be specific to their geographical niche. PMID:24016099

Thornburg, Christopher C; Cowley, Elise S; Sikorska, Justyna; Shaala, Lamiaa A; Ishmael, Jane E; Youssef, Diaa T A; McPhail, Kerry L

2013-09-09

197

Anilofos tolerance and its mineralization by the cyanobacterium Synechocystis sp. strain PUPCCC 64.  

PubMed

This study deals with anilofos tolerance and its mineralization by the common rice field cyanobacterium Synechocystis sp. strain PUPCCC 64. The organism tolerated anilofos up to 25 mg L(-1). The herbicide caused inhibitory effects on photosynthetic pigments of the test organism in a dose-dependent manner. The organism exhibited 60, 89, 96, 85 and 79% decrease in chlorophyll a, carotenoids, phycocyanin, allophycocyanin and phycoerythrin, respectively, in 20 mg L(-1) anilofos on day six. Activities of superoxide dismutase, catalase and peroxidase increased by 1.04 to 1.80 times over control cultures in presence of 20 mg L(-1) anilofos. Glutathione content decreased by 26% while proline content was unaffected by 20 mg L(-1) anilofos. The test organism showed intracellular uptake and metabolized the herbicide. Uptake of herbicide by test organism was fast during initial six hours followed by slow uptake until 120 hours. The organism exhibited maximum anilofos removal at 100 mg protein L(-1), pH 8.0 and 30°C. Its growth in phosphate deficient basal medium in the presence of anilofos (2.5 mg L(-1)) indicated that herbicide was used by the strain PUPCCC 64 as a source of phosphate. PMID:23382844

Singh, D P; Khattar, J I S; Kaur, Mandeep; Kaur, Gurdeep; Gupta, Meenu; Singh, Yadvinder

2013-01-31

198

The amino acid sequence of low-potential cytochrome c550 from the cyanobacterium Microcystis aeruginosa.  

PubMed

The low-potential cytochrome c550 has been purified from the cyanobacterium Microcystis aeruginosa and its amino acid sequence has been determined. The protein contains 135 amino acid residues with the Cys-X-X-Cys-His heme binding site at residues 37 to 41. The sequence from residue 28 to 45 shows similarity to cytochrome c553 residues 1 to 18 when the heme binding sites are aligned. Another region of similarity is in the carboxyl-terminal regions of these two proteins. The two aligning regions of cytochrome c553 correspond to helical segments in other related cytochromes. A partial sequence of cytochrome c550 from Aphanizomenon flos-aquae was obtained and showed a 48% identity to the sequence of the M. aeruginosa cytochrome. The single methionine residue in cytochrome c550 of M. aeruginosa occurs at position 119 but there is no methionine in this region in the A. flos-aquae cytochrome, indicating that methionine is not the sixth ligand to the heme iron atom. Histidine 92 is a possible sixth ligand in M. aeruginosa cytochrome c550. The far-uv circular dichroism spectrum indicates that this protein is approximately 17% alpha helix, 42% beta-pleated sheet, and 41% random coil. PMID:2539046

Cohn, C L; Sprinkle, J R; Alam, J; Hermodson, M; Meyer, T; Krogmann, D W

1989-04-01

199

Malyngolide from the cyanobacterium Lyngbya majuscula interferes with quorum sensing circuitry.  

PubMed

Extracts of several cyanobacterial species collected from different marine and estuarine locations predominately in Florida (USA), with one sample each from Belize and Oman, were screened for their ability to disrupt quorum sensing (QS) in the reporter strain Chromobacterium violaceum CV017. Inhibitory activities were detected in the ethyl acetate?:?methanol (1:1) extracts of several Lyngbya spp., and extracts of Lyngbya majuscula contained the strongest QS inhibitory activities. Extracts of L. majuscula from the Indian River Lagoon, FL, USA, were further purified by bioassay-guided fractionation. The antibiotic malyngolide (MAL) was identified as a QS inhibitor. Activity of MAL was investigated using N-acyl homoserine lactone (AHL) reporters based on the LasR receptor of Pseudomonas aeruginosa. MAL at concentrations ranging from 3.57?µM to 57?µM (EC50 ?=?12.2?±?1.6?µM) inhibited responses of the LasR reporters without affecting bacterial growth. MAL inhibited (EC50 ?=? 10.6?±?1.8?µM) Las QS-dependent production of elastase by P. aeruginosa PAO1. We propose that this QS inhibitor plays a role in controlling interactions of heterotrophic bacteria associated with the cyanobacterium L. majuscula. PMID:23766278

Dobretsov, Sergey; Teplitski, Max; Alagely, Ali; Gunasekera, Sarath P; Paul, Valerie J

2010-12-01

200

An experimentally anchored map of transcriptional start sites in the model cyanobacterium Synechocystis sp. PCC6803  

PubMed Central

There has been an increasing interest in cyanobacteria because these photosynthetic organisms convert solar energy into biomass and because of their potential for the production of biofuels. However, the exploitation of cyanobacteria for bioengineering requires knowledge of their transcriptional organization. Using differential RNA sequencing, we have established a genome-wide map of 3,527 transcriptional start sites (TSS) of the model organism Synechocystis sp. PCC6803. One-third of all TSS were located upstream of an annotated gene; another third were on the reverse complementary strand of 866 genes, suggesting massive antisense transcription. Orphan TSS located in intergenic regions led us to predict 314 noncoding RNAs (ncRNAs). Complementary microarray-based RNA profiling verified a high number of noncoding transcripts and identified strong ncRNA regulations. Thus, ?64% of all TSS give rise to antisense or ncRNAs in a genome that is to 87% protein coding. Our data enhance the information on promoters by a factor of 40, suggest the existence of additional small peptide-encoding mRNAs, and provide corrected 5? annotations for many genes of this cyanobacterium. The global TSS map will facilitate the use of Synechocystis sp. PCC6803 as a model organism for further research on photosynthesis and energy research.

Mitschke, Jan; Georg, Jens; Scholz, Ingeborg; Sharma, Cynthia M.; Dienst, Dennis; Bantscheff, Jens; Voss, Bjorn; Steglich, Claudia; Wilde, Annegret; Vogel, Jorg; Hess, Wolfgang R.

2011-01-01

201

Surplus Photosynthetic Antennae Complexes Underlie Diagnostics of Iron Limitation in a Cyanobacterium  

PubMed Central

Chlorophyll fluorescence from phytoplankton provides a tool to assess iron limitation in the oceans, but the physiological mechanism underlying the fluorescence response is not understood. We examined fluorescence properties of the model cyanobacterium Synechocystis PCC6803 and a ?isiA knock-out mutant of the same species grown under three culture conditions which simulate nutrient conditions found in the open ocean: (1) nitrate and iron replete, (2) limiting-iron and high-nitrate, representative of natural high-nitrate, low-chlorophyll regions, and (3) iron and nitrogen co-limiting. We show that low variable fluorescence, a key diagnostic of iron limitation, results from synthesis of antennae complexes far in excess of what can be accommodated by the iron-restricted pool of photosynthetic reaction centers. Under iron and nitrogen co-limiting conditions, there are no excess antennae complexes and variable fluorescence is high. These results help to explain the well-established fluorescence characteristics of phytoplankton in high-nutrient, low-chlorophyll ocean regions, while also accounting for the lack of these properties in low-iron, low-nitrogen regions. Importantly, our results complete the link between unique molecular consequences of iron stress in phytoplankton and global detection of iron stress in natural populations from space.

Schrader, Paul S.; Milligan, Allen J.; Behrenfeld, Michael J.

2011-01-01

202

Glycosylated Porphyra-334 and Palythine-Threonine from the Terrestrial Cyanobacterium Nostoc commune  

PubMed Central

Mycosporine-like amino acids (MAAs) are water-soluble UV-absorbing pigments, and structurally different MAAs have been identified in eukaryotic algae and cyanobacteria. In this study novel glycosylated MAAs were found in the terrestrial cyanobacterium Nostoc commune (N. commune). An MAA with an absorption maximum at 334 nm was identified as a hexose-bound porphyra-334 derivative with a molecular mass of 508 Da. Another MAA with an absorption maximum at 322 nm was identified as a two hexose-bound palythine-threonine derivative with a molecular mass of 612 Da. These purified MAAs have radical scavenging activities in vitro, which suggests multifunctional roles as sunscreens and antioxidants. The 612-Da MAA accounted for approximately 60% of the total MAAs and contributed approximately 20% of the total radical scavenging activities in a water extract, indicating that it is the major water-soluble UV-protectant and radical scavenger component. The hexose-bound porphyra-334 derivative and the glycosylated palythine-threonine derivatives were found in a specific genotype of N. commune, suggesting that glycosylated MAA patterns could be a chemotaxonomic marker for the characterization of the morphologically indistinguishable N. commune. The glycosylation of porphyra-334 and palythine-threonine in N. commune suggests a unique adaptation for terrestrial environments that are drastically fluctuating in comparison to stable aquatic environments.

Nazifi, Ehsan; Wada, Naoki; Yamaba, Minami; Asano, Tomoya; Nishiuchi, Takumi; Matsugo, Seiichi; Sakamoto, Toshio

2013-01-01

203

Amino acid availability determines the ratio of microcystin variants in the cyanobacterium Planktothrix agardhii.  

PubMed

Cyanobacteria are capable of producing multiple microcystin variants simultaneously. The mechanisms that determine the composition of microcystin variants in cyanobacteria are still debated. [Asp(3)]microcystin-RR contains arginine at the position where the more toxic [Asp(3)]microcystin-LR incorporates leucine. We cultured the filamentous cyanobacterium Planktothrix agardhii strain 126/3 with and without external addition of leucine and arginine. Addition of leucine to the growth medium resulted in a strong increase of the [Asp(3)]microcystin LR/RR ratio, while addition of arginine resulted in a decrease. This demonstrates that amino acid availability plays a role in the synthesis of different microcystin variants. Environmental changes affecting cell metabolism may cause differences in the intracellular availability of leucine and arginine, which can thus affect the production of microcystin variants. Because leucine contains one nitrogen atom while arginine contains four nitrogen atoms, we hypothesized that low nitrogen availability might shift the amino acid composition in favor of leucine, which might explain seasonal increases in the [Asp(3)]microcystin LR/RR ratio in natural populations. However, when a continuous culture of P. agardhii was shifted from nitrogen-saturated to a nitrogen-limited mineral medium, leucine and arginine concentrations decreased, but the leucine/arginine ratio did not change. Accordingly, while the total microcystin concentration of the cells decreased, we did not observe changes in the [Asp(3)]microcystin LR/RR ratio in response to nitrogen limitation. PMID:18616588

Tonk, Linda; van de Waal, Dedmer B; Slot, Pieter; Huisman, Jef; Matthijs, Hans C P; Visser, Petra M

2008-07-08

204

Microcystin production by a freshwater spring cyanobacterium of the genus Fischerella.  

PubMed

We investigated the production of a hepatotoxic, cyclic heptapeptide, microcystin, by a filamentous branched cyanobacterium belonging to the order Stigonematales, genus Fischerella. The freshwater Fischerella sp. strain CENA161 was isolated from spring water in a small concrete dam in Piracicaba, São Paulo State, Brazil, and identified by combining a morphological description with 16S rRNA gene sequencing and phylogenetic analysis. Microcystin (MCYST) analysis performed using an ELISA assay on cultured cells gave positive results. High performance liquid chromatography-mass spectrometry (HPLC-MS) analysis detected 33.6microg MCYST-LR per gram dry weight of cyanobacterial cells. Microcystin profile revealed by quadrupole time-of-flight tandem mass spectrometry (Q-TOF-MS/MS) analysis confirmed the production of MCYST-LR. Furthermore, genomic DNA was analyzed by PCR for sequences similar to the ketosynthase (KS) domain of the type I polyketide synthase gene, which is involved in microcystin biosynthesis. This revealed the presence of a KS nucleotide fragment similar to the mcyD and ndaD genes of the microcystin and nodularin synthetase complexes. Phylogenetic analysis grouped the Fischerella KS sequence together with mcyD sequences of the three known microcystin synthetase operon (Microcystis, Planktothrix and Anabaena) and ndaD of the nodularin synthetase operon, with 100% bootstrap support. Our findings demonstrate that Fischerella sp. CENA161 produces MYCST-LR and for the first time identify a nucleotide sequence putatively involved in microcystin synthesis in this genus. PMID:19233225

Fiore, Marli Fátima; Genuário, Diego Bonaldo; da Silva, Caroline Souza Pamplona; Shishido, Tânia Keiko; Moraes, Luiz Alberto Beraldo; Cantúsio Neto, Romeu; Silva-Stenico, Maria Estela

2009-02-21

205

P-type ATPase from the cyanobacterium Synechococcus 7942 related to the human Menkes and Wilson disease gene products.  

PubMed

DNA encoding a P-type ATPase was cloned from the cyanobacterium Synechococcus 7942. The cloned ctaA gene encodes a 790-amino acid polypeptide related to the CopA Cu(2+)-uptake ATPase of Enterococcus hirae, to other known P-type ATPases, and to the candidate gene products for the human diseases of copper metabolism, Menkes disease and Wilson disease. Disruption of the single chromosomal gene in Synechococcus 7942 by insertion of an antibiotic-resistance cassette results in a mutant cell line with increased tolerance to Cu2+ compared with the wild type. PMID:7937823

Phung, L T; Ajlani, G; Haselkorn, R

1994-09-27

206

Organization of the genes for protein synthesis elongation factors Tu and G in the cyanobacterium Anacystis nidulans.  

PubMed Central

The genes for protein synthesis elongation factors Tu and G were cloned from the cyanobacterium Anacystis nidulans. The locations of these genes were mapped within the cloned DNA fragment by hybridization with Escherichia coli probes. The organization of the cloned fragment and the DNA flanking it in the A. nidulans chromosome was also determined. The elongation factor Tu and G genes are adjacent to one another and in the same 5'-to-3' orientation. In contrast to other gram-negative bacteria, A. nidulans contains only one gene for elongation factor Tu. Images

Mickel, F S; Spremulli, L L

1986-01-01

207

Identification of a carotenoid-binding protein in the cytoplasmic membrane from the heterotrophic cyanobacterium Synechocystis sp. strain PCC6714.  

PubMed Central

We isolated a carotenoid-binding protein from the cytoplasmic membrane of the cyanobacterium Synechocystis sp. strain PCC6714. The polypeptide demonstrated a characteristic mobility shift when electrophoresed in lithium dodecyl sulfate-polyacrylamide gels. The protein migrated with an apparent molecular mass of 35 kilodaltons when solubilized at 0 degrees C, but after solubilization at 70 degrees C, the protein migrated as a 45-kilodalton species. The carotenoid-binding protein accumulated only in autotrophically grown cells; cytoplasmic membranes prepared from photoheterotrophically grown cells lacked this component. Images

Bullerjahn, G S; Sherman, L A

1986-01-01

208

Fermentation and Sulfur Reduction in the Mat-Building Cyanobacterium Microcoleus chthonoplastes.  

PubMed

The mat-building cyanobacterium Microcoleus chthonoplastes carried out a mixed-acid fermentation when incubated under anoxic conditions in the dark. Endogenous storage carbohydrate was fermented to acetate, ethanol, formate, lactate, H(inf2), and CO(inf2). Cells with a low glycogen content (about 0.3 (mu)mol of glucose per mg of protein) produced acetate and ethanol in equimolar amounts. In addition to glycogen, part of the osmoprotectant, glucosyl-glycerol, was degraded. The glucose component of glucosyl-glycerol was fermented, whereas glycerol was released into the medium. Cells with a high content of glycogen (about 2 (mu)mol of glucose per mg of protein) did not utilize glucosyl-glycerol. These cells produced more acetate than ethanol. M. chthonoplastes was also capable of using elemental sulfur as the electron acceptor during fermentation, resulting in the production of sulfide. With sulfur present, acetate production increased whereas ethanol production decreased. Also, less formate was produced and the evolution of hydrogen ceased completely. In general, the carbon recoveries were satisfactory but the oxidation-reduction balances were too high. The latter could be explained by assuming the reduction of ferric iron, which is associated with the cells, mediated by the oxidation of formate. The switch from photoautotrophic to fermentative metabolism did not require de novo protein synthesis, and fermentation started immediately upon transfer to dark anoxic conditions. From the molar ratios of the fermentation products and from measurement of enzyme activities in cell extracts, we concluded that glucose derived from glycogen and glucosyl-glycerol is degraded via the Embden-Meyerhof-Parnas pathway. PMID:16535319

Moezelaar, R; Bijvank, S M; Stal, L J

1996-05-01

209

Effects of a simulated martian UV flux on the cyanobacterium, Chroococcidiopsis sp. 029.  

PubMed

Dried monolayers of Chroococcidiopsis sp. 029, a desiccation-tolerant, endolithic cyanobacterium, were exposed to a simulated martian-surface UV and visible light flux, which may also approximate to the worst-case scenario for the Archean Earth. After 5 min, there was a 99% loss of cell viability, and there were no survivors after 30 min. However, this survival was approximately 10 times higher than that previously reported for Bacillus subtilis. We show that under 1 mm of rock, Chroococcidiopsis sp. could survive (and potentially grow) under the high martian UV flux if water and nutrient requirements for growth were met. In isolated cells, phycobilisomes and esterases remained intact hours after viability was lost. Esterase activity was reduced by 99% after a 1-h exposure, while 99% loss of autofluorescence required a 4-h exposure. However, cell morphology was not changed, and DNA was still detectable by 4',6-diamidino-2-phenylindole staining after an 8-h exposure (equivalent to approximately 1 day on Mars at the equator). Under 1 mm of simulant martian soil or gneiss, the effect of UV radiation could not be detected on esterase activity or autofluorescence after 4 h. These results show that under the intense martian UV flux the morphological signatures of life can persist even after viability, enzymatic activity, and pigmentation have been destroyed. Finally, the global dispersal of viable, isolated cells of even this desiccation-tolerant, ionizing-radiation-resistant microorganism on Mars is unlikely as they are killed quickly by unattenuated UV radiation when in a desiccated state. These findings have implications for the survival of diverse microbial contaminants dispersed during the course of human exploratory class missions on the surface of Mars. PMID:15815164

Cockell, Charles S; Schuerger, Andrew C; Billi, Daniela; Friedmann, E Imre; Panitz, Corinna

2005-04-01

210

Characterization of the response to zinc deficiency in the cyanobacterium Anabaena sp. strain PCC 7120.  

PubMed

Zur regulators control zinc homeostasis by repressing target genes under zinc-sufficient conditions in a wide variety of bacteria. This paper describes how part of a survey of duplicated genes led to the identification of the open reading frame all2473 as the gene encoding the Zur regulator of the cyanobacterium Anabaena sp. strain PCC 7120. All2473 binds to DNA in a zinc-dependent manner, and its DNA-binding sequence was characterized, which allowed us to determine the relative contribution of particular nucleotides to Zur binding. A zur mutant was found to be impaired in the regulation of zinc homeostasis, showing sensitivity to elevated concentrations of zinc but not other metals. In an effort to characterize the Zur regulon in Anabaena, 23 genes containing upstream putative Zur-binding sequences were identified and found to be regulated by Zur. These genes are organized in six single transcriptional units and six operons, some of them containing multiple Zur-regulated promoters. The identities of genes of the Zur regulon indicate that Anabaena adapts to conditions of zinc deficiency by replacing zinc metalloproteins with paralogues that fulfill the same function but presumably with a lower zinc demand, and with inducing putative metallochaperones and membrane transport systems likely being involved in the scavenging of extracellular zinc, including plasma membrane ABC transport systems and outer membrane TonB-dependent receptors. Among the Zur-regulated genes, the ones showing the highest induction level encode proteins of the outer membrane, suggesting a primary role for components of this cell compartment in the capture of zinc cations from the extracellular medium. PMID:22389488

Napolitano, Mauro; Rubio, Miguel Ángel; Santamaría-Gómez, Javier; Olmedo-Verd, Elvira; Robinson, Nigel J; Luque, Ignacio

2012-03-02

211

DL-7-azatryptophan and citrulline metabolism in the cyanobacterium Anabaena sp. strain 1F  

SciTech Connect

An alternative route for the primary assimilation of ammonia proceeds via glutamine synthetase-carbamyl phosphate synthetase and its inherent glutaminase activity in Anabaena sp. strain 1F, a marine filamentous, heterocystous cyanobacterium. Evidence for the presence of this possible alternative route to glutamate was provided by the use of amino acid analogs as specific enzyme inhibitors, enzymological studies, and radioistopic labeling experiments. The amino acid pool patterns of continuous cultures of Anabaena sp. strain 1F were markedly influenced by the nitrogen source. A relatively high concentration of glutamate was maintained in the amino acid pools of all cultures irrespective of the nitrogen source, reflecting the central role of glutamate in nitrogen metabolism. The addition of 1.0 microM azaserine increased the intracellular pools of glutamate and glutamine. All attempts to detect any enzymatic activity for glutamate synthase by measuring the formation of L-(/sup 14/C)glutamate from 2-keto-(1-/sup 14/C)glutarate and glutamine failed. The addition of 10 microM DL-7-azatryptophan caused a transient accumulation of intracellular citrulline and alanine which was not affected by the presence of chloramphenicol. The in vitro activity of carbamyl phosphate synthetase and glutaminase increased severalfold in the presence of azatryptophan. Results from radioisotopic labeling experiments with (/sup 14/C)bicarbonate and L-(1-/sup 14/C)ornithine also indicated that citrulline was formed via carbamyl phosphate synthetase and ornithine transcarbamylase. In addition to its effects on nitrogen metabolism, azatryptophan also affected carbon metabolism by inhibiting photosynthetic carbon assimilation and photosynthetic oxygen evolution.

Chen, C.H.; Van Baalen, C.; Tabita, F.R.

1987-03-01

212

A Novel Nitrate/Nitrite Permease in the Marine Cyanobacterium Synechococcus sp. Strain PCC 7002  

PubMed Central

The nrtP and narB genes, encoding nitrate/nitrite permease and nitrate reductase, respectively, were isolated from the marine cyanobacterium Synechococcus sp. strain PCC 7002 and characterized. NrtP is a member of the major facilitator superfamily and is unrelated to the ATP-binding cassette-type nitrate transporters that previously have been described for freshwater strains of cyanobacteria. However, NrtP is similar to the NRT2-type nitrate transporters found in diverse organisms. An nrtP mutant strain consumes nitrate at a 4.5-fold-lower rate than the wild type, and this mutant grew exponentially on a medium containing 12 mM nitrate at a rate approximately 2-fold lower than that of the wild type. The nrtP mutant cells could not consume nitrite as rapidly as the wild type at pH 10, suggesting that NrtP also functions in nitrite uptake. A narB mutant was unable to grow on a medium containing nitrate as a nitrogen source, although this mutant could grow on media containing urea or nitrite with rates similar to those of the wild type. Exogenously added nitrite enhanced the in vivo activity of nitrite reductase in the narB mutant; this suggests that nitrite acts as a positive effector of nitrite reductase. Transcripts of the nrtP and narB genes were detected in cells grown on nitrate but were not detected in cells grown on urea or ammonia. Transcription of the nrtP and narB genes is probably controlled by the NtcA transcription factor for global nitrogen control. The discovery of a nitrate/nitrite permease in Synechococcus sp. strain PCC 7002 suggests that significant differences in nutrient transporters may occur in marine and freshwater cyanobacteria.

Sakamoto, Toshio; Inoue-Sakamoto, Kaori; Bryant, Donald A.

1999-01-01

213

Dependence of the cyanobacterium Prochlorococcus on hydrogen peroxide scavenging microbes for growth at the ocean's surface.  

PubMed

The phytoplankton community in the oligotrophic open ocean is numerically dominated by the cyanobacterium Prochlorococcus, accounting for approximately half of all photosynthesis. In the illuminated euphotic zone where Prochlorococcus grows, reactive oxygen species are continuously generated via photochemical reactions with dissolved organic matter. However, Prochlorococcus genomes lack catalase and additional protective mechanisms common in other aerobes, and this genus is highly susceptible to oxidative damage from hydrogen peroxide (HOOH). In this study we showed that the extant microbial community plays a vital, previously unrecognized role in cross-protecting Prochlorococcus from oxidative damage in the surface mixed layer of the oligotrophic ocean. Microbes are the primary HOOH sink in marine systems, and in the absence of the microbial community, surface waters in the Atlantic and Pacific Ocean accumulated HOOH to concentrations that were lethal for Prochlorococcus cultures. In laboratory experiments with the marine heterotroph Alteromonas sp., serving as a proxy for the natural community of HOOH-degrading microbes, bacterial depletion of HOOH from the extracellular milieu prevented oxidative damage to the cell envelope and photosystems of co-cultured Prochlorococcus, and facilitated the growth of Prochlorococcus at ecologically-relevant cell concentrations. Curiously, the more recently evolved lineages of Prochlorococcus that exploit the surface mixed layer niche were also the most sensitive to HOOH. The genomic streamlining of these evolved lineages during adaptation to the high-light exposed upper euphotic zone thus appears to be coincident with an acquired dependency on the extant HOOH-consuming community. These results underscore the importance of (indirect) biotic interactions in establishing niche boundaries, and highlight the impacts that community-level responses to stress may have in the ecological and evolutionary outcomes for co-existing species. PMID:21304826

Morris, J Jeffrey; Johnson, Zackary I; Szul, Martin J; Keller, Martin; Zinser, Erik R

2011-02-03

214

Dependence of the Cyanobacterium Prochlorococcus on Hydrogen Peroxide Scavenging Microbes for Growth at the Ocean's Surface  

PubMed Central

The phytoplankton community in the oligotrophic open ocean is numerically dominated by the cyanobacterium Prochlorococcus, accounting for approximately half of all photosynthesis. In the illuminated euphotic zone where Prochlorococcus grows, reactive oxygen species are continuously generated via photochemical reactions with dissolved organic matter. However, Prochlorococcus genomes lack catalase and additional protective mechanisms common in other aerobes, and this genus is highly susceptible to oxidative damage from hydrogen peroxide (HOOH). In this study we showed that the extant microbial community plays a vital, previously unrecognized role in cross-protecting Prochlorococcus from oxidative damage in the surface mixed layer of the oligotrophic ocean. Microbes are the primary HOOH sink in marine systems, and in the absence of the microbial community, surface waters in the Atlantic and Pacific Ocean accumulated HOOH to concentrations that were lethal for Prochlorococcus cultures. In laboratory experiments with the marine heterotroph Alteromonas sp., serving as a proxy for the natural community of HOOH-degrading microbes, bacterial depletion of HOOH from the extracellular milieu prevented oxidative damage to the cell envelope and photosystems of co-cultured Prochlorococcus, and facilitated the growth of Prochlorococcus at ecologically-relevant cell concentrations. Curiously, the more recently evolved lineages of Prochlorococcus that exploit the surface mixed layer niche were also the most sensitive to HOOH. The genomic streamlining of these evolved lineages during adaptation to the high-light exposed upper euphotic zone thus appears to be coincident with an acquired dependency on the extant HOOH-consuming community. These results underscore the importance of (indirect) biotic interactions in establishing niche boundaries, and highlight the impacts that community-level responses to stress may have in the ecological and evolutionary outcomes for co-existing species.

Morris, J. Jeffrey; Johnson, Zackary I.; Szul, Martin J.; Keller, Martin; Zinser, Erik R.

2011-01-01

215

Lyngbyoic acid, a "tagged" fatty acid from a marine cyanobacterium, disrupts quorum sensing in Pseudomonas aeruginosa†  

PubMed Central

Quorum sensing (QS) is a mechanism of bacterial gene regulation in response to increases in population density. Perhaps most studied are QS pathways mediated by acylhomoserine lactones (AHLs) in Gram-negative bacteria. Production of small molecule QS signals, their accumulation within a diffusion-limited environment and their binding to a LuxR-type receptor trigger QS-controlled gene regulatory cascades. In Pseudomonas aeruginosa, for example, binding of AHLs to their cognate receptors (LasR, RhlR) controls production of virulence factors, pigments, antibiotics and other behaviors important for its interactions with eukaryotic hosts and other bacteria. We have previously shown that marine cyanobacteria produce QS-inhibitory molecules, including 8-epi-malyngamide C (1), malyngamide C (2) and malyngolide (3). Here we isolated a new small cyclopropane-containing fatty acid, lyngbyoic acid (4), as a major metabolite of the marine cyanobacterium, Lyngbya cf. majuscula, collected at various sites in Florida. We screened 4 against four reporters based on different AHL receptors (LuxR, AhyR, TraR and LasR) and found that 4 most strongly affected LasR. We also show that 4 reduces pyocyanin and elastase (LasB) both on the protein and transcript level in wild-type P. aeruginosa, and that 4 directly inhibits LasB enzymatic activity. Conversely, dodecanoic acid (9) increased pyocyanin and LasB, demonstrating that the fused cyclopropane “tag” is functionally relevant and potentially confers resistance to ?-oxidation. Global transcriptional effects of 4 in some ways replicate the gene expression changes of P. aeruginosa during chronic lung infections of cystic fibrosis patients, with reduced lasR signaling, increased biofilm and expression of the virulence locus HSI-I. Compound 4 may therefore prove to be a useful tool in the study of P. aeruginosa adaption during such chronic infections.

Kwan, Jason Christopher; Meickle, Theresa; Ladwa, Dheran; Teplitski, Max; Paul, Valerie; Luesch, Hendrik

2013-01-01

216

The Biosynthetic Pathway for Myxol-2? Fucoside (Myxoxanthophyll) in the Cyanobacterium Synechococcus sp. Strain PCC 7002? †  

PubMed Central

Synechococcus sp. strain PCC 7002 produces a variety of carotenoids, which comprise predominantly dicylic ?-carotene and two dicyclic xanthophylls, zeaxanthin and synechoxanthin. However, this cyanobacterium also produces a monocyclic myxoxanthophyll, which was identified as myxol-2? fucoside. Compared to the carotenoid glycosides produced by diverse microorganisms, cyanobacterial myxoxanthophyll and closely related compounds are unusual because they are glycosylated on the 2?-OH rather than on the 1?-OH position of the ? end of the molecule. In this study, the genes encoding two enzymes that modify the ? end of myxoxanthophyll in Synechococcus sp. strain PCC 7002 were identified. Mutational and biochemical studies showed that open reading frame SynPCC7002_A2032, renamed cruF, encodes a 1?-hydroxylase and that open reading frame SynPCC7002_A2031, renamed cruG, encodes a 2?-O-glycosyltransferase. The enzymatic activity of CruF was verified by chemical characterization of the carotenoid products synthesized when cruF was expressed in a lycopene-producing strain of Escherichia coli. Database searches showed that homologs of cruF and cruG occur in the genomes of all sequenced cyanobacterial strains that are known to produce myxol or the acylic xanthophyll oscillaxanthin. The genomes of many other bacteria that produce hydroxylated carotenoids but do not contain crtC homologs also contain cruF orthologs. Based upon observable intermediates, a complete biosynthetic pathway for myxoxanthophyll is proposed. This study expands the suite of enzymes available for metabolic engineering of carotenoid biosynthetic pathways for biotechnological applications.

Graham, Joel E.; Bryant, Donald A.

2009-01-01

217

Posttranslational Regulation of Nitrate Assimilation in the Cyanobacterium Synechocystis sp. Strain PCC 6803  

PubMed Central

Posttranslational regulation of nitrate assimilation was studied in the cyanobacterium Synechocystis sp. strain PCC 6803. The ABC-type nitrate and nitrite bispecific transporter encoded by the nrtABCD genes was completely inhibited by ammonium as in Synechococcus elongatus strain PCC 7942. Nitrate reductase was insensitive to ammonium, while it is inhibited in the Synechococcus strain. Nitrite reductase was also insensitive to ammonium. The inhibition of nitrate and nitrite transport required the PII protein (glnB gene product) and the C-terminal domain of NrtC, one of the two ATP-binding subunits of the transporter, as in the Synechococcus strain. Mutants expressing the PII derivatives in which Ala or Glu is substituted for the conserved Ser49, which has been shown to be the phosphorylation site in the Synechococcus strain, showed ammonium-promoted inhibition of nitrate uptake like that of the wild-type strain. The S49A and S49E substitutions in GlnB did not affect the regulation of the nitrate and nitrite transporter in Synechococcus either. These results indicated that the presence or absence of negative electric charge at the 49th position does not affect the activity of the PII protein to regulate the cyanobacterial ABC-type nitrate and nitrite transporter according to the cellular nitrogen status. This finding suggested that the permanent inhibition of nitrate assimilation by an S49A derivative of PII, as was previously reported for Synechococcus elongatus strain PCC 7942, is likely to have resulted from inhibition of nitrate reductase rather than the nitrate and nitrite transporter.

Kobayashi, Masaki; Takatani, Nobuyuki; Tanigawa, Mari; Omata, Tatsuo

2005-01-01

218

Gramine-induced growth inhibition, oxidative damage and antioxidant responses in freshwater cyanobacterium Microcystis aeruginosa.  

PubMed

In recent years, the exploration and development of the effective methods of treatment and prevention to algal blooms, especially Microcystis aeruginosa blooms has been an important issue in the field of water environment protection. Allelochemicals (natural plant toxins) are considered promising sources of algicides to control algal blooms. The objective of this study is to determine the inhibitory effects and potential mechanisms of a well-known allelochemical gramine (N,N-dimethyl-3-amino-methylindole) on bloom-forming cyanobacterium M. aeruginosa. The results showed that this indole alkaloid effectively inhibited the growth of M. aeruginosa. The effective concentration causing a 50% inhibition at 3 d (EC(50, 3 d)) increased with the initial algal density (IAD) increasing. When IAD increased from 5x10(4) to 5x10(5)cellsmL(-1), the values of EC(50, 3 d) increased from 0.5 to 2.1mgL(-1). In the cells of M. aeruginosa, gramine caused an obvious increase in the level of reactive oxygen species (ROS). The lipid-peroxidation product malondialdehyde (MDA) increased significantly in gramine-treated cells. The effects of gramine on enzymatic and non-enzymatic antioxidants were in different manners. The activity of superoxide dismutase (SOD) was decreased after gramine exposure. The catalase (CAT) activity was increased after 4h but decreased from 60h. Both the contents and the regeneration rates of ascorbic acid (AsA) and reduced glutathione (GSH) were increased after 4h of exposure to gramine. However, only GSH content was still increased after 40h of exposure. These results suggested that the activation of antioxidants in M. aeruginosa played an important role to resist the stress from gramine at initial time, the inactivation of SOD is crucial to the growth inhibition of M. aeruginosa by gramine, and the phytotoxicity of gramine on M. aeruginosa may be due to oxidative damage via oxidation of ROS. PMID:19131120

Hong, Yu; Hu, Hong-Ying; Xie, Xing; Sakoda, Akiyoshi; Sagehashi, Masaki; Li, Feng-Min

2008-11-27

219

Photosynthesis and Inorganic Carbon Usage by the Marine Cyanobacterium, Synechococcus sp  

PubMed Central

The marine cyanobacterium, Synechococcus sp. Nägeli (strain RRIMP N1) changes its affinity for external inorganic carbon used in photosynthesis, depending on the concentration of CO2 provided during growth. The high affinity for CO2 + HCO3? of air-grown cells (K½ < 80 nanomoles [pH 8.2]) would seem to be the result of the presence of an inducible mechanism which concentrates inorganic carbon (and thus CO2) within the cells. Silicone-oil centrifugation experiments indicate that the inorganic carbon concentration inside suitably induced cells may be in excess of 1,000-fold greater than that in the surrounding medium, and that this accumulation is dependent upon light energy. The quantum requirements for O2 evolution appear to be some 2-fold greater for low CO2-grown cells, compared with high CO2-grown cells. This presumably is due to the diversion of greater amounts of light energy into inorganic carbon transport in these cells. A number of experimental approaches to the question of whether CO2 or HCO3? is primarily utilized by the inorganic carbon transport system in these cells show that in fact both species are capable of acting as substrate. CO2, however, is more readily taken up when provided at an equivalent concentration to HCO3?. This discovery suggests that the mechanistic basis for the inorganic carbon concentrating system may not be a simple HCO3? pump as has been suggested. It is clear, however, that during steady-state photosynthesis in seawater equilibrated with air, HCO3? uptake into the cell is the primary source of internal inorganic carbon.

Badger, Murray R.; Andrews, T. John

1982-01-01

220

Low temperature delays timing and enhances the cost of nitrogen fixation in the unicellular cyanobacterium Cyanothece.  

PubMed

Marine nitrogen-fixing cyanobacteria are largely confined to the tropical and subtropical ocean. It has been argued that their global biogeographical distribution reflects the physiologically feasible temperature range at which they can perform nitrogen fixation. In this study we refine this line of argumentation for the globally important group of unicellular diazotrophic cyanobacteria, and pose the following two hypotheses: (i) nitrogen fixation is limited by nitrogenase activity at low temperature and by oxygen diffusion at high temperature, which is manifested by a shift from strong to weak temperature dependence of nitrogenase activity, and (ii) high respiration rates are required to maintain very low levels of oxygen for nitrogenase, which results in enhanced respiratory cost per molecule of fixed nitrogen at low temperature. We tested these hypotheses in laboratory experiments with the unicellular cyanobacterium Cyanothece sp. BG043511. In line with the first hypothesis, the specific growth rate increased strongly with temperature from 18 to 30?°C, but leveled off at higher temperature under nitrogen-fixing conditions. As predicted by the second hypothesis, the respiratory cost of nitrogen fixation and also the cellular C:N ratio rose sharply at temperatures below 21?°C. In addition, we found that low temperature caused a strong delay in the onset of the nocturnal nitrogenase activity, which shortened the remaining nighttime available for nitrogen fixation. Together, these results point at a lower temperature limit for unicellular nitrogen-fixing cyanobacteria, which offers an explanation for their (sub)tropical distribution and suggests expansion of their biogeographical range by global warming. PMID:23823493

Brauer, Verena S; Stomp, Maayke; Rosso, Camillo; van Beusekom, Sebastiaan Am; Emmerich, Barbara; Stal, Lucas J; Huisman, Jef

2013-07-04

221

Optimal conditions for genetic transformations of the cyanobacterium Anacystis nidulans R2  

SciTech Connect

Under optimal conditions, the cyanobacterium Anacystis nidulans R2 was transformed to ampicillin resistance at frequencies of >10/sup 7/ transformants per ..mu..g of plasmid (pCH1) donor DNA. No stringent period of competency was detected, and high frequencies of transformation were achieved with cultures at various growth stages. Transformation increased with time after addition of donor DNA up to 15 to 18 h. The peak of transformation efficiency (transformants/donor molecule) occurred at plasmid concentrations of 125 to 325 ng/ml with an ampicillin resistance donor plasmid (pCH1) and 300 to 625 ng/ml for chloramphenicol resistance conferred by plasmid pSG111. The efficiency of transformation was enhanced by excluding light during the incubation or by blocking photosynthesis with the electron transport inhibitor 3-(3, 4-dichlorophenyl)-1, 1-dimethylurea (DCMU) or the uncoupler carbonyl cyanide-m-chlorophenyl hydrazone. Preincubation of cells in darkness for 15 to 18 h before addition of donor DNA significantly decreased transformation efficiency. Growth of cells in iron-deficient medium before transformation enhanced efficiency fourfold. These results were obtained with selection for ampicillin (pCH1 donor plasmid)- or chloramphenicol (pSG111 donor plasmid)-resistant transformants. Approximately 1000 transformants per ..mu..g were obtained when chromosomal DNA from a herbicide (DCMU)-resistant mutant was used as donor DNA. DCMU resistance was also transferred to recipient cells by using restriction fragments of chromosomal DNA from DCMU-resistant mutants. This procedure allowed size classes of fragments to be assayed for the presence of the DCMU resistance gene.

Golden, S.S.; Sherman, L.A.

1984-04-01

222

Stable transformation of the cyanobacterium Synechocystis sp. PCC 6803 induced by UV irradiation  

SciTech Connect

Irradiation of the photoheterotrophic cyanobacterium Synechocystis sp. PCC 6803 with low levels of UV light allows for stable, integrative transformation of these cells by heterologous DNA. In this system, transformation does not rely on an autonomously replicating plasmid and is independent of homologous recombination. Cells treated with UV light in the absence of DNA and cells given DNA but not exposed to UV do not yield antibiotic-resistant colonies in platings of up to 2 x 10/sup 8/ cells. Optimal conditions for this UV-induced transformation are described. Analysis of the transformants indicates that (i) only a segment of the introduced plasmid is found in the DNA of the transformed cells; (ii) in independently isolated clones, DNA insertion apparently occurs at different sites in the chromosome; and (iii) hybridization data suggest that insertion in one of the transformants may have occurred into a region of the chromosome that is repeated or that integration of plasmid DNA may have been accompanied by a rearrangement or duplication of DNA sequences near the insertion site. DNA isolated from the primary transformants as well as a cloned fragment containing the UV- inserted plasmid sequence and flanking cyanobacterial DNA transform wild-type cells at a high frequency (5.0 x 10/sup -4/ and 1.5 x 10/sup -5/, respectively). Possible mechanisms of this transformation system are discussed, as are the potential uses of this system as an integrative cloning-complementation vector and as a mutagenic agent in which the genetic lesion is already tagged with a selectable marker.

Dzelzkalns, V.A.; Bogorad, L.

1986-03-01

223

Highly plastic genome of Microcystis aeruginosa PCC 7806, a ubiquitous toxic freshwater cyanobacterium  

PubMed Central

Background The colonial cyanobacterium Microcystis proliferates in a wide range of freshwater ecosystems and is exposed to changing environmental factors during its life cycle. Microcystis blooms are often toxic, potentially fatal to animals and humans, and may cause environmental problems. There has been little investigation of the genomics of these cyanobacteria. Results Deciphering the 5,172,804 bp sequence of Microcystis aeruginosa PCC 7806 has revealed the high plasticity of its genome: 11.7% DNA repeats containing more than 1,000 bases, 6.8% putative transposases and 21 putative restriction enzymes. Compared to the genomes of other cyanobacterial lineages, strain PCC 7806 contains a large number of atypical genes that may have been acquired by lateral transfers. Metabolic pathways, such as fermentation and a methionine salvage pathway, have been identified, as have genes for programmed cell death that may be related to the rapid disappearance of Microcystis blooms in nature. Analysis of the PCC 7806 genome also reveals striking novel biosynthetic features that might help to elucidate the ecological impact of secondary metabolites and lead to the discovery of novel metabolites for new biotechnological applications. M. aeruginosa and other large cyanobacterial genomes exhibit a rapid loss of synteny in contrast to other microbial genomes. Conclusion Microcystis aeruginosa PCC 7806 appears to have adopted an evolutionary strategy relying on unusual genome plasticity to adapt to eutrophic freshwater ecosystems, a property shared by another strain of M. aeruginosa (NIES-843). Comparisons of the genomes of PCC 7806 and other cyanobacterial strains indicate that a similar strategy may have also been used by the marine strain Crocosphaera watsonii WH8501 to adapt to other ecological niches, such as oligotrophic open oceans.

Frangeul, Lionel; Quillardet, Philippe; Castets, Anne-Marie; Humbert, Jean-Francois; Matthijs, Hans CP; Cortez, Diego; Tolonen, Andrew; Zhang, Cheng-Cai; Gribaldo, Simonetta; Kehr, Jan-Christoph; Zilliges, Yvonne; Ziemert, Nadine; Becker, Sven; Talla, Emmanuel; Latifi, Amel; Billault, Alain; Lepelletier, Anthony; Dittmann, Elke; Bouchier, Christiane; Tandeau de Marsac, Nicole

2008-01-01

224

Transcription of a 'photosynthetic' T4-type phage during infection of a marine cyanobacterium.  

PubMed

The transcription of S-PM2 phage following infection of Synechococcus sp. WH7803, a marine cyanobacterium, was analysed by quantitative real-time PCR. Unlike the distantly related coliphage T4, there were only two (early and late) instead of three (early, middle and late) classes of transcripts during the developmental cycle of the phage. This difference is consistent with the absence from the S-PM2 genome of T4-like middle mode promoter sequences and the transcription factors associated with their recognition. Phage S-PM2 carries the 'photosynthetic' genes psbA and psbD that encode homologues of the host photosystem II proteins D1 and D2. Transcripts of the phage psbA gene appeared soon after infection and remained at high levels until lysis. Throughout the course of infection, the photosynthetic capacity of the cells remained constant. A considerable transient increase in the abundance of the host psbA transcripts occurred shortly after infection, suggesting that the host responds to the trauma of phage infection in a similar way as it does to a variety of other environmental stresses. The very substantial transcription of the phage psbA gene during the latter phase of phage infection suggests that S-PM2 has acquired this cellular gene to ensure that D1 levels and thus photosynthesis are fully maintained until the infected cell finally lyses. Unexpectedly, transcripts of a phage-encoded S-layer protein gene were among the earliest and most abundant detected, suggesting that this partial homologue of a host protein plays an important role in the S-PM2 infection process. PMID:16623740

Clokie, Martha R J; Shan, Jinyu; Bailey, Shaun; Jia, Ying; Krisch, Henry M; West, Stephen; Mann, Nicholas H

2006-05-01

225

Ultrafast dynamics of phytochrome from the cyanobacterium synechocystis, reconstituted with phycocyanobilin and phycoerythrobilin.  

PubMed Central

Femtosecond time-resolved transient absorption spectroscopy was employed to characterize for the first time the primary photoisomerization dynamics of a bacterial phytochrome system in the two thermally stable states of the photocycle. The 85-kDa phytochrome Cph1 from the cyanobacterium Synechocystis PCC 6803 expressed in Escherichia coli was reconstituted with phycocyanobilin (Cph1-PCB) and phycoerythrobilin (Cph1-PEB). The red-light-absorbing form Pr of Cph1-PCB shows an approximately 150 fs relaxation in the S(1) state after photoexcitation at 650 nm. The subsequent Z-E isomerization between rings C and D of the linear tetrapyrrole-chromophore is best described by a distribution of rate constants with the first moment at (16 ps)(-1). Excitation at 615 nm leads to a slightly broadened distribution. The reverse E-Z isomerization, starting from the far-red-absorbing form Pfr, is characterized by two shorter time constants of 0.54 and 3.2 ps. In the case of Cph1-PEB, double-bond isomerization does not take place, and the excited-state lifetime extends into the nanosecond regime. Besides a stimulated emission rise time between 40 and 150 fs, no fast relaxation processes are observed. This suggests that the chromophore-protein interaction along rings A, B, and C does not contribute much to the picosecond dynamics observed in Cph1-PCB but rather the region around ring D near the isomerizing C(15) [double bond] C(16) double bond. The primary reaction dynamics of Cph1-PCB at ambient temperature is found to exhibit very similar features as those described for plant type A phytochrome, i.e., a relatively slow Pr, and a fast Pfr, photoreaction. This suggests that the initial reactions were established already before evolution of plant phytochromes began.

Heyne, Karsten; Herbst, Johannes; Stehlik, Dietmar; Esteban, Berta; Lamparter, Tilman; Hughes, Jon; Diller, Rolf

2002-01-01

226

Characterization of the Response to Zinc Deficiency in the Cyanobacterium Anabaena sp. Strain PCC 7120  

PubMed Central

Zur regulators control zinc homeostasis by repressing target genes under zinc-sufficient conditions in a wide variety of bacteria. This paper describes how part of a survey of duplicated genes led to the identification of the open reading frame all2473 as the gene encoding the Zur regulator of the cyanobacterium Anabaena sp. strain PCC 7120. All2473 binds to DNA in a zinc-dependent manner, and its DNA-binding sequence was characterized, which allowed us to determine the relative contribution of particular nucleotides to Zur binding. A zur mutant was found to be impaired in the regulation of zinc homeostasis, showing sensitivity to elevated concentrations of zinc but not other metals. In an effort to characterize the Zur regulon in Anabaena, 23 genes containing upstream putative Zur-binding sequences were identified and found to be regulated by Zur. These genes are organized in six single transcriptional units and six operons, some of them containing multiple Zur-regulated promoters. The identities of genes of the Zur regulon indicate that Anabaena adapts to conditions of zinc deficiency by replacing zinc metalloproteins with paralogues that fulfill the same function but presumably with a lower zinc demand, and with inducing putative metallochaperones and membrane transport systems likely being involved in the scavenging of extracellular zinc, including plasma membrane ABC transport systems and outer membrane TonB-dependent receptors. Among the Zur-regulated genes, the ones showing the highest induction level encode proteins of the outer membrane, suggesting a primary role for components of this cell compartment in the capture of zinc cations from the extracellular medium.

Napolitano, Mauro; Rubio, Miguel Angel; Santamaria-Gomez, Javier; Olmedo-Verd, Elvira; Robinson, Nigel J.

2012-01-01

227

Lack of Phylogeographic Structure in the Freshwater Cyanobacterium Microcystis aeruginosa Suggests Global Dispersal  

PubMed Central

Background Free-living microorganisms have long been assumed to have ubiquitous distributions with little biogeographic signature because they typically exhibit high dispersal potential and large population sizes. However, molecular data provide contrasting results and it is far from clear to what extent dispersal limitation determines geographic structuring of microbial populations. We aimed to determine biogeographical patterns of the bloom-forming freshwater cyanobacterium Microcystis aeruginosa. Being widely distributed on a global scale but patchily on a regional scale, this prokaryote is an ideal model organism to study microbial dispersal and biogeography. Methodology/Principal Findings The phylogeography of M. aeruginosa was studied based on a dataset of 311 rDNA internal transcribed spacer (ITS) sequences sampled from six continents. Richness of ITS sequences was high (239 ITS types were detected). Genetic divergence among ITS types averaged 4% (maximum pairwise divergence was 13%). Preliminary analyses revealed nearly completely unresolved phylogenetic relationships and a lack of genetic structure among all sequences due to extensive homoplasy at multiple hypervariable sites. After correcting for this, still no clear phylogeographic structure was detected, and no pattern of isolation by distance was found on a global scale. Concomitantly, genetic differentiation among continents was marginal, whereas variation within continents was high and was mostly shared with all other continents. Similarly, no genetic structure across climate zones was detected. Conclusions/Significance The high overall diversity and wide global distribution of common ITS types in combination with the lack of phylogeographic structure suggest that intercontinental dispersal of M. aeruginosa ITS types is not rare, and that this species might have a truly cosmopolitan distribution.

van Gremberghe, Ineke; Vanormelingen, Pieter; Van der Gucht, Katleen; Debeer, Ann-Eline; Lacerot, Gissell; De Meester, Luc; Vyverman, Wim

2011-01-01

228

Sustained H2 Production Driven by Photosynthetic Water Splitting in a Unicellular Cyanobacterium  

PubMed Central

ABSTRACT The relationship between dinitrogenase-driven H2 production and oxygenic photosynthesis was investigated in a unicellular cyanobacterium, Cyanothece sp. ATCC 51142, using a novel custom-built photobioreactor equipped with advanced process control. Continuously illuminated nitrogen-deprived cells evolved H2 at rates up to 400 µmol ? mg Chl?1 ? h?1 in parallel with uninterrupted photosynthetic O2 production. Notably, sustained coproduction of H2 and O2 occurred over 100 h in the presence of CO2, with both gases displaying inverse oscillations which eventually dampened toward stable rates of 125 and 90 µmol ? mg Chl?1 ? h?1, respectively. Oscillations were not observed when CO2 was omitted, and instead H2 and O2 evolution rates were positively correlated. The sustainability of the process was further supported by stable chlorophyll content, maintenance of baseline protein and carbohydrate levels, and an enhanced capacity for linear electron transport as measured by chlorophyll fluorescence throughout the experiment. In situ light saturation analyses of H2 production displayed a strong dose dependence and lack of O2 inhibition. Inactivation of photosystem II had substantial long-term effects but did not affect short-term H2 production, indicating that the process is also supported by photosystem I activity and oxidation of endogenous glycogen. However, mass balance calculations suggest that carbohydrate consumption in the light may, at best, account for no more than 50% of the reductant required for the corresponding H2 production over that period. Collectively, our results demonstrate that uninterrupted H2 production in unicellular cyanobacteria can be fueled by water photolysis without the detrimental effects of O2 and have important implications for sustainable production of biofuels.

Melnicki, Matthew R.; Pinchuk, Grigoriy E.; Hill, Eric A.; Kucek, Leo A.; Fredrickson, Jim K.; Konopka, Allan; Beliaev, Alexander S.

2012-01-01

229

Evidence for HCO3? Transport by the Blue-Green Alga (Cyanobacterium) Coccochloris peniocystis1  

PubMed Central

The possibility of HCO3? transport in the blue-green alga (cyanobacterium) Coccochloris peniocystis has been investigated. Coccochloris photosynthesized most rapidly in the pH range 8 to 10, where most of the inorganic C exists as HCO3?. If photosynthesis used only CO2 from the external solution the rate of photosynthesis would be limited by the rate of HCO3? dehydration to CO2. Observed rates of photosynthesis at alkaline pH were as much as 48-fold higher than could be supported by spontaneous dehydration of HCO3? in the external solution. Assays for extracellular carbonic anhydrase were negative. The evidence strongly suggests that HCO3? was a direct C source for photosynthesis. Weakly buffered solutions became alkaline during photosynthesis with a one-to-one stoichiometry between OH? appearance in the medium and HCO3? initially added. Alkalization occurred only during photosynthesis and was blocked by 3-(3,4-dichlorophenyl)-1, 1-dimethylurea, diuron. It is suggested that HCO3? was transported into cells of Coccochloris in exchange for OH? produced as a result of HCO3? fixation in photosynthesis. The inorganic C concentration required to support a rate of photosynthesis of half the maximum rate (Km) was 6 micromolar at pH 8.0 or, in terms of available CO2, a Km of 0.16 micromolar. This value is two orders of magnitude lower than reported Km values for the d-ribulose-1,5-bisphosphate carboxylase for blue-green algae. It is suggested that the putative HCO3? transport by Coccochloris serves to raise the CO2 concentration around the carboxylase to levels high enough for effective fixation.

Miller, Anthony G.; Colman, Brian

1980-01-01

230

Protein expression profiles in an endosymbiotic cyanobacterium revealed by a proteomic approach.  

PubMed

Molecular mechanisms behind adaptations in the cyanobacterium (Nostoc sp.) to a life in endosymbiosis with plants are still not clarified, nor are the interactions between the partners. To get further insights, the proteome of a Nostoc strain, freshly isolated from the symbiotic gland tissue of the angiosperm Gunnera manicata Linden, was analyzed and compared with the proteome of the same strain when free-living. Extracted proteins were separated by two-dimensional gel electrophoresis and were identified by matrix-assisted laser desorption/ionization-time of flight mass spectrometry combined with tandem mass spectrometry. Even when the higher percentage of differentiated cells (heterocysts) in symbiosis was compensated for, the majority of the proteins detected in the symbiotic cyanobacteria were present in the free-living counterpart, indicating that most cellular processes were common for both stages. However, differential expression profiling revealed a significant number of proteins to be down-regulated or missing in the symbiotic stage, while others were more abundant or only expressed in symbiosis. The differential protein expression was primarily connected to i) cell envelope-associated processes, including proteins involved in exopolysaccharide synthesis and surface and membrane associated proteins, ii) to changes in growth and metabolic activities (C and N), including upregulation of nitrogenase and proteins involved in the oxidative pentose phosphate pathway and downregulation of Calvin cycle enzymes, and iii) to the dark, microaerobic conditions offered inside the Gunnera gland cells, including changes in relative phycobiliprotein concentrations. This is the first comprehensive analysis of proteins in the symbiotic state. PMID:17073307

Ekman, Martin; Tollbäck, Petter; Klint, Johan; Bergman, Birgitta

2006-11-01

231

The genome sequence of the cyanobacterium Oscillatoria sp. PCC 6506 reveals several gene clusters responsible for the biosynthesis of toxins and secondary metabolites.  

PubMed

We report a draft sequence of the genome of Oscillatoria sp. PCC 6506, a cyanobacterium that produces anatoxin-a and homoanatoxin-a, two neurotoxins, and cylindrospermopsin, a cytotoxin. Beside the clusters of genes responsible for the biosynthesis of these toxins, we have found other clusters of genes likely involved in the biosynthesis of not-yet-identified secondary metabolites. PMID:20675499

Méjean, Annick; Mazmouz, Rabia; Mann, Stéphane; Calteau, Alexandra; Médigue, Claudine; Ploux, Olivier

2010-07-30

232

Growth of Daphnia magna males and females fed with the cyanobacterium Microcystis aeruginosa and the green alga Scenedesmus obliquus in different proportions  

Microsoft Academic Search

Laboratory experiments were used to study the sensitivity of both male and female Daphnia magna to a toxic cyanobacterium, Microcystis aeruginosa. Male and female D. magna were fed with M. aeruginosa and a green alga (Scenedesmus obliquus) in different mixtures that included 0%, 25%, 50%, 75% and 100% Microcystis. Growth of both males and females declined with increasing proportion of

M. F. L. L. W. Lürling; Wendy Beekman

2006-01-01

233

Viability of the cyanobacterium Planktothrix rubescens in the cold and dark, related to over-winter survival and summer recruitment in Lake Zürich  

Microsoft Academic Search

Over the winter many phytoplankton in deep lakes encounter prolonged periods of dark and cold. Survival times of the planktonic cyanobacterium Planktothrix rubescens were assessed by isolating single filaments of six strains in separate tubes that were stored at 4–5° C in the dark for 2–16 weeks and then incubating them at 20° C in the light. Viable filaments grew

Daryl P. Holland; Anthony E. Walsby

2008-01-01

234

EPR study of electron transport in the cyanobacterium Synechocystis sp. PCC 6803: Oxygen-dependent interrelations between photosynthetic and respiratory electron transport chains  

Microsoft Academic Search

In this work, we investigated electron transport processes in the cyanobacterium Synechocystis sp. PCC 6803, with a special emphasis focused on oxygen-dependent interrelations between photosynthetic and respiratory electron transport chains. Redox transients of the photosystem I primary donor P700 and oxygen exchange processes were measured by the EPR method under the same experimental conditions. To discriminate between the factors controlling

Boris V. Trubitsin; Vasilii V. Ptushenko; Olga A. Koksharova; Mahir D. Mamedov; Liya A. Vitukhnovskaya; Igor A. Grigor'ev; Alexey Yu. Semenov; Alexander N. Tikhonov

2005-01-01

235

Insights into the complex 3-D architecture of thylakoid membranes in unicellular cyanobacterium Cyanothece sp. ATCC 51142.  

PubMed

In cyanobacteria and chloroplasts, thylakoids are the complex internal membrane system where the light reactions of oxygenic photosynthesis occur. In plant chloroplasts, thylakoids are differentiated into a highly interconnected system of stacked grana and unstacked stroma membranes. In contrast, in cyanobacteria, the evolutionary progenitors of chloroplasts, thylakoids do not routinely form stacked and unstacked regions, and the architecture of the thylakoid membrane systems is only now being described in detail in these organisms. We used electron tomography to examine the thylakoid membrane systems in one cyanobacterium, Cyanothece sp. ATCC 51142. Our data showed that thylakoids form a complicated branched network with a rudimentary quasi-helical architecture in this organism. A well accepted helical model of grana-stroma architecture of plant thylakoids describes an organization in which stroma thylakoids wind around stacked granum in right-handed spirals. Here we present data showing that the simplified helical architecture in Cyanothece 51142 is left-handed in nature. We propose a model comparing the thylakoid membranes in plants and this cyanobacterium in which the system in Cyanothece 51142 is composed of non-stacked membranes linked by fret-like connections to other membrane components of the system in a limited left-handed arrangement. PMID:21445014

Liberton, Michelle; Austin, Jotham R; Berg, R Howard; Pakrasi, Himadri B

2011-04-01

236

Effects of the cyanobacterium Cylindrospermopsis raciborskii on feeding and life-history characteristics of the grazer Daphnia magna.  

PubMed

Laboratory experiments were used to test the hypothesis that feeding and growth of the zooplankton grazer Daphnia magna will decrease with increasing proportions of the cyanobacterium Cylindrospermopsis raciborskii in the diet (mixed feeds with the green alga Scenedesmus obliquus). A strain of C. raciborskii, which does not produce cylindrospermopsin but contains saxitoxins and gonyautoxins, was not acutely toxic to Daphnia, as the daphnids survived slightly longer in suspensions with the cyanobacterium as the sole feed than in medium without food. Daphnia growth rates were only depressed at feeds comprised of 75% C. raciborskii or more. Daphnids were larger with increased proportions of Scenedesmus in the food, but there was no difference between animals reared on mixed feeds and those grown on different proportions of a pure diet of Scenedesmus. Daphnia clearance rates on feeds with a high share of C. raciborskii were significantly lower than on mixtures with a low share of C. raciborskii. Consequently, in cylindrospermopsin-free strains, chemotypes that have been observed so far in Europe and Brazil, feeding inhibition and the resulting energy limitation might be the dominant factor affecting growth of large-bodied cladocerans. PMID:18951629

Soares, Maria Carolina S; Lürling, Miquel; Panosso, Renata; Huszar, Vera

2008-10-31

237

Insights into the complex 3-D architecture of thylakoid membranes in the unicellular cyanobacterium Cyanothece sp. ATCC 51142  

PubMed Central

In cyanobacteria and chloroplasts, thylakoids are the complex internal membrane system where the light reactions of oxygenic photosynthesis occur. In plant chloroplasts, thylakoids are differentiated into a highly interconnected system of stacked grana and unstacked stroma membranes. In contrast, in cyanobacteria, the evolutionary progenitors of chloroplasts, thylakoids do not routinely form stacked and unstacked regions, and the architecture of the thylakoid membrane systems is only now being described in detail in these organisms. We used electron tomography to examine the thylakoid membrane systems in one cyanobacterium, Cyanothece sp. ATCC 51142. Our data showed that thylakoids form a complicated branched network with a rudimentary quasi-helical architecture in this organism. A well accepted helical model of grana-stroma architecture of plant thylakoids describes an organization in which stroma thylakoids wind around stacked granum in right-handed spirals. Here we present data showing that the simplified helical architecture in Cyanothece 51142 is lefthanded in nature. We propose a model comparing the thylakoid membranes in plants and this cyanobacterium in which the system in Cyanothece 51142 is composed of non-stacked membranes linked by fret-like connections to other membrane components of the system in a limited left-handed arrangement.

Liberton, Michelle; Austin, Jotham R; Berg, R Howard

2011-01-01

238

Unique Thylakoid Membrane Architecture of a Unicellular N2-Fixing Cyanobacterium Revealed by Electron Tomography1[W][OA  

PubMed Central

Cyanobacteria, descendants of the endosymbiont that gave rise to modern-day chloroplasts, are vital contributors to global biological energy conversion processes. A thorough understanding of the physiology of cyanobacteria requires detailed knowledge of these organisms at the level of cellular architecture and organization. In these prokaryotes, the large membrane protein complexes of the photosynthetic and respiratory electron transport chains function in the intracellular thylakoid membranes. Like plants, the architecture of the thylakoid membranes in cyanobacteria has direct impact on cellular bioenergetics, protein transport, and molecular trafficking. However, whole-cell thylakoid organization in cyanobacteria is not well understood. Here we present, by using electron tomography, an in-depth analysis of the architecture of the thylakoid membranes in a unicellular cyanobacterium, Cyanothece sp. ATCC 51142. Based on the results of three-dimensional tomographic reconstructions of near-entire cells, we determined that the thylakoids in Cyanothece 51142 form a dense and complex network that extends throughout the entire cell. This thylakoid membrane network is formed from the branching and splitting of membranes and encloses a single lumenal space. The entire thylakoid network spirals as a peripheral ring of membranes around the cell, an organization that has not previously been described in a cyanobacterium. Within the thylakoid membrane network are areas of quasi-helical arrangement with similarities to the thylakoid membrane system in chloroplasts. This cyanobacterial thylakoid arrangement is an efficient means of packing a large volume of membranes in the cell while optimizing intracellular transport and trafficking.

Liberton, Michelle; Austin, Jotham R.; Berg, R. Howard; Pakrasi, Himadri B.

2011-01-01

239

Fur regulates the expression of iron-stress genes in the cyanobacterium Synechococcus sp. strain PCC 7942.  

PubMed

A homologue of the 'ferric uptake regulation' gene (fur) was isolated from the cyanobacterium Synechococcus sp. strain PCC 7942 by an Escherichia coli-based 'in vivo repression assay'. The assay uses a reporter-gene construct containing the promoter region of the iron-regulated cyanobacterial gene isiA, fused to the coding region for chloramphenicol acetyltransferase. The isolated gene codes for a protein that has 41% sequence similarity (36% identity) to Fur from E. coli and contains the putative iron-binding motif found in the Fur proteins of purple bacteria. No significant similarity was found to the DxtR repressor that regulates the expression of toxin and siderophore production in Gram-positive bacteria. Insertional mutagenesis of the cloned cyanobacterial fur gene led to the creation of heteroallelic mutants that showed iron-deficiency symptoms in iron-replete medium, including the constitutive production of flavodoxin and of hydroxamate siderophores. Failure to eliminate wild-type copies of the fur gene from the polyploid genome of Synechococcus 7942 implies that in this cyanobacterium Fur may have essential functions in addition to the regulation of genes involved in iron scavenging or photosynthetic electron transport. PMID:8704986

Ghassemian, M; Straus, N A

1996-06-01

240

Localization of Membrane Proteins in the Cyanobacterium Synechococcus sp. PCC7942 (Radial Asymmetry in the Photosynthetic Complexes).  

PubMed Central

Localization of membrane proteins in the cyanobacterium Synechococcus sp. PCC7942 was determined by transmission electron microscopy utilizing immunocytochemistry with cells prepared by freeze-substitution. This preparation procedure maintained cellular morphology and permitted detection of cellular antigens with high sensitivity and low background. Synechococcus sp. PCC7942 is a unicellular cyanobacterium with thylakoids organized in concentric layers toward the periphery of the cell. Cytochrome oxidase was localized almost entirely in the cytoplasmic membrane, whereas a carotenoprotein (P35) was shown to be a cell wall component. The major photosystem II (PSII) proteins (D1, D2 CP43, and CP47) were localized throughout the thylakoids. Proteins of the Cyt b6/f complex were found to have a similar distribution. Thylakoid luminal proteins, such as the Mn-stabilizing protein, were located primarily in the thylakoid, but a small, reproducible fraction was found in the outer compartment. The photosystem I (PSI) reaction center proteins and the ATP synthase proteins were found associated mostly with the outermost thylakoid and with the cytoplasmic membrane. These results indicated that the photosynthetic apparatus is not evenly distributed throughout the thylakoids. Rather, there is a radial asymmetry such that much of the PSI and the ATPase synthase is located in the outermost thylakoid. The relationship of this structure to the photosynthetic mechanism is discussed. It is suggested that the photosystems are separated because of kinetic differences between PSII and PSI, as hypothesized by H.-W. Trissl and C. Wilhelm (Trends Biochem Sci [1993] 18:415-419).

Sherman, D. M.; Troyan, T. A.; Sherman, L. A.

1994-01-01

241

Novel Insights into the Regulation of LexA in the Cyanobacterium Synechocystis sp. Strain PCC 6803 ? †  

PubMed Central

The transcription factor LexA in the cyanobacterium Synechocystis sp. strain PCC 6803 has been shown to regulate genes that are not directly involved in DNA repair but instead in several different metabolic pathways. However, the signal transduction pathways remain largely uncharacterized. The present work gives novel insights into the regulation of LexA in this unicellular cyanobacterium. A combination of Northern and Western blotting, using specific antibodies against the cyanobacterial LexA, was employed to show that this transcription regulator is under posttranscriptional control, in addition to the classical and already-described transcriptional regulation. Moreover, detailed two-dimensional (2D) electrophoresis analyses of the protein revealed that LexA undergoes posttranslational modifications. Finally, a fully segregated LexA::GFP (green fluorescent protein) fusion-modified strain was produced to image LexA's spatial distribution in live cells. The fusion protein retains DNA binding capabilities, and the GFP fluorescence indicates that LexA is localized in the innermost region of the cytoplasm, decorating the DNA in an evenly distributed pattern. The implications of these findings for the overall role of LexA in Synechocystis sp. strain PCC 6803 are further discussed.

Oliveira, Paulo; Lindblad, Peter

2011-01-01

242

Towards a proteome project of cyanobacterium Synechocystis sp. strain PCC6803: linking 130 protein spots with their respective genes.  

PubMed

Following the complete sequencing of the genome of the univellular cyanobacterium, Synechocystis sp. strain PCC6803 within our institute, a protein-gene linkage map of this photosynthetic microorganism was successfully constructed for 130 high abundance proteins present on two-dimensional gels. An additional six proteins were analyzed, but were probably encoded extrachromosomally. In order to demonstrate the usefulness of this protein-gene linkage map, we analyzed the changes that occur in cellular proteins after illumination of PCC6803 cells. The results indicate that this protein-gene linkage map greatly simplifies the identification process of such modulated genes. After illumination, at least three distinctive spots with reduced intensity were detected on two-dimensional gels and the corresponding genes of two of these were successfully identified as chaperonin 2 and a Tortula ruralis rehydrin-related gene. Thus, the combination of the protein-gene linkage map and two-dimensional gel electrophoresis should permit a comprehensive analyses of the proteins encoded by the genome (i.e., "proteome") of this photosynthetic autotroph. This post-genome project represents a productive way of exploiting the information obtained from the sequencing of the cyanobacterium genome. PMID:9298645

Sazuka, T; Ohara, O

1997-08-01

243

Application of real-time PCR to estimate toxin production by the cyanobacterium Planktothrix sp.  

PubMed

Quantitative real-time PCR methods are increasingly being applied for the enumeration of toxic cyanobacteria in the environment. However, to justify the use of real-time PCR quantification as a monitoring tool, significant correlations between genotype abundance and actual toxin concentrations are required. In the present study, we aimed to explain the concentrations of three structural variants of the hepatotoxin microcystin (MC) produced by the filamentous cyanobacterium Planktothrix sp., [Asp, butyric acid (Dhb)]-microcystin-RR (where RR means two arginines), [Asp, methyl-dehydro-alanine (Mdha)]-microcystin-RR, and [Asp, Dhb]-microcystin-homotyrosine-arginine (HtyR), by the abundance of the microcystin genotypes encoding their synthesis. Three genotypes of microcystin-producing cyanobacteria (denoted the Dhb, Mdha, and Hty genotypes) in 12 lakes of the Alps in Austria, Germany, and Switzerland from 2005 to 2007 were quantified by means of real-time PCR. Their absolute and relative abundances were related to the concentration of the microcystin structural variants in aliquots determined by high-performance liquid chromatography (HPLC). The total microcystin concentrations varied from 0 to 6.2 microg liter(-1) (mean +/- standard error [SE] of 0.6 +/- 0.1 microg liter(-1)) among the samples, in turn resulting in an average microcystin content in Planktothrix of 3.1 +/- 0.7 microg mm(-3) biovolume. Over a wide range of the population density (0.001 to 3.6 mm(3) liter(-1) Planktothrix biovolume), the Dhb genotype and [Asp, Dhb]-MC-RR were most abundant, while the Hty genotype and MC-HtyR were found to be in the lowest proportion only. In general, there was a significant linear relationship between the abundance/proportion of specific microcystin genotypes and the concentration/proportion of the respective microcystin structural variants on a logarithmic scale. We conclude that estimating the abundance of specific microcystin genotypes by quantitative real-time PCR is useful for predicting the concentration of microcystin variants in water. PMID:20363794

Ostermaier, Veronika; Kurmayer, Rainer

2010-04-02

244

Application of Real-Time PCR To Estimate Toxin Production by the Cyanobacterium Planktothrix sp.? †  

PubMed Central

Quantitative real-time PCR methods are increasingly being applied for the enumeration of toxic cyanobacteria in the environment. However, to justify the use of real-time PCR quantification as a monitoring tool, significant correlations between genotype abundance and actual toxin concentrations are required. In the present study, we aimed to explain the concentrations of three structural variants of the hepatotoxin microcystin (MC) produced by the filamentous cyanobacterium Planktothrix sp., [Asp, butyric acid (Dhb)]-microcystin-RR (where RR means two arginines), [Asp, methyl-dehydro-alanine (Mdha)]-microcystin-RR, and [Asp, Dhb]-microcystin-homotyrosine-arginine (HtyR), by the abundance of the microcystin genotypes encoding their synthesis. Three genotypes of microcystin-producing cyanobacteria (denoted the Dhb, Mdha, and Hty genotypes) in 12 lakes of the Alps in Austria, Germany, and Switzerland from 2005 to 2007 were quantified by means of real-time PCR. Their absolute and relative abundances were related to the concentration of the microcystin structural variants in aliquots determined by high-performance liquid chromatography (HPLC). The total microcystin concentrations varied from 0 to 6.2 ?g liter?1 (mean ± standard error [SE] of 0.6 ± 0.1 ?g liter?1) among the samples, in turn resulting in an average microcystin content in Planktothrix of 3.1 ± 0.7 ?g mm?3 biovolume. Over a wide range of the population density (0.001 to 3.6 mm3 liter?1 Planktothrix biovolume), the Dhb genotype and [Asp, Dhb]-MC-RR were most abundant, while the Hty genotype and MC-HtyR were found to be in the lowest proportion only. In general, there was a significant linear relationship between the abundance/proportion of specific microcystin genotypes and the concentration/proportion of the respective microcystin structural variants on a logarithmic scale. We conclude that estimating the abundance of specific microcystin genotypes by quantitative real-time PCR is useful for predicting the concentration of microcystin variants in water.

Ostermaier, Veronika; Kurmayer, Rainer

2010-01-01

245

Sanctolide A, a 14-membered PK-NRP hybrid macrolide from the cultured cyanobacterium Oscillatoria sancta (SAG 74.79).  

PubMed

Sanctolide A (1), a 14-membered polyketide-nonribosomal peptide (PK-NRP) hybrid macrolide, was isolated from the cultured cyanobacterium Oscillatoria sancta (SAG 74.79). The planar structure was determined using various spectroscopic techniques including HRESIMS, and 1D and 2D NMR analyses. The relative configuration was assigned by J-based configurational analysis in combination with NOE correlations. The absolute configuration was determined by Mosher ester and enantioselective HPLC analyses. The structure of sanctolide A (1) features a rare N-methyl enamide and a 2-hydroxyisovaleric acid, which are incorporated to form a 14-membered macrolide ring structure, comprising a new type of cyanobacterial macrolides derived from a PKS-NRPS hybrid biosynthetic pathway. PMID:22711943

Kang, Hahk-Soo; Krunic, Aleksej; Orjala, Jimmy

2012-05-01

246

Reduction of exogenous ketones depends upon NADPH generated photosynthetically in cells of the cyanobacterium Synechococcus PCC 7942  

PubMed Central

Effective utilization of photosynthetic microorganisms as potential biocatalysts is favorable for the production of useful biomaterials and the reduction of atmospheric CO2. For example, biocatalytic transformations are used in the synthesis of optically active alcohols. We previously found that ketone reduction in cells of the cyanobacterium Synechococcus PCC 7942 is highly enantioselective and remarkably enhanced under light illumination. In this study, the mechanism of light-enhanced ketone reduction was investigated in detail using several inhibitors of photosynthetic electron transport and of enzymes of the Calvin cycle. It is demonstrated that light intensity and photosynthesis inhibitors significantly affect the ketone reduction activity in Synechococcus. This indicates that the reduction correlates well with photosynthetic activity. Moreover, ketone reduction in Synechococcus specifically depends upon NADPH and not NADH. These results also suggest that cyanobacteria have the potential to be utilized as biocatalytic systems for direct usage of light energy in various applications such as syntheses of useful compounds and remediation of environmental pollutants.

2011-01-01

247

Dark hexose metabolism by photoautotrophically and heterotrophically grown cells of the blue-green alga (Cyanobacterium) Nostoc sp. strain Mac.  

PubMed Central

Photoautotrophically grown cells of the blue-green alga (cyanobacterium) Nostoc sp. strain Mac assimilated and oxidized both glucose and fructose in the dark at different rates. The rate of fructose metabolism in these cells could be stimulated by casein hydrolysate, the effect being most pronounced at low sugar concentrations. This stimulation was not seen in cells grown heterotrophically in the dark, suggesting that it is a transitory phenomenon which disappears during the autotrophy-heterotrophy growth transition. The stimulation of fructose assimilation by casein hydrolysate was abolished by chloramphenicol or streptomycin, suggesting there are rate-limiting steps in protein biosynthesis in the dark that ultimately lead to inhibition of fructose uptake. Glucose metabolism did not show these phenomena, indicating there are differences in the metabolism of the two sugars.

Bottomley, P J; van Baalen, C

1978-01-01

248

Malyngamide 2, an oxidized lipopeptide with nitric oxide inhibiting activity from a Papua New Guinea marine cyanobacterium.  

PubMed

A Papua New Guinea collection of the marine cyanobacterium cf. Lyngbya sordida yielded three known compounds as well as a new PKS-NRPS-derived malyngamide with anti-inflammatory and cytotoxic activity. Malyngamide 2 features an extensively oxidized cyclohexanone ring. Resolution of the ring core as a 6,8,9-triol rather then a 7,8,9-triol and relative configuration was based on chemical shift and bond geometry modeling in conjunction with homonuclear and heteronuclear coupling constants, NOE and ROE correlations, and other structural information. Malyngamide 2 exhibited anti-inflammatory activity in LPS-induced RAW macrophage cells (IC(50) = 8.0 ?M) with only modest cytotoxicity to the mammalian cell line. PMID:21155594

Malloy, Karla L; Villa, Francisco A; Engene, Niclas; Matainaho, Teatulohi; Gerwick, Lena; Gerwick, William H

2010-12-14

249

Phosphorus addition reverses the positive effect of zebra mussels (Dreissena polymorpha) on the toxic cyanobacterium, Microcystis aeruginosa.  

PubMed

We tested the hypothesis that zebra mussels (Dreissena polymorpha) have positive effects on the toxin-producing cyanobacterium, Microcystis aeruginosa, at low phosphorus (P) concentrations, but negative effects on M. aeruginosa at high P, with a large-scale enclosure experiment in an oligotrophic lake. After three weeks, mussels had a significantly positive effect on M. aeruginosa at ambient P (total phosphorus, TP ?10 ?g L?¹), and a significantly negative effect at high P (simulating a TP of ?40 ?g L?¹ in lakes). Positive and negative effects were strong and very similar in magnitude. Thus, we were able to ameliorate a negative effect of Dreissena invasion on water quality (i.e., promotion of Microcystis) by adding P to water from an oligotrophic lake. Our results are congruent with many field observations of Microcystis response to Dreissena invasion across ecosystems of varying P availability. PMID:22507249

Sarnelle, Orlando; White, Jeffrey D; Horst, Geoffrey P; Hamilton, Stephen K

2012-04-01

250

Establishment of a Pure Culture of the Hitherto Uncultured Unicellular Cyanobacterium Aphanothece sacrum, and Phylogenetic Position of the Organism  

PubMed Central

Aphanothece sacrum, an edible freshwater unicellular cyanobacterium, was isolated by using novel synthetic media (designated AST and AST-5xNP). The media were designed on the basis of the ratio of inorganic elements contained in A. sacrum cells cultured in a natural pond. The isolated strain exhibits unicellular rod-shaped cells ?6 ?m in length that are scattered in an exopolysaccharide matrix, a feature similar to that of natural A. sacrum. DNA analysis of the isolated strain revealed that it carried two ferredoxin genes whose deduced amino acid sequences were almost identical to previously published sequences of ferredoxins from natural A. sacrum. Analysis of the 16S rRNA gene and ferredoxin genes revealed that A. sacrum occupies a phylogenetically unique position among the cyanobacteria.

Fujishiro, Tsuneo; Ogawa, Takahira; Matsuoka, Masayoshi; Nagahama, Kazuhiro; Takeshima, Yasunobu; Hagiwara, Hideaki

2004-01-01

251

The Biosynthetic Pathway for Synechoxanthin, an Aromatic Carotenoid Synthesized by the Euryhaline, Unicellular Cyanobacterium Synechococcus sp. Strain PCC 7002? †  

PubMed Central

The euryhaline, unicellular cyanobacterium Synechococcus sp. strain PCC 7002 produces the dicyclic aromatic carotenoid synechoxanthin (?,?-caroten-18,18?-dioic acid) as a major pigment (>15% of total carotenoid) and when grown to stationary phase also accumulates small amounts of renierapurpurin (?,?-carotene) (J. E. Graham, J. T. J. Lecomte, and D. A. Bryant, J. Nat. Prod. 71:1647-1650, 2008). Two genes that were predicted to encode enzymes involved in the biosynthesis of synechoxanthin were identified by comparative genomics, and these genes were insertionally inactivated in Synechococcus sp. strain PCC 7002 to verify their function. The cruE gene (SYNPCC7002_A1248) encodes ?-carotene desaturase/methyltransferase, which converts ?-carotene to renierapurpurin. The cruH gene (SYNPCC7002_A2246) encodes an enzyme that is minimally responsible for the hydroxylation/oxidation of the C-18 and C-18? methyl groups of renierapurpurin. Based on observed and biochemically characterized intermediates, a complete pathway for synechoxanthin biosynthesis is proposed.

Graham, Joel E.; Bryant, Donald A.

2008-01-01

252

Protein synthesis and proteolysis in immobilized cells of the cyanobacterium Nostoc commune UTEX 584 exposed to matric water stress.  

PubMed Central

Cells of the cyanobacterium Nostoc commune UTEX 584 in exponential growth were subjected to acute water stress by immobilizing them on solid supports and drying them at a matric water potential (psi m) of -99.5 MPa. Cells which had been grown in the presence of Na235SO4 before immobilization and rapid drying continued to incorporate 35S into protein for 90 min. This incorporation was inhibited by chloramphenicol. No unique proteins appeared to be synthesized during this time. Upon further drying, the level of incorporation of 35S in protein began to decrease. In contrast, there was an apparent increase in the level of certain phycobiliprotein subunits in solubilized protein extracts of these cells. Extensive proteolysis was detected after prolonged desiccation (17 days) of the cells in the light, although they still remained intact. Phycobilisomes became dissociated in both light- and dark-stored desiccated material. Images

Potts, M

1985-01-01

253

Trimeric forms of the photosystem I reaction center complex pre-exist in the membranes of the cyanobacterium Spirulina platensis.  

PubMed

Oligomeric and monomeric forms of chlorophyll-protein complexes of photosystem I (PSI) have been isolated from the mesophilic cyanobacterium Spirulina [(1992) FEBS Lett. 309, 340-342]. Electron microscopic analysis of the complexes showed that the oligomeric form is a trimer of the shape and dimensions similar to those of the trimer from thermophilic cyanobacteria. The chlorophyl ratio in the isolated trimer and monomer was found to be 7:3. The trimeric form of PSI complex in contrast to the monomeric one contains the chlorophyll emitting at 760 nm (77K), which is also found in Spirulina membranes and therefore could be used as an intrinsic probe for the trimeric complex. The 77K circular dichroism spectrum of the trimeric form is much more similar to that of Spirulina membranes than the spectrum of the monomer. Thus, the trimeric PSI complexes exist and dominate in the Spirulina membranes. PMID:8224233

Shubin, V V; Tsuprun, V L; Bezsmertnaya, I N; Karapetyan, N V

1993-11-01

254

Structure of Trichamide, a Cyclic Peptide from the Bloom-Forming Cyanobacterium Trichodesmium erythraeum, Predicted from the Genome Sequence†  

PubMed Central

A gene cluster for the biosynthesis of a new small cyclic peptide, dubbed trichamide, was discovered in the genome of the global, bloom-forming marine cyanobacterium Trichodesmium erythraeum ISM101 because of striking similarities to the previously characterized patellamide biosynthesis cluster. The tri cluster consists of a precursor peptide gene containing the amino acid sequence for mature trichamide, a putative heterocyclization gene, an oxidase, two proteases, and hypothetical genes. Based upon detailed sequence analysis, a structure was predicted for trichamide and confirmed by Fourier transform mass spectrometry. Trichamide consists of 11 amino acids, including two cysteine-derived thiazole groups, and is cyclized by an N—C terminal amide bond. As the first natural product reported from T. erythraeum, trichamide shows the power of genome mining in the prediction and discovery of new natural products.

Sudek, Sebastian; Haygood, Margo G.; Youssef, Diaa T. A.; Schmidt, Eric W.

2006-01-01

255

Organization and transcription of the class I phycoerythrin genes of the marine cyanobacterium Synechococcus sp. WH7803.  

PubMed

The nucleotide sequences of the class I phycoerythrin (PE) alpha- and beta-subunit genes (cpeA and cpeB) from the marine cyanobacterium Synechococcus sp. WH7803 are reported. The cpeB gene is located upstream of cpeA with a separation of 56 nucleotides and the two genes are co-transcribed as a transcript of 1.3 kb, with the transcription startpoint being localized to 110-111 bp upstream of cpeB. The sequence of the promoter region bears no similarity to promoters reported for other cyanobacterial PE genes. Pentanucleotide repeats found upstream of some PE operons, particularly in the case of cyanobacterial strains capable of chromatic adaption, are not found in Synechococcus sp. WH7803; instead the sequence 5'-CGGTT-3' is repeated three times in the promoter region. PMID:7512390

Newman, J; Mann, N H; Carr, N G

1994-02-01

256

Species-specific real-time PCR cell number quantification of the bloom-forming cyanobacterium Planktothrix agardhii.  

PubMed

A species-specific method to detect and quantify Planktothrix agardhii was developed by combining the SYBR Green I real-time polymerase chain reaction technique with a simplified DNA extraction procedure for standard curve preparation. Newly designed PCR primers were used to amplify a specific fragment within the rpoC1 gene. Since this gene exists in single copy in the genome, it allows the direct achievement of cell concentrations. The cell concentration determined by real-time PCR showed a linear correlation with the cell concentration determined from direct microscopic counts. The detection limit for cell quantification of the method was 8 cells ?L(-1), corresponding to 32 cells per reaction. Furthermore, the real-time qPCR method described in this study allowed a successful quantification of P. agardhii from environmental water samples, showing that this protocol is an accurate and economic tool for a rapid absolute quantification of the potentially toxic cyanobacterium P. agardhii. PMID:22484452

Churro, Catarina; Pereira, Paulo; Vasconcelos, Vitor; Valério, Elisabete

2012-04-07

257

High-Titer Heterologous Production in E. coli of Lyngbyatoxin, a Protein Kinase C Activator from an Uncultured Marine Cyanobacterium.  

PubMed

Many chemically complex cyanobacterial polyketides and nonribosomal peptides are of great pharmaceutical interest, but the levels required for exploitation are difficult to achieve from native sources. Here we develop a framework for the expression of these multifunctional cyanobacterial assembly lines in Escherichia coli using the lyngbyatoxin biosynthetic pathway, derived from a marine microbial assemblage dominated by the cyanobacterium Moorea producens. Heterologous expression of this pathway afforded high titers of both lyngbyatoxin A (25.6 mg L(-1)) and its precursor indolactam-V (150 mg L(-1)). Production, isolation, and identification of all expected chemical intermediates of lyngbyatoxin biosynthesis in E. coli also confirmed the previously proposed biosynthetic route, setting a solid chemical foundation for future pathway engineering. The successful production of the nonribosomal peptide lyngbyatoxin A in E. coli also opens the possibility for future heterologous expression, characterization, and exploitation of other cyanobacterial natural product pathways. PMID:23751865

Ongley, Sarah E; Bian, Xiaoying; Zhang, Youming; Chau, Rocky; Gerwick, William H; Müller, Rolf; Neilan, Brett A

2013-06-17

258

Photosystem trap energies and spectrally-dependent energy-storage efficiencies in the Chl d-utilizing cyanobacterium, Acaryochloris marina.  

PubMed

Acaryochloris marina is the only species known to utilize chlorophyll (Chl) d as a principal photopigment. The peak absorption wavelength of Chl d is redshifted ?40nm in vivo relative to Chl a, enabling this cyanobacterium to perform oxygenic phototrophy in niche environments enhanced in far-red light. We present measurements of the in vivo energy-storage (E-S) efficiency of photosynthesis in A. marina, obtained using pulsed photoacoustics (PA) over a 90-nm range of excitation wavelengths in the red and far-red. Together with modeling results, these measurements provide the first direct observation of the trap energies of PSI and PSII, and also the photosystem-specific contributions to the total E-S efficiency. We find the maximum observed efficiency in A. marina (40±1% at 735nm) is higher than in the Chl a cyanobacterium Synechococcus leopoliensis (35±1% at 690nm). The efficiency at peak absorption wavelength is also higher in A. marina (36±1% at 710nm vs. 31±1% at 670nm). In both species, the trap efficiencies are ?40% (PSI) and ?30% (PSII). The PSI trap in A. marina is found to lie at 740±5nm, in agreement with the value inferred from spectroscopic methods. The best fit of the model to the PA data identifies the PSII trap at 723±3nm, supporting the view that the primary electron-donor is Chl d, probably at the accessory (Chl(D1)) site. A decrease in efficiency beyond the trap wavelength, consistent with uphill energy transfer, is clearly observed and fit by the model. These results demonstrate that the E-S efficiency in A. marina is not thermodynamically limited, suggesting that oxygenic photosynthesis is viable in even redder light environments. PMID:23159726

Mielke, Steven P; Kiang, Nancy Y; Blankenship, Robert E; Mauzerall, David

2012-11-15

259

Global transcriptional responses of the toxic cyanobacterium, Microcystis aeruginosa, to nitrogen stress, phosphorus stress, and growth on organic matter.  

PubMed

Whole transcriptome shotgun sequencing (RNA-seq) was used to assess the transcriptomic response of the toxic cyanobacterium Microcystis aeruginosa during growth with low levels of dissolved inorganic nitrogen (low N), low levels of dissolved inorganic phosphorus (low P), and in the presence of high levels of high molecular weight dissolved organic matter (HMWDOM). Under low N, one third of the genome was differentially expressed, with significant increases in transcripts observed among genes within the nir operon, urea transport genes (urtBCDE), and amino acid transporters while significant decreases in transcripts were observed in genes related to photosynthesis. There was also a significant decrease in the transcription of the microcystin synthetase gene set under low N and a significant decrease in microcystin content per Microcystis cell demonstrating that N supply influences cellular toxicity. Under low P, 27% of the genome was differentially expressed. The Pho regulon was induced leading to large increases in transcript levels of the alkaline phosphatase phoX, the Pst transport system (pstABC), and the sphX gene, and transcripts of multiple sulfate transporter were also significantly more abundant. While the transcriptional response to growth on HMWDOM was smaller (5-22% of genes differentially expressed), transcripts of multiple genes specifically associated with the transport and degradation of organic compounds were significantly more abundant within HMWDOM treatments and thus may be recruited by Microcystis to utilize these substrates. Collectively, these findings provide a comprehensive understanding of the nutritional physiology of this toxic, bloom-forming cyanobacterium and the role of N in controlling microcystin synthesis. PMID:23894552

Harke, Matthew J; Gobler, Christopher J

2013-07-23

260

Microenvironmental Ecology of the Chlorophyll b-Containing Symbiotic Cyanobacterium Prochloron in the Didemnid Ascidian Lissoclinum patella.  

PubMed

The discovery of the cyanobacterium Prochloron was the first finding of a bacterial oxyphototroph with chlorophyll (Chl) b, in addition to Chl a. It was first described as Prochloron didemni but a number of clades have since been described. Prochloron is a conspicuously large (7-25??m) unicellular cyanobacterium living in a symbiotic relationship, primarily with (sub-) tropical didemnid ascidians; it has resisted numerous cultivation attempts and appears truly obligatory symbiotic. Recently, a Prochloron draft genome was published, revealing no lack of metabolic genes that could explain the apparent inability to reproduce and sustain photosynthesis in a free-living stage. Possibly, the unsuccessful cultivation is partly due to a lack of knowledge about the microenvironmental conditions and ecophysiology of Prochloron in its natural habitat. We used microsensors, variable chlorophyll fluorescence imaging and imaging of O(2) and pH to obtain a detailed insight to the microenvironmental ecology and photobiology of Prochloron in hospite in the didemnid ascidian Lissoclinum patella. The microenvironment within ascidians is characterized by steep gradients of light and chemical parameters that change rapidly with varying irradiances. The interior zone of the ascidians harboring Prochloron thus became anoxic and acidic within a few minutes of darkness, while the same zone exhibited O(2) super-saturation and strongly alkaline pH after a few minutes of illumination. Photosynthesis showed lack of photoinhibition even at high irradiances equivalent to full sunlight, and photosynthesis recovered rapidly after periods of anoxia. We discuss these new insights on the ecological niche of Prochloron and possible interactions with its host and other microbes in light of its recently published genome and a recent study of the overall microbial diversity and metagenome of L. patella. PMID:23226144

Kühl, Michael; Behrendt, Lars; Trampe, Erik; Qvortrup, Klaus; Schreiber, Ulrich; Borisov, Sergey M; Klimant, Ingo; Larkum, Anthony W D

2012-11-30

261

Halotolerant cyanobacterium Aphanothece halophytica contains a betaine transporter active at alkaline pH and high salinity.  

PubMed

Aphanothece halophytica is a halotolerant alkaliphilic cyanobacterium which can grow in media of up to 3.0 M NaCl and pH 11. This cyanobacterium can synthesize betaine from glycine by three-step methylation using S-adenosylmethionine as a methyl donor. To unveil the mechanism of betaine uptake and efflux in this alkaliphile, we isolated and characterized a betaine transporter. A gene encoding a protein (BetT(A. halophytica)) that belongs to the betaine-choline-carnitine transporter (BCCT) family was isolated. Although the predicted isoelectric pH of a typical BCCT family transporter, OpuD of Bacillus subtilis, is basic, 9.54, that of BetT(A. halophytica) is acidic, 4.58. BetT(A. halophytica) specifically catalyzed the transport of betaine. Choline, gamma-aminobutyric acid, betaine aldehyde, sarcosine, dimethylglycine, and amino acids such as proline did not compete for the uptake of betaine by BetT(A. halophytica). Sodium markedly enhanced betaine uptake rates, whereas potassium and other cations showed no effect, suggesting that BetT(A. halophytica) is a Na(+)-betaine symporter. Betaine uptake activities of BetT(A. halophytica) were high at alkaline pH values, with the optimum pH around 9.0. Freshwater Synechococcus cells overexpressing BetT(A. halophytica) showed NaCl-activated betaine uptake activities with enhanced salt tolerance, allowing growth in seawater supplemented with betaine. Kinetic properties of betaine uptake in Synechococcus cells overexpressing BetT(A. halophytica) were similar to those in A. halophytica cells. These findings indicate that A. halophytica contains a Na(+)-betaine symporter that contributes to the salt stress tolerance at alkaline pH. BetT(A. halophytica) is the first identified transporter for compatible solutes in cyanobacteria. PMID:16957224

Laloknam, Surasak; Tanaka, Kimihiro; Buaboocha, Teerapong; Waditee, Rungaroon; Incharoensakdi, Aran; Hibino, Takashi; Tanaka, Yoshito; Takabe, Teruhiro

2006-09-01

262

Halotolerant Cyanobacterium Aphanothece halophytica Contains a Betaine Transporter Active at Alkaline pH and High Salinity  

PubMed Central

Aphanothece halophytica is a halotolerant alkaliphilic cyanobacterium which can grow in media of up to 3.0 M NaCl and pH 11. This cyanobacterium can synthesize betaine from glycine by three-step methylation using S-adenosylmethionine as a methyl donor. To unveil the mechanism of betaine uptake and efflux in this alkaliphile, we isolated and characterized a betaine transporter. A gene encoding a protein (BetTA. halophytica) that belongs to the betaine-choline-carnitine transporter (BCCT) family was isolated. Although the predicted isoelectric pH of a typical BCCT family transporter, OpuD of Bacillus subtilis, is basic, 9.54, that of BetTA. halophytica is acidic, 4.58. BetTA. halophytica specifically catalyzed the transport of betaine. Choline, ?-aminobutyric acid, betaine aldehyde, sarcosine, dimethylglycine, and amino acids such as proline did not compete for the uptake of betaine by BetTA. halophytica. Sodium markedly enhanced betaine uptake rates, whereas potassium and other cations showed no effect, suggesting that BetTA. halophytica is a Na+-betaine symporter. Betaine uptake activities of BetTA. halophytica were high at alkaline pH values, with the optimum pH around 9.0. Freshwater Synechococcus cells overexpressing BetTA. halophytica showed NaCl-activated betaine uptake activities with enhanced salt tolerance, allowing growth in seawater supplemented with betaine. Kinetic properties of betaine uptake in Synechococcus cells overexpressing BetTA. halophytica were similar to those in A. halophytica cells. These findings indicate that A. halophytica contains a Na+-betaine symporter that contributes to the salt stress tolerance at alkaline pH. BetTA. halophytica is the first identified transporter for compatible solutes in cyanobacteria.

Laloknam, Surasak; Tanaka, Kimihiro; Buaboocha, Teerapong; Waditee, Rungaroon; Incharoensakdi, Aran; Hibino, Takashi; Tanaka, Yoshito; Takabe, Teruhiro

2006-01-01

263

Reduction of photosystem I reaction center by recombinant DrgA protein in isolated thylakoid membranes of the cyanobacterium Synechocystis sp. PCC 6803.  

PubMed

To study the function of soluble NAD(P)H:quinone oxidoreductase of the cyanobacterium Synechocystis sp. PCC 6803 encoded by drgA gene, recombinant DrgA protein carrying 12 histidine residues on the C-terminal end was expressed in Escherichia coli and purified. Recombinant DrgA is a flavoprotein that exhibits quinone reductase and nitroreductase activities with NAD(P)H as the electron donor. Using EPR spectroscopy, it was demonstrated that addition of recombinant DrgA protein and NADPH to DCMU-treated isolated thylakoid membranes of the cyanobacterium increased the dark re-reduction rate of the photosystem I reaction center (P700(+)). Thus, DrgA can participate in electron transfer from NADPH to the electron transport chain of the Synechocystis sp. PCC 6803 thylakoid membrane. PMID:19916920

Elanskaya, I V; Toporova, V A; Grivennikova, V G; Muronets, E M; Lukashev, E P; Timofeev, K N

2009-10-01

264

An Alkaline Phosphatase/Phosphodiesterase, PhoD, Induced by Salt Stress and Secreted Out of the Cells of Aphanothece halophytica, a Halotolerant Cyanobacterium ? †  

PubMed Central

Alkaline phosphatases (APases) are important enzymes in organophosphate utilization. Three prokaryotic APase gene families, PhoA, PhoX, and PhoD, are known; however, their functional characterization in cyanobacteria largely remains to be clarified. In this study, we cloned the phoD gene from a halotolerant cyanobacterium, Aphanothece halophytica (phoDAp). The deduced protein, PhoDAp, contains Tat consensus motifs and a peptidase cleavage site at the N terminus. The PhoDAp enzyme was activated by Ca2+ and exhibited APase and phosphodiesterase (APDase) activities. Subcellular localization experiments revealed the secretion and processing of PhoDAp in a transformed cyanobacterium. Expression of the phoDAp gene in A. halophytica cells was upregulated not only by phosphorus (P) starvation but also under salt stress conditions. Our results suggest that A. halophytica cells possess a PhoD that participates in the assimilation of P under salinity stress.

Kageyama, Hakuto; Tripathi, Keshawanand; Rai, Ashwani K.; Cha-um, Suriyan; Waditee-Sirisattha, Rungaroon; Takabe, Teruhiro

2011-01-01

265

Nitrate\\/Nitrite Assimilation System of the Marine Picoplanktonic Cyanobacterium Synechococcus sp. Strain WH 8103: Effect of Nitrogen Source and Availability on Gene Expression  

Microsoft Academic Search

The genes encoding the structural components of the nitrate\\/nitrite assimilation system of the oceanic cyanobacterium Synechococcus sp. strain WH 8103 were cloned and characterized. The genes encoding nitrate reductase (narB) and nitrite reductase (nirA) are clustered on the chromosome but are organized in separate transcriptional units. Upstream of narB is a homologue of nrtP that encodes a nitrate\\/nitrite-bispecific permease rather

Clare Bird; Michael Wyman

2003-01-01

266

Two Functionally Distinct Forms of the Photosystem II Reaction-Center Protein D1 in the Cyanobacterium Synechococcus sp. PCC 7942  

Microsoft Academic Search

The cyanobacterium Synechococcus sp. PCC 7942 possesses a small psbA multigene family that codes for two distinct forms of the photosystem II reaction-center protein D1 (D1:1 and D1:2). We showed previously that the normally predominant D1 form (D1:1) was rapidly replaced with the alternative D1:2 when cells adapted to a photon irradiance of 50 mu mol\\\\cdotm-2\\\\cdots-1 are shifted to 500

Adrian K. Clarke; Vaughan M. Hurry; Petter Gustafsson; Gunnar Oquist

1993-01-01

267

The heat shock response in the cyanobacterium Synechocystis sp. Strain PCC 6803 and regulation of gene expression by HrcA and SigB  

Microsoft Academic Search

We report on the genome-wide response, based on DNA microarrays, of the cyanobacterium Synechocystis sp. PCC 6803 wild type and ?sigB to a 15 min heat shock. Approximately 9% of the genes in wild type and ?sigB were significantly regulated (P sigB had no dramatic effect on specific genes induced by heat shock, but did affect the level of transcription of the

Abhay K. Singh; Tina C. Summerfield; Hong Li; Louis A. Sherman

2006-01-01

268

Roles for heme–copper oxidases in extreme high-light and oxidative stress response in the cyanobacterium Synechococcus sp. PCC 7002  

Microsoft Academic Search

The ctaCIDIEI and ctaCIIDIIEII gene clusters that encode heme–copper cytochrome oxidases have been characterized in the marine cyanobacterium Synechococcus sp. PCC 7002 and the inactivation of ctaDI was shown to affect high-light adaptation. In this study, Synechococcus sp. PCC 7002 wild-type, ctaDI, ctaDII, and ctaDI–ctaDII double mutants were grown under extreme high-light and oxidative stress to further assess the roles

Christopher T. Nomura; Toshio Sakamoto; Donald A. Bryant

2006-01-01

269

Expression of photosynthesis genes in the cyanobacteriumSynechocystis sp. PCC 6803:psaA-psaB andpsbA transcripts accumulate in dark-grown cells  

Microsoft Academic Search

We have cloned and sequenced thepsaA andpsaB genes from the unicellular cyanobacteriumSynechocystis sp. PCC 6803. These genes are arranged in tandem, are co-transcribed, and are highly homologous to thepsaA andpsaB genes previously characterized. RNA was isolated from light-grown cells, from cells put in total darkness with and without glucose, and from cells grown under light-activated heterotrophic growth (LAHG) conditions. Quantitation

Lawrence B. Smart; Lee McIntosh

1991-01-01

270

Evolutionary Changes in Growth Rate and Toxin Production in the Cyanobacterium Microcystis aeruginosa Under a Scenario of Eutrophication and Temperature Increase  

Microsoft Academic Search

Toxic blooms of the cyanobacterium Microcystis aeruginosa affect humans and animals in inland water systems worldwide, and it has been hypothesized that the development of these blooms\\u000a will increase under the future scenario of global change, considering eutrophication and temperature increase as two important\\u000a consequences. The importance of genetic adaptation, chance and history on evolution of growth rate, and toxin

Mónica Rouco; Victoria López-Rodas; Antonio Flores-Moya; Eduardo Costas

271

Transcription and regulation of the hydrogenase(s) accessory genes, hypFCDEAB , in the cyanobacterium Lyngbya majuscula CCAP 1446\\/4  

Microsoft Academic Search

Lyngbya majuscula CCAP 1446\\/4 is a filamentous cyanobacterium possessing both an uptake and a bi-directional hydrogenase. The presence of a\\u000a single copy of the hyp operon in the cyanobacterial genomes suggests that these accessory genes might be responsible for the maturation of both\\u000a hydrogenases. We investigated the concomitant transcription of hypFCDEAB with the hydrogenases structural genes—hup and hox. RT-PCRs performed

Daniela Ferreira; Elsa Leitão; Johannes Sjöholm; Paulo Oliveira; Peter Lindblad; Pedro Moradas-Ferreira; Paula Tamagnini

2007-01-01

272

Calcium-dependent protease of the cyanobacterium Anabaena : molecular cloning and expression of the gene in Escherichia coli , sequencing and site-directed mutagenesis  

Microsoft Academic Search

It has been suggested that a calcium-dependent intracellular protease of the cyanobacterium, Anabaena sp., participates in the differentiation of heterocysts, cells that are specialized for fixation of N2. Clones of the structural gene (designated prcA) for this protease from Anabaena variabilis strain ATCC 29413 and Anabaena sp. strain PCC 7120 were identified via their expression in Escherichia coli. The prcA

Iris Maldener; Wolfgang Lockau; Yuping Cai; C. Peter Wolk

1991-01-01

273

Directed Mutagenesis of an Iron-Sulfur Protein of the Photosystem I Complex in the Filamentous Cyanobacterium Anabaena variabilis ATCC 29413  

Microsoft Academic Search

In oxygenic photosynthetic organisms the PSI-C polypeptide, encoded by the psaC gene, provides the ligands for two [4Fe-4S] centers, F_A and F_B, the terminal electron acceptors in the photosystem I (PSI) complex. An insertion mutation introduced in the psaC locus of the filamentous cyanobacterium Anabaena variabilis ATCC 29413 resulted in the creation of a mutant strain, T398-1, that lacks the

R. Mannar Mannan; John Whitmarsh; Philip Nyman; Himadri B. Pakrasi

1991-01-01

274

Accumulation of nodularin-like compounds from the cyanobacterium Nodularia spumigena and changes in acetylcholinesterase activity in the clam Macoma balthica during short-term laboratory exposure  

Microsoft Academic Search

In this laboratory study the effects of the cyanobacterium Nodularia spumigena (strain AV1) that produces hepatotoxic nodularin (NODLN), non-toxic Nodularia sphaerocarpa (strain UP16f) and purified NODLN on the infaunal clam Macoma balthica from the Baltic Sea were examined. N. sphaerocarpa (2.4 and 12.5 mg dw l?1), N. spumigena (4 and 20 mg dw l?1, intracellular NODLN content ca. 4 and

Kari K Lehtonen; Harri Kankaanpää; Sari Leiniö; Vesa O Sipiä; Stephan Pflugmacher; Eva Sandberg-Kilpi

2003-01-01

275

Genetic and biochemical evidence for distinct key functions of two highly divergent GAPDH genes in catabolic and anabolic carbon flow of the cyanobacterium Synechocystis sp. PCC 6803  

Microsoft Academic Search

Cyanobacterial genomes harbour two separate highly divergent glyceraldehyde-3-phosphate dehydrogenase (GAPDH) genes, gap1 and gap2, which are closely related at the sequence level to the nuclear genes encoding cytosolic and chloroplast GAPDH of higher plants, respectively. Genes gap1 and gap2 of the unicellular cyanobacterium Synechocystis sp. PCC 6803 were cloned and sequenced and subsequently inactivated by insertional mutagenesis to understand their

Olga Koksharova; Milada Schubert; Sergey Shestakov; Rüdiger Cerff

1998-01-01

276

Theophylline-Dependent Riboswitch as a Novel Genetic Tool for Strict Regulation of Protein Expression in Cyanobacterium Synechococcus elongatus PCC 7942.  

PubMed

The cyanobacterium Synechococcus elongatus PCC 7942 is a major model species for studies of photosynthesis. It is are also a potential cell factory for the production of renewable biofuels and valuable chemicals. We employed engineered riboswitches to control translational initiation of target genes in this cyanobacterium. A firefly luciferase reporter assay revealed that three theophylline riboswitches performed as expected in the cyanobacterium. Riboswitch-E* exhibited very low leaky expression of luciferase and superior and dose-dependent on/off regulation of protein expression by theophylline. The maximum magnitude of the induction vs. basal level was ?190-fold. Furthermore, the induction level was responsive to a wide range of theophylline concentrations in the medium, from 0 to 2 mM, facilitating the fine-tuning of luciferase expression. We adapted this riboswitch to another gene regulation system, in which expression of the circadian clock kaiC gene product is controlled by the theophylline concentration in the culture medium. The results demonstrated that the adequately adjusted expression level of KaiC restored complete circadian rhythm in the kaiC-deficient arrhythmic mutant. This theophylline-dependent riboswitch system has potential for various applications as a useful genetic tool in cyanobacteria. PMID:23969558

Nakahira, Yoichi; Ogawa, Atsushi; Asano, Hiroyuki; Oyama, Tokitaka; Tozawa, Yuzuru

2013-08-21

277

The 2.15 A crystal structure of a triple mutant plastocyanin from the cyanobacterium Synechocystis sp. PCC 6803.  

PubMed

The crystal structure of the triple mutant A42D/D47P/A63L plastocyanin from the cyanobacterium Synechocystis sp. PCC 6803 has been determined by Patterson search methods using the known structure of the poplar protein. Crystals of the triple mutant A42D/D47P/A63L, which are stable for days in its oxidized form, were grown from ammonium sulfate, with the cell constants a = b = 34.3 A and c = 111.8 A belonging to space group P3(2)21. The structure was refined using restrained crystallographic refinement to an R-factor of 16.7% for 4070 independent reflections between 8.0 and 2.15 A with intensities greater than 2 sigma (I), with root mean square deviations of 0.013 A and 1.63 degrees from ideal bond lengths and bond angles, respectively. The final model comprises 727 non-hydrogen protein atoms within 98 residues, 75 water molecules and a single copper ion. The overall tertiary fold of Synechocystis plastocyanin consists of a compact ellipsoidal beta-sandwich structure made up of two beta-sheets embracing a hydrophobic core. Each sheet contains parallel and antiparallel beta-strands. In addition to the beta-sheets, the structure contains an alpha-helix from Pro47 to Lys54 that follows beta-strand 4. The three-dimensional structure of Synechocystis plastocyanin is thus similar to those reported for the copper protein isolated from eukaryotic organisms and, in particular, from the cyanobacterium Anabaena variabilis, the only cyanobacterial plastocyanin structure available so far. The molecule holds an hydrophobic region surrounding His87, as do other plastocyanins, but the lack of negatively charged residues at the putative distant remote site surrounding Tyr83 could explain why the Synechocystis protein exhibits a collisional reaction mechanism for electron transfer to photosystem I (PSI), which involves no formation of the transient plastocyanin-PSI complex kinetically observed in green algae and higher plants. PMID:9466912

Romero, A; De la Cerda, B; Varela, P F; Navarro, J A; Hervás, M; De la Rosa, M A

1998-01-16

278

Engineering a cyanobacterium as the catalyst for the photosynthetic conversion of CO2 to 1,2-propanediol  

PubMed Central

Background The modern society primarily relies on petroleum and natural gas for the production of fuels and chemicals. One of the major commodity chemicals 1,2-propanediol (1,2-PDO), which has an annual production of more than 0.5 million tons in the United States, is currently produced by chemical processes from petroleum derived propylene oxide, which is energy intensive and not sustainable. In this study, we sought to achieve photosynthetic production of 1,2-PDO from CO2 using a genetically engineered cyanobacterium Synechococcus elongatus PCC 7942. Compared to the previously reported biological 1,2-PDO production processes which used sugar or glycerol as the substrates, direct chemical production from CO2 in photosynthetic organisms recycles the atmospheric CO2 and will not compete with food crops for arable land. Results In this study, we reported photosynthetic production of 1,2-PDO from CO2 using a genetically engineered cyanobacterium Synechococcus elongatus PCC 7942. Introduction of the genes encoding methylglyoxal synthase (mgsA), glycerol dehydrogenase (gldA), and aldehyde reductase (yqhD) resulted in the production of ~22mg/L 1,2-PDO from CO2. However, a comparable amount of the pathway intermediate acetol was also produced, especially during the stationary phase. The production of 1,2-PDO requires a robust input of reducing equivalents from cellular metabolism. To take advantage of cyanobacteria’s NADPH pool, the synthetic pathway of 1,2-PDO was engineered to be NADPH-dependent by exploiting the NADPH-specific secondary alcohol dehydrogenases which have not been reported for 1,2-PDO production previously. This optimization strategy resulted in the production of ~150mg/L 1,2-PDO and minimized the accumulation of the incomplete reduction product, acetol. Conclusion This work demonstrated that cyanobacteria can be engineered as a catalyst for the photosynthetic conversion of CO2 to 1,2-PDO. This work also characterized two NADPH-dependent sADHs for their catalytic capacity in 1,2-PDO formation, and suggested that they may be useful tools for renewable production of reduced chemicals in photosynthetic organisms.

2013-01-01

279

Insertional mutagenesis of a peptide synthetase gene that is responsible for hepatotoxin production in the cyanobacterium Microcystis aeruginosa PCC 7806.  

PubMed

Several bloom-forming cyanobacterial genera produce potent inhibitors of eukaryotic protein phosphatases called microcystins. Microcystins are hepatotoxic cyclic heptapeptides and are presumed to be synthesized non-ribosomally by peptide synthetases. We identified putative peptide synthetase genes in the microcystin-producing strain Microcystis aeruginosa PCC 7806. Non-hepatotoxic strains of M. aeruginosa lack these genes. Strain PCC 7806 was transformed to chloramphenicol resistance. The antibiotic resistance cassette insertionally inactivated a peptide synthetase gene of strain PCC 7806 as revealed by Southern hybridization and DNA amplification. This is the first report of genetic transformation and mutation, by homologous recombination, of a bloom-forming cyanobacterium. Chemical and enzymatic analyses, including high-performance liquid chromatography (HPLC), mass spectrometry, amino acid activation, and protein phosphatase inhibition, revealed the inability of derived mutant cells to produce any variant of microcystin while maintaining their ability to synthesize other small peptides. The disrupted gene therefore encodes a peptide synthetase (microcystin synthetase) that is specifically involved in the biosynthesis of microcystins. Our results confirm that microcystins are synthesized non-ribosomally and that a basic difference between toxic and non-toxic strains of M. aeruginosa is the presence of one or more genes coding for microcystin synthetases. PMID:9427407

Dittmann, E; Neilan, B A; Erhard, M; von Döhren, H; Börner, T

1997-11-01

280

Decoupling of ammonium regulation and ntcA transcription in the diazotrophic marine cyanobacterium Trichodesmium sp. IMS101.  

PubMed

Nitrogen (N) physiology in the marine cyanobacterium Trichodesmium IMS101 was studied along with transcript accumulation of the N-regulatory gene ntcA and of two of its target genes: napA (nitrate assimilation) and nifH (N(2) fixation). N(2) fixation was impaired in the presence of nitrite, nitrate and urea. Strain IMS101 was capable of growth on these combined N sources at <2 ?M but growth rates declined at elevated concentrations. Assimilation of nitrate and urea was impaired in the presence of ammonium. Whereas ecologically relevant N concentrations (2-20 ?M) suppressed growth and assimilation, much higher concentrations were required to affect transcript levels. Transcripts of nifH accumulated under nitrogen-fixing conditions; these transcript levels were maintained in the presence of nitrate (100 ?M) and ammonium (20 ?M). However, nifH transcript levels were below detection at ammonium concentrations >20 ?M. napA mRNA was found at low levels in both N(2)-fixing and ammonium-utilizing filaments, and it accumulated in filaments grown with nitrate. The positive effect of nitrate on napA transcription was abolished by ammonium additions of >200 ?M. This effect was restored upon addition of the glutamine synthetase inhibitor L-methionin-DL-sulfoximine. Surprisingly, ntcA transcript levels remained high in the presence of ammonium, even at elevated concentrations. These findings indicate that ammonium repression is decoupled from transcriptional activation of ntcA in Trichodesmium IMS101. PMID:21938021

Post, Anton F; Rihtman, Branko; Wang, Qingfeng

2011-09-22

281

ArsH from the cyanobacterium Synechocystis sp. PCC 6803 is an efficient NADPH-dependent quinone reductase.  

PubMed

The cyanobacterium Synechocystis sp. PCC 6803 possesses an arsenic resistance operon that encodes, among others, an ArsH protein. ArsH is a flavin mononucleotide (FMN)-containing protein of unknown function and a member of the family of NADPH-dependent FMN reductases. The nature of its final electron acceptor and the role of ArsH in the resistance to arsenic remained to be clarified. Here we have expressed and purified Synechocystis ArsH and conducted an intensive biochemical study. We present kinetic evidence supporting a quinone reductase activity for ArsH, with a preference for quinones with hydrophobic substituents. By using steady-state activity measurements, as well as stopped-flow and laser-flash photolysis kinetic analyses, it has been possible to establish the mechanism of the process and estimate the values of the kinetic constants. Although the enzyme is able to stabilize the anionic semiquinone form of the FMN, reduction of quinones involves the hydroquinone form of the flavin cofactor, and the enzymatic reaction occurs through a ping-pong-type mechanism. ArsH is able to catalyze one-electron reactions (oxygen and cytocrome c reduction), involving the FMN semiquinone form, but with lower efficiency. In addition, arsH mutants are sensitive to the oxidizing agent menadione, suggesting that ArsH plays a role in the response to oxidative stress caused by arsenite. PMID:22304305

Hervás, Manuel; López-Maury, Luis; León, Pilar; Sánchez-Riego, Ana M; Florencio, Francisco J; Navarro, José A

2012-02-03

282

Posttranscriptional regulation of glutamine synthetase in the filamentous Cyanobacterium Anabaena sp. PCC 7120: differential expression between vegetative cells and heterocysts.  

PubMed

Genes homologous to those implicated in glutamine synthetase (GS) regulation by protein-protein interaction in the cyanobacterium Synechocystis sp. strain PCC 6803 are conserved in several cyanobacterial sequenced genomes. We investigated this GS regulatory mechanism in Anabaena sp. strain PCC 7120. In this strain the system operates with only one GS inactivation factor (inactivation factor 7A [IF7A]), encoded by open reading frame (ORF) asl2329 (gifA). Following addition of ammonium, expression of gifA is derepressed, leading to the synthesis of IF7A, and consequently, GS is inactivated. Upon ammonium removal, the GS activity returns to the initial level and IF7A becomes undetectable. The global nitrogen control protein NtcA binds to the gifA promoter. Constitutive high expression levels of gifA were found in an Anabaena ntcA mutant (CSE2), indicating a repressor role for NtcA. In vitro studies demonstrate that Anabaena GS is not inactivated by Synechocystis IFs (IF7 and IF17), indicating the specificity of the system. We constructed an Anabaena strain expressing a second inactivating factor, containing the amino-terminal part of IF17 from Synechocystis fused to IF7A. GS inactivation in this strain is more effective than that in the wild type (WT) and resembles that observed in Synechocystis. Finally we found differential expression of the IF system between heterocysts and vegetative cells of Anabaena. PMID:20639319

Galmozzi, Carla V; Saelices, Lorena; Florencio, Francisco J; Muro-Pastor, M Isabel

2010-07-16

283

Posttranscriptional Regulation of Glutamine Synthetase in the Filamentous Cyanobacterium Anabaena sp. PCC 7120: Differential Expression between Vegetative Cells and Heterocysts ?  

PubMed Central

Genes homologous to those implicated in glutamine synthetase (GS) regulation by protein-protein interaction in the cyanobacterium Synechocystis sp. strain PCC 6803 are conserved in several cyanobacterial sequenced genomes. We investigated this GS regulatory mechanism in Anabaena sp. strain PCC 7120. In this strain the system operates with only one GS inactivation factor (inactivation factor 7A [IF7A]), encoded by open reading frame (ORF) asl2329 (gifA). Following addition of ammonium, expression of gifA is derepressed, leading to the synthesis of IF7A, and consequently, GS is inactivated. Upon ammonium removal, the GS activity returns to the initial level and IF7A becomes undetectable. The global nitrogen control protein NtcA binds to the gifA promoter. Constitutive high expression levels of gifA were found in an Anabaena ntcA mutant (CSE2), indicating a repressor role for NtcA. In vitro studies demonstrate that Anabaena GS is not inactivated by Synechocystis IFs (IF7 and IF17), indicating the specificity of the system. We constructed an Anabaena strain expressing a second inactivating factor, containing the amino-terminal part of IF17 from Synechocystis fused to IF7A. GS inactivation in this strain is more effective than that in the wild type (WT) and resembles that observed in Synechocystis. Finally we found differential expression of the IF system between heterocysts and vegetative cells of Anabaena.

Galmozzi, Carla V.; Saelices, Lorena; Florencio, Francisco J.; Muro-Pastor, M. Isabel

2010-01-01

284

Spatiotemporal changes in the genetic diversity in French bloom-forming populations of the toxic cyanobacterium, Microcystis aeruginosa.  

PubMed

Microcystis aeruginosa is a toxic cyanobacterium, which is able to bloom in a wide range of freshwater ecosystems. By sequencing the Internal Transcribed Spacer (ITS) of the ribosomal operon, we compared the genetic composition of several French bloom-forming M. aeruginosa populations from two reservoirs located on the Loire River, at two sampling points located between these reservoirs, and finally in two ponds closely linked to this river. No significant difference was found in the genetic diversity of the six Microcystis populations but we evidenced a strong genetic differentiation between most of these populations. Indeed, the Microcystis population in the Grangent reservoir was genetically differentiated from the other three populations sampled further downstream, implying that no massive transfer of population occurs from this reservoir to downstream segments. We also found genetic differentiation between the populations from the two ponds, and between these populations and those from the Loire River. On the other hand, the same dominant genotype was found in the populations sampled both in the river and in the Villerest reservoir, suggesting the selection of a distinct genotype adapted to river conditions and also an accumulation of this genotype in the downstream reservoir. Finally, by comparing our ITS sequences with those available in the GenBank, no biogeographical differentiation could be detected at a global scale, suggesting that most of the Microcystis genotypes seem to be ubiquitous. PMID:23765856

Sabart, Marion; Pobel, David; Latour, Delphine; Robin, Joel; Salençon, Marie-J; Humbert, Jean-F

2009-07-01

285

Electrostatic strain and concerted motions in the transient complex between plastocyanin and cytochrome f from the cyanobacterium Phormidium laminosum.  

PubMed

Many fleeting macromolecular interactions, like those being involved in electron transport, are essential in biology. However, little is known about the behaviour of the partners and their dynamics within their short-lived complex. To tackle such issue, we have performed molecular dynamics simulations on an electron transfer complex formed by plastocyanin and cytochrome f from the cyanobacterium Phormidium laminosum. Besides simulations of the isolated partners, two independent trajectories of the complex were calculated, starting from the two different conformations in the NMR ensemble. The first one leads to a more stable ensemble with a shorter distance between the metal sites of the two partners. The second experiences a significant drift of the complex conformation. Analyses of the distinct calculations show that the conformation of cytochrome f is strained upon binding of its partner, and relaxes upon its release. Interestingly, the principal component analysis of the trajectories indicates that plastocyanin displays a concerted motion with the small domain of cytochrome f that can be attributed to electrostatic interactions between the two proteins. PMID:19616485

Díaz-Moreno, Irene; Muñoz-López, Francisco J; Frutos-Beltrán, Estrella; De la Rosa, Miguel A; Díaz-Quintana, Antonio

2009-06-16

286

Viequeamide A, a Cytotoxic Member of the Kulolide Superfamily of Cyclic Depsipeptides from a Marine Button Cyanobacterium  

PubMed Central

The viequeamides, a family of 2,2-dimethyl-3-hydroxy-7-octynoic acid (Dhoya) containing cyclic depsipeptides, were isolated from a shallow subtidal collection of a ‘button’ cyanobacterium (Rivularia sp.) from near the island of Vieques, Puerto Rico. Planar structures of the two major compounds, viequeamide A (1) and viequeamide B (2), were elucidated by 2D-NMR spectroscopy and mass spectrometry, whereas absolute configurations were determined by traditional hydrolysis, derivative formation, and chromatography in comparison with standards. In addition, a series of related minor metabolites, viequeamide C–F (3–6), were characterized by high resolution mass spectroscopic (HRMS) fragmentation methods. Viequeamide A was found to be highly toxic to H460 human lung cancer cells (IC50 = 60 ± 10 nM), whereas the mixture of B–F was inactive. From a broader perspective, the viequeamides help to define a “superfamily” of related cyanobacterial natural products, the first of which to be discovered was ‘kulolide’. Within the kulolide superfamily, a wide variation in biological properties is observed, and the reported producing strains are also highly divergent, giving rise to several intriguing questions about structure-activity relationships and the evolutionary origins of this metabolite class.

Boudreau, Paul D.; Byrum, Tara; Liu, Wei-Ting; Dorrestein, Pieter C.; Gerwick, William H.

2012-01-01

287

Reduction of exogenous ketones depends upon NADPH generated photosynthetically in cells of the cyanobacterium Synechococcus PCC 7942.  

PubMed

Effective utilization of photosynthetic microorganisms as potential biocatalysts is favorable for the production of useful biomaterials and the reduction of atmospheric CO2. For example, biocatalytic transformations are used in the synthesis of optically active alcohols. We previously found that ketone reduction in cells of the cyanobacterium Synechococcus PCC 7942 is highly enantioselective and remarkably enhanced under light illumination. In this study, the mechanism of light-enhanced ketone reduction was investigated in detail using several inhibitors of photosynthetic electron transport and of enzymes of the Calvin cycle. It is demonstrated that light intensity and photosynthesis inhibitors significantly affect the ketone reduction activity in Synechococcus. This indicates that the reduction correlates well with photosynthetic activity. Moreover, ketone reduction in Synechococcus specifically depends upon NADPH and not NADH. These results also suggest that cyanobacteria have the potential to be utilized as biocatalytic systems for direct usage of light energy in various applications such as syntheses of useful compounds and remediation of environmental pollutants. PMID:21906270

Yamanaka, Rio; Nakamura, Kaoru; Murakami, Akio

2011-09-01

288

Oscillating behavior of carbohydrate granule formation and dinitrogen fixation in the cyanobacterium Cyanothece sp. strain ATCC 51142.  

PubMed Central

It has been shown that some aerobic, unicellular, diazotrophic cyanobacteria temporally separate photosynthetic O2 evolution and oxygen-sensitive N2 fixation. Cyanothece sp. ATCC strain 51142 is an aerobic, unicellular, diazotrophic cyanobacterium that fixes N2 during discrete periods of its cell cycle. When the bacteria are maintained under diurnal light-dark cycles, N2 fixation occurs in the dark. Similar cycling is observed in continuous light, implicating a circadian rhythm. Under N2-fixing conditions, large inclusion granules form between the thylakoid membranes. Maximum granulation, as observed by electron microscopy, occurs before the onset of N2 fixation, and the granules decrease in number during the period of N2 fixation. The granules can be purified from cell homogenates by differential centrifugation. Biochemical analyses of the granules indicate that these structures are primarily carbohydrate, with some protein. Further analyses of the carbohydrate have shown that it is a glucose polymer with some characteristics of glycogen. It is proposed that N2 fixation is driven by energy and reducing power stored in these inclusion granules. Cyanothece sp. strain ATCC 51142 represents an excellent experimental organism for the study of the protective mechanisms of nitrogenase, metabolic events in cyanobacteria under normal and stress conditions, the partitioning of resources between growth and storage, and biological rhythms. Images

Schneegurt, M A; Sherman, D M; Nayar, S; Sherman, L A

1994-01-01

289

Acute Exposure to Microcystin-Producing Cyanobacterium Microcystis aeruginosa Alters Adult Zebrafish (Danio rerio) Swimming Performance Parameters  

PubMed Central

Microcystins (MCs) are toxins produced by cyanobacteria (blue-green algae), primarily Microcystis aeruginosa, forming water blooms worldwide. When an organism is exposed to environmental perturbations, alterations in normal behavioral patterns occur. Behavioral repertoire represents the consequence of a diversity of physiological and biochemical alterations. In this study, we assessed behavioral patterns and whole-body cortisol levels of adult zebrafish (Danio rerio) exposed to cell culture of the microcystin-producing cyanobacterium M. aeruginosa (MC-LR, strain RST9501). MC-LR exposure (100??g/L) decreased by 63% the distance traveled and increased threefold the immobility time when compared to the control group. Interestingly, no significant alterations in the number of line crossings were found at the same MC-LR concentration and time of exposure. When animals were exposed to 50 and 100??g/L, MC-LR promoted a significant increase (around 93%) in the time spent in the bottom portion of the tank, suggesting an anxiogenic effect. The results also showed that none of the MC-LR concentrations tested promoted significant alterations in absolute turn angle, path efficiency, social behavior, or whole-body cortisol level. These findings indicate that behavior is susceptible to MC-LR exposure and provide evidence for a better understanding of the ecological consequences of toxic algal blooms.

Kist, Luiza Wilges; Piato, Angelo Luis; da Rosa, Joao Gabriel Santos; Koakoski, Gessi; Barcellos, Leonardo Jose Gil; Yunes, Joao Sarkis; Bonan, Carla Denise; Bogo, Mauricio Reis

2011-01-01

290

?-Tocopherol Is Essential for Acquired Chill-Light Tolerance in the Cyanobacterium Synechocystis sp. Strain PCC 6803? †  

PubMed Central

Unlike Escherichia coli, the cyanobacterium Synechocystis sp. strain PCC 6803 is insensitive to chill (5°C) in the dark but rapidly losses viability when exposed to chill in the light (100 ?mol photons m?2 s?1). Preconditioning at a low temperature (15°C) greatly enhances the chill-light tolerance of Synechocystis sp. strain PCC 6803. This phenomenon is called acquired chill-light tolerance (ACLT). Preconditioned wild-type cells maintained a substantially higher level of ?-tocopherol after exposure to chill-light stress. Mutants unable to synthesize ?-tocopherol, such as slr1736, slr1737, slr0089, and slr0090 mutants, almost completely lost ACLT. When exposed to chill without light, these mutants showed no or a slight difference from the wild type. When complemented, the slr0089 mutant regained its ACLT. Copper-regulated expression of slr0090 from PpetE controlled the level of ?-tocopherol and ACLT. We conclude that ?-tocopherol is essential for ACLT of Synechocystis sp. strain PCC 6803. The role of ?-tocopherol in ACLT may be based largely on a nonantioxidant activity that is not possessed by other tocopherols or pathway intermediates.

Yang, Yang; Yin, Chuntao; Li, Weizhi; Xu, Xudong

2008-01-01

291

The blooms of a cyanobacterium, Microcystis cf. aeruginosa in a severely polluted estuary, the Golden Horn, Turkey  

NASA Astrophysics Data System (ADS)

The distribution of toxic cyanobacterium Microcystis cf. aeruginosa in the severely polluted Golden Horn Estuary was studied from 1998 to 2000. Microcystis persisted at the upper estuary where the water circulation was poor and values ranged between 2.9 × 104 and 2.7 × 106 cells ml-1 throughout the study. Simultaneously measured physical (salinity, temperature, rainfall and secchi disc) and chemical parameters (nutrients and dissolved oxygen) were evaluated together with Microcystis data. Although the Microcystis blooms generally occur in summer due to the increase in temperature, the blooms were recorded in winter in the present study. The abundance of Microcystis depended on the variations in salinity and both blooms were recorded below S = 2. A moderate partial correlation between Microcystis abundance and salinity was detected in the presence of temperature, dissolved oxygen and precipitation data (r = -0.561, p = 0.002). The M. cf. aeruginosa abundance was low in the summer when the salinity was higher than winter. A remarkable increase in the eukaryotic phytoplankton abundance following the improvements in the water quality of the estuary occurred, whilst the Microcystis abundance remained below bloom level.

Ta?, Seyfettin; Oku?, Erdo?an; Aslan-Yilmaz, Asli

2006-07-01

292

Photoacclimation of cultured strains of the cyanobacterium Microcystis aeruginosa to high-light and low-light conditions.  

PubMed

The cyanobacterium Microcystis aeruginosa forms blooms that can consist of colonies. We have investigated how M. aeruginosa acclimatizes to changing light conditions such as can occur during blooms. Three different strains were exposed to two irradiance levels: lower (LL) and higher (HL) than the irradiance-onset saturation parameter. We measured the photosynthetic pigment concentrations, PSII photochemical efficiency, electron transport rate (ETR), irradiance-saturated ETR and ETR efficiency. The relationship between ETR and photosynthetic oxygen production and the excess in PSII capacity were also studied for one strain. Higher values of chlorophyll a and phycocyanin and lower values of total carotenoids were found under LL conditions in the three strains. The strains showed clear differences in the irradiance-saturated ETR and in ETR efficiency under both LL and HL treatments. No differences were found in the linear relationship between ETR and photosynthetic oxygen production under both irradiance treatments. LL-acclimated cells showed higher PSII excess capacity than HL ones, possibly because their higher pigment content could result in a higher light stress than HL cells when forming surface blooms. The fact that the genetically different strains show different photosynthetic physiologies suggests that the very dynamic light climate observed in lakes may allow their coexistence. PMID:23057858

Bañares-España, Elena; Kromkamp, Jacco C; López-Rodas, Victoria; Costas, Eduardo; Flores-Moya, Antonio

2012-11-21

293

An extracellular glycoprotein is implicated in cell-cell contacts in the toxic cyanobacterium Microcystis aeruginosa PCC 7806.  

PubMed

Microcystins are the most common cyanobacterial toxins found in freshwater lakes and reservoirs throughout the world. They are frequently produced by the unicellular, colonial cyanobacterium Microcystis aeruginosa; however, the role of the peptide for the producing organism is poorly understood. Differences in the cellular aggregation of M. aeruginosa PCC 7806 and a microcystin-deficient Delta mcyB mutant guided the discovery of a surface-exposed protein that shows increased abundance in PCC 7806 mutants deficient in microcystin production compared to the abundance of this protein in the wild type. Mass spectrometric and immunoblot analyses revealed that the protein, designated microcystin-related protein C (MrpC), is posttranslationally glycosylated, suggesting that it may be a potential target of a putative O-glycosyltransferase of the SPINDLY family encoded downstream of the mrpC gene. Immunofluorescence microscopy detected MrpC at the cell surface, suggesting an involvement of the protein in cellular interactions in strain PCC 7806. Further analyses of field samples of Microcystis demonstrated a strain-specific occurrence of MrpC possibly associated with distinct Microcystis colony types. Our results support the implication of microcystin in the colony specificity of and colony formation by Microcystis. PMID:18281396

Zilliges, Yvonne; Kehr, Jan-Christoph; Mikkat, Stefan; Bouchier, Christiane; de Marsac, Nicole Tandeau; Börner, Thomas; Dittmann, Elke

2008-02-15

294

Circadian expression of genes involved in the purine biosynthetic pathway of the cyanobacterium Synechococcus sp. strain PCC 7942.  

PubMed

Extensive circadian (daily) control over gene expression in the cyanobacterium Synechococcus sp. strain PCC 7942 is programmed into at least two differentially phased groups. The transcriptional activity of the smaller group of genes is maximal at about dawn and minimal at about dusk. We identified one of the genes belonging to this latter group as purF, which encodes the key regulatory enzyme in the de novo purine synthetic pathway, glutamine PRPP amidotransferase (also known as amidophosphoribosyltransferase). Its expression pattern as a function of circadian time was confirmed by both luminescence from a purF::luxAB reporter strain and the abundance of purF mRNA. By fusing sequences upstream of the purF coding region to promoterless luxAB genes, we identified a limited upstream region, which potentially regulates purF circadian expression patterns in vivo. We also identified the purL gene immediately upstream of purF. The purL gene encodes FGAM synthetase, the fourth enzyme in the purine nucleotide biosynthesis pathway. Although these genes are expressed as part of a larger operon in other bacteria, reporter gene fusions revealed that purF and purL are transcribed independently in Synechococcus and that they are expressed at different phases of the circadian cycle. This differential expression pattern may be related to the oxygen sensitivity of amidophosphoribosyltransferase. PMID:8809759

Liu, Y; Tsinoremas, N F; Golden, S S; Kondo, T; Johnson, C H

1996-06-01

295

Nitrate Transport and Not Photoinhibition Limits Growth of the Freshwater Cyanobacterium Synechococcus Species PCC 6301 at Low Temperature1  

PubMed Central

The effect of low temperature on cell growth, photosynthesis, photoinhibition, and nitrate assimilation was examined in the cyanobacterium Synechococcus sp. PCC 6301 to determine the factor that limits growth. Synechococcus sp. PCC 6301 grew exponentially between 20°C and 38°C, the growth rate decreased with decreasing temperature, and growth ceased at 15°C. The rate of photosynthetic oxygen evolution decreased more slowly with temperature than the growth rate, and more than 20% of the activity at 38°C remained at 15°C. Oxygen evolution was rapidly inactivated at high light intensity (3 mE m?2 s?1) at 15°C. Little or no loss of oxygen evolution was observed under the normal light intensity (250 ?E m?2 s?1) for growth at 15°C. The decrease in the rate of nitrate consumption by cells as a function of temperature was similar to the decrease in the growth rate. Cells could not actively take up nitrate or nitrite at 15°C, although nitrate reductase and nitrite reductase were still active. These data demonstrate that growth at low temperature is not limited by a decrease in the rate of photosynthetic electron transport or by photoinhibition, but that inactivation of the nitrate/nitrite transporter limits growth at low temperature.

Sakamoto, Toshio; Bryant, Donald A.

1999-01-01

296

Characterization of native and histidine-tagged deoxyxylulose 5-phosphate reductoisomerase from the cyanobacterium Synechocystis sp. PCC6803.  

PubMed

The dxr gene encoding the 1-deoxy-D-xylulose 5-phosphate reductoisomerase (DXR) from the cyanobacterium Synechocystis sp. PCC6803 was expressed in Escherichia coli to produce both the native and N-terminal histidine-tagged forms of DXR. The enzymes were purified from the cell extracts using either anion exchange chromatography or metal affinity chromatography and gel filtration. The purified recombinant native and histidine-tagged enzymes each displayed a single band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) gels, corresponding to the calculated subunit molecular weights of 42,500 and 46,700, respectively. By native PAGE, both enzymes were dimers under reducing conditions. The kinetic properties for the enzymes were characterized and only minor variations were observed, demonstrating that the N-terminal histidine tag does not greatly affect the activity of the enzyme. Both enzymes had similar properties to previously characterized reductoisomerases from other sources. The K(m)'s for the metal ions Mn(2+), Mg(2+), and Co(2+) were determined for native DXR for the first time, with the K(m) for Mg(2+) being approximately 200-fold higher than the K(m)'s for Mn(2+) and Co(2+). PMID:14580998

Yin, Xihou; Proteau, Philip J

2003-11-01

297

Cyclic depsipeptides, grassypeptolides D and E and Ibu-epidemethoxylyngbyastatin 3, from a Red Sea Leptolyngbya cyanobacterium.  

PubMed

Two new grassypeptolides and a lyngbyastatin analogue, together with the known dolastatin 12, have been isolated from field collections and laboratory cultures of the marine cyanobacterium Leptolyngbya sp. collected from the SS Thistlegorm shipwreck in the Red Sea. The overall stereostructures of grassypeptolides D (1) and E (2) and Ibu-epidemethoxylyngbyastatin 3 (3) were determined by a combination of 1D and 2D NMR experiments, MS analysis, Marfey's methodology, and HPLC-MS. Compounds 1 and 2 contain 2-methyl-3-aminobutyric acid and 2-aminobutyric acid, while biosynthetically distinct 3 contains 3-amino-2-methylhexanoic acid and the ?-keto amino acid 4-amino-2,2-dimethyl-3-oxopentanoic acid (Ibu). Grassypeptolides D (1) and E (2) showed significant cytotoxicity to HeLa (IC?? = 335 and 192 nM, respectively) and mouse neuro-2a blastoma cells (IC?? = 599 and 407 nM, respectively), in contrast to Ibu-epidemethoxylyngbyastatin 3 (neuro-2a cells, IC?? > 10 ?M) and dolastatin 12 (neuro-2a cells, IC?? > 1 ?M). PMID:21806012

Thornburg, Christopher C; Thimmaiah, Muralidhara; Shaala, Lamiaa A; Hau, Andrew M; Malmo, Jay M; Ishmael, Jane E; Youssef, Diaa T A; McPhail, Kerry L

2011-08-01

298

Active transport of glucosylglycerol is involved in salt adaptation of the cyanobacterium Synechocystis sp. strain PCC 6803.  

PubMed

An active-transport system for the osmoprotective compound glucosylglycerol (GG) was found in the cyanobacterium Synechocystis sp. strain PCC 6803. Uptake assays with 14C-labelled GG showed that the GG transport was enhanced in cells adapted to increasing concentrations of NaCl. Kinetic studies indicated a Michaelis-Menten relationship. The uptake of GG was energy dependent and occurred against a steep concentration gradient. It was inhibited by uncouplers as well as by a combination of darkness and KCN. The affinity of the transporter seems to be restricted to osmoprotective compounds of cyanobacteria; from a variety of compounds tested only sucrose and trehalose competed with GG for uptake. A salt-sensitive mutant of Synechocystis 6803 unable to synthesize GG could be complemented to salt resistance by exogenous GG. Accumulation of GG from the medium was essential for the restoration of photosynthesis and growth in mutant cells under high-salt conditions. In wild-type cells, the GG transporter probably serves to prevent GG leaking out of salt-stressed cells. PMID:8757737

Mikkat, S; Hagemann, M; Schoor, A

1996-07-01

299

Effects of phosphorus starvation versus limitation on the marine cyanobacterium Prochlorococcus?MED4 II: gene expression.  

PubMed

Phosphorus (P) availability drives niche differentiation in the most abundant phytoplankter in the oceans, the marine cyanobacterium Prochlorococcus. We compared the molecular response of Prochlorococcus strain MED4 to P starvation in batch culture to P-limited growth in chemostat culture. We also identified an outer membrane porin, PMM0709, which may allow transport of organic phosphorous compounds, rather than phosphate as previously suggested. The expression of three P uptake genes, pstS, the high-affinity phosphate-binding component of the phosphate transporter, phoA, an alkaline phosphatase, and porin PMM0709, were strongly upregulated (between 10- and 700-fold) under both P starvation and limitation. pstS exhibits high basal expression under P-replete conditions and is likely necessary for P uptake regardless of P availability. A P-stress regulatory gene, ptrA, was upregulated in response to both P starvation and limitation although a second regulatory gene, phoB, was not. Elevated expression levels (>?10-fold) of phoR, a P-sensing histidine kinase, were only observed under conditions of P limitation. We suggest Prochlorococcus in P-limited systems are physiologically distinct from cells subjected to abrupt P depletion. Detection of expression of both pstS and phoR in field populations will enable discernment of the present P status of Prochlorococcus in the oligotrophic oceans. PMID:23647921

Reistetter, Emily Nahas; Krumhardt, Kristen; Callnan, Kate; Roache-Johnson, Kathryn; Saunders, Jaclyn K; Moore, Lisa R; Rocap, Gabrielle

2013-05-06

300

Effect of Nitrogen on Cellular Production and Release of the Neurotoxin Anatoxin-A in a Nitrogen-Fixing Cyanobacterium  

PubMed Central

Anatoxin-a (ANTX) is a neurotoxin produced by several freshwater cyanobacteria and implicated in lethal poisonings of domesticated animals and wildlife. The factors leading to its production in nature and in culture are not well understood. Resource availability may influence its cellular production as suggested by the carbon-nutrient hypothesis, which links the amount of secondary metabolites produced by plants or microbes to the relative abundance of nutrients. We tested the effects of nitrogen supply (as 1, 5, and 100% N of standard cyanobacterial medium corresponding to 15, 75, and 1500?mg?L?1 of NaNO3 respectively) on ANTX production and release in a toxic strain of the planktonic cyanobacterium Aphanizomenon issatschenkoi (Nostocales). We hypothesized that nitrogen deficiency might constrain the production of ANTX. However, the total concentration and more significantly the cellular content of anatoxin-a peaked (max. 146??g/L and 1683??g?g?1 dry weight) at intermediate levels of nitrogen supply when N-deficiency was evident based on phycocyanin to chlorophyll a and carbon to nitrogen ratios. The results suggest that the cellular production of anatoxin-a may be stimulated by moderate nitrogen stress. Maximal cellular contents of other cyanotoxins have recently been reported under severe stress conditions in another Nostocales species.

Gagnon, Alexis; Pick, Frances R.

2012-01-01

301

Purification and Biochemical Analysis of the Cytoplasmic Membrane from the Desiccation-Tolerant Cyanobacterium Nostoc commune UTEX 584  

PubMed Central

The cytoplasmic membrane of the heterocystous cyanobacterium Nostoc commune UTEX 584 was isolated free of thylakoids and phycobiliprotein-membrane complexes by flotation centrifugation. Purified membranes had a buoyant density of 1.07 g cm?3 and were bright orange. Twelve major proteins were detected in the membrane, and of these, the most abundant had molecular masses of 83, 71, 68, 51, and 46 kilodaltons. The ester-linked fatty acids of the methanol fraction contained 16:0, 18:0, 18:1?9c, 20:0, and 20:3?3 with no traces of hydroxy fatty acids. Compound 20:3?3 represented 56.8% of the total fatty acid methyl esters, a feature which distinguishes the cell membrane of N. commune UTEX 584 from those of all other cyanobacteria which have been characterized to date. Fatty acid 18:3 was not detected. Carotenoids were analyzed by highperformance liquid chromatography. The cytoplasmic membrane contained ?-carotene and echinenone as the dominant carotenoids and lacked chlorophyll a and pheophytin a. Whole cells contained ?-carotene and echinenone, and lesser amounts of zeaxanthin and (3R)-cryptoxanthin. Images

Olie, Jaap J.; Potts, Malcolm

1986-01-01

302

Sulfate-Driven Elemental Sparing Is Regulated at the Transcriptional and Posttranscriptional Levels in a Filamentous Cyanobacterium? †  

PubMed Central

Sulfur is an essential nutrient that can exist at growth-limiting concentrations in freshwater environments. The freshwater cyanobacterium Fremyella diplosiphon (also known as Tolypothrix sp. PCC 7601) is capable of remodeling the composition of its light-harvesting antennae, or phycobilisomes, in response to changes in the sulfur levels in its environment. Depletion of sulfur causes these cells to cease the accumulation of two forms of a major phycobilisome protein called phycocyanin and initiate the production of a third form of phycocyanin, which possesses a minimal number of sulfur-containing amino acids. Since phycobilisomes make up approximately 50% of the total protein in these cells, this elemental sparing response has the potential to significantly influence the fitness of this species under low-sulfur conditions. This response is specific for sulfate and occurs over the physiological range of sulfate concentrations likely to be encountered by this organism in its natural environment. F. diplosiphon has two separate sulfur deprivation responses, with low sulfate levels activating the phycobilisome remodeling response and low sulfur levels activating the chlorosis or bleaching response. The phycobilisome remodeling response results from changes in RNA abundance that are regulated at both the transcriptional and posttranscriptional levels. The potential of this response, and the more general bleaching response of cyanobacteria, to provide sulfur-containing amino acids during periods of sulfur deprivation is examined.

Gutu, Andrian; Alvey, Richard M.; Bashour, Sami; Zingg, Daniel; Kehoe, David M.

2011-01-01

303

Contribution of a sodium ion gradient to energy conservation during fermentation in the cyanobacterium Arthrospira (Spirulina) maxima CS-328.  

PubMed

Sodium gradients in cyanobacteria play an important role in energy storage under photoautotrophic conditions but have not been well studied during autofermentative metabolism under the dark, anoxic conditions widely used to produce precursors to fuels. Here we demonstrate significant stress-induced acceleration of autofermentation of photosynthetically generated carbohydrates (glycogen and sugars) to form excreted organic acids, alcohols, and hydrogen gas by the halophilic, alkalophilic cyanobacterium Arthrospira (Spirulina) maxima CS-328. When suspended in potassium versus sodium phosphate buffers at the start of autofermentation to remove the sodium ion gradient, photoautotrophically grown cells catabolized more intracellular carbohydrates while producing 67% higher yields of hydrogen, acetate, and ethanol (and significant amounts of lactate) as fermentative products. A comparable acceleration of fermentative carbohydrate catabolism occurred upon dissipating the sodium gradient via addition of the sodium-channel blocker quinidine or the sodium-ionophore monensin but not upon dissipating the proton gradient with the proton-ionophore dinitrophenol (DNP). The data demonstrate that intracellular energy is stored via a sodium gradient during autofermentative metabolism and that, when this gradient is blocked, the blockage is compensated by increased energy conversion via carbohydrate catabolism. PMID:21890670

Carrieri, Damian; Ananyev, Gennady; Lenz, Oliver; Bryant, Donald A; Dismukes, G Charles

2011-09-02

304

Effect of tryptophan on 2,4-dichlorophenoxyacetic acid toxicity in the nitrogen-fixing-cyanobacterium Nostoc linckia.  

PubMed

The combined effect of a hormone weed killer 2,4-dichlorophenoxyacetic acid (2,4-D) and an amino acid (tryptophan) has been studied on growth and heterocyst differentiation in the cyanobacterium Nostoc linckia. 2.4-D at 100 micrograms/ml stimulated growth and heterocyst frequency in combined nitrogen-free medium while its higher concentrations inhibited both. Tryptophan under similar conditions promoted much growth yield with 3-4 fold enhanced heterocyst frequency than the control. Such heterocysts were immature and showed germination under in situ condition. The concentrations of 2,4-D (100 micrograms/ml) and tryptophan (50 micrograms/ml), stimulatory to growth and heterocyst formation, caused additive effect while herbicide inhibition of nitrogen-fixing growth at higher doses was partially relieved by tryptophan but tryptophan-induced heterocyst frequency was completely suppressed under this condition. The possible role of interaction of these two chemicals on growth and heterocyst formation has been discussed. PMID:3083089

Mishra, A K; Tiwari, D N

1986-01-01

305

Mutational analysis of the structure and biogenesis of the photosystem I reaction center in the cyanobacterium Synechocystis sp. PCC 6803  

SciTech Connect

The authors have utilized the unicellular cyanobacterium Synechocystis sp. PCC 6803 to incorporate site-directed amino acid substitutions into the photosystem I (PSI) reaction-center protein PsaB. A cysteine residue (position 565 of PSaB) proposed to serve as a ligand to the [4Fe-4S] center F[sub x] was changed to serine, histidine, and aspartate. These three mutants - C565S, C565H, and C565D - all exhibited greatly reduced accumulation of PSI reaction-center proteins and failed to grow autotrophically, indicating that this cysteine most likely does coordinate F[sub x], which is crucial for PSI biogenesis. Interestingly, the strain C565S accumulated significantly more PSI than the other two cysteine mutants and displayed photoreduction of the [4Fe-4S] terminal electron acceptors F[sub A] and F[sub B]. Mutations were also introduced into a leucine zipper motif of PsaB, proposed to participate in reaction-center dimerization. The mutants L522V, L536M, and L522V/L536M all exhibited wild-type characteristics and grew autotrophically, whereas the L522P mutation prevented PSI accumulation. These data do not provide support for a major structural role of the leucine zipper in reaction-center dimerization or in assembly of F[sub x]. However, the amino acid substitutions incorporated were conservative and might not have perturbed the leucine zipper. 31 refs., 4 figs., 1 tab.

Smart, L.B.; McIntosh, L. (Michigan State Univ., East Lansing (United States)); Warren, P.V.; Golbeck, J.H. (Univ. of Nebraska, Lincoln (United States))

1993-02-01

306

Transcriptional analysis of the isiAB operon in salt-stressed cells of the cyanobacterium Synechocystis sp. PCC 6803.  

PubMed

Expression of the isiA and isiB genes was analysed in the cyanobacterium Synechocystis sp. PCC 6803 grown in high salt or in iron-deficient medium. The detection of a 2.3-knt transcript in Northern blot experiments indicated cotranscription of isiAB in an operon, which was confirmed by reverse transcriptase PCR. The abundance of a monocistronic 1.25-knt isiA-specific mRNA was about 10-fold higher than the dicistronic message. The isiAB-specific transcripts were most abundant in cells adapted to 342 mM NaCl and under iron deficiency. The promoter of the operon was mapped to 211 bp upstream of the translational start. A putative Fur binding site was detected immediately upstream of the GTG start codon. A preliminary transcript of about 0.2 knt was detected in cells grown in conditions in which the isiAB operon was not transcribed. This indicates that a repressor binds to the identified Fur binding site and thus inhibits isiAB transcription under low salt and iron replete conditions. PMID:9868777

Vinnemeier, J; Kunert, A; Hagemann, M

1998-12-15

307

Iron limitation in the marine cyanobacterium Trichodesmium reveals new insights into regulation of photosynthesis and nitrogen fixation.  

PubMed

* As iron (Fe) deficiency is a main limiting factor of ocean productivity, its effects were investigated on interactions between photosynthesis and nitrogen fixation in the marine nonheterocystous diazotrophic cyanobacterium Trichodesmium IMS101. * Biophysical methods such as fluorescence kinetic microscopy, fast repetition rate (FRR) fluorimetry, and in vivo and in vitro spectroscopy of pigment composition were used, and nitrogenase activity and the abundance of key proteins were measured. * Fe limitation caused a fast down-regulation of nitrogenase activity and protein levels. By contrast, the abundance of Fe-requiring photosystem I (PSI) components remained constant. Total levels of phycobiliproteins remained unchanged according to single-cell in vivo spectra. However, the regular 16-kDa phycoerythrin band decreased and finally disappeared 16-20 d after initiation of Fe limitation, concomitant with the accumulation of a 20-kDa protein cross-reacting with the phycoerythrin antibody. Concurrently, nitrogenase expression and activity increased. Fe limitation dampened the daily cycle of photosystem II (PSII) activity characteristic of diazotrophic Trichodesmium cells. Further, it increased the number and prolonged the time period of occurrence of cells with elevated basic fluorescence (F(0)). Additionally, it increased the effective cross-section of PSII, probably as a result of enhanced coupling of phycobilisomes to PSII, and led to up-regulation of the Fe stress protein IsiA. * Trichodesmium survives short-term Fe limitation by selectively down-regulating nitrogen fixation while maintaining but re-arranging the photosynthetic apparatus. PMID:18513224

Küpper, Hendrik; Setlík, Ivan; Seibert, Sven; Prásil, Ondrej; Setlikova, Eva; Strittmatter, Martina; Levitan, Orly; Lohscheider, Jens; Adamska, Iwona; Berman-Frank, Ilana

2008-05-30

308

Localization and function of the IdiA homologue Slr1295 in the cyanobacterium Synechocystis sp. strain PCC 6803.  

PubMed

Slr1295 (and Slr0513) in the cyanobacterium Synechocystis sp. PCC 6803 has amino acid similarity to the bacterial FbpA protein family and also to IdiA of Synechococcus PCC 6301/PCC 7942. To determine whether Slr1295 is the periplasm-located component of an iron transporter, or has a function in protecting photosystem (PS) II, subcellular localization and Deltaslr1295 mutant characterization studies were performed. Localization of Slr1295 provided evidence that it has an intracellular function, since virtually no Slr1295 was detected in the soluble protein fraction of the periplasm or in the cytoplasmic membrane. Characterization of a Deltaslr1295 Synechocystis PCC 6803 mutant indicated that PS II is more susceptible to inactivation in the mutant than in the wild-type (WT). Under mild iron limitation, modification of PS I to the PS I-IsiA complex is more advanced in the Deltaslr1295 mutant, indicating that iron deficiency leads more rapidly to changes in the photosynthetic apparatus in the mutant than in the WT. Biochemical fractionation procedures provide evidence that Slr1295 co-purifies with PS II. These results suggest a function of Slr1295 that is comparable to the function of IdiA in Synechococcus PCC 6301/PCC 7942 being a protein that protects PS II under iron limitation in an as yet unknown way. PMID:12368463

Tölle, Jörg; Michel, Klaus-Peter; Kruip, Jochen; Kahmann, Uwe; Preisfeld, Angelika; Pistorius, Elfriede K

2002-10-01

309

Contribution of a Sodium Ion Gradient to Energy Conservation during Fermentation in the Cyanobacterium Arthrospira (Spirulina) maxima CS-328 ? †  

PubMed Central

Sodium gradients in cyanobacteria play an important role in energy storage under photoautotrophic conditions but have not been well studied during autofermentative metabolism under the dark, anoxic conditions widely used to produce precursors to fuels. Here we demonstrate significant stress-induced acceleration of autofermentation of photosynthetically generated carbohydrates (glycogen and sugars) to form excreted organic acids, alcohols, and hydrogen gas by the halophilic, alkalophilic cyanobacterium Arthrospira (Spirulina) maxima CS-328. When suspended in potassium versus sodium phosphate buffers at the start of autofermentation to remove the sodium ion gradient, photoautotrophically grown cells catabolized more intracellular carbohydrates while producing 67% higher yields of hydrogen, acetate, and ethanol (and significant amounts of lactate) as fermentative products. A comparable acceleration of fermentative carbohydrate catabolism occurred upon dissipating the sodium gradient via addition of the sodium-channel blocker quinidine or the sodium-ionophore monensin but not upon dissipating the proton gradient with the proton-ionophore dinitrophenol (DNP). The data demonstrate that intracellular energy is stored via a sodium gradient during autofermentative metabolism and that, when this gradient is blocked, the blockage is compensated by increased energy conversion via carbohydrate catabolism.

Carrieri, Damian; Ananyev, Gennady; Lenz, Oliver; Bryant, Donald A.; Dismukes, G. Charles

2011-01-01

310

Identification of a benthic microcystin-producing filamentous cyanobacterium (Oscillatoriales) associated with a dog poisoning in New Zealand.  

PubMed

In November 2008 a dog died soon after ingesting benthic "algal" mat material from the Waitaki River, New Zealand. Based on a morphological examination of environmental material, the causative organism was putatively identified as the filamentous cyanobacterium Phormidium sp. Two strains (VUW25 and CYN61) were isolated and cultured to enable further taxonomic and cyanotoxin characterisation. Phylogenetic analyses based on a region of the 16S rRNA gene sequence, intergenic spacer (ITS) region and the mcyE gene demonstrated that the species was likely to be a new Planktothrix species that is either benthic or has a biphasic life cycle. Using liquid chromatography-mass spectrometry (LC-MS), microcystin-LR, [D-Asp(3), Dha(7)] microcystin-LR, [D-Asp(3)] microcystin-LR, and minor proportions of [D-Asp(3), ADMAdda(5)] microcystin-LhR were identified. This is the first report of [D-Asp(3)] microcystin-LR, [D-Asp(3), Dha(7)] microcystin-LR and an ADMAadda variant in New Zealand. No cylindrospermopsins, saxitoxins or anatoxins were detected. Dog deaths caused by the consumption of cyanobacterial mats containing anatoxins have previously been reported in New Zealand. To our knowledge, however, this is the first instance of a benthic microcystin-producing species causing an animal death in New Zealand. PMID:20043936

Wood, Susanna A; Heath, Mark W; Holland, Patrick T; Munday, Rex; McGregor, Glenn B; Ryan, Ken G

2010-01-04

311

Pulsed nitrogen supply induces dynamic changes in the amino acid composition and microcystin production of the harmful cyanobacterium Planktothrix agardhii.  

PubMed

Planktothrix agardhii is a widespread harmful cyanobacterium of eutrophic waters, and can produce the hepatotoxins [Asp(3)]microcystin-LR and [Asp(3)]microcystin-RR. These two microcystin variants differ in their first variable amino acid position, which is occupied by either leucine (L) or arginine (R). Although microcystins are extensively investigated, little is known about the mechanisms that determine the production of different microcystin variants. We hypothesize that enhanced nitrogen availability will increase the intracellular content of the nitrogen-rich amino acid arginine, and thereby promote the production of the variant [Asp(3)]microcystin-RR. To test this hypothesis, we transferred P. agardhii strain 126/3 from nitrogen-replete to nitrogen-deficient conditions, and after 2 weeks of growth under nitrogen deficiency, we added a nitrogen pulse. We found a rapid increase in the cellular nitrogen to carbon ratio and the amino acids aspartic acid and arginine, indicative of cyanophycin synthesis. This was followed by a more gradual increase of the total amino acid content connected to balanced growth. As expected, the [Asp(3)]microcystin-RR variant increased strongly after the nitrogen pulse, while the [Asp(3)]microcystin-LR increased to a much lesser extent. We conclude that sudden nitrogen enrichment affects the amino acid composition of harmful cyanobacteria, which, in turn, affects the production and composition of their microcystins. PMID:20735475

Van de Waal, Dedmer B; Ferreruela, Gonzalo; Tonk, Linda; Van Donk, Ellen; Huisman, Jef; Visser, Petra M; Matthijs, Hans C P

2010-08-23

312

Adaptation to High-Intensity, Low-Wavelength Light among Surface Blooms of the Cyanobacterium Microcystis aeruginosa  

PubMed Central

Natural populations of the nuisance bloom cyanobacterium Microcystis aeruginosa obtained from the eutrophic Neuse River, N.C., revealed optimal chlorophyll a-normalized photosynthetic rates and resistance to photoinhibition at surface photosynthetically active radiation (PAR) intensities. At saturating PAR levels these populations exhibited higher photosynthetic rates in quartz than in Pyrex vessels. Eucaryotic algal populations obtained from the same river failed to counteract photoinhibition. At saturating PAR levels, such populations generally yielded lower photosynthetic rates in quartz containers than they did in Pyrex containers. Cultivation of natural Microcystis populations under laboratory conditions led to physiologically distinct populations which had photoinhibitory characteristics similar to those of other cultured cyanobacterial and eucaryotic algae. Our findings indicate that (i) photosynthetic production among natural surface populations is best characterized and quantified in quartz rather than Pyrex incubation vessels; (ii) extrapolation of natural photoinhibitory trends from laboratory populations is highly subjective to culture and PAR histories and may yield contradictory results; and (iii) buoyant surface-dwelling populations, rather than exhibiting senescence, are poised at optimizing PAR utilization, thereby maintaining numerical dominance in eutrophic waters when physico-chemical conditions favor bloom formation.

Paerl, Hans W.; Bland, Patricia T.; Bowles, N. Dean; Haibach, Mark E.

1985-01-01

313

Two functionally distinct forms of the photosystem II reaction-center protein D1 in the cyanobacterium Synechococcus sp. PCC 7942  

Microsoft Academic Search

The cyanobacterium Synechococcus sp. PCC 7942 possesses a small psbA multigene family that codes for two distinct forms of the photosystem II reaction-center protein D1 (D1:1 and D1:2). The authors showed previously that the normally predominant D1 form (D1:1) was rapidly replaced with the alternative D1:2 when cells adapted to a photon irradiance of 50 [mu]mol[center dot]m[sup [minus]2][center dot]s[sup [minus]1

A. K. Clarke; V. M. Hurry; P. Gustafsson; G. Oquist

1993-01-01

314

Oxidative stress response and fatty acid changes associated with bioaccumulation of chromium [Cr(VI)] by a fresh water cyanobacterium Chroococcus sp.  

PubMed

Cr(VI) at 2.5, 5, 7.5 and 10 mg/l was removed over 1-5 days by a freshwater cyanobacterium, Chroococcus sp. 2.5 mg Cr(VI)/l gave the optimum rate. With 5 mg Cr(VI)/l, activities of superoxide dismutase and catalase were increased. Amounts of palmitic (16:0), stearic (18:0) and oleic acid (18:1) in the cell also increased after exposure to Cr(VI). PMID:22002251

Kumar, Muthukannan Satheesh; Praveenkumar, Ramasamy; Ilavarasi, Asokraja; Rajeshwari, Kamaraj; Thajuddin, Nooruddin

2011-10-16

315

Organization of a large gene cluster encoding ribosomal proteins in the cyanobacterium S ynechococcus sp. strain PCC 6301: comparison of gene clusters among cyanobacteria, eubacteria and chloroplast genomes  

Microsoft Academic Search

The structure of a large gene cluster containing 22 ribosomal protein (r-protein) genes of the cyanobacterium Synechococcus sp. strain PCC6301 is presented. Based on DNA and protein sequence analyses, genes encoding r-proteins L3, L4, L23, L2, S19, L22, S3, L16, L29, S17, L14, L24, L5, S8, L6, L18, S5, L15, L36, S13, S11, L17, SecY, adenylate kinase (AK) and the

Mamoru Sugita; Hiroyuki Sugishita; Tsuneo Fujishiro; Mari Tsuboi; Chieko Sugita; Toshiya Endo; Masahiro Sugiura

1997-01-01

316

Wastewater Utilization for Poly-?-Hydroxybutyrate Production by the Cyanobacterium Aulosira fertilissima in a Recirculatory Aquaculture System?  

PubMed Central

Intensive aquaculture releases large quantities of nutrients into aquatic bodies, which can lead to eutrophication. The objective of this study was the development of a biological recirculatory wastewater treatment system with a diazotrophic cyanobacterium, Aulosira fertilissima, and simultaneous production of valuable product in the form of poly-?-hydroxybutyrate (PHB). To investigate this possible synergy, batch scale tests were conducted under a recirculatory aquaculture system in fiber-reinforced plastic tanks enhanced by several manageable parameters (e.g., sedimentation, inoculum size, depth, turbulence, and light intensity), an adequate combination of which showed better productivity. The dissolved-oxygen level increased in the range of 3.2 to 6.9 mg liter?1 during the culture period. Nutrients such as ammonia, nitrite, and phosphate decreased to as low as zero within 15 days of incubation, indicating the system's bioremediation capability while yielding valuable cyanobacterial biomass for PHB production. Maximum PHB accumulation in A. fertilissima was found in sedimented fish pond discharge at 20-cm culture depth with stirring and an initial inoculum size of 80 mg dry cell weight (dcw) liter?1. Under optimized conditions, the PHB yield was boosted to 92, 89, and 80 g m?2, respectively for the summer, rainy, and winter seasons. Extrapolation of the result showed that a hectare of A. fertilissima cultivation in fish pond discharge would give an annual harvest of ?17 tons dry biomass, consisting of 14 tons of PHB with material properties comparable to those of the bacterial polymer, with simultaneous treatment of 32,640 m3 water discharge.

Samantaray, Shilalipi; Nayak, Jitendra Kumar; Mallick, Nirupama

2011-01-01

317

Differential Transcriptional Analysis of the Cyanobacterium Cyanothece sp. Strain ATCC 51142 during Light-Dark and Continuous-Light Growth? †  

PubMed Central

We analyzed the metabolic rhythms and differential gene expression in the unicellular, diazotrophic cyanobacterium Cyanothece sp. strain ATCC 51142 under N2-fixing conditions after a shift from normal 12-h light-12-h dark cycles to continuous light. We found that the mRNA levels of ?10% of the genes in the genome demonstrated circadian behavior during growth in free-running (continuous light) conditions. The genes for N2 fixation displayed a strong circadian behavior, whereas photosynthesis and respiration genes were not as tightly regulated. One of our main objectives was to determine the strategies used by these cells to perform N2 fixation under normal day-night conditions, as well as under the greater stress caused by continuous light. We determined that N2 fixation cycled in continuous light but with a lower N2 fixation activity. Glycogen degradation, respiration, and photosynthesis were also lower; nonetheless, O2 evolution was about 50% of the normal peak. We also demonstrated that nifH (encoding the nitrogenase Fe protein), nifB, and nifX were strongly induced in continuous light; this is consistent with the role of these proteins during the assembly of the enzyme complex and suggested that the decreased N2 fixation activity was due to protein-level regulation or inhibition. Many soluble electron carriers (e.g., ferredoxins), as well as redox carriers (e.g., thioredoxin and glutathione), were strongly induced during N2 fixation in continuous light. We suggest that these carriers are required to enhance cyclic electron transport and phosphorylation for energy production and to maintain appropriate redox levels in the presence of elevated O2, respectively.

Toepel, Jorg; Welsh, Eric; Summerfield, Tina C.; Pakrasi, Himadri B.; Sherman, Louis A.

2008-01-01

318

THE TOXIC CYANOBACTERIUM NOSTOC SP. STRAIN 152 PRODUCES HIGHEST AMOUNTS OF MICROCYSTIN AND NOSTOPHYCIN UNDER STRESS CONDITIONS  

PubMed Central

The understanding of how environmental factors regulate toxic secondary metabolite production in cyanobacteria is important to guarantee water quality. Very little is known on the regulation of toxic secondary metabolite production in benthic cyanobacteria. In this study the physiological regulation of the production of the toxic heptapeptide microcystin (MC) and the non-toxic related peptide nostophycin (NP) in the benthic cyanobacterium Nostoc sp. strain 152 was studied under contrasting environmental conditions. I used a 2k levels factorial design, where k is the number of four factors that have been tested: Reduction in temperature (20 vs. 12°C), irradiance (50 vs. 1 ?mol · m?2 · s?1), P-PO4 (144 vs. 0.14 ?M P-PO4), N-NO3 (5.88 mM vs. N-NO3 free). While the growth rate was reduced more than hundred fold under most severe conditions of temperature, irradiance, and phosphate reduction the production of MC and NP never ceased. The MC and NP contents per cell varied at maximum 5- and 10.6-fold each, however the physiological variation did not outweigh the highly significant linear relationship between the daily cell division rate and the MC and NP net production rates. Surprisingly the MC and NP contents per cell showed a maximum under P-PO4 reduced and irradiance reduced conditions. Both intra- and extracellular MC and NP concentrations were negatively related to P-PO4 and irradiance. It is concluded that the proximate factor behind maximal cellular MC and NP contents is physiological stress.

Kurmayer, Rainer

2012-01-01

319

THE TOXIC CYANOBACTERIUM NOSTOC SP. STRAIN 152 PRODUCES HIGHEST AMOUNTS OF MICROCYSTIN AND NOSTOPHYCIN UNDER STRESS CONDITIONS.  

PubMed

The understanding of how environmental factors regulate toxic secondary metabolite production in cyanobacteria is important to guarantee water quality. Very little is known on the regulation of toxic secondary metabolite production in benthic cyanobacteria. In this study the physiological regulation of the production of the toxic heptapeptide microcystin (MC) and the non-toxic related peptide nostophycin (NP) in the benthic cyanobacterium Nostoc sp. strain 152 was studied under contrasting environmental conditions. I used a 2(k) levels factorial design, where k is the number of four factors that have been tested: Reduction in temperature (20 vs. 12°C), irradiance (50 vs. 1 ?mol · m(-2) · s(-1)), P-PO(4) (144 vs. 0.14 ?M P-PO(4)), N-NO(3) (5.88 mM vs. N-NO(3) free). While the growth rate was reduced more than hundred fold under most severe conditions of temperature, irradiance, and phosphate reduction the production of MC and NP never ceased. The MC and NP contents per cell varied at maximum 5- and 10.6-fold each, however the physiological variation did not outweigh the highly significant linear relationship between the daily cell division rate and the MC and NP net production rates. Surprisingly the MC and NP contents per cell showed a maximum under P-PO(4) reduced and irradiance reduced conditions. Both intra- and extracellular MC and NP concentrations were negatively related to P-PO(4) and irradiance. It is concluded that the proximate factor behind maximal cellular MC and NP contents is physiological stress. PMID:22723716

Kurmayer, Rainer

2011-02-01

320

Combined effects of CO2 and light on the N2-fixing cyanobacterium Trichodesmium IMS101: a mechanistic view.  

PubMed

The marine diazotrophic cyanobacterium Trichodesmium responds to elevated atmospheric CO(2) partial pressure (pCO(2)) with higher N(2) fixation and growth rates. To unveil the underlying mechanisms, we examined the combined influence of pCO(2) (150 and 900 microatm) and light (50 and 200 micromol photons m(-2) s(-1)) on Trichodesmium IMS101. We expand on a complementary study that demonstrated that while elevated pCO(2) enhanced N(2) fixation and growth, oxygen evolution and carbon fixation increased mainly as a response to high light. Here, we investigated changes in the photosynthetic fluorescence parameters of photosystem II, in ratios of the photosynthetic units (photosystem I:photosystem II), and in the pool sizes of key proteins involved in the fixation of carbon and nitrogen as well as their subsequent assimilation. We show that the combined elevation in pCO(2) and light controlled the operation of the CO(2)-concentrating mechanism and enhanced protein activity without increasing their pool size. Moreover, elevated pCO(2) and high light decreased the amounts of several key proteins (NifH, PsbA, and PsaC), while amounts of AtpB and RbcL did not significantly change. Reduced investment in protein biosynthesis, without notably changing photosynthetic fluxes, could free up energy that can be reallocated to increase N(2) fixation and growth at elevated pCO(2) and light. We suggest that changes in the redox state of the photosynthetic electron transport chain and posttranslational regulation of key proteins mediate the high flexibility in resources and energy allocation in Trichodesmium. This strategy should enable Trichodesmium to flourish in future surface oceans characterized by elevated pCO(2), higher temperatures, and high light. PMID:20625002

Levitan, Orly; Kranz, Sven A; Spungin, Dina; Prásil, Ondrej; Rost, Björn; Berman-Frank, Ilana

2010-07-12

321

Salinity tolerance of Picochlorum atomus and the use of salinity for contamination control by the freshwater cyanobacterium Pseudanabaena limnetica.  

PubMed

Microalgae are ideal candidates for waste-gas and -water remediation. However, salinity often varies between different sites. A cosmopolitan microalga with large salinity tolerance and consistent biochemical profiles would be ideal for standardised cultivation across various remediation sites. The aims of this study were to determine the effects of salinity on Picochlorum atomus growth, biomass productivity, nutrient uptake and biochemical profiles. To determine if target end-products could be manipulated, the effects of 4-day nutrient limitation were also determined. Culture salinity had no effect on growth, biomass productivity, phosphate, nitrate and total nitrogen uptake at 2, 8, 18, 28 and 36 ppt. 11 ppt, however, initiated a significantly higher total nitrogen uptake. While salinity had only minor effects on biochemical composition, nutrient depletion was a major driver for changes in biomass quality, leading to significant increases in total lipid, fatty acid and carbohydrate quantities. Fatty acid composition was also significantly affected by nutrient depletion, with an increased proportion of saturated and mono-unsaturated fatty acids. Having established that P. atomus is a euryhaline microalga, the effects of culture salinity on the development of the freshwater cyanobacterial contaminant Pseudanabaena limnetica were determined. Salinity at 28 and 36 ppt significantly inhibited establishment of P. limnetica in P. atomus cultures. In conclusion, P. atomus can be deployed for bioremediation at sites with highly variable salinities without effects on end-product potential. Nutrient status critically affected biochemical profiles--an important consideration for end-product development by microalgal industries. 28 and 36 ppt slow the establishment of the freshwater cyanobacterium P. limnetica, allowing for harvest of low contaminant containing biomass. PMID:23667639

von Alvensleben, Nicolas; Stookey, Katherine; Magnusson, Marie; Heimann, Kirsten

2013-05-07

322

Structural and functional characterization of a macrophage migration inhibitory factor homologue from the marine cyanobacterium Prochlorococcus marinus .  

PubMed

Macrophage migration inhibitory factor (MIF) is a multifunctional mammalian cytokine, which exhibits tautomerase and oxidoreductase activity. MIF homologues with pairwise sequence identities to human MIF ranging from 31% to 41% have been detected in various cyanobacteria. The gene encoding the MIF homologue from the marine cyanobacterium Prochlorococcus marinus strain MIT9313 has been cloned and the corresponding protein (PmMIF) overproduced, purified, and subjected to functional and structural characterization. Kinetic and (1)H NMR spectroscopic studies show that PmMIF tautomerizes phenylenolpyruvate and (p-hydroxyphenyl)enolpyruvate at low levels. The N-terminal proline of PmMIF is critical for these reactions because the P1A mutant has strongly reduced tautomerase activities. PmMIF shows high structural homology with mammalian MIFs as revealed by a crystal structure of PmMIF at 1.63 A resolution. MIF contains a Cys-X-X-Cys motif that mediates oxidoreductase activity, which is lacking from PmMIF. Engineering of the motif into PmMIF did not result in oxidoreductase activity but increased the tautomerase activity 8-fold. The shared tautomerase activities and the conservation of the beta-alpha-beta structural fold and key functional groups suggest that eukaryotic MIFs and cyanobacterial PmMIF are related by divergent evolution from a common ancestor. While several MIF homologues have been identified in eukaryotic parasites, where they are thought to play a role in modulating the host immune response, PmMIF is the first nonparasitic, bacterial MIF-like protein characterized in detail. This work sets the stage for future studies which could address the question whether a MIF-like protein from a free-living bacterium possesses immunostimulatory features similar to those of mammalian MIFs and MIF-like proteins found in parasitic nematodes and protozoa. PMID:20715791

Wasiel, Anna A; Rozeboom, Henriëtte J; Hauke, Doreen; Baas, Bert-Jan; Zandvoort, Ellen; Quax, Wim J; Thunnissen, Andy-Mark W H; Poelarends, Gerrit J

2010-09-01

323

Reversal in competitive dominance of a toxic versus non-toxic cyanobacterium in response to rising CO2.  

PubMed

Climate change scenarios predict a doubling of the atmospheric CO(2) concentration by the end of this century. Yet, how rising CO(2) will affect the species composition of aquatic microbial communities is still largely an open question. In this study, we develop a resource competition model to investigate competition for dissolved inorganic carbon in dense algal blooms. The model predicts how dynamic changes in carbon chemistry, pH and light conditions during bloom development feed back on competing phytoplankton species. We test the model predictions in chemostat experiments with monocultures and mixtures of a toxic and non-toxic strain of the freshwater cyanobacterium Microcystis aeruginosa. The toxic strain was able to reduce dissolved CO(2) to lower concentrations than the non-toxic strain, and became dominant in competition at low CO(2) levels. Conversely, the non-toxic strain could grow at lower light levels, and became dominant in competition at high CO(2) levels but low light availability. The model captured the observed reversal in competitive dominance, and was quantitatively in good agreement with the results of the competition experiments. To assess whether microcystins might have a role in this reversal of competitive dominance, we performed further competition experiments with the wild-type strain M. aeruginosa PCC 7806 and its mcyB mutant impaired in microcystin production. The microcystin-producing wild type had a strong selective advantage at low CO(2) levels but not at high CO(2) levels. Our results thus demonstrate both in theory and experiment that rising CO(2) levels can alter the community composition and toxicity of harmful algal blooms. PMID:21390081

Van de Waal, Dedmer B; Verspagen, Jolanda M H; Finke, Jan F; Vournazou, Vasiliki; Immers, Anne K; Kardinaal, W Edwin A; Tonk, Linda; Becker, Sven; Van Donk, Ellen; Visser, Petra M; Huisman, Jef

2011-03-10

324

Gene transfer in Leptolyngbya sp. strain BL0902, a cyanobacterium suitable for production of biomass and bioproducts.  

PubMed

Current cyanobacterial model organisms were not selected for their growth traits or potential for the production of renewable biomass, biofuels, or other products. The cyanobacterium strain BL0902 emerged from a search for strains with superior growth traits. Morphology and 16S rRNA sequence placed strain BL0902 in the genus Leptolyngbya. Leptolyngbya sp. strain BL0902 (hereafter Leptolyngbya BL0902) showed robust growth at temperatures from 22°C to 40°C and tolerated up to 0.5 M NaCl, 32 mM urea, high pH, and high solar irradiance. Its growth rate under outdoor conditions rivaled Arthrospira ("pirulina" strains. Leptolyngbya BL0902 accumulated higher lipid content and a higher proportion of monounsaturated fatty acids than Arthrospira strains. In addition to these desirable qualities, Leptolyngbya BL0902 is amenable to genetic engineering that is reliable, efficient, and stable. We demonstrated conjugal transfer from Escherichia coli of a plasmid based on RSF1010 and expression of spectinomycin/streptomycin resistance and yemGFP reporter transgenes. Conjugation efficiency was investigated in biparental and triparental matings with and without a "elper"plasmid that carries DNA methyltransferase genes, and with two different conjugal plasmids. We also showed that Leptolyngbya BL0902 is amenable to transposon mutagenesis with a Tn5 derivative. To facilitate genetic manipulation of Leptolyngbya BL0902, a conjugal plasmid vector was engineered to carry a trc promoter upstream of a Gateway recombination cassette. These growth properties and genetic tools position Leptolyngbya BL0902 as a model cyanobacterial production strain. PMID:22292073

Taton, Arnaud; Lis, Ewa; Adin, Dawn M; Dong, Guogang; Cookson, Scott; Kay, Steve A; Golden, Susan S; Golden, James W

2012-01-24

325

Heterocyst Development and Diazotrophic Metabolism in Terminal Respiratory Oxidase Mutants of the Cyanobacterium Anabaena sp. Strain PCC 7120?  

PubMed Central

Heterocyst development was analyzed in mutants of the heterocyst-forming cyanobacterium Anabaena sp. strain PCC 7120 bearing inactivated cox2 and/or cox3 genes, encoding heterocyst-specific terminal respiratory oxidases. At the morphological level, the cox2 cox3 double mutant (strain CSAV141) was impaired in membrane reorganization involving the so-called honeycomb system that in the wild-type strain is largely or exclusively devoted to respiration, accumulated glycogen granules at conspicuously higher levels than the wild type (in both vegetative cells and heterocysts), and showed a delay in carboxysome degradation upon combined nitrogen deprivation. Consistently, chemical analysis confirmed higher accumulation of glycogen in strain CSAV141 than in the wild type. No impairment was observed in the formation of the glycolipid or polysaccharide layers of the heterocyst envelope, consistent with the chemical detection of heterocyst-specific glycolipids, or in the expression of the heterocyst-specific genes nifHDK and fdxH. However, nitrogenase activity under oxic conditions was impaired in strain CSAV135 (cox3) and undetectable in strain CSAV141 (cox2 cox3). These results show that these dedicated oxidases are required for normal development and performance of the heterocysts and indicate a central role of Cox2 and, especially, of Cox3 in the respiratory activity of the heterocysts, decisively contributing to protection of the N2 fixation machinery against oxygen. However, in contrast to the case for other diazotrophic bacteria, expression of nif genes in Anabaena seems not to be affected by oxygen.

Valladares, Ana; Maldener, Iris; Muro-Pastor, Alicia M.; Flores, Enrique; Herrero, Antonia

2007-01-01

326

Effects of UV-B radiation and periodic desiccation on the morphogenesis of the edible terrestrial cyanobacterium Nostoc flagelliforme.  

PubMed

The terrestrial cyanobacterium Nostoc flagelliforme Berk. et M. A. Curtis has been a popular food and herbal ingredient for hundreds of years. To meet great market demand and protect the local ecosystem, for decades researchers have tried to cultivate N. flagelliforme but have failed to get macroscopic filamentous thalli. In this study, single trichomes with 50 to 200 vegetative cells were induced from free-living cells by low light and used to investigate the morphogenesis of N. flagelliforme under low UV-B radiation and periodic desiccation. Low-fluence-rate UV-B (0.1 W m(-2)) did not inhibit trichome growth; however, it significantly increased the synthesis of extracellular polysaccharides and mycosporine-like amino acids and promoted sheath formation outside the trichomes. Under low UV-B radiation, single trichomes developed into filamentous thalli more than 1 cm long after 28 days of cultivation, most of which grew separately in liquid BG11 medium. With periodic desiccation treatment, the single trichomes formed flat or banded thalli that grew up to 2 cm long after 3 months on solid BG11 medium. When trichomes were cultivated on solid BG11 medium with alternate treatments of low UV-B and periodic desiccation, dark and scraggly filamentous thalli that grew up to about 3 cm in length after 40 days were obtained. In addition, the cultivation of trichomes on nitrogen-deficient solid BG11 medium (BG11(0)) suggested that nitrogen availability could affect the color and lubricity of newly developed thalli. This study provides promising techniques for artificial cultivation of N. flagelliforme in the future. PMID:22865081

Feng, Yan-Na; Zhang, Zhong-Chun; Feng, Jun-Li; Qiu, Bao-Sheng

2012-08-03

327

Novel Derivatives of 9,10-Anthraquinone Are Selective Algicides against the Musty-Odor Cyanobacterium Oscillatoria perornata  

PubMed Central

Musty “off-flavor” in pond-cultured channel catfish (Ictalurus punctatus) costs the catfish production industry in the United States at least $30 million annually. The cyanobacterium Oscillatoria perornata (Skuja) is credited with being the major cause of musty off-flavor in farm-raised catfish in Mississippi. The herbicides diuron and copper sulfate, currently used by catfish producers as algicides to help mitigate musty off-flavor problems, have several drawbacks, including broad-spectrum toxicity towards the entire phytoplankton community that can lead to water quality deterioration and subsequent fish death. By use of microtiter plate bioassays, a novel group of compounds derived from the natural compound 9,10-anthraquinone have been found to be much more selectively toxic towards O. perornata than diuron and copper sulfate. In efficacy studies using limnocorrals placed in catfish production ponds, application rates of 0.3 ?M (125 ?g/liter) of the most promising anthraquinone derivative, 2-[methylamino-N-(1?-methylethyl)]-9,10-anthraquinone monophosphate (anthraquinone-59), dramatically reduced the abundance of O. perornata and levels of 2-methylisoborneol, the musty compound produced by O. perornata. The abundance of green algae and diatoms increased dramatically 2 days after application of a 0.3 ?M concentration of anthraquinone-59 to pond water within the limnocorrals. The half-life of anthraquinone-59 in pond water was determined to be 19 h, making it much less persistent than diuron. Anthraquinone-59 appears to be promising for use as a selective algicide in catfish aquaculture.

Schrader, Kevin K.; Dhammika Nanayakkara, N. P.; Tucker, Craig S.; Rimando, Agnes M.; Ganzera, Markus; Schaneberg, Brian T.

2003-01-01

328

Optimization of Metabolic Capacity and Flux through Environmental Cues To Maximize Hydrogen Production by the Cyanobacterium "Arthrospira (Spirulina) maxima"? †  

PubMed Central

Environmental and nutritional conditions that optimize the yield of hydrogen (H2) from water using a two-step photosynthesis/fermentation (P/F) process are reported for the hypercarbonate-requiring cyanobacterium “Arthrospira maxima.” Our observations lead to four main conclusions broadly applicable to fermentative H2 production by bacteria: (i) anaerobic H2 production in the dark from whole cells catalyzed by a bidirectional [NiFe] hydrogenase is demonstrated to occur in two temporal phases involving two distinct metabolic processes that are linked to prior light-dependent production of NADPH (photosynthetic) and dark/anaerobic production of NADH (fermentative), respectively; (ii) H2 evolution from these reductants represents a major pathway for energy production (ATP) during fermentation by regenerating NAD+ essential for glycolysis of glycogen and catabolism of other substrates; (iii) nitrate removal during fermentative H2 evolution is shown to produce an immediate and large stimulation of H2, as nitrate is a competing substrate for consumption of NAD(P)H, which is distinct from its slower effect of stimulating glycogen accumulation; (iv) environmental and nutritional conditions that increase anaerobic ATP production, prior glycogen accumulation (in the light), and the intracellular reduction potential (NADH/NAD+ ratio) are shown to be the key variables for elevating H2 evolution. Optimization of these conditions and culture age increases the H2 yield from a single P/F cycle using concentrated cells to 36 ml of H2/g (dry weight) and a maximum 18% H2 in the headspace. H2 yield was found to be limited by the hydrogenase-mediated H2 uptake reaction.

Ananyev, Gennady; Carrieri, Damian; Dismukes, G. Charles

2008-01-01

329

Enzymatic synthesis of a bicyclobutane fatty acid by a hemoprotein lipoxygenase fusion protein from the cyanobacterium Anabaena PCC 7120.  

PubMed

Biological transformations of polyunsaturated fatty acids often lead to chemically unstable products, such as the prostaglandin endoperoxides and leukotriene A(4) epoxide of mammalian biology and the allene epoxides of plants. Here, we report on the enzymatic production of a fatty acid containing a highly strained bicyclic four-carbon ring, a moiety known previously only as a model compound for mechanistic studies in chemistry. Starting from linolenic acid (C18.3omega3), a dual function protein from the cyanobacterium Anabaena PCC 7120 forms 9R-hydroperoxy-C18.3omega3 in a lipoxygenase domain, then a catalase-related domain converts the 9R-hydroperoxide to two unstable allylic epoxides. We isolated and identified the major product as 9R,10R-epoxy-11trans-C18.1 containing a bicyclo[1.1.0]butyl ring on carbons 13-16, and the minor product as 9R,10R-epoxy-11trans,13trans,15cis-C18.omega3, an epoxide of the leukotriene A type. Synthesis of both epoxides can be understood by initial transformation of the hydroperoxide to an epoxy allylic carbocation. Rearrangement to an intermediate bicyclobutonium ion followed by deprotonation gives the bicyclobutane fatty acid. This enzymatic reaction has no parallel in aqueous or organic solvent, where ring-opened cyclopropanes, cyclobutanes, and homoallyl products are formed. Given the capability shown here for enzymatic formation of the highly strained and unstable bicyclobutane, our findings suggest that other transformations involving carbocation rearrangement, in both chemistry and biology, should be examined for the production of the high energy bicyclobutanes. PMID:18025466

Schneider, Claus; Niisuke, Katrin; Boeglin, William E; Voehler, Markus; Stec, Donald F; Porter, Ned A; Brash, Alan R

2007-11-19

330

A LOV-domain-mediated blue-light-activated adenylate (adenylyl) cyclase from the cyanobacterium Microcoleus chthonoplastes PCC 7420.  

PubMed

Genome screening of the cyanobacterium Microcoleus chthonoplastes PCC 7420 identified a gene encoding a protein (483 amino acids, 54.2 kDa in size) characteristic of a BL (blue light)-regulated adenylate (adenylyl) cyclase function. The photoreceptive part showed signatures of a LOV (light, oxygen, voltage) domain. The gene product, mPAC (Microcoleus photoactivated adenylate cyclase), exhibited the LOV-specific three-peaked absorption band (?max=450 nm) and underwent conversion into the photoadduct form (?max=390 nm) upon BL-irradiation. The lifetime for thermal recovery into the parent state was determined as 16 s at 20°C (25 s at 11°C). The adenylate cyclase function showed a constitutive activity (in the dark) that was in-vitro-amplified by a factor of 30 under BL-irradiation. Turnover of the purified protein at saturating light and pH 8 is estimated to 1 cAMP/mPAC per s at 25°C (2 cAMP/mPAC per s at 35°C). The lifetime of light-activated cAMP production after a BL flash was ~14 s at 20°C. The temperature optimum was determined to 35°C and the pH optimum to 8.0. The value for half-maximal activating light intensity is 6 W/m2 (at 35°C). A comparison of mPAC and the BLUF (BL using FAD) protein bPAC (Beggiatoa PAC), as purified proteins and expressed in Xenopus laevis oocytes, yielded higher constitutive activity for mPAC in the dark, but also when illuminated with BL. PMID:24112109

Raffelberg, Sarah; Wang, Linzhu; Gao, Shiqiang; Losi, Aba; Gärtner, Wolfgang; Nagel, Georg

2013-11-01

331

Induction, isolation, and some properties of the NADPH-dependent glutamate dehydrogenase from the nonheterocystous cyanobacterium Phormidium laminosum.  

PubMed Central

The level of the NADPH-dependent glutamate dehydrogenase activity (EC 1.4.1.4) from nitrate-grown cells of the thermophilic non-N2-fixing cyanobacterium Phormidium laminosum OH-1-p.Cl1 could be significantly enhanced by the presence of ammonium or nitrite, as well as by L-methionine-DL-sulfoximine and other sources of organic nitrogen (L-Glu, L-Gln, and methylamine). The enzyme was purified more than 4,400-fold by ultracentrifugation, ion-exchange chromatography, and affinity chromatography, and at 30 degrees C it showed a specific activity of 32.9 mumol of NADPH oxidized per min per mg of protein. The purified enzyme showed no aminotransferase activity and catalyzed the amination of 2-oxoglutarate preferentially to the reverse catabolic reaction. The enzyme was very specific for its substrates 2-oxoglutarate (Km = 1.25 mM) and NADPH (Km = 64 microM), for which hyperbolic kinetics were obtained. However, negative cooperativity (Hill coefficient h = 0.89) and [S]0.5 of 18.2 mM were observed for ammonium. The mechanism of the aminating reaction was of a random type with independent sites. The purified enzyme showed its maximal activity at 60 degrees C (Ea = 5.1 kcal/mol [21.3 kJ/mol]) and optimal pH values of 8.0 and 7.5 when assayed in Tris hydrochloride and potassium phosphate buffers, respectively. The native molecular mass of the enzyme was about 280 kilodaltons. The possible physiological role of the enzyme in ammonia assimilation is discussed.

Martinez-Bilbao, M; Martinez, A; Urkijo, I; Llama, M J; Serra, J L

1988-01-01

332

Detection of microcystin synthetase genes in health food supplements containing the freshwater cyanobacterium Aphanizomenon flos-aquae.  

PubMed

In this study we investigated the presence of toxin-producing cyanobacterial contaminants in food supplements manufactured from blooms of the non-toxic freshwater cyanobacterium Aphanizomenon flos-aquae. Previous reports investigating the contamination of health food supplements with toxin-producing cyanobacteria have used chemical and or biochemical methods such as HPLC, ELISA and protein phosphatase assays. Whilst these studies have drawn attention to the presence of hepatotoxic microcystins in some commercially available food supplements, the methods used do not provide any information on the source of the contaminant. Such information would be useful for the quality control of food supplements produced for human consumption. In this study we applied a molecular technique, involving the amplification of the 16s rRNA gene, the phycocyanin operon, and two genes of the microcystin synthetase gene cluster to show that all 12 food supplement samples, sourced from various internet distributors and containing non-toxic A. flos-aquae, also contained toxigenic cyanobacteria. Sequencing of the microcystin synthetase genes detected in all of the food supplements showed that M. aeruginosa was the organism responsible for the production of microcystins in the samples. The presence of microcystins in the food supplements was confirmed by ELISA, with concentrations within the range of 0.1--4.72 microgg(-1) (microcystin-LR equivalents). Given that the molecular methods applied here are highly sensitive, and show good agreement with the results obtained from ELISA, we believe that they could potentially be used as a quality control technique for food products that contain cyanobacteria. PMID:16098554

Saker, M L; Jungblut, A-D; Neilan, B A; Rawn, D F K; Vasconcelos, V M

2005-10-01

333

Reversal in competitive dominance of a toxic versus non-toxic cyanobacterium in response to rising CO2  

PubMed Central

Climate change scenarios predict a doubling of the atmospheric CO2 concentration by the end of this century. Yet, how rising CO2 will affect the species composition of aquatic microbial communities is still largely an open question. In this study, we develop a resource competition model to investigate competition for dissolved inorganic carbon in dense algal blooms. The model predicts how dynamic changes in carbon chemistry, pH and light conditions during bloom development feed back on competing phytoplankton species. We test the model predictions in chemostat experiments with monocultures and mixtures of a toxic and non-toxic strain of the freshwater cyanobacterium Microcystis aeruginosa. The toxic strain was able to reduce dissolved CO2 to lower concentrations than the non-toxic strain, and became dominant in competition at low CO2 levels. Conversely, the non-toxic strain could grow at lower light levels, and became dominant in competition at high CO2 levels but low light availability. The model captured the observed reversal in competitive dominance, and was quantitatively in good agreement with the results of the competition experiments. To assess whether microcystins might have a role in this reversal of competitive dominance, we performed further competition experiments with the wild-type strain M. aeruginosa PCC 7806 and its mcyB mutant impaired in microcystin production. The microcystin-producing wild type had a strong selective advantage at low CO2 levels but not at high CO2 levels. Our results thus demonstrate both in theory and experiment that rising CO2 levels can alter the community composition and toxicity of harmful algal blooms.

Van de Waal, Dedmer B; Verspagen, Jolanda MH; Finke, Jan F; Vournazou, Vasiliki; Immers, Anne K; Kardinaal, W Edwin A; Tonk, Linda; Becker, Sven; Van Donk, Ellen; Visser, Petra M; Huisman, Jef

2011-01-01

334

Combined Effects of CO2 and Light on the N2-Fixing Cyanobacterium Trichodesmium IMS101: A Mechanistic View1  

PubMed Central

The marine diazotrophic cyanobacterium Trichodesmium responds to elevated atmospheric CO2 partial pressure (pCO2) with higher N2 fixation and growth rates. To unveil the underlying mechanisms, we examined the combined influence of pCO2 (150 and 900 ?atm) and light (50 and 200 ?mol photons m?2 s?1) on Trichodesmium IMS101. We expand on a complementary study that demonstrated that while elevated pCO2 enhanced N2 fixation and growth, oxygen evolution and carbon fixation increased mainly as a response to high light. Here, we investigated changes in the photosynthetic fluorescence parameters of photosystem II, in ratios of the photosynthetic units (photosystem I:photosystem II), and in the pool sizes of key proteins involved in the fixation of carbon and nitrogen as well as their subsequent assimilation. We show that the combined elevation in pCO2 and light controlled the operation of the CO2-concentrating mechanism and enhanced protein activity without increasing their pool size. Moreover, elevated pCO2 and high light decreased the amounts of several key proteins (NifH, PsbA, and PsaC), while amounts of AtpB and RbcL did not significantly change. Reduced investment in protein biosynthesis, without notably changing photosynthetic fluxes, could free up energy that can be reallocated to increase N2 fixation and growth at elevated pCO2 and light. We suggest that changes in the redox state of the photosynthetic electron transport chain and posttranslational regulation of key proteins mediate the high flexibility in resources and energy allocation in Trichodesmium. This strategy should enable Trichodesmium to flourish in future surface oceans characterized by elevated pCO2, higher temperatures, and high light.

Levitan, Orly; Kranz, Sven A.; Spungin, Dina; Prasil, Ondrej; Rost, Bjorn; Berman-Frank, Ilana

2010-01-01

335

Differential Transcriptional Analysis of the Cyanobacterium Cyanothece sp. Strain ATCC 51142 during Light-Dark and Continuous-Light Growth  

SciTech Connect

We analyzed the metabolic rhythms and differential gene transcription in the unicellular, diazotrophic cyanobacterium Cyanothece sp. ATCC51142 under N?-fixing conditions with 12h light-12h dark cycles followed by 36 h continuous light. Cultures were grown in a 6-L bioreactor that was specially designed for photosynthetic microorganisms and that permitted continuous monitoring of parameters such as pH and dissolved oxygen. Our main objective was to determine the strategies used by these cells to perform N? fixation under normal day-night conditions, as well as under greater stress caused by continuous light. Our results strongly suggested that the level of N? fixation is dependent upon respiration for energy production and for removal of intracellular O?. We determined that N? fixation cycled in continuous light, but that the N? fixation peak was lower and that glycogen degradation and respiration were also lower under these conditions. We also demonstrated that nifH (the gene encoding the Fe protein) and nifB and nifX were strongly induced in the continuous light; this is consistent with the mode of operation of these proteins relative to the MoFe protein and suggested that any regulation of N? fixation was at a posttranscriptional level. Also, many soluble electron carriers (e.g., ferredoxins), as well as redox carriers (e.g., thioredoxin and glutathione) were strongly induced during N? fixation in continuous light. We suggest that these carriers were required to generate enhanced cyclic electron transport and phosphorylation for energy production and to maintain appropriate redox levels in the presence of enhanced O?, respectively.

Toepel, Jorg; Welsh, Eric A.; Summerfield, Tina; Pakrasi, Himadri B.; Sherman, Louis A.

2008-06-01

336

Transcriptional analysis of the unicellular, diazotrophic cyanobacterium Cyanothece sp. ATCC 51142 grown under short day/night cycles  

SciTech Connect

Cyanothece sp. strain ATCC 51142 is a unicellular, diazotrophic cyanobacterium that demonstrates extensive metabolic periodicities of photosynthesis, respiration and nitrogen fixation when grown under N2-fixing conditions. We have performed a global transcription analysis of this organism using 6 h light/dark cycles in order to determine the response of the cell to these conditions and to differentiate between diurnal and circadian regulated genes. In addition, we used a context-likelihood of relatedness (CLR) analysis with this data and those from two-day light/dark and light-dark plus continuous light experiments to better differentiate between diurnal and circadian regulated genes. Cyanothece sp. adapted in several ways to growth under short light/dark conditions. Nitrogen was fixed in every second dark period and only once in each 24 h period. Nitrogen fixation was strongly correlated to the energy status of the cells and glycogen breakdown and high respiration rates were necessary to provide appropriate energy and anoxic conditions for this process. We conclude that glycogen breakdown is a key regulatory step within these complex processes. Our results demonstrated that the main metabolic genes involved in photosynthesis, respiration, nitrogen fixation and central carbohydrate metabolism have strong (or total) circadian-regulated components. The short light/dark cycles enable us to identify transcriptional differences among the family of psbA genes, as well as the differing patterns of the hup genes, which follow the same pattern as nitrogenase genes, relative to the hox genes which displayed a diurnal, dark-dependent gene expression.

Toepel, Jorg; McDermott, Jason E.; Summerfield, Tina; Sherman, Louis A.

2009-06-01

337

Cluster of genes that encode positive and negative elements influencing filament length in a heterocyst-forming cyanobacterium.  

PubMed

The filamentous, heterocyst-forming cyanobacteria perform oxygenic photosynthesis in vegetative cells and nitrogen fixation in heterocysts, and their filaments can be hundreds of cells long. In the model heterocyst-forming cyanobacterium Anabaena sp. strain PCC 7120, the genes in the fraC-fraD-fraE operon are required for filament integrity mainly under conditions of nitrogen deprivation. The fraC operon transcript partially overlaps gene all2395, which lies in the opposite DNA strand and ends 1 bp beyond fraE. Gene all2395 produces transcripts of 1.35 kb (major transcript) and 2.2 kb (minor transcript) that overlap fraE and whose expression is dependent on the N-control transcription factor NtcA. Insertion of a gene cassette containing transcriptional terminators between fraE and all2395 prevented production of the antisense RNAs and resulted in an increased length of the cyanobacterial filaments. Deletion of all2395 resulted in a larger increase of filament length and in impaired growth, mainly under N2-fixing conditions and specifically on solid medium. We denote all2395 the fraF gene, which encodes a protein restricting filament length. A FraF-green fluorescent protein (GFP) fusion protein accumulated significantly in heterocysts. Similar to some heterocyst differentiation-related proteins such as HglK, HetL, and PatL, FraF is a pentapeptide repeat protein. We conclude that the fraC-fraD-fraE?fraF gene cluster (where the arrow indicates a change in orientation), in which cis antisense RNAs are produced, regulates morphology by encoding proteins that influence positively (FraC, FraD, FraE) or negatively (FraF) the length of the filament mainly under conditions of nitrogen deprivation. This gene cluster is often conserved in heterocyst-forming cyanobacteria. PMID:23813733

Merino-Puerto, Victoria; Herrero, Antonia; Flores, Enrique

2013-09-01

338

Bentazon triggers the promotion of oxidative damage in the Portuguese ricefield cyanobacterium Anabaena cylindrica: response of the antioxidant system.  

PubMed

Rice fields are frequently exposed to environmental contamination by herbicides and cyanobacteria, as primary producers of these aquatic ecosystems, are adversely affected. Anabaena cylindrica is a cyanobacterium with a significantly widespread occurrence in Portuguese rice fields. This strain was studied throughout 72 h in laboratory conditions for its stress responses to sublethal concentrations (0.75-2 mM) of bentazon, a selective postemergence herbicide recommended for integrated weed management in rice, with special reference to oxidative stress, role of proline and intracellular antioxidant enzymes in herbicide-induced free radicals detoxification. Activities of antioxidant enzymes such as superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), and glutathione S-transferase (GST) increased in a time- and herbicide dose-response manner and were higher than those in the control samples after 72 h. A time- and concentration-dependent increase of malondialdehyde (MDA) levels and the enhanced cell membrane leakage following bentazon exposure are indicative of lipid peroxidation, free radicals formation, and oxidative damage, while increased amounts of SOD, CAT, APX, GST, and proline indicated their involvement in free radical scavenging mechanisms. The appreciable decline in the reduced glutathione (GSH) pool after 72 h at higher bentazon concentrations could be explained by the reduction of the NADPH-dependent glutathione reductase (GR) activity. The obtained results suggested that the alterations of antioxidant systems in A. cylindrica might be useful biomarkers of bentazon exposure. As the toxic mechanism of bentazon is a complex phenomenon, this study also adds relevant findings to explain the oxidative stress pathways of bentazon promoting oxidative stress in cyanobacteria. PMID:20549627

Galhano, Victor; Peixoto, Francisco; Gomes-Laranjo, José

2010-10-01

339

The NADPH thioredoxin reductase C functions as an electron donor to 2-Cys peroxiredoxin in a thermophilic cyanobacterium Thermosynechococcus elongatus BP-1  

SciTech Connect

An NADPH thioredoxin reductase C was co-purified with a 2-Cys peroxiredoxin by the combination of anion exchange chromatography and electroelution from gel slices after native PAGE from a thermophilic cyanobacterium Thermosynechococcus elongatus as an NAD(P)H oxidase complex induced by oxidative stress. The result provided a strong evidence that the NADPH thioredoxin reductase C interacts with the 2-Cys peroxiredoxin in vivo. An in vitro reconstitution assay with purified recombinant proteins revealed that both proteins were essential for an NADPH-dependent reduction of H{sub 2}O{sub 2}. These results suggest that the reductase transfers the reducing power from NADPH to the peroxiredoxin, which reduces peroxides in the cyanobacterium under oxidative stress. In contrast with other NADPH thioredoxin reductases, the NADPH thioredoxin reductase C contains a thioredoxin-like domain in addition to an NADPH thioredoxin reductase domain in the same polypeptide. Each domain contains a conserved CXYC motif. A point mutation at the CXYC motif in the NADPH thioredoxin reductase domain resulted in loss of the NADPH oxidation activity, while a mutation at the CXYC motif in the thioredoxin-like domain did not affect the electron transfer, indicating that this motif is not essential in the electron transport from NADPH to the 2-Cys peroxiredoxin.

Sueoka, Keigo [Department of Molecular Biology, Graduate School of Science, Nagoya University, Chikusa-ku, Nagoya 464-8602 (Japan); Department of Biochemistry and Molecular Biology, Saitama University, 255 Shimo-Okubo, Sakura-ku, Saitama 338-8570 (Japan); Yamazaki, Teruaki [Department of Biochemistry and Molecular Biology, Saitama University, 255 Shimo-Okubo, Sakura-ku, Saitama 338-8570 (Japan); JASCO International Co., Ltd., 4-21 Sennin-cho 2-chome, Hachioji, Tokyo 193-0835 (Japan); Hiyama, Tetsuo [Department of Biochemistry and Molecular Biology, Saitama University, 255 Shimo-Okubo, Sakura-ku, Saitama 338-8570 (Japan); Nakamoto, Hitoshi [Department of Biochemistry and Molecular Biology, Saitama University, 255 Shimo-Okubo, Sakura-ku, Saitama 338-8570 (Japan)], E-mail: nakamoto@mail.saitama-u.ac.jp

2009-03-13

340

The coxBAC Operon Encodes a Cytochrome c Oxidase Required for Heterotrophic Growth in the Cyanobacterium Anabaena variabilis Strain ATCC 29413  

PubMed Central

Three genes, coxB, coxA, and coxC, found in a clone from a gene library of the cyanobacterium Anabaena variabilis strain ATCC 29413, were identified by hybridization with an oligonucleotide specific for aa3-type cytochrome c oxidases. Deletion of these genes from the genome of A. variabilis strain ATCC 29413 FD yielded strain CSW1, which displayed no chemoheterotrophic growth and an impaired cytochrome c oxidase activity. Photoautotrophic growth of CSW1, however, was unchanged, even with dinitrogen as the nitrogen source. A higher cytochrome c oxidase activity was detected in membrane preparations from dinitrogen-grown CSW1 than from nitrate-grown CSW1, but comparable activities of respiratory oxygen uptake were found in the wild type and in CSW1. Our data indicate that the identified cox gene cluster is essential for fructose-dependent growth in the dark, but not for growth on dinitrogen, and that other terminal respiratory oxidases are expressed in this cyanobacterium. Transcription analysis showed that coxBAC constitutes an operon which is expressed from two transcriptional start points. The use of one of them was stimulated by fructose.

Schmetterer, Georg; Valladares, Ana; Pils, Dietmar; Steinbach, Susanne; Pacher, Margit; Muro-Pastor, Alicia M.; Flores, Enrique; Herrero, Antonia

2001-01-01

341

Oligonucleotide-directed mutagenesis of psbB, the gene encoding CP47, employing a deletion mutant strain of the cyanobacterium Synechocystis sp. PCC 6803.  

PubMed

A mutant strain of the cyanobacterium Synechocystis sp. PCC (Pasteur Culture Collection) 6803 has been developed in which psbB, the gene coding for the chlorophyl alpha-binding protein CP47 in Photosystem II (PSII), has been deleted. This deletion mutant can be used for the reintroduction of modified psbB into the cyanobacterium. To study the role of a large hydrophilic region in CP47, presumably located on the lumenal side of the thylakoid membrane between the fifth and sixth membrane-spanning regions, specific deletions have been introduced in psbB coding for regions within this domain. One psbB mutation leads to deletion of Gly-351 to Thr-365 in CP47, another psbB mutation was targeted towards deletion of Arg-384 to Val-392 in this protein. The deletion from Gly-351 to Thr-365 results in a loss of PSII activity and of photoautotrophic growth of the mutant, but the deletion between Arg-384 and Val-392 retains PSII activity and the ability to grow photoautotrophically. The mutant strain with the deletion from Gly-351 to Thr-365 does not assemble a stable PSII reaction center complex in its thylakoid membranes, and exhibits diminished levels of CP47 and of the reaction center proteins D1 and D2. In contrast to the Arg-384 to Val-392 portion of this domain, the region between Gly-351 and Thr-365 appears essential for the normal structure and function of photosystem II. PMID:1932693

Eaton-Rye, J J; Vermaas, W F

1991-12-01

342

Insights into the Physiology and Ecology of the Brackish-Water-Adapted Cyanobacterium Nodularia spumigena CCY9414 Based on a Genome-Transcriptome Analysis  

PubMed Central

Nodularia spumigena is a filamentous diazotrophic cyanobacterium that dominates the annual late summer cyanobacterial blooms in the Baltic Sea. But N. spumigena also is common in brackish water bodies worldwide, suggesting special adaptation allowing it to thrive at moderate salinities. A draft genome analysis of N. spumigena sp. CCY9414 yielded a single scaffold of 5,462,271 nucleotides in length on which genes for 5,294 proteins were annotated. A subsequent strand-specific transcriptome analysis identified more than 6,000 putative transcriptional start sites (TSS). Orphan TSSs located in intergenic regions led us to predict 764 non-coding RNAs, among them 70 copies of a possible retrotransposon and several potential RNA regulators, some of which are also present in other N2-fixing cyanobacteria. Approximately 4% of the total coding capacity is devoted to the production of secondary metabolites, among them the potent hepatotoxin nodularin, the linear spumigin and the cyclic nodulapeptin. The transcriptional complexity associated with genes involved in nitrogen fixation and heterocyst differentiation is considerably smaller compared to other Nostocales. In contrast, sophisticated systems exist for the uptake and assimilation of iron and phosphorus compounds, for the synthesis of compatible solutes, and for the formation of gas vesicles, required for the active control of buoyancy. Hence, the annotation and interpretation of this sequence provides a vast array of clues into the genomic underpinnings of the physiology of this cyanobacterium and indicates in particular a competitive edge of N. spumigena in nutrient-limited brackish water ecosystems.

Voss, Bjorn; Bolhuis, Henk; Fewer, David P.; Kopf, Matthias; Moke, Fred; Haas, Fabian; El-Shehawy, Rehab; Hayes, Paul; Bergman, Birgitta; Sivonen, Kaarina; Dittmann, Elke; Scanlan, Dave J.; Hagemann, Martin; Stal, Lucas J.; Hess, Wolfgang R.

2013-01-01

343

Ciliate Nassula sp. grazing on a microcystin-producing cyanobacterium (Planktothrix agardhii): impact on cell growth and in the microcystin fractions.  

PubMed

The proliferation of microcystins (MCs)-producing cyanobacteria (MCs) can have detrimental effects on the food chain in aquatic environments. Until recently, few studies had focused on the fate of MCs in exposed organisms, such as primary consumers of cyanobacteria. In this study, we investigate the impact of an MC-producing strain of the cyanobacterium Planktothrix agardhii on the growth and physiology of a Nassula sp. ciliate isolated from a non-toxic cyanobacterial bloom. We show that this Nassula sp. strain was able to consume and grow while feeding exclusively on an MC-producing cyanobacterium over a prolonged period of time (8 months). In short-term exposure experiments (8 days), ciliates consuming an MC-producing cyanobacterial strain displayed slower growth rate and higher levels of antioxidant enzymes than ciliates feeding on two non-MC-producing strains. Three high-performance methods (LC/MS, LC/MS-MS and ELISA) were used to quantify the free and bound MCs in the culture medium and in the cells. We show that ciliate grazing led to a marked decrease in free MCs (methanol extractable) in cells, the MCs were therefore no longer found in the surrounding culture medium. These findings suggest that MCs may have undergone redistribution (free vs bound MCs) or chemical degradation within the ciliates. PMID:23010390

Combes, Audrey; Dellinger, Marc; Cadel-six, Sabrina; Amand, Severine; Comte, Katia

2012-08-31

344

The coxBAC operon encodes a cytochrome c oxidase required for heterotrophic growth in the cyanobacterium Anabaena variabilis strain ATCC 29413.  

PubMed

Three genes, coxB, coxA, and coxC, found in a clone from a gene library of the cyanobacterium Anabaena variabilis strain ATCC 29413, were identified by hybridization with an oligonucleotide specific for aa(3)-type cytochrome c oxidases. Deletion of these genes from the genome of A. variabilis strain ATCC 29413 FD yielded strain CSW1, which displayed no chemoheterotrophic growth and an impaired cytochrome c oxidase activity. Photoautotrophic growth of CSW1, however, was unchanged, even with dinitrogen as the nitrogen source. A higher cytochrome c oxidase activity was detected in membrane preparations from dinitrogen-grown CSW1 than from nitrate-grown CSW1, but comparable activities of respiratory oxygen uptake were found in the wild type and in CSW1. Our data indicate that the identified cox gene cluster is essential for fructose-dependent growth in the dark, but not for growth on dinitrogen, and that other terminal respiratory oxidases are expressed in this cyanobacterium. Transcription analysis showed that coxBAC constitutes an operon which is expressed from two transcriptional start points. The use of one of them was stimulated by fructose. PMID:11591688

Schmetterer, G; Valladares, A; Pils, D; Steinbach, S; Pacher, M; Muro-Pastor, A M; Flores, E; Herrero, A

2001-11-01

345

NADP(+)-isocitrate dehydrogenase from the cyanobacterium Anabaena sp. strain PCC 7120: purification and characterization of the enzyme and cloning, sequencing, and disruption of the icd gene.  

PubMed Central

NADP(+)-isocitrate dehydrogenase (NADP(+)-IDH) from the dinitrogen-fixing filamentous cyanobacterium Anabaena sp. strain PCC 7120 was purified to homogeneity. The native enzyme is composed of two identical subunits (M(r), 57,000) and cross-reacts with antibodies obtained against the previously purified NADP(+)-IDH from the unicellular cyanobacterium Synechocystis sp. strain PCC 6803. Anabaena NADP(+)-IDH resembles in its physicochemical and kinetic parameters the typical dimeric IDHs from prokaryotes. The gene encoding Anabaena NADP(+)-IDH was cloned by complementation of an Escherichia coli icd mutant with an Anabaena genomic library. The complementing DNA was located on a 6-kb fragment. It encodes an NADP(+)-IDH that has the same mobility as that of Anabaena NADP(+)-IDH on nondenaturing polyacrylamide gels. The icd gene was subcloned and sequenced. Translation of the nucleotide sequence gave a polypeptide of 473 amino acids that showed high sequence similarity to the E. coli enzyme (59% identity) and with IDH1 and IDH2, the two subunits of the heteromultimeric NAD(+)-IDH from Saccharomyces cerevisiae (30 to 35% identity); however, a low level of similarity to NADP(+)-IDHs of eukaryotic origin was found (23% identity). Furthermore, Anabaena NADP(+)-IDH contains a 44-residue amino acid sequence in its central region that is absent in the other IDHs so far sequenced. Attempts to generate icd mutants by insertional mutagenesis were unsuccessful, suggesting an essential role of IDH in Anabaena sp. strain PCC 7120. Images

Muro-Pastor, M I; Florencio, F J

1994-01-01

346

PSP toxin release from the cyanobacterium Raphidiopsis brookii D9 (Nostocales) can be induced by sodium and potassium ions.  

PubMed

Paralytic shellfish poisoning (PSP) toxins are a group of naturally occurring neurotoxic alkaloids produced among several genera of primarily freshwater cyanobacteria and marine dinoflagellates. Although saxitoxin (STX) and analogs are all potent Na(+) channel blockers in vertebrate cells, the functional role of these compounds for the toxigenic microorganisms is unknown. Based upon the known importance of monovalent cations (such as sodium) in the maintenance of cellular homeostasis and ion channel function, we examined the effect of high extracellular concentrations of these ions on growth, cellular integrity, toxin production and release to the external medium in the filamentous freshwater cyanobacterium, Raphidiopsis brookii D9; a gonyautoxins (GTX2/3) and STX producing toxigenic strain. We observed a toxin export in response to high (17 mM) NaCl and KCl concentrations in the growth medium that was not primarily related to osmotic stress effects, compared to the osmolyte mannitol. Addition of exogenous PSP toxins with the same compositional profile as the one produced by R. brookii D9 was able to partially mitigate this effect of high Na? (17 mM). The PSP toxin biosynthetic gene cluster (sxt) in D9 has two genes (sxtF and sxtM) that encode for a MATE (multidrug and toxic compound extrusion) transporter. This protein family, represented by NorM in the bacterium Vibrio parahaemolyticus, confers resistance to multiple cationic toxic agents through Na?/drug antiporters. Conserved domains for Na? and drug recognition have been described in NorM. For the D9 sxt cluster, the Na? recognition domain is conserved in both SxtF and SxtM, but the drug recognition domain differs between them. These results suggest that PSP toxins are exported directly in response to the presence of monovalent cations (Na?, K?) at least at elevated concentrations. Thus, the presence of both genes in the sxt cluster from strain D9 can be explained as a selective recognition mechanism by the SxtF/M transporters for GTX2/3 and STX. We propose that these toxins in cyanobacteria could act extracellularly as a protective mechanism to ensure homeostasis against extreme salt variation in the environment. PMID:22983012

Soto-Liebe, Katia; Méndez, Marco A; Fuenzalida, Loreto; Krock, Bernd; Cembella, Allan; Vásquez, Mónica

2012-09-13

347

Analysis of a genomic DNA region from the cyanobacterium Synechococcus sp. strain PCC7942 involved in carboxysome assembly and function.  

PubMed Central

We report on the sequencing and analysis of a 3,557-bp genomic DNA clone that is located between 4.8 and 1.2 kilobase pairs (kb) upstream of the rbcL gene and is capable of complementing a class of cyanobacterium Synechococcus sp. strain PCC7942 mutants requiring a high level of CO2. The upstream 2,704 bp of this sequence is novel, the remaining 852 bp having been reported by other workers. Four new open reading frames (ORFs) have been identified along with putative promoter elements. These ORFs, which could code for proteins of 7, 10.9, 11, and 58 kDa in size, have been named ORF 64, ccmK, ccmL, and ccmM, respectively. The last three have been named ccm genes on the basis that insertional mutagenesis of each produces a phenotype requiring a high level of CO2 (i.e., each produces a lesion in the CO2 concentrating mechanism). The putative gene product for the large ccmM ORF has three internally repeated regions and also has two possible DNA binding motifs. Two defined mutants in the 3,557-bp region, mutants PVU and P-N, have been more fully characterized. The PVU mutant has a drug marker inserted into the ccmL gene, and it possesses abnormal rod-shaped carboxysomes. The P-N mutant is a 2.64-kb deletion of DNA from the same position in ccmL to a region closer to rbcL. This mutant, which has previously been shown to lack carboxysomes and have soluble ribulosebiphosphate carboxylase/oxygenase activity, has now been shown to have a predominantly soluble carboxysomal carbonic anhydrase activity. Both mutants were found to possess carboxysomal carbonic anhydrase activities which are below wild-type levels, and in the P-N mutant this activity appears to be unstable. The results are discussed in terms of the possible interactions of putative ccm gene products in the process of carboxysome assembly and function. Images

Price, G D; Howitt, S M; Harrison, K; Badger, M R

1993-01-01

348

Construction of shuttle plasmids which can be efficiently mobilized from Escherichia coli into the chromatically adapting cyanobacterium, Fremyella diplosiphon.  

PubMed

In some strains of cyanobacteria the composition of the light-harvesting antennae is determined by the color of available light. The mechanism of this chromatic adaptation involves the regulation of gene expression by red and green light and has been most studied in Fremyella diplosiphon (Calothrix sp. PCC 7601), a filamentous cyanobacterium for which there has been no reported means of genetic manipulation. We have constructed shuttle plasmids which can be efficiently mobilized by RP4 from Escherichia coli into F. diplosiphon and which can be recovered from transconjugant F. diplosiphon and returned to E. coli by transformation. The ability of these plasmids to replicate in F. diplosiphon is conferred by an 8.0-kb DNA fragment isolated from pFDA, a plasmid native to F. diplosiphon. To create these shuttle plasmids the 8.0-kb fragment was cloned into pJCF22, a mobilizable plasmid constructed from oriV and bom from pBR322, cat from pACYC184 and aphA from pACYC177.pJCF22 lacks sites for the restriction enzymes FdiI and II. Transconjugant F. diplosiphon containing shuttle plasmid pJCF62 are resistant to chloramphenicol and highly resistant to the aminoglycosides, G418 and neomycin. When aadA from the omega interposon was incorporated into a shuttle plasmid transconjugant F. diplosiphon could also be selected with streptomycin or spectinomycin. In F. diplosiphon shuttle plasmid pJCF62 replicates with a minimum copy number of seven. The oriV for replication in F. diplosiphon was localized to a 2.8-kb region within the cyanobacterial part of pJCF62. The presence on a shuttle plasmid of a single recognition site for FdiI reduced the efficiency of mobilization into F. diplosiphon by 5- to 10-fold. Restriction at this site was prevented when the E. coli donor strain in the mating contained the enzyme Eco47II methylase. PMID:8234495

Cobley, J G; Zerweck, E; Reyes, R; Mody, A; Seludo-Unson, J R; Jaeger, H; Weerasuriya, S; Navankasattusas, S

1993-09-01

349

Ultraviolet stress delays chromosome replication in light/dark synchronized cells of the marine cyanobacterium Prochlorococcus marinus PCC9511  

PubMed Central

Background The marine cyanobacterium Prochlorococcus is very abundant in warm, nutrient-poor oceanic areas. The upper mixed layer of oceans is populated by high light-adapted Prochlorococcus ecotypes, which despite their tiny genome (~1.7 Mb) seem to have developed efficient strategies to cope with stressful levels of photosynthetically active and ultraviolet (UV) radiation. At a molecular level, little is known yet about how such minimalist microorganisms manage to sustain high growth rates and avoid potentially detrimental, UV-induced mutations to their DNA. To address this question, we studied the cell cycle dynamics of P. marinus PCC9511 cells grown under high fluxes of visible light in the presence or absence of UV radiation. Near natural light-dark cycles of both light sources were obtained using a custom-designed illumination system (cyclostat). Expression patterns of key DNA synthesis and repair, cell division, and clock genes were analyzed in order to decipher molecular mechanisms of adaptation to UV radiation. Results The cell cycle of P. marinus PCC9511 was strongly synchronized by the day-night cycle. The most conspicuous response of cells to UV radiation was a delay in chromosome replication, with a peak of DNA synthesis shifted about 2 h into the dark period. This delay was seemingly linked to a strong downregulation of genes governing DNA replication (dnaA) and cell division (ftsZ, sepF), whereas most genes involved in DNA repair (such as recA, phrA, uvrA, ruvC, umuC) were already activated under high visible light and their expression levels were only slightly affected by additional UV exposure. Conclusions Prochlorococcus cells modified the timing of the S phase in response to UV exposure, therefore reducing the risk that mutations would occur during this particularly sensitive stage of the cell cycle. We identified several possible explanations for the observed timeshift. Among these, the sharp decrease in transcript levels of the dnaA gene, encoding the DNA replication initiator protein, is sufficient by itself to explain this response, since DNA synthesis starts only when the cellular concentration of DnaA reaches a critical threshold. However, the observed response likely results from a more complex combination of UV-altered biological processes.

2010-01-01

350

Spectroscopic study of the CP43' complex and the PSI-CP43' supercomplex of the cyanobacterium Synechocystis PCC 6803.  

PubMed

The PSI-CP43' supercomplex of the cyanobacterium Synechocystis PCC 6803, grown under iron-starvation conditions, consists of a trimeric core Photosystem I (PSI) complex and an outer ring of 18 CP43' light-harvesting complexes. We have investigated the electronic structure and excitation energy transfer (EET) pathways within the CP43' (also known as the isiA gene product) ring using low-temperature absorption, fluorescence, fluorescence excitation, and hole-burning (HB) spectroscopies. Analysis of the absorption spectra of PSI, CP43', and PSI-CP43' complexes suggests that there are 13 chlorophylls (Chls) per CP43' monomer, i.e., a number that was observed in the CP43 complex of Photosystem II (PSII) (Umena, Y. et al. Nature 2011, 473, 55-60). This is in contrast with the recent modeling studies of Zhang et al. (Biochim. Biophys. Acta 2010, 1797, 457-465), which suggested that IsiA likely contains 15 Chls. Modeling studies of various optical spectra of the CP43' ring using the uncorrelated EET model (Zazubovich, V.; Jankowiak, R. J. Lumin. 2007, 127, 245-250) suggest that CP43' monomers (in analogy to the CP43 complexes of the PSII core) also possess two quasi-degenerate low-energy states, A' and B'. The site distribution functions of states A' and B' maxima/full width at half-maximum (fwhm) are at 684 nm/180 cm(-1) and 683 nm/80 cm(-1), respectively. Our analysis shows that pigments mostly contributing to the lowest-energy A' and B' states must be located on the side of the CP43' complex facing the PSI core, a finding that contradicts the model of Zhang et al. but is in agreement with the model suggested by Nield et al. (Biochemistry2003, 42, 3180-3188). We demonstrate that the A'-A' and B'-B' EET between different monomers is possible, though with a slower rate than intramonomer A'-B' and/or B'-A' energy transfer. PMID:21978372

Feng, Ximao; Neupane, Bhanu; Acharya, Khem; Zazubovich, Valter; Picorel, Rafael; Seibert, Michael; Jankowiak, Ryszard

2011-10-26

351

Combined Effects of CO2 and Light on the N2-Fixing Cyanobacterium Trichodesmium IMS101: Physiological Responses1[OA  

PubMed Central

Recent studies on the diazotrophic cyanobacterium Trichodesmium erythraeum (IMS101) showed that increasing CO2 partial pressure (pCO2) enhances N2 fixation and growth. Significant uncertainties remain as to the degree of the sensitivity to pCO2, its modification by other environmental factors, and underlying processes causing these responses. To address these questions, we examined the responses of Trichodesmium IMS101 grown under a matrix of low and high levels of pCO2 (150 and 900 ?atm) and irradiance (50 and 200 ?mol photons m?2 s?1). Growth rates as well as cellular carbon and nitrogen contents increased with increasing pCO2 and light levels in the cultures. The pCO2-dependent stimulation in organic carbon and nitrogen production was highest under low light. High pCO2 stimulated rates of N2 fixation and prolonged the duration, while high light affected maximum rates only. Gross photosynthesis increased with light but did not change with pCO2. HCO3? was identified as the predominant carbon source taken up in all treatments. Inorganic carbon uptake increased with light, but only gross CO2 uptake was enhanced under high pCO2. A comparison between carbon fluxes in vivo and those derived from 13C fractionation indicates high internal carbon cycling, especially in the low-pCO2 treatment under high light. Light-dependent oxygen uptake was only detected under low pCO2 combined with high light or when low-light-acclimated cells were exposed to high light, indicating that the Mehler reaction functions also as a photoprotective mechanism in Trichodesmium. Our data confirm the pronounced pCO2 effect on N2 fixation and growth in Trichodesmium and further show a strong modulation of these effects by light intensity. We attribute these responses to changes in the allocation of photosynthetic energy between carbon acquisition and the assimilation of carbon and nitrogen under elevated pCO2. These findings are supported by a complementary study looking at photosynthetic fluorescence parameters of photosystem II, photosynthetic unit stoichiometry (photosystem I:photosystem II), and pool sizes of key proteins in carbon and nitrogen acquisition.

Kranz, Sven A.; Levitan, Orly; Richter, Klaus-Uwe; Prasil, Ondrej; Berman-Frank, Ilana; Rost, Bjorn

2010-01-01

352

Laboratory Simulation of Biogeochemical Interactions Between Cyanobacterium-Growth and CaCO3 Deposition: Implications for Carbon Accumulation Under Extreme Atmospheric Conditions of Precambrian Earth  

NASA Astrophysics Data System (ADS)

The atmosphere of Precambrian Earth was characterized by high PCO2, low PO2, and high violent UV radiation. To better understand the interaction between cyanobacterium-growth and CaCO3 deposition in such extreme environments, we grew Oscillatoria tenuis, a prokaryotic alga that is morphologically similar to micro-fossils found in Precambrian chert, in the laboratory under controlled temperature and patial presure of CO2. During algal cell growth, oxygen was absorbed continously by chromous chloride oxygen-absorbent and the levels of PCO2 were controlled by adding different amounts of HCO3- (NaHCO3) in culture medium with initial pH 7.4. Our observation indicates that PCO2 excerises the first order of control on the accumulation of cyanobaterium biomass. Under 100,000 Pa of PCO2, the growth rate of cyanobaterium increases along with the elevation of CO2 partial pressure; however, when PCO2 is higher than 100,000 Pa, the increase of PCO2 results in the decrease of cyanobacterium biomass. On the other hand, photosynthesis of cyanobacteria controls CaCO3 deposition via the function of adjusting pH in the solution. In a 5 day cell growth experiment with PCO2 controlled at about 50,000 Pa and additional 0.0001, 0.001, 0.01, 0.1 and 1.0 M Ca2+ input separately at speed of 2.5 ml/h, the largest total biomass of cyanobacterium (896 mg/L) including living suspension cells and deposited cells was obtained when Ca2+ input was maintained at 0.01 M with 2.5 ml/h. Otherwise, less Ca2+ input resulted in more living suspension cells and less deposited cells. More Ca2+ input resulted in less living suspension cells and more deposited cells. At last both conditions were not good for cell growth and accumulation of organic matter in carbonate deposition in long term. Our laboratory simulation illustrates that the Ca2+ input is critical to CaCO3 deposition and such controls are indirectly enforced through the accumulation of cyanobacteria biomass under a warm, anoxic and high pCO2 atmospheric condition during a part of Precambrian time.

Wu, Q.; Chen, L.; Chen, G.; Yang, H.

2004-05-01

353

Looking at the stability of life-support microorganisms in space : the MELGEN activity highlights the cyanobacterium Arthrospira sp. PCC8005  

NASA Astrophysics Data System (ADS)

The MELGEN activity (MELiSSA Genetic Stability Study) mainly covers the molecular aspects of the regenerative life-support system MELiSSA (Micro-Ecological Life Support System Alternative) of the European Space Agency (ESA). The general objective of MELGEN is to establish and validate methods and the related hardware in order to detect genetic instability and microbial contaminants in the MELISSA compartments. This includes (1) a genetic description of the MELISSA strains, (2) studies of microbial behavior and genetic stability in bioreactors and (3) the detection of chemical, genetical and biological contamination and their effect on microbial metabolism. Selected as oxygen producer and complementary food source, the cyanobacterium Arthrospira sp. PCC8005 plays a major role within the MELiSSA loop. As the genomic information on this organism was insufficient, sequencing of its genome was proposed at the French National Sequencing Center, Genoscope, as a joint effort between ESA and different laboratories. So far, a preliminary assembly of 16 contigs representing circa 6.3 million basepairs was obtained. Even though the finishing of the genome is on its way, automatic annotation of the contigs has already been performed on the MaGe annotation platform, and curation of the sequence is currently being carried out, with a special focus on biosynthesis pathways, photosynthesis, and maintenance processes of the cell. According to the index of repetitiveness described by Haubold and Wiehe (2006), we discovered that the genome of Arthrospira sp. is among the 50 most repeated bacterial genomes sequenced to date. Thanks to the sequencing project, we have identified and catalogued mobile genetics elements (MGEs) dispersed throughout the unique chromosome of this cyanobacterium. They represent a quite large proportion of the genome, as genes identified as putative transposases are indeed found in circa 5 Results : We currently have a first draft of the complete genome of Arthrospira sp. PCC 8005, fully annotated. This genomic information opens the gates to a better understanding of the biology of this cyanobacterium and will be a key to the development of appropriate derivatives that provide enhanced performances (e.g. radiation resistance, genetic stability, photosynthesis and nutritive properties).

Morin, Nicolas

354

Efficiency of photosynthesis in a Chl d-utilizing cyanobacterium is comparable to or higher than that in Chl a-utilizing oxygenic species.  

PubMed

The cyanobacterium Acaryochloris marina uses chlorophyll d to carry out oxygenic photosynthesis in environments depleted in visible and enhanced in lower-energy, far-red light. However, the extent to which low photon energies limit the efficiency of oxygenic photochemistry in A. marina is not known. Here, we report the first direct measurements of the energy-storage efficiency of the photosynthetic light reactions in A. marina whole cells, and find it is comparable to or higher than that in typical, chlorophyll a-utilizing oxygenic species. This finding indicates that oxygenic photosynthesis is not fundamentally limited at the photon energies employed by A. marina, and therefore is potentially viable in even longer-wavelength light environments. PMID:21708123

Mielke, S P; Kiang, N Y; Blankenship, R E; Gunner, M R; Mauzerall, D

2011-06-25

355

Role of Two NtcA-Binding Sites in the Complex ntcA Gene Promoter of the Heterocyst-Forming Cyanobacterium Anabaena sp. Strain PCC 7120? †  

PubMed Central

Anabaena sp. strain PCC 7120 is a filamentous cyanobacterium that fixes N2 in specialized cells called heterocysts, which differentiate from vegetative cells in a process that requires the nitrogen control transcription factor NtcA. 2-Oxoglutarate-stimulated binding of purified NtcA to wild-type and modified versions of the ntcA gene promoter from Anabaena sp. was analyzed by mobility shift and DNase I footprinting assays, and the role of NtcA-binding sites in the expression of the ntcA gene during heterocyst differentiation was studied in vivo by using an ntcA-gfp translational fusion and primer extension analysis. Mutation of neither of the two identified NtcA-binding sites eliminated localized expression of ntcA in proheterocysts, but mutation of both sites led to very low, nonlocalized expression.

Olmedo-Verd, Elvira; Valladares, Ana; Flores, Enrique; Herrero, Antonia; Muro-Pastor, Alicia M.

2008-01-01

356

Symplocin A, a Linear Peptide from the Bahamian Cyanobacterium Symploca sp. Configurational Analysis of N,N-Dimethylamino Acids by Chiral-Phase HPLC of Naphthacyl Esters†  

PubMed Central

The absolute stereostructures of the components of symplocin A (3), a new N,N-dimethyl-terminated peptide from the Bahamian cyanobacterium, Symploca sp., were assigned from spectroscopic analysis, including MS and 2D NMR and Marfey’s analysis. The complete absolute configuration of symplocin A, including the unexpected D-configurations of the terminal N,N-dimethylisoleucine and valic acid residues, were assigned by chiral-phase HPLC of the corresponding 2-naphthacyl esters, a highly sensitive, complementary strategy for assignment of N-blocked peptide residues where Marfey’s method is ineffectual, or other methods fall short. Symplocin A exhibited potent activity as an inhibitor of cathepsin E (IC50 300 pM).

Molinski, Tadeusz F.; Reynolds, Kirk A.; Morinaka, Brandon I.

2012-01-01

357

Ability to use the diazo dye, C.I. Acid Black 1 as a nitrogen source by the marine cyanobacterium Oscillatoria curviceps BDU92191.  

PubMed

Ten different strains of marine cyanobacteria were tested for their ability to decolourise and degrade a recalcitrant diazo dye, C.I. Acid Black 1. Of them, Oscillatoria curvicepsBDU92191 was able to grow up to a tested concentration of 500 mG L(-1). The organism degraded 84% of the dye at 100 mG L(-1) in 8 days in a medium free of combined nitrogen. The dye degrading ability is attributed to the activities of the enzymes: laccase, polyphenol oxidase and azoreductase. The absence of the doublet amine peak in addition to the overall reduction of absorption in the IR spectra confirmed the mineralisation of the tested azo dye. The nitrogen assimilating enzyme studies along with nitrogenase assay strongly suggested the ability of the non-heterocystous, filamentous marine cyanobacterium, O. curvicepsBDU92191 to use C.I. Acid Black 1 as a nitrogen source in an oligotrophic environment. PMID:21571528

Priya, Balakrishnan; Uma, Lakshmanan; Ahamed, Abdul Khaleel; Subramanian, Gopalakrishnan; Prabaharan, Dharmar

2011-04-02

358

Symplocin A, a linear peptide from the Bahamian cyanobacterium Symploca sp. Configurational analysis of N,N-dimethylamino acids by chiral-phase HPLC of naphthacyl esters.  

PubMed

The absolute stereostructures of the components of symplocin A (3), a new N,N-dimethyl-terminated peptide from the Bahamian cyanobacterium Symploca sp., were assigned from spectroscopic analysis, including MS, 2D NMR, and Marfey's analysis. The complete absolute configuration of symplocin A, including the unexpected D-configurations of the terminal N,N-dimethylisoleucine and valic acid residues, was assigned by chiral-phase HPLC of the corresponding 2-naphthacyl esters, a highly sensitive, complementary strategy for assignment of N-blocked peptide residues where Marfey's method is ineffectual or other methods fall short. Symplocin A exhibited potent activity as an inhibitor of cathepsin E (IC(50) 300 pM). PMID:22360587

Molinski, Tadeusz F; Reynolds, Kirk A; Morinaka, Brandon I

2012-02-23

359

Characterization of the genes encoding a phosphate-regulated two component sensory system in the marine cyanobacterium Synechococcus sp. WH7803.  

PubMed

An oligomer probe was designed to detect the presence of a putative phoB gene in the genome of the marine, phycoerythrin-containing cyanobacterium Synechococcus sp. WH7803. A 2.2 kb PstI fragment, identified using this probe, was cloned and the complete nucleotide sequence determined. The fragment contained two open reading frames encoding polypeptides which display all the sequence features expected of the response regulator and histidine protein kinase elements of a two component sensory system. Northern analysis confirmed that transcription of these genes was induced by phosphate limitation. On the basis of the sequence similarities and the regulation of their transcription by the availability of inorganic phosphate (Pi) these open reading frames were designated as phoB and phoR, respectively. PMID:8759795

Watson, G M; Scanlan, D J; Mann, N H

1996-08-15

360

A role for osmotic stress-induced proteins in the osmotolerance of a nitrogen-fixing cyanobacterium, Anabaena sp. strain L-31.  

PubMed Central

The molecular basis of tolerance to osmotic stress was investigated with a cyanobacterium, Anabaena sp. strain L-31. The inherent osmotolerance of this strain (50% growth inhibition at 350 mM sucrose) was enhanced by adaptation with 100 mM sucrose for 30 min. Addition of 10 mM KNO3 during growth also conferred significant osmoprotection, but addition of 3 mM NH4Cl did not. Exposure of cells to 350 mM sucrose induced the expression of at least 12 osmotic-stress-induced proteins (OSPs) within 30 min, in the molecular mass range of 11.5 to 84 kDa. Exposure of cells to 100 mM sucrose or to 10 mM nitrate also induced all the OSPs, but addition of ammonium did not. The observed correspondence between the presence of OSPs and osmotolerance strongly suggests a role for OSPs in osmotolerance of Anabaena sp. strain L-31. Images

Iyer, V; Fernandes, T; Apte, S K

1994-01-01

361

Negative Regulation of Expression of the Nitrate Assimilation nirA Operon in the Heterocyst-Forming Cyanobacterium Anabaena sp. Strain PCC 7120?  

PubMed Central

In the filamentous, heterocyst-forming cyanobacterium Anabaena sp. strain PCC 7120, expression of the nitrate assimilation nirA operon takes place in the absence of ammonium and the presence of nitrate or nitrite. Several positive-action proteins that are required for expression of the nirA operon have been identified. Whereas NtcA and NtcB exert their action by direct binding to the nirA operon promoter, CnaT acts by an as yet unknown mechanism. In the genome of this cyanobacterium, open reading frame (ORF) all0605 (the nirB gene) is found between the nirA (encoding nitrite reductase) and ntcB genes. A nirB mutant was able to grow at the expense of nitrate as a nitrogen source and showed abnormally high levels of nirA operon mRNA both in the presence and in the absence of nitrate. This mutant showed increased nitrate reductase activity but decreased nitrite reductase activity, an imbalance that resulted in excretion of nitrite, which accumulated in the extracellular medium, when the nirB mutant was grown in the presence of nitrate. A nirA in-frame deletion mutant also showed a phenotype of increased expression of the nirA operon in the absence of ammonium, independent of the presence of nitrate in the medium. Both NirB and NirA are therefore needed to keep low levels of expression of the nirA operon in the absence of an inducer. Because NirB is also needed to attain high levels of nitrite reductase activity, NirA appears to be a negative element in the nitrate regulation of expression of the nirA operon in Anabaena sp. strain PCC 7120.

Frias, Jose Enrique; Flores, Enrique

2010-01-01

362

Fluorescence changes accompanying short-term light adaptations in photosystem I and photosystem II of the cyanobacterium Synechocystis sp. PCC 6803 and phycobiliprotein-impaired mutants: State 1\\/State 2 transitions and carotenoid-induced quenching of phycobilisomes  

Microsoft Academic Search

The features of the two types of short-term light-adaptations of photosynthetic apparatus, State 1\\/State 2 transitions, and\\u000a non-photochemical fluorescence quenching of phycobilisomes (PBS) by orange carotene-protein (OCP) were compared in the cyanobacterium\\u000a Synechocystis sp. PCC 6803 wild type, CK pigment mutant lacking phycocyanin, and PAL mutant totally devoid of phycobiliproteins. The permanent\\u000a presence of PBS-specific peaks in the in situ

Igor N. Stadnichuk; Evgeny P. Lukashev; Irina V. Elanskaya

2009-01-01

363

Desensitization Mechanism in Prokaryotic Ligand-gated Ion Channel  

PubMed Central

Crystal structures of Gloeobacter violaceus ligand-gated ion channel (GLIC), a proton-gated prokaryotic homologue of pentameric ligand-gated ion channel (LGIC) from G. violaceus, have provided high-resolution models of the channel architecture and its role in selective ion conduction and drug binding. However, it is still unclear which functional states of the LGIC gating scheme these crystal structures represent. Much of this uncertainty arises from a lack of thorough understanding of the functional properties of these prokaryotic channels. To elucidate the molecular events that constitute gating, we have carried out an extensive characterization of GLIC function and dynamics in reconstituted proteoliposomes by patch clamp measurements and EPR spectroscopy. We find that GLIC channels show rapid activation upon jumps to acidic pH followed by a time-dependent loss of conductance because of desensitization. GLIC desensitization is strongly coupled to activation and is modulated by voltage, permeant ions, pore-blocking drugs, and membrane cholesterol. Many of these properties are parallel to functions observed in members of eukaryotic LGIC. Conformational changes in loop C, measured by site-directed spin labeling and EPR spectroscopy, reveal immobilization during desensitization analogous to changes in LGIC and acetylcholine binding protein. Together, our studies suggest conservation of mechanistic aspects of desensitization among LGICs of prokaryotic and eukaryotic origin.

Velisetty, Phanindra; Chakrapani, Sudha

2012-01-01

364

Role of a NifS-like protein from the cyanobacterium Synechocystis PCC 6803 in the maturation of FeS proteins.  

PubMed

In Azotobacter vinelandii and Escherichia coli NifS or NifS-like proteins are involved in FeS protein assembly by mobilizing sulfur from free cysteine. This sulfur together with Fe(2+) is then incorporated into apo-FeS proteins to form an FeS center. A different activity termed C-DES [for cyst(e)ine desulfurylase] was recently isolated from the cyanobacterium Synechocystis PCC 6714 which also mobilized sulfur and which was able to incorporate the FeS center into apoferredoxin. In the genome of the cyanobacterium Synechocystis PCC 6803, there are three open reading frames (orfs) that are similar to NifS and one that is similar to C-DES, indicating that this bacterium might contain both activities, NifS and C-DES. One orf from Synechocystis PCC 6803 encoding a NifS-like protein, slr0387, was overexpressed in E. coli and purified. The molecular mass of the recombinant protein was determined to be about 82 kDa, indicating that it is a homodimer. The absorption spectrum was typical for PLP-containing proteins with an absorption maximum at 390 nm at pH 9.0 and at 425 nm at pH 6.5. The pH dependence of the absorption spectrum correlated with enzyme activity. Maximal activity measured as sulfide production was observed between pH 8.5 and 10. The activity decreased at lower pH values and was undetectable at pH 5.5. pH-dependent changes in the absorption spectrum and activity were attributed to protonation of the Schiff base formed by a lysine side chain and the PLP cofactor. Studies on substrate specificity demonstrated that cysteine derivatives other than cysteine methyl ester and cysteine-sulfinic acid could not serve as substrates for this enzyme. In particular, cystine was not a substrate for the Synechocystis NifS-like protein, whereas it is the best substrate for C-DES. In the presence of Fe(2+), cysteine, and a reductant, the NifS-like protein was able to produce holoferredoxin from apoferredoxin. The implications of two different activities for FeS center biosynthesis in Synechocystis are discussed. PMID:10727236

Jaschkowitz, K; Seidler, A

2000-03-28

365

The hglK gene is required for localization of heterocyst-specific glycolipids in the cyanobacterium Anabaena sp. strain PCC 7120.  

PubMed

Mutant strain 543 of the cyanobacterium Anabaena sp. strain PCC 7120 was originally isolated as a Fox- mutant following chemical mutagenesis. Ultrastructural analysis shows that in nitrogen-replete media the vegetative cells of the mutant are more cylindrical and have thicker septa than those of the wild type, while in nitrogen-free media the mutant heterocysts lack the normal glycolipid layer external to the cell wall. Although this layer is absent, strain 543 heterocysts nevertheless contain heterocyst-specific glycolipids, as determined by thin-layer chromatography. The mutation in strain 543 is in a gene we have named hglK, encoding a protein of 727 amino acids. The wild-type HglK protein appears to contain four membrane-spanning regions followed by 36 repeats of a degenerate pentapeptide sequence, AXLXX. The mutation in strain 543 introduces a termination codon immediately upstream of the pentapeptide repeat region. A mutant constructed by insertion of an antibiotic resistance cassette near the beginning of the hglK gene has the same phenotype as strain 543. We propose that hglK encodes a protein necessary for the localization of heterocyst glycolipids and that this function requires the pentapeptide repeats of the HglK protein. PMID:7592418

Black, K; Buikema, W J; Haselkorn, R

1995-11-01

366

A TPR-family membrane protein gene is required for light-activated heterotrophic growth of the cyanobacterium Synechocystis sp. PCC 6803.  

PubMed

The unicellular cyanobacterium Synechocystis sp. PCC6803 can grow heterotrophically in complete darkness, given that a brief period of illumination is supplemented every day (light-activated heterotrophic growth, LAHG), or under very weak (<0.5 micromol m(-2) s(-1)) but continuous light. By random insertion of the genome with an antibiotic resistance cassette, mutants defective in LAHG were generated. In two identical mutants, sll0886, a tetratricopeptide repeat (TPR)-family membrane protein gene, was disrupted. Targeted insertion of sll0886 and three downstream genes showed that the phenotype was not due to a polar effect. The sll0886 mutant shows normal photoheterotrophic growth when the light intensity is at 2.5 micromol m(-2) s(-1) or above, but no growth at 0.5 micromol m(-2) s(-1). Homologs to sll0886 are also present in cyanobacteria that are not known of LAHG. sll0886 and homologs may be involved in controlling different physiological processes that respond to light of low fluence. PMID:12594026

Kong, Renqiu; Xu, Xudong; Hu, Zhengyu

2003-02-14

367

Fra proteins influencing filament integrity, diazotrophy and localization of septal protein SepJ in the heterocyst-forming cyanobacterium Anabaena sp.  

PubMed

Anabaena sp. strain PCC 7120 is a filamentous cyanobacterium that can fix N(2) in differentiated cells called heterocysts, which exchange nutritional and regulatory compounds with the neighbouring photosynthetic vegetative cells. The cells in the filament appear to be joined by some protein structures, of which SepJ (FraG) that is located at the cell poles in the intercellular septa and is needed for filament integrity seems to be a component. Other known proteins required for filament integrity include FraC and FraH. Whereas fraC (alr2392) was constitutively expressed as an operon together with two downstream genes, alr2393 (fraD) and alr2394 (fraE), fraH (alr1603) was induced under nitrogen deprivation. Single mutants of these genes showed filament fragmentation under nitrogen deprivation and did not grow diazotrophically, although they formed heterocysts. The fraC and fraD mutants showed an impaired localization of SepJ at the intercellular septa and were hampered in the intercellular transfer of the fluorescent probe calcein. As shown with GFP fusions, FraC and FraD are also located at the intercellular septa. Therefore, at least three different proteins, SepJ, FraC and FraD, influence the architecture and function of the intercellular septa in the Anabaena filaments. PMID:20487302

Merino-Puerto, Victoria; Mariscal, Vicente; Mullineaux, Conrad W; Herrero, Antonia; Flores, Enrique

2010-03-01

368

Organization of a large gene cluster encoding ribosomal proteins in the cyanobacterium Synechococcus sp. strain PCC 6301: comparison of gene clusters among cyanobacteria, eubacteria and chloroplast genomes.  

PubMed

The structure of a large gene cluster containing 22 ribosomal protein (r-protein) genes of the cyanobacterium Synechococcus sp. strain PCC6301 is presented. Based on DNA and protein sequence analyses, genes encoding r-proteins L3, L4, L23, L2, S19, L22, S3, L16, L29, S17, L14, L24, L5, S8, L6, L18, S5, L15, L36, S13, S11, L17, SecY, adenylate kinase (AK) and the alpha subunit of RNA polymerase were identified. The gene order is similar to that of the E. coli S10, spc and alpha operons. Unlike the corresponding E. coli operons, the genes for r-proteins S4, S10, S14 and L30 are not present in this cluster. The organization of Synechococcus r-protein genes also resembles that of chloroplast (cp) r-protein genes of red and brown algal species. This strongly supports the endosymbiotic theory that the cp genome evolved from an ancient photosynthetic bacterium. PMID:9300823

Sugita, M; Sugishita, H; Fujishiro, T; Tsuboi, M; Sugita, C; Endo, T; Sugiura, M

1997-08-11

369

The PsbH protein is associated with the inner antenna CP47 and facilitates D1 processing and incorporation into PSII in the cyanobacterium Synechocystis PCC 6803.  

PubMed

Analysis of a number of PSII complexes detectable in the wild-type and mutant cells of the cyanobacterium Synechocystis sp. PCC 6803 showed that the PsbH protein is present in the complexes containing CP47, including unassembled CP47. In a mutant lacking CP47, in which the PSII assembly is stopped at the level of the D1-D2-cytochrome b-559 reaction centre complex, a negligible amount of the PsbH protein was not bound to this complex but was detected in the free form. The results indicate that the PsbH protein has a high affinity for CP47 and during PSII assembly most probably first associates with CP47 and this pair is subsequently attached to the reaction centre complex. Similarly to CP47, the PsbH protein exhibits a slow light-induced degradation in the presence of protein synthesis inhibitor. The absence of the PsbH protein leads to a greatly increased D1 pool that is not associated with other PSII proteins or it is present as a part of the reaction centre complex. We conclude that PsbH is important for the prompt incorporation of the newly synthesized D1 protein into PSII complexes and for the fast D1 maturation. PMID:15970599

Komenda, Josef; Tichý, Martin; Eichacker, Lutz A

2005-06-20

370

Natural osmolytes are much less effective substrates than glycogen for catabolic energy production in the marine cyanobacterium Synechococcus sp. strain PCC 7002.  

PubMed

ADP-glucose pyrophosphorylase, encoded by glgC, catalyzes the first step of glycogen and glucosylglycer(ol/ate) biosynthesis. Here we report the construction of the first glgC null mutant of a marine cyanobacterium (Synechococcus sp. PCC 7002) and investigate its impact on dark anoxic metabolism (autofermentation). The glgC mutant had 98% lower ADP-glucose, synthesized no glycogen and produced appreciably more soluble sugars (mainly sucrose) than wild type (WT). Some glucosylglycerol was still observed, which suggests that the mutant has another, inefficient ADP-glucose synthesis pathway. In contrast, hypersaline conditions (1M NaCl) were lethal to the mutant strain, indicating that, unlike other strains, the elevated sucrose does not compensate for the reduced GG as osmolyte. In contrast to WT, nitrate limitation did not cause bleaching of N-containing pigments or carbohydrate accumulation in the glgC mutant, indicating impaired recycling of nitrogen stores. Despite the 2-fold increase in osmolytes, both the respiration and autofermentation rates of the glgC mutant were appreciably slower (2-4-fold) and correlated quantitatively with the lower fraction of insoluble carbohydrates relative to WT (85% vs. 12%). However, the remaining insoluble carbohydrates still accounted for a high fraction of the carbohydrate catabolized (38%), indicating that insoluble carbohydrates rather than osmolytes were the preferred substrate for autofermentation. PMID:23608552

Guerra, L Tiago; Xu, Yu; Bennette, Nicholas; McNeely, Kelsey; Bryant, Donald A; Dismukes, G Charles

2013-04-19

371

Global Gene Expression Profiles of the Cyanobacterium Synechocystis sp. Strain PCC 6803 in Response to Irradiation with UV-B and White Light  

PubMed Central

We developed a transcript profiling methodology to elucidate expression patterns of the cyanobacterium Synechocystis sp. strain PCC 6803 and used the technology to investigate changes in gene expression caused by irradiation with either intermediate-wavelength UV light (UV-B) or high-intensity white light. Several families of transcripts were altered by UV-B treatment, including mRNAs specifying proteins involved in light harvesting, photosynthesis, photoprotection, and the heat shock response. In addition, UV-B light induced the stringent response in Synechocystis, as indicated by the repression of ribosomal protein transcripts and other mRNAs involved in translation. High-intensity white light- and UV-B-mediated expression profiles overlapped in the down-regulation of photosynthesis genes and induction of heat shock response but differed in several other transcriptional processes including those specifying carbon dioxide uptake and fixation, the stringent response, and the induction profile of the high-light-inducible proteins. These two profile comparisons not only corroborated known physiological changes but also suggested coordinated regulation of many pathways, including synchronized induction of D1 protein recycling and a coupling between decreased phycobilisome biosynthesis and increased phycobilisome degradation. Overall, the gene expression profile analysis generated new insights into the integrated network of genes that adapts rapidly to different wavelengths and intensities of light.

Huang, Lixuan; McCluskey, Michael P.; Ni, Hao; LaRossa, Robert A.

2002-01-01

372

An alternative methionine aminopeptidase, MAP-A, is required for nitrogen starvation and high-light acclimation in the cyanobacterium Synechocystis sp. PCC 6803.  

PubMed

Methionine aminopeptidases (MetAPs or MAPs, encoded by map genes) are ubiquitous and pivotal enzymes for protein maturation in all living organisms. Whereas most bacteria harbour only one map gene, many cyanobacterial genomes contain two map paralogues, the genome of Synechocystis sp. PCC 6803 even three. The physiological function of multiple map paralogues remains elusive so far. This communication reports for the first time differential MetAP function in a cyanobacterium. In Synechocystis sp. PCC 6803, the universally conserved mapC gene (sll0555) is predominantly expressed in exponentially growing cells and appears to be a housekeeping gene. By contrast, expression of mapA (slr0918) and mapB (slr0786) genes increases during stress conditions. The mapB paralogue is only transiently expressed, whereas the widely distributed mapA gene appears to be the major MetAP during stress conditions. A mapA-deficient Synechocystis mutant shows a subtle impairment of photosystem II properties even under non-stressed conditions. In particular, the binding site for the quinone Q(B) is affected, indicating specific N-terminal methionine processing requirements of photosystem II components. MAP-A-specific processing becomes essential under certain stress conditions, since the mapA-deficient mutant is severely impaired in surviving conditions of prolonged nitrogen starvation and high light exposure. PMID:19359320

Drath, Miriam; Baier, Kerstin; Forchhammer, Karl

2009-04-09

373

CRISPR-Cas Systems in the Cyanobacterium Synechocystis sp. PCC6803 Exhibit Distinct Processing Pathways Involving at Least Two Cas6 and a Cmr2 Protein  

PubMed Central

The CRISPR-Cas (Clustered Regularly Interspaced Short Palindrome Repeats – CRISPR associated proteins) system provides adaptive immunity in archaea and bacteria. A hallmark of CRISPR-Cas is the involvement of short crRNAs that guide associated proteins in the destruction of invading DNA or RNA. We present three fundamentally distinct processing pathways in the cyanobacterium Synechocystis sp. PCC6803 for a subtype I-D (CRISPR1), and two type III systems (CRISPR2 and CRISPR3), which are located together on the plasmid pSYSA. Using high-throughput transcriptome analyses and assays of transcript accumulation we found all CRISPR loci to be highly expressed, but the individual crRNAs had profoundly varying abundances despite single transcription start sites for each array. In a computational analysis, CRISPR3 spacers with stable secondary structures displayed a greater ratio of degradation products. These structures might interfere with the loading of the crRNAs into RNP complexes, explaining the varying abundancies. The maturation of CRISPR1 and CRISPR2 transcripts depends on at least two different Cas6 proteins. Mutation of gene sll7090, encoding a Cmr2 protein led to the disappearance of all CRISPR3-derived crRNAs, providing in vivo evidence for a function of Cmr2 in the maturation, regulation of expression, Cmr complex formation or stabilization of CRISPR3 transcripts. Finally, we optimized CRISPR repeat structure prediction and the results indicate that the spacer context can influence individual repeat structures.

Hein, Stephanie; Hess, Wolfgang R.; Backofen, Rolf

2013-01-01

374

Inactivation of the petE gene for plastocyanin lowers photosynthetic capacity and exacerbates chilling-induced photoinhibition in the cyanobacterium Synechococcus.  

PubMed Central

We describe the identification and expression of a petE gene in Synechococcus sp. PCC 7942, a cyanobacterium previously thought to lack plastocyanin. The petE gene is a 420-bp open reading frame that encodes a protein 70 to 75% similar to plastocyanins from other cyanobacteria. Synechococcus possesses a single genomic copy of petE located immediately upstream of the clpB gene. It is transcribed as a single mRNA (550 bases) and, in contrast to most other photobionts, the level of petE expression in Synechococcus is unaffected by variable copper concentrations during acclimated growth. Inactivation of petE does not prevent photoautotrophic growth, but does induce a dramatic increase in mRNA for the alternative electron carrier cytochrome C6. Despite this adjustment, loss of plastocyanin results in slower growth, lower photosystem I content, and a decreased maximum capacity for photosynthetic electron transport. The mutant is also more susceptible to chilling-induced photoinhibition during a shift from 37 to 25 degrees C, at which temperature its inherently lower photosynthetic capacity exacerbates the normal slowing of electron transfer reactions at low temperatures. Under similar conditions, the amount of petE message in the wild type decreases by 50% in the 1st h, but then increases dramatically to almost three times the 37 degrees C level by 9 h.

Clarke, A K; Campbell, D

1996-01-01

375

Inhibition of chlorophyll biosynthesis at the protochlorophyllide reduction step results in the parallel depletion of Photosystem I and Photosystem II in the cyanobacterium Synechocystis PCC 6803.  

PubMed

In most oxygenic phototrophs, including cyanobacteria, two independent enzymes catalyze the reduction of protochlorophyllide to chlorophyllide, which is the penultimate step in chlorophyll (Chl) biosynthesis. One is light-dependent NADPH:protochlorophyllide oxidoreductase (LPOR) and the second type is dark-operative protochlorophyllide oxidoreductase (DPOR). To clarify the roles of both enzymes, we assessed synthesis and accumulation of Chl-binding proteins in mutants of cyanobacterium Synechocystis PCC 6803 that either completely lack LPOR or possess low levels of the active enzyme due to its ectopic regulatable expression. The LPOR-less mutant grew photoautotrophically in moderate light and contained a maximum of 20 % of the wild-type (WT) Chl level. Both Photosystem II (PSII) and Photosystem I (PSI) were reduced to the same degree. Accumulation of PSII was mostly limited by the synthesis of antennae CP43 and especially CP47 as indicated by the accumulation of reaction center assembly complexes. The phenotype of the LPOR-less mutant was comparable to the strain lacking DPOR that also contained <25 % of the wild-type level of PSII and PSI when cultivated under light-activated heterotrophic growth conditions. However, in the latter case, we detected no reaction center assembly complexes, indicating that synthesis was almost completely inhibited for all Chl-proteins, including the D1 and D2 proteins. PMID:23011568

Kope?ná, Jana; Sobotka, Roman; Komenda, Josef

2012-09-26

376

Overexpression of serine hydroxymethyltransferase from halotolerant cyanobacterium in Escherichia coli results in increased accumulation of choline precursors and enhanced salinity tolerance.  

PubMed

Serine hydroxymethyltransferase (SHMT) is a key enzyme in cellular one-carbon pathway and has been studied in many living organisms from bacteria to higher plants and mammals. However, biochemical and molecular characterization of SHMT from photoautotrophic microorganisms remains a challenge. Here, we isolated the SHMT gene from a halotolerant cyanobacterium Aphanothece halophytica (ApSHMT) and expressed it in Escherichia coli. Purified recombinant ApSHMT protein exhibited catalytic reactions for dl-threo-3-phenylserine as well as for l-serine. Catalytic reaction for l-serine was strongly inhibited by NaCl, but not to that level with glycine betaine. Overexpression of ApSHMT in E. coli resulted in the increased accumulation of glycine and serine. Choline and glycine betaine levels were also significantly increased. Under high salinity, the growth rate of ApSHMT-expressing cells was faster compared to its respective control. High salinity also strongly induced the transcript level of ApSHMT in A. halophytica. Our results indicate the importance of a novel pathway; salt-induced ApSHMT increased the level of glycine betaine via serine and choline and conferred the tolerance to salinity stress. PMID:22587350

Waditee-Sirisattha, Rungaroon; Sittipol, Daungjai; Tanaka, Yoshito; Takabe, Teruhiro

2012-06-11

377

Genome-Scale Modeling of Light-Driven Reductant Partitioning and Carbon Fluxes in Diazotrophic Unicellular Cyanobacterium Cyanothece sp. ATCC 51142  

PubMed Central

Genome-scale metabolic models have proven useful for answering fundamental questions about metabolic capabilities of a variety of microorganisms, as well as informing their metabolic engineering. However, only a few models are available for oxygenic photosynthetic microorganisms, particularly in cyanobacteria in which photosynthetic and respiratory electron transport chains (ETC) share components. We addressed the complexity of cyanobacterial ETC by developing a genome-scale model for the diazotrophic cyanobacterium, Cyanothece sp. ATCC 51142. The resulting metabolic reconstruction, iCce806, consists of 806 genes associated with 667 metabolic reactions and includes a detailed representation of the ETC and a biomass equation based on experimental measurements. Both computational and experimental approaches were used to investigate light-driven metabolism in Cyanothece sp. ATCC 51142, with a particular focus on reductant production and partitioning within the ETC. The simulation results suggest that growth and metabolic flux distributions are substantially impacted by the relative amounts of light going into the individual photosystems. When growth is limited by the flux through photosystem I, terminal respiratory oxidases are predicted to be an important mechanism for removing excess reductant. Similarly, under photosystem II flux limitation, excess electron carriers must be removed via cyclic electron transport. Furthermore, in silico calculations were in good quantitative agreement with the measured growth rates whereas predictions of reaction usage were qualitatively consistent with protein and mRNA expression data, which we used to further improve the resolution of intracellular flux values.

Pinchuk, Grigoriy E.; Hill, Eric A.; Kucek, Leo A.; Brown, Roslyn N.; Lipton, Mary S.; Osterman, Andrei; Fredrickson, Jim K.; Konopka, Allan E.; Beliaev, Alexander S.; Reed, Jennifer L.

2012-01-01

378

Modification of energy-transfer processes in the cyanobacterium, Arthrospira platensis, to adapt to light conditions, probed by time-resolved fluorescence spectroscopy.  

PubMed

In cyanobacteria, the interactions among pigment-protein complexes are modified in response to changes in light conditions. In the present study, we analyzed excitation energy transfer from the phycobilisome and photosystem II to photosystem I in the cyanobacterium Arthrospira (Spirulina) platensis. The cells were grown under lights with different spectral profiles and under different light intensities, and the energy-transfer characteristics were evaluated using steady-state absorption, steady-state fluorescence, and picosecond time-resolved fluorescence spectroscopy techniques. The fluorescence rise and decay curves were analyzed by global analysis to obtain fluorescence decay-associated spectra. The direct energy transfer from the phycobilisome to photosystem I and energy transfer from photosystem II to photosystem I were modified depending on the light quality, light quantity, and cultivation period. However, the total amount of energy transferred to photosystem I remained constant under the different growth conditions. We discuss the differences in energy-transfer processes under different cultivation and light conditions. PMID:23605291

Akimoto, Seiji; Yokono, Makio; Aikawa, Shimpei; Kondo, Akihiko

2013-04-21

379

The AplI restriction-modification system in an edible cyanobacterium, Arthrospira (Spirulina) platensis NIES-39, recognizes the nucleotide sequence 5'-CTGCAG-3'.  

PubMed

The degradation of foreign DNAs by restriction enzymes in an edible cyanobacterium, Arthrospira platensis, is a potential barrier for gene-transfer experiments in this economically valuable organism. We overproduced in Escherichia coli the proteins involved in a putative restriction-modification system of A. platensis NIES-39. The protein produced from the putative type II restriction enzyme gene NIES39_K04640 exhibited an endonuclease activity that cleaved DNA within the sequence 5'-CTGCAG-3' between the A at the fifth position and the G at the sixth position. We designated this enzyme AplI. The protein from the adjacent gene NIES39_K04650, which encodes a putative DNA (cytosine-5-)-methyltransferase, rendered DNA molecules resistant to AplI by modifying the C at the fourth position (but not the C at the first position) in the recognition sequence. This modification enzyme, M.AplI, should be useful for converting DNA molecules into AplI-resistant forms for use in gene-transfer experiments. A summary of restriction enzymes in various Arthrospira strains is also presented in this paper. PMID:23563565

Shiraishi, Hideaki; Tabuse, Yosuke

2013-04-07

380

The Presence of Glutamate Dehydrogenase Is a Selective Advantage for the Cyanobacterium Synechocystis sp. Strain PCC 6803 under Nonexponential Growth Conditions  

PubMed Central

The unicellular cyanobacterium Synechocystis sp. strain PCC 6803 has two putative pathways for ammonium assimilation: the glutamine synthetase-glutamate synthase cycle, which is the main one and is finely regulated by the nitrogen source; and a high NADP-dependent glutamate dehydrogenase activity (NADP-GDH) whose contribution to glutamate synthesis is uncertain. To investigate the role of the latter, we used two engineered mutants, one lacking and another overproducing NADP-GDH. No major disturbances in the regulation of nitrogen-assimilating enzymes or in amino acids pools were detected in the null mutant, but phycobiline content, a sensitive indicator of the nutritional state of cyanobacterial cells, was significantly reduced, indicating that NADP-GDH plays an auxiliary role in ammonium assimilation. This effect was already prominent in the initial phase of growth, although differences in growth rate between the wild type and the mutants were observed at this stage only at low light intensities. However, the null mutant was unable to sustain growth at the late stage of the culture at the point when the wild type showed the maximum NADP-GDH activity, and died faster in ammonium-containing medium. Overexpression of NADP-GDH improved culture proliferation under moderate ammonium concentrations. Competition experiments between the wild type and the null mutant confirmed that the presence of NADP-GDH confers a selective advantage to Synechocystis sp. strain PCC 6803 in late stages of growth.

Chavez, S.; Lucena, J. M.; Reyes, J. C.; Florencio, F. J.; Candau, P.

1999-01-01

381

A Novel Mechanism of Osmosensing, a Salt-dependent Protein-Nucleic Acid Interaction in the Cyanobacterium Synechocystis Species PCC 6803  

PubMed Central

The de novo synthesis of compatible solutes is an essential part of the cellular osmotic stress response. Upon an osmotic challenge, it is regulated by the immediate biochemical activation of preformed enzymes and by activation of gene expression. Whereas the transcriptional response has been investigated intensively, the mechanisms by which enzymes are activated in osmotic stress situations are still elusive. Here, we address this topic for the moderately halotolerant cyanobacterium Synechocystis sp. PCC 6803, which synthesizes glucosylglycerol as a compatible solute. The key enzyme of the glucosylglycerol pathway (GgpS) is inhibited by nucleic acids in a sequence- and length-independent manner. The protein binds DNA, RNA, and heparin via a salt-dependent electrostatic interaction with the negatively charged backbone of the polyanions. Mechanistically, DNA binding to the enzyme causes noncompetitive inhibition of GgpS activity. The interaction of the enzyme and nucleic acids under in vivo conditions is indicated by the co-purification of both after cross-linking in Synechocystis cells. We propose a novel mechanism of activity regulation by the nonspecific salt-dependent binding of an enzyme to nucleic acids.

Novak, Jens F.; Stirnberg, Marit; Roenneke, Benjamin; Marin, Kay

2011-01-01

382

Long-term microclimatic stress causes rapid adaptive radiation of kaiABC clock gene family in a cyanobacterium, Nostoc linckia, from "Evolution Canyons" I and II, Israel.  

PubMed

Cyanobacteria are the only prokaryotes known thus far possessing regulation of physiological functions with approximate daily periodicity, or circadian rhythms, that are controlled by a cluster of three genes, kaiA, kaiB, and kaiC. Here we demonstrate considerably higher genetic polymorphism and extremely rapid evolution of the kaiABC gene family in a filamentous cyanobacterium, Nostoc linckia, permanently exposed to the acute natural environmental stress in the two microsite evolutionary models known as "Evolution Canyons," I (Mount Carmel) and II (Upper Galilee) in Israel. The family consists of five distinct subfamilies (kaiI-kaiV) comprising at least 20 functional genes and pseudogenes. The obtained data suggest that the duplications of kai genes have adaptive significance, and some of them are evolutionarily quite recent (approximately 80,000 years ago). The observed patterns of within- and between-subfamily polymorphisms indicate that positive diversifying, balancing, and purifying selections are the principal driving forces of the kai gene family's evolution. PMID:11842226

Dvornyk, Volodymyr; Vinogradova, Oxana; Nevo, Eviatar

2002-02-12

383

Three-dimensional model and characterization of the iron stress-induced CP43'-photosystem I supercomplex isolated from the cyanobacterium Synechocystis PCC 6803.  

PubMed

The cyanobacterium Synechocystis PCC 6803 has been subjected to growth under iron-deficient conditions. As a consequence, the isiA gene is expressed, and its product, the chlorophyll a-binding protein CP43', accumulates in the cell. Recently, we have shown for the first time that 18 copies of this photosystem II (PSII)-like chlorophyll a-binding protein forms a ring around the trimeric photosystem I (PSI) reaction center (Bibby, T. S., Nield, J., and Barber, J. (2001) Nature, 412, 743-745). Here we further characterize the biochemical and structural properties of this novel CP43'-PSI supercomplex confirming that it is a functional unit of approximately 1900 kDa where the antenna size of PSI is increased by 70% or more. Using electron microscopy and single particle analysis, we have constructed a preliminary three-dimensional model of the CP43'-PSI supercomplex and used it as a framework to incorporate higher resolution structures of PSI and CP43 recently derived from x-ray crystallography. Not only does this work emphasize the flexibility of cyanobacterial light-harvesting systems in response to the lowering of phycobilisome and PSI levels under iron-deficient conditions, but it also has implications for understanding the organization of the related chlorophyll a/b-binding Pcb proteins of oxychlorobacteria, formerly known as prochlorophytes. PMID:11518716

Bibby, T S; Nield, J; Barber, J

2001-08-22

384

Identification of salt-regulated genes in the genome of the cyanobacterium Synechocystis sp. strain PCC 6803 by subtractive RNA hybridization.  

PubMed

To identify genes transcribed preferentially under salt stress, a subtractive RNA hybridization procedure was applied to the cyanobacterium Synechocystis sp. PCC 6803. The screening of a genomic library led to the identification of several RNA species that were more abundant in salt-stressed cells than in control cells. Salt-dependent transcription of the identified genes was verified in Northern blot experiments. In addition to the previously characterized genes cpn60 (encoding GroEL; a molecular chaperone) and isiA (encoding a chlorophyll-binding protein), genes encoding a protein of unknown function (slr0082) and a putative RNA helicase (slr0083) were identified as salt-regulated genes in Synechocystis. Genes slr0082 and slr0083, located at sites adjacent to each other on the Synechocystis chromosome, were transcribed from separate promoters and showed the most significant induction 1-3 h after salt shock. The salt-regulated promoters of these two genes were mapped. Genes cpn60, slr0082, and slr0083 were also found to be induced by a cold shock. The possible role of the identified gene products for salt adaptation of Synechocystis is discussed. PMID:10591847

Vinnemeier, J; Hagemann, M

1999-12-01

385

The small CAB-like proteins of the cyanobacterium Synechocystis sp. PCC 6803: their involvement in chlorophyll biogenesis for Photosystem II.  

PubMed

The five small CAB-like proteins (ScpA-E) of the cyanobacterium Synechocystis sp. PCC 6803 belong to the family of stress-induced light-harvesting-like proteins, but are constitutively expressed in a mutant deficient of Photosystem I (PSI). Using absorption, fluorescence and thermoluminescence measurements this PSI-less strain was compared with a mutant, in which all SCPs were additionally deleted. Depletion of SCPs led to structural rearrangements in Photosystem II (PSII): less photosystems were assembled; and in these, the Q(B) site was modified. Despite the lower amount of PSII, the SCP-deficient cells contained the same amount of phycobilisomes (PBS) as the control. Although the excess PBS were functionally disconnected, their fluorescence was quenched under high irradiance by the activated Orange Carotenoid Protein (OCP). Additionally the amount of OCP, but not of the iron-stress induced protein (isiA), was higher in this SCP-depleted mutant compared with the control. As previously described, the lack of SCPs affects the chlorophyll biosynthesis (Vavilin, D., Brune, D. C., Vermaas, W. (2005) Biochim Biophys Acta 1708, 91-101). We demonstrate that chlorophyll synthesis is required for efficient PSII repair and that it is partly impaired in the absence of SCPs. At the same time, the amount of chlorophyll also seems to influence the expression of ScpC and ScpD. PMID:21605542

Hernandez-Prieto, Miguel A; Tibiletti, Tania; Abasova, Leyla; Kirilovsky, Diana; Vass, Imre; Funk, Christiane

2011-05-14

386

CP43', the isiA gene product, functions as an excitation energy dissipator in the cyanobacterium Synechococcus sp. PCC 7942.  

PubMed

Under conditions of iron deficiency certain cyanobacteria induce a chlorophyll (Chl)-binding protein, CP43', which is encoded by the isiA gene. We have previously suggested that CP43' functions as a nonradiative dissipator of light energy. To further substantiate its functional role an isiA overexpression construct was introduced into the genome of a cyanobacterium Synechococcus sp. PCC 7942 (giving isiAoe cells). The presence of functional CP43' in isiAoe cells was confirmed by Western blot as well as by the presence of a characteristic blueshift of the red Chl a absorption peak and a notable increase in the 77 K fluorescence peak at 685 nm. Compared to wild-type cells isiAoe cells, with induced CP43', had both smaller functional antenna size and decreased yields of room temperature Chl fluorescence at various light irradiances. These observations strongly suggest that isiAoe cells, with induced CP43', have an increased capacity for dissipating light energy as heat. In agreement with this hypothesis isiAoe cells were also more resistant to photoinhibition of photosynthesis than wild-type cells. Based on these results we have further strengthened the hypothesis that CP43' functions as a nonradiative dissipator of light energy, thus protecting photosystem II from excessive excitation under iron-deficient conditions. PMID:11594057

Sandström, S; Park, Y I; Oquist, G; Gustafsson, P

2001-09-01

387

Expression, purification, crystallization and preliminary X-ray crystallographic studies of alkyl hydroperoxide reductase (AhpC) from the cyanobacterium Anabaena sp. PCC 7120  

PubMed Central

Alkyl hydroperoxide reductase (AhpC) is a key component of a large family of thiol-specific antioxidant (TSA) proteins distributed among prokaryotes and eukaryotes. AhpC is involved in the detoxification of reactive oxygen species (ROS) and reactive sulfur species (RSS). Sequence analysis of AhpC from the cyanobacterium Anabaena sp. PCC 7120 shows that this protein belongs to the 1-­Cys class of peroxiredoxins (Prxs). It has recently been reported that enhanced expression of this protein in Escherichia coli offers tolerance to multiple stresses such as heat, salt, copper, cadmium, pesticides and UV-B. However, the structural features and the mechanism behind this process remain unclear. To provide insights into its biochemical function, AhpC was expressed, purified and crystallized by the hanging-drop vapour-diffusion method. Diffraction data were collected to a maximum d-spacing of 2.5?Å using synchrotron radiation. The crystal belonged to space group P212121, with unit-cell parameters a = 80, b = 102, c = 109.6?Å. The structure of AhpC from Anabaena sp. PCC 7120 was determined by molecular-replacement methods using the human Prx enzyme hORF6 (PDB entry 1prx) as the template.

Mishra, Yogesh; Hall, Michael; Chaurasia, Neha; Rai, Lal Chand; Jansson, Stefan; Schroder, Wolfgang P.; Sauer, Uwe H.

2011-01-01

388

Long-term microclimatic stress causes rapid adaptive radiation of kaiABC clock gene family in a cyanobacterium, Nostoc linckia, from "Evolution Canyons" I and II, Israel  

PubMed Central

Cyanobacteria are the only prokaryotes known thus far possessing regulation of physiological functions with approximate daily periodicity, or circadian rhythms, that are controlled by a cluster of three genes, kaiA, kaiB, and kaiC. Here we demonstrate considerably higher genetic polymorphism and extremely rapid evolution of the kaiABC gene family in a filamentous cyanobacterium, Nostoc linckia, permanently exposed to the acute natural environmental stress in the two microsite evolutionary models known as “Evolution Canyons,” I (Mount Carmel) and II (Upper Galilee) in Israel. The family consists of five distinct subfamilies (kaiI–kaiV) comprising at least 20 functional genes and pseudogenes. The obtained data suggest that the duplications of kai genes have adaptive significance, and some of them are evolutionarily quite recent (?80,000 years ago). The observed patterns of within- and between-subfamily polymorphisms indicate that positive diversifying, balancing, and purifying selections are the principal driving forces of the kai gene family's evolution.

Dvornyk, Volodymyr; Vinogradova, Oxana; Nevo, Eviatar

2002-01-01

389

A Zinc(II)\\/Lead(II)\\/Cadmium(II)Inducible Operon from the Cyanobacterium Anabaena Is Regulated by AztR, an ?3N ArsR\\/SmtB Metalloregulator †  

Microsoft Academic Search

A novel Zn(II)\\/Pb(II)\\/Cd(II)-responsive operon that consists of genes encoding a Zn(II)\\/Pb- (II) CPx-ATPase efflux pump (aztA) and a Zn(II)\\/Cd(II)\\/Pb(II)-specific SmtB\\/ArsR family repressor ( aztR) has been identified and characterized from the cyanobacterium Anabaena PCC 7120. In vivo real time quantitative RT-PCR assays reveal that both aztR and aztA expression are induced by divalent metal ions Zn(II), Cd(II), and Pb(II) but

Tong Liu; James W. Golden; David P. Giedroc

2005-01-01

390

Effect of Metal Cations on the Viscosity of a Pectin-Like Capsular Polysaccharide from the Cyanobacterium Microcystis flos-aquae C3-40  

PubMed Central

The properties of purified capsular polysaccharide from the cyanobacterium Microcystis flos-aquae C3-40 were examined by capillary viscometry. Capsule suspensions exhibited similar viscosities between pH 6 and 10 but were more viscous at pH <=4 than at pH 6 to 11. At pH 7, a biphasic effect of metal ion concentration on capsule viscosity was observed: (i) capsule viscosity increased with increasing metal ion concentration until a maximal viscosity occurred at a specific concentration that was a reproducible characteristic of each metal ion, and (ii) the viscosity decreased with further addition of that ion. Because the latter part of the biphasic curve was complicated by additional factors (especially the precipitation or gelation of capsule by divalent metal ions), the effects of various metal chlorides were compared for the former phase in which capsule viscosity increased in the presence of metal ions. Equivalent increases in capsule viscosity were observed with micromolar concentrations of divalent metal ions but only with 10 to 20 times greater concentrations of Na(sup+). The relative abilities of various metal salts to increase capsule viscosity were as follows: CdCl(inf2), Pb(NO(inf3))(inf2), FeCl(inf2) > MnCl(inf2) > CuCl(inf2), CaCl(inf2) >> NaCl. This pattern of metal efficacy resembles known cation influences on the structural integrity of capsule in naturally occurring and cultured M. flos-aquae colonies. The data are the first direct demonstration of an interaction between metal ions and purified M. flos-aquae capsule, which has previously been proposed to play a role in the environmental cycling of certain multivalent metals, especially manganese. The M. flos-aquae capsule and the plant polysaccharide pectin have similar sugar compositions but differ in their relative responses to various metals, suggesting that capsular polysaccharide could be a preferable alternative to pectin for certain biotechnological applications.

Parker, D. L.; Schram, B. R.; Plude, J. L.; Moore, R. E.

1996-01-01

391

Ethoxyzolamide Differentially Inhibits CO2 Uptake and Na+-Independent and Na+-Dependent HCO3- Uptake in the Cyanobacterium Synechococcus sp. UTEX 625.  

PubMed Central

The effects of ethoxyzolamide (EZ), a carbonic anhydrase inhibitor, on the active CO2 and Na+-independent and Na+-dependent HCO3- transport systems of the unicellular cyanobacterium Synechococcus sp. UTEX 625 were examined. Measurements of transport and accumulation using radiochemical, fluorometric, and mass spectrometric assays indicated that active CO2 transport and active Na+-independent HCO3- transport were inhibited by EZ. However, Na+-independent HCO3- transport was about 1 order of magnitude more sensitive to EZ inhibition than was CO2 transport (50% inhibition = 12 [mu]M versus 80 [mu]M). The data suggest that both the active CO2 (G.D. Price, M.R. Badger [1989] Plant Physiol 89: 37-43) and the Na+ -independent HCO3 - transport systems possessed carbonic anhydrase-like activity as part of their mechanism of action. In contrast, Na+-dependent HCO3- transport was only partially (50% inhibition = 230 [mu]M) and noncompetitively inhibited by EZ. The collective evidence suggested that EZ inhibition of Na+ -dependent HCO3- transport was an indirect consequence of the action of EZ on the CO2 transport system, rather than a direct effect on HCO3- transport. A model is presented in which the core of the inorganic carbon translocating system is formed by Na+-dependent HCO3- transport and the CO2 transport system. It is argued that the Na+-independent HCO3 - utilizing system was not directly involved in translocation, but converted HCO3- to CO2 for use in CO2 transport.

Tyrrell, P. N.; Kandasamy, R. A.; Crotty, C. M.; Espie, G. S.

1996-01-01

392

DNA sequence and regulation of the gene (cbpA) encoding the 42-kilodalton cytoplasmic membrane carotenoprotein of the cyanobacterium Synechococcus sp. strain PCC 7942  

SciTech Connect

The gene (cbpA) coding for a carotenoid-binding protein of the cyanobacterium Synechococcus sp. strain PCC 7942 (Anacystis nidulans R2) has been cloned and sequenced. A polyclonal antibody against the protein was used to identify immunoreactive clones from a {lambda}gt11 expression library of Synechococcus strain PCC 7942. The initial positive clone ({lambda}gtAN42) contained a 0.9-kilobase (kb) chromosomal fragment, which was used to detect a larger chromosomal fragment from a {lambda}EMBL3 library. The {lambda}EMBL3 recombinant, {lambda}EM109, contained an 18-kb portion of the Synechococcus strain PCC 7942 chromosome. The open reading frame of cbpA encoded 450 amino acids which give rise to a protein of 49,113 daltons. The hydrophobicity plot indicates that the protein may have a 49-residue signal sequence which is cleaved to yield a mature protein of 43,709 daltons. The protein has been localized in the cytoplasmic membrane by biochemical procedures as well as by electron microscopic immunocytochemistry. Northern (RNA) blot analysis indicates that transcription of cbpA is tightly regulated by DNA topology, light intensity, and iron concentration. Transcription is greatly induced by growth under high light intensities and repressed during growth under iron-deficient conditions. The DNA gyrase inhibitor novobiocin specifically inhibited the light-induced transcription. In Northern blots, the gene-specific probe hybridized to two size classes of RNA, with lengths of 2.0 and 6.2 kb. Since cbpA appears to be a component of the 6.2-kb transcript, it is likely part of a larger operon.

Reddy, K.J.; Masamoto, K.; Sherman, D.M.; Sherman, L.A. (Univ. of Missouri-Columbia (USA))

1989-06-01

393

The NtcA-Regulated amtB Gene Is Necessary for Full Methylammonium Uptake Activity in the Cyanobacterium Synechococcus elongatus?  

PubMed Central

The Amt proteins constitute a ubiquitous family of transmembrane ammonia channels that permit the net uptake of ammonium by cells. In many organisms, there is more than one amt gene, and these genes are subjected to nitrogen control. The mature Amt protein is a homo- or heterooligomer of three Amt subunits. We previously characterized an amt1 gene in the unicellular cyanobacterium Synechococcus elongatus strain PCC 7942. In this work, we describe the presence in this organism of a second amt gene, amtB, which encodes a protein more similar to the bacterial AmtB proteins than to any other characterized cyanobacterial Amt protein. The expression of amtB took place in response to nitrogen step-down, required the NtcA transcription factor, and occurred parallel to the expression of amt1. However, the transcript levels of amtB measured after 2 h of nitrogen deprivation were about 100-fold lower than those of amt1. An S. elongatus amtB insertional mutant exhibited an activity for uptake of [14C]methylammonium that was about 55% of that observed in the wild type, but inactivation of amtB had no noticeable effect on the uptake of ammonium when it was supplied at a concentration of 100 ?M or more. Because an S. elongatus amt1 mutant is essentially devoid of [14C]methylammonium uptake activity, the mature Amt transporter is functional in the absence of AmtB subunits but not in the absence of Amt1 subunits. However, the S. elongatus amtB mutant could not concentrate [14C]methylammonium within the cells to the same extent as the wild type. Therefore, AmtB is necessary for full methylammonium uptake activity in S. elongatus.

Paz-Yepes, Javier; Herrero, Antonia; Flores, Enrique

2007-01-01

394

Expression and Mutational Analysis of the glnB Genomic Region in the Heterocyst-Forming Cyanobacterium Anabaena sp. Strain PCC 7120?  

PubMed Central

In the filamentous, heterocyst-forming cyanobacterium Anabaena sp. strain PCC 7120, the glnB gene is expressed at considerable levels both in the presence and in the absence of combined nitrogen, although induction, influenced by NtcA, takes place upon combined-nitrogen deprivation likely localized to vegetative cells. In spite of extensive efforts, a derivative of PCC 7120 lacking a functional glnB gene could be obtained only with constructs that lead to overexpression of a downstream open reading frames (ORF), particularly all2318. Strain CSP10 [glnB all2318(Con)] exhibited growth rates similar to those of the wild type when it was using nitrate or ammonium, but its diazotrophic growth was impaired. However, it differentiated heterocysts with a time course and distribution pattern similar to those of the wild type, although with no cyanophycin-containing polar granules, and exhibited impaired nitrogenase activity under oxic conditions, but not under microoxic conditions. In the mutant, NtcA-dependent inducion of the hetC and nifH genes was unaltered, but induction of the urtA gene and urea transport activity were increased. Active uptake of nitrite was also increased and insensitive to the ammonium-promoted inhibition observed for the wild type. Thus, regulation of the nitrite transport activity requires the glnB gene product. In the presence of a wild-type glnB gene, neither inactivation nor overexpression of all2318 produced an apparent phenotype. Thus, in an otherwise wild-type background, the glnB gene appears to be essential for growth of strain PCC 7120. For growth with combined nitrogen but not for diazotrophic growth, the requirement for glnB can be overridden by increasing the expression of all2318 (and/or ORFs downstream of it).

Paz-Yepes, Javier; Flores, Enrique; Herrero, Antonia

2009-01-01

395

Identification of Homophenylalanine Biosynthetic Genes from the Cyanobacterium Nostoc punctiforme PCC73102 and Application to Its Microbial Production by Escherichia coli  

PubMed Central

l-Homophenylalanine (l-Hph) is a useful chiral building block for synthesis of several drugs, including angiotensin-converting enzyme inhibitors and the novel proteasome inhibitor carfilzomib. While the chemoenzymatic route of synthesis is fully developed, we investigated microbial production of l-Hph to explore the possibility of a more efficient and sustainable approach to l-Hph production. We hypothesized that l-Hph is synthesized from l-Phe via a mechanism homologous to 3-methyl-2-oxobutanoic acid conversion to 4-methyl-2-oxopentanoic acid during leucine biosynthesis. Based on bioinformatics analysis, we found three putative homophenylalanine biosynthesis genes, hphA (Npun_F2464), hphB (Npun_F2457), and hphCD (Npun_F2458), in the cyanobacterium Nostoc punctiforme PCC73102, located around the gene cluster responsible for anabaenopeptin biosynthesis. We constructed Escherichia coli strains harboring hphABCD-expressing plasmids and achieved the fermentative production of l-Hph from l-Phe. To our knowledge, this is the first identification of the genes responsible for homophenylalanine synthesis in any organism. Furthermore, to improve the low conversion efficiency of the initial strain, we optimized the expression of hphA, hphB, and hphCD, which increased the yield to ?630 mg/liter. The l-Hph biosynthesis and l-Leu biosynthesis genes from E. coli were also compared. This analysis revealed that HphB has comparatively relaxed substrate specificity and can perform the function of LeuB, but HphA and HphCD show tight substrate specificity and cannot complement the LeuA and LeuC/LeuD functions, and vice versa. Finally, the range of substrate tolerance of the l-Hph-producing strain was examined, which showed that m-fluorophenylalanine, o-fluorophenylalanine, and l-tyrosine were accepted as substrates and that the corresponding homoamino acids were generated.

Mitsuhashi, Satoshi; Tabata, Kazuhiko

2013-01-01

396

?-Tocopherol Plays a Role in Photosynthesis and Macronutrient Homeostasis of the Cyanobacterium Synechocystis sp. PCC 6803 That Is Independent of Its Antioxidant Function1  

PubMed Central

?-Tocopherol is synthesized exclusively in oxygenic phototrophs and is known to function as a lipid-soluble antioxidant. Here, we report that ?-tocopherol also has a novel function independent of its antioxidant properties in the cyanobacterium Synechocystis sp. PCC 6803. The photoautotrophic growth rates of wild type and mutants impaired in ?-tocopherol biosynthesis are identical, but the mutants exhibit elevated photosynthetic activities and glycogen levels. When grown photomixotrophically with glucose (Glc), however, these mutants cease growth within 24 h and exhibit a global macronutrient starvation response associated with nitrogen, sulfur, and carbon, as shown by decreased phycobiliprotein content (35% of the wild-type level) and accumulation of the nblA1-nblA2, sbpA, sigB, sigE, and sigH transcripts. Photosystem II activity and carboxysome synthesis are lost in the tocopherol mutants within 24 h of photomixotrophic growth, and the abundance of carboxysome gene (rbcL, ccmK1, ccmL) and ndhF4 transcripts decreases to undetectable levels. These results suggest that ?-tocopherol plays an important role in optimizing photosynthetic activity and macronutrient homeostasis in Synechocystis sp. PCC 6803. Several lines of evidence indicate that increased oxidative stress in the tocopherol mutants is unlikely to be the underlying cause of photosystem II inactivation and Glc-induced lethality. Interestingly, insertional inactivation of the pmgA gene, which encodes a putative serine-threonine kinase similar to RsbW and RsbT in Bacillus subtilis, results in a similar increase in glycogen and Glc-induced lethality. Based on these results, we propose that ?-tocopherol plays a nonantioxidant regulatory role in photosynthesis and macronutrient homeostasis through a signal transduction pathway that also involves PmgA.

Sakuragi, Yumiko; Maeda, Hiroshi; DellaPenna, Dean; Bryant, Donald A.

2006-01-01

397

Investigation of the excited states dynamics in the Chl d- containing cyanobacterium Acaryochloris marina by time and wavelength correlated single-photon counting  

NASA Astrophysics Data System (ADS)

The phototrophic cyanobacterium Acaryochloris marina discovered in 1996 has a unique composition of the light harvesting system. The chlorophyll (Chl) antenna contains mainly Chl d instead of the usually dominant Chl a and the Phycobiliprotein (PBP) antenna has a simpler rod shaped structure than in typical cynobacteria [1]. The interaction of the photosynthetic subunits and especially the mechanisms regulating the energy transfer under different stress conditions are presently interesting and open fields in photosynthesis research. In this study we use time- and wavelength-resolved single photon counting to investigate the excited states dynamics in living cells of A.marina. The fluorescence dynamics is synchronistically monitored in the visible and near infrared spectrum with high signal to noise ratio and short data acquisition times while using low excitation light intensities. These attributes are necessary to investigate photosynthetic processes in sensitive biological samples, when the light emission varies due to metabolic changes. The results suggest a fast excitation energy transfer kinetics of 20-30 ps along the PBP antenna of A.marina followed by a transfer of about 60 ps to the Chl d antenna. Cells of A. marina which are stored at 0°C for some time show a decoupling of the PBP antenna, which is partially reversible when the sample is kept at 25 °C for a short time. Decoupling effects appearing after strong illumination with white light (1600 W/m2) suggest a mechanism which removes the PBP antenna at different stress conditions to avoid photo damage of the reaction center of Photosystem II (PS II).

Schmitt, Franz-Josef; Theiss, Christoph; Wache, Karin; Fuesers, Justus; Andree, Stefan; Handojo, Andrianto; Karradt, Anne; Kiekebusch, Daniela; Eichler, Hans Joachim; Eckert, Hann-Jörg

2006-11-01

398

Microarray Analysis of the Genome-Wide Response to Iron Deficiency and Iron Reconstitution in the Cyanobacterium Synechocystis sp. PCC 68031[w  

PubMed Central

A full-genome microarray of the (oxy)photosynthetic cyanobacterium Synechocystis sp. PCC 6803 was used to identify genes that were transcriptionally regulated by growth in iron (Fe)-deficient versus Fe-sufficient media. Transcript accumulation for 3,165 genes in the genome was analyzed using an analysis of variance model that accounted for slide and replicate (random) effects and dye (a fixed) effect in testing for differences in the four time periods. We determined that 85 genes showed statistically significant changes in the level of transcription (P ? 0.05/3,165 = 0.0000158) across the four time points examined, whereas 781 genes were characterized as interesting (P ? 0.05 but greater than 0.0000158; 731 of these had a fold change >1.25×). The genes identified included those known previously to be Fe regulated, such as isiA that encodes a novel chlorophyll-binding protein responsible for the pigment characteristics of low-Fe (LoFe) cells. ATP synthetase and phycobilisome genes were down-regulated in LoFe, and there were interesting changes in the transcription of genes involved in chlorophyll biosynthesis, in photosystem I and II assembly, and in energy metabolism. Hierarchical clustering demonstrated that photosynthesis genes, as a class, were repressed in LoFe and induced upon the re-addition of Fe. Specific regulatory genes were transcriptionally active in LoFe, including two genes that show homology to plant phytochromes (cph1 and cph2). These observations established the existence of a complex network of regulatory interactions and coordination in response to Fe availability.

Singh, Abhay K.; McIntyre, Lauren M.; Sherman, Louis A.

2003-01-01

399

Mutagenesis of hetR Reveals Amino Acids Necessary for HetR Function in the Heterocystous Cyanobacterium Anabaena sp. Strain PCC 7120?  

PubMed Central

HetR is the master regulator of heterocyst differentiation in the filamentous cyanobacterium Anabaena sp. strain PCC 7120. Genetic selection was used to identify 33 amino acid substitutions in HetR that reduced the proportion of cells undergoing heterocyst differentiation to less than 2%. Conservative substitutions in the wild-type HetR protein revealed three mutations that dramatically reduced the amount of heterocyst differentiation when the mutant allele was present in place of the wild-type allele on a replicating plasmid in a mutant lacking hetR on the chromosome. An H69Y substitution resulted in heterocyst formation among less than 0.1% of cells, and D17E and G36A substitutions resulted in a Het? phenotype, compared to heterocyst formation among approximately 25% of cells with the wild-type hetR under the same conditions. The D17E substitution prevented DNA binding activity exhibited by wild-type HetR in mobility shift assays, whereas G36A and H69Y substitutions had no affect on DNA binding. D17E, G36A, and H69Y substitutions also resulted in higher levels of the corresponding HetR protein than of the wild-type protein when each was expressed from an inducible promoter in a hetR deletion strain, suggesting an effect on HetR protein turnover. Surprisingly, C48A and S152A substitutions, which were previously reported to result in a Het? phenotype, were found to have no effect on heterocyst differentiation or patterning when the corresponding mutations were introduced into an otherwise wild-type genetic background in Anabaena sp. strain PCC 7120. The clustering of mutations that satisfied the positive selection near the amino terminus suggests an important role for this part of the protein in HetR function.

Risser, Douglas D.; Callahan, Sean M.

2007-01-01

400

Mutagenesis of hetR reveals amino acids necessary for HetR function in the heterocystous cyanobacterium Anabaena sp. strain PCC 7120.  

PubMed

HetR is the master regulator of heterocyst differentiation in the filamentous cyanobacterium Anabaena sp. strain PCC 7120. Genetic selection was used to identify 33 amino acid substitutions in HetR that reduced the proportion of cells undergoing heterocyst differentiation to less than 2%. Conservative substitutions in the wild-type HetR protein revealed three mutations that dramatically reduced the amount of heterocyst differentiation when the mutant allele was present in place of the wild-type allele on a replicating plasmid in a mutant lacking hetR on the chromosome. An H69Y substitution resulted in heterocyst formation among less than 0.1% of cells, and D17E and G36A substitutions resulted in a Het- phenotype, compared to heterocyst formation among approximately 25% of cells with the wild-type hetR under the same conditions. The D17E substitution prevented DNA binding activity exhibited by wild-type HetR in mobility shift assays, whereas G36A and H69Y substitutions had no affect on DNA binding. D17E, G36A, and H69Y substitutions also resulted in higher levels of the corresponding HetR protein than of the wild-type protein when each was expressed from an inducible promoter in a hetR deletion strain, suggesting an effect on HetR protein turnover. Surprisingly, C48A and S152A substitutions, which were previously reported to result in a Het- phenotype, were found to have no effect on heterocyst differentiation or patterning when the corresponding mutations were introduced into an otherwise wild-type genetic background in Anabaena sp. strain PCC 7120. The clustering of mutations that satisfied the positive selection near the amino terminus suggests an important role for this part of the protein in HetR function. PMID:17220221

Risser, Douglas D; Callahan, Sean M

2007-01-12

401

Independent regulation of nifHDK operon transcription and DNA rearrangement during heterocyst differentiation in the cyanobacterium Anabaena sp. strain PCC 7120.  

PubMed Central

The filamentous cyanobacterium Anabaena sp. strain PCC 7120 expresses the genes required for nitrogen fixation in terminally differentiated cells called heterocysts. The nifHDK operon encodes the nitrogenase polypeptides and is expressed at high levels in heterocysts. During heterocyst differentiation, an 11-kb DNA element is excised from the nifD gene by site-specific recombination. The xisA gene, located on the 11-kb element, is required for the excision of the element. Transcription and DNA rearrangement of the nifHDK operon both occur late during heterocyst differentiation, about 18 to 24 h after induction, suggesting that the regulation of these events might be coupled. We show that heterocyst-specific transcription and DNA rearrangement of the nifHDK operon are independent of one another. Northern (RNA) analysis of the xisA mutant strain DW12-2.2, which cannot excise the nifD 11-kb element or fix nitrogen, showed that the nifH and nifD genes are transcribed on unrearranged chromosomes. The nifK gene was not transcribed in DW12-2.2, indicating that its expression is dependent on the nifH promoter and excision of the 11-kb element from the operon. A 1.68-kb DNA fragment containing the nifH promoter was deleted from the chromosome to produce the mutant strain LW1. LW1 formed heterocysts but did not grow on nitrogen-free medium and showed no transcription through nifD. Southern analysis of LW1 showed normal excision of the 11-kb element from the nifHDK operon, indicating that transcription from the nifH promoter is not required for the developmentally regulated DNA rearrangement. Images FIG. 2 FIG. 3 FIG. 5 FIG. 6 FIG. 7

Golden, J W; Whorff, L L; Wiest, D R

1991-01-01

402

Group 2 Sigma Factor Mutant ?sigCDE of the Cyanobacterium Synechocystis sp. PCC 6803 Reveals Functionality of Both Carotenoids and Flavodiiron Proteins in Photoprotection of Photosystem II.  

PubMed

Adjustment of gene expression during acclimation to stress conditions, such as bright light, in the cyanobacterium Synechocystis sp. PCC 6803 depends on four group 2 ? factors (SigB, SigC, SigD, SigE). A ?sigCDE strain containing the stress-responsive SigB as the only functional group 2 ? factor appears twice as resistant to photoinhibition of photosystem II (PSII) as the control strain. Microarray analyses of the ?sigCDE strain indicated that 77 genes in standard conditions and 79 genes in high light were differently expressed compared with the control strain. Analysis of possible photoprotective mechanisms revealed that high carotenoid content and up-regulation of the photoprotective flavodiiron operon flv4-sll0218-flv2 protected PSII in ?sigCDE, while up-regulation of pgr5-like, hlipB or isiA genes in the mutant strain did not offer particular protection against photoinhibition. Photoinhibition resistance was lost if ?sigCDE was grown in high CO2, where carotenoid and Flv4, Sll0218, and Flv2 contents were low. Additionally, photoinhibition resistance of the ?rpoZ strain (lacking the omega subunit of RNA polymerase), with high carotenoid but low Flv4-Sll0218-Flv2 content, supported the importance of carotenoids in PSII protection. Carotenoids likely protect mainly by quenching of singlet oxygen, but efficient nonphotochemical quenching in ?sigCDE might offer some additional protection. Comparison of photoinhibition kinetics in control, ?sigCDE, and ?rpoZ strains showed that protection by the flavodiiron operon was most efficient during the first minutes of high-light illumination. PMID:24009334

Hakkila, Kaisa; Antal, Taras; Gunnelius, Liisa; Kurkela, Juha; Matthijs, Hans C P; Tyystjärvi, Esa; Tyystjärvi, Taina

2013-09-04

403

A mutant lacking the glutamine synthetase gene (glnA) is impaired in the regulation of the nitrate assimilation system in the cyanobacterium Synechocystis sp. strain PCC 6803.  

PubMed Central

The existence in the unicellular cyanobacterium Synechocystis sp. strain PCC 6803 of two genes (glnA and glnN) coding for glutamine synthetase (GS) has been recently reported (J.C. Reyes and F.J. Florencio, J. Bacteriol. 176:1260-1267, 1994). In the current work, the regulation of the nitrate assimilation system was studied with a glnA-disrupted Synechocystis mutant (strain SJCR3) in which the only GS activity is that corresponding to the glnN product. This mutant was unable to grow in ammonium-containing medium because of its very low levels of GS activity. In the SJCR3 strain, nitrate and nitrite reductases were not repressed by ammonium, and short-term ammonium-promoted inhibition of nitrate uptake was impaired. In Synechocystis sp. strain PCC 6803, nitrate seems to act as a true inducer of its assimilation system, in a way similar to that proposed for the dinitrogen-fixing cyanobacteria. A spontaneous derivative strain from SJCR3 (SJCR3.1), was able to grow in ammonium-containing medium and exhibited a fourfold-higher level of GS activity than but the same amount of glnN transcript as its parental strain (SJCR3). Taken together, these finding suggest that SJCR3.1 is a mutant affected in the posttranscriptional regulation of the GS encoded by glnN. This strain recovered regulation by ammonium of nitrate assimilation. SJCR3 cells were completely depleted of intracellular glutamine shortly after addition of ammonium to cells growing with nitrate, while SJCR3.1 cells maintained glutamine levels similar to that reached in the wild-type Synechocystis sp. strain PCC 6803. Our results indicate that metabolic signals that control the nitrate assimilation system in Synechocystis sp. strain PCC 6803 require ammonium metabolism through GS. Images

Reyes, J C; Florencio, F J

1994-01-01

404

Distribution of the mevalonate and glyceraldehyde phosphate/pyruvate pathways for isoprenoid biosynthesis in unicellular algae and the cyanobacterium Synechocystis PCC 6714.  

PubMed

Isopentenyl diphosphate, the universal isoprenoid precursor, can be produced by two different biosynthetic routes: either via the acetate/mevalonate (MVA) pathway, or via the more recently identified MVA-independent glyceraldehyde phosphate/pyruvate pathway. These two pathways are easily differentiated by incorporation of [1-13C]glucose and analysis of the resulting labelling patterns found in the isoprenoids. This method was successfully applied to several unicellular algae raised under heterotrophic growth conditions and allowed for the identification of the pathways that were utilized for isoprenoid biosynthesis. All isoprenoids examined (sterols, phytol, carotenoids) of the green algae Chlorella fusca and Chlamydomonas reinhardtii were synthesized via the GAP/pyruvate pathway, as in another previously investigated green alga, Scenedesmus obliquus, which was also shown in this study to synthesize ubiquinone by the same MVA-independent route. In the red alga Cyanidium caldarium and in the Chrysophyte Ochromonas danica a clear dichotomy was observed: as in higher plants, sterols were formed via the MVA route, whereas chloroplast isoprenoids (phytol in Cy. caldarium and O. danica and beta-carotene in O. danica) were synthesized via the GAP/pyruvate route. In contrast, the Euglenophyte Euglena gracilis synthesized ergosterol, as well as phytol, via the acetate/MVA route. Similar feeding experiments were performed with the cyanobacterium Synechocystis PCC 6714 using [1-13C]- and [6-13C]-glucose. The two isoprenoids examined, phytol and beta-carotene, were shown to have the typical labelling pattern derived from the GAP/pyruvate route. PMID:9657979

Disch, A; Schwender, J; Müller, C; Lichtenthaler, H K; Rohmer, M

1998-07-15

405

Spectral inhomogeneity of photosystem I and its influence on excitation equilibration and trapping in the cyanobacterium Synechocystis sp. PCC6803 at 77 K.  

PubMed Central

Ultrafast transient absorption spectroscopy was used to probe excitation energy transfer and trapping at 77 K in the photosystem I (PSI) core antenna from the cyanobacterium Synechocystis sp. PCC 6803. Excitation of the bulk antenna at 670 and 680 nm induces a subpicosecond energy transfer process that populates the Chl a spectral form at 685--687 nm within few transfer steps (300--400 fs). On a picosecond time scale equilibration with the longest-wavelength absorbing pigments occurs within 4-6 ps, slightly slower than at room temperature. At low temperatures in the absence of uphill energy transfer the energy equilibration processes involve low-energy shifted chlorophyll spectral forms of the bulk antenna participating in a 30--50-ps process of photochemical trapping of the excitation by P(700). These spectral forms might originate from clustered pigments in the core antenna and coupled chlorophylls of the reaction center. Part of the excitation is trapped on a pool of the longest-wavelength absorbing pigments serving as deep traps at 77 K. Transient hole burning of the ground-state absorption of the PSI with excitation at 710 and 720 nm indicates heterogeneity of the red pigment absorption band with two broad homogeneous transitions at 708 nm and 714 nm (full-width at half-maximum (fwhm) approximately 200--300 cm(-1)). The origin of these two bands is attributed to the presence of two chlorophyll dimers, while the appearance of the early time bleaching bands at 683 nm and 678 nm under excitation into the red side of the absorption spectrum (>690 nm) can be explained by borrowing of the dipole strength by the ground-state absorption of the chlorophyll a monomers from the excited-state absorption of the dimeric red pigments.

Melkozernov, A N; Lin, S; Blankenship, R E; Valkunas, L

2001-01-01

406

Directed mutagenesis of an iron-sulfur protein of the photosystem I complex in the filamentous cyanobacterium Anabaena variabilis ATCC 29413.  

PubMed Central

In oxygenic photosynthetic organisms the PSI-C polypeptide, encoded by the psaC gene, provides the ligands for two [4Fe-4S] centers, FA and FB, the terminal electron acceptors in the photosystem I (PSI) complex. An insertion mutation introduced in the psaC locus of the filamentous cyanobacterium Anabaena variabilis ATCC 29413 resulted in the creation of a mutant strain, T398-1, that lacks the PSI-C polypeptide. In medium supplemented with 5 mM fructose, the mutant cells grew well in the dark. However, when grown in the same medium under light, the doubling rate of T398-1 cells was significantly decreased. In intact cells of T398-1, bicarbonate-dependent whole-chain electron transport (PSII and PSI) could not be detected, although partial electron transport reactions involving either one of the two photosystems could be measured at significant rates. The low-temperature EPR signals attributed to the [4Fe-4S] centers FA and FB were absent in the mutant cells. Chemical titration measurements indicated that the ratios of chlorophyll to the primary donor P700 were virtually identical in membranes from the wild-type and mutant cells. Moreover, room-temperature optical spectroscopic analysis of the thylakoid membranes isolated from T398-1 showed flash-induced P700 oxidation followed by dark rereduction, indicating primary photochemistry in PSI. Thus stable assembly of the reaction center of PSI can occur in the absence of the Fe-S cluster cofactors FA and FB. These studies demonstrate that Anabaena 29413 offers a useful genetic system for targeted mutagenesis of the PSI complex. Images

Mannan, R M; Whitmarsh, J; Nyman, P; Pakrasi, H B

1991-01-01

407

Proteomic analysis of a highly active photosystem II preparation from the cyanobacterium Synechocystis sp. PCC 6803 reveals the presence of novel polypeptides.  

PubMed

A highly active oxygen-evolving photosystem II (PSII) complex was purified from the HT-3 strain of the widely used cyanobacterium Synechocystis sp. PCC 6803, in which the CP47 polypeptide has been genetically engineered to contain a polyhistidine tag at its carboxyl terminus [Bricker, T. M., Morvant, J., Masri, N., Sutton, H. M., and Frankel, L. K. (1998) Biochim. Biophys. Acta 1409, 50-57]. These purified PSII centers had four manganese atoms, one calcium atom, and two cytochrome b(559) hemes each. Optical absorption and fluorescence emission spectroscopy as well as western immunoblot analysis demonstrated that the purified PSII preparation was devoid of any contamination with photosystem I and phycobiliproteins. A comprehensive proteomic analysis using a system designed to enhance resolution of low-molecular-weight polypeptides, followed by MALDI mass spectrometry and N-terminal amino acid sequencing, identified 31 distinct polypeptides in this PSII preparation. We propose a new nomenclature for the polypeptide components of PSII identified after PsbZ, which proceeds sequentially from Psb27. During this study, the polypeptides PsbJ, PsbM, PsbX, PsbY, PsbZ, Psb27, and Psb28 proteins were detected for the first time in a purified PSII complex from Synechocystis 6803. Five novel polypeptides were also identified in this preparation. They included the Sll1638 protein, which shares significant sequence similarity to PsbQ, a peripheral protein of PSII that was previously thought to be present only in chloroplasts. This work describes newly identified proteins in a highly purified cyanobacterial PSII preparation that is being widely used to investigate the structure, function, and biogenesis of this photosystem. PMID:12069591

Kashino, Yasuhiro; Lauber, Wendy M; Carroll, James A; Wang, Qingjun; Whitmarsh, John; Satoh, Kazuhiko; Pakrasi, Himadri B

2002-06-25

408

Co-ordinated expression of phycobiliprotein operons in the chromatically adapting cyanobacterium Calothrix PCC 7601: a role for RcaD and RcaG.  

PubMed

In the cyanobacterium Calothrix sp. PCC 7601 the cpc2 operon encoding phycocyanin 2 (PC2) is expressed if red radiations are available. RcaD was previously identified in extracts from red-light-grown cells as an alkaline phosphatase-sensitive protein that binds upstream of the transcription start point (TSP) of the cpc2 operon. In this work, RcaD was purified, and the corresponding gene cloned with a PCR probe obtained using degenerated primers based on RcaD peptide sequences (accession no. AJ319541). Purified RcaD binds to the cpc2 promoter region and also to those of the constitutive cpc1 and apc1 operons that encode phycocyanin 1 and allophycocyanin. Escherichia coli-overexpressed RcaD can bind to the cpc2 promoter region. The rcaD gene is upstream of an open reading frame (ORF) termed rcaG. Co-transcription of both genes was demonstrated by reverse transcription (RT)-PCR experiments, and found to be independent of the light wavelengths. A single TSP was mapped. Se