RÉSUMÉ
Some cyanobacteria produce a UVA-absorbing pigment, scytonemin, at extracellular sheaths. Although scytonemin-containing dark sheaths are recognizable through optical microscopes and its redox changes have been known for decades, there has been no report to obtain images of both oxidized and reduced scytonemins at subcellular resolution. Here, we show that a spontaneous Raman scattering spectral microscopy based on an excitation-laser-line-scanning method unveil 3D subcellular distributions of both the oxidized and reduced scytonemins in a filamentous cyanobacterium. The redox changes of scytonemin were supported by comparison in the Raman spectra between the cyanobacterial cells, solid-state scytonemin and quantum chemical normal mode analysis. Distributions of carotenoids, phycobilins, and the two redox forms of scytonemin were simultaneously visualized with an excitation wavelength at 1064 nm that is virtually free from the optical screening by the dark sheaths. The redox differentiation of scytonemin will advance our understanding of the redox homeostasis and secretion mechanisms of individual cyanobacteria as well as microscopic chemical environments in various microbial communities. The line-scanning Raman microscopy based on the 1064 nm excitation is thus a promising tool for exploring hitherto unreported Raman spectral features and distribution of nonfluorescent molecules embedded below nontransparent layers for visible light, while avoiding interference by autofluorescence.
Sujet(s)
Cyanobactéries , Analyse spectrale Raman , Cyanobactéries/composition chimique , Lumière , OxydoréductionRÉSUMÉ
Cyanobacteria are some of the primary producers in extremely cold biospheres such as the Arctic, Antarctic, and vast ice sheets. Many genera of cyanobacteria are identified from these harsh environments, but their specific mechanisms for cold adaptation are not fully understood. Nostoc sp. strain SO-36 is a cyanobacterium isolated in Antarctica more than 30 years ago and regarded as a psychrotolelant species. To determine whether the strain is psychrotolelant or psychrophilic, it was first grown at 30 °C and 10 °C. The cells grew exponentially at 30 °C, but their growth stopped at 10 °C, indicating that the strain is only psychrotolerant. Microscopic analysis revealed that the morphology of the cells grown at 30 °C was filamentous and differentiated heterocysts, which are specialized cells for gaseous nitrogen fixation under nitrogen-deprived conditions, indicating that the strain can grow diazotrophically. The cells grown at 10 °C have a smaller size, shortened filament length and decreased chlorophyll content per cell. At 10 °C, the cells are aggregated with extracellular polymeric substrates (EPSs), which is a common mechanism to protect cells from ultraviolet light. These results imply that segmentation into short filaments was induced by photodamage at low temperatures. To fully understand the adaptation mechanisms of Nostoc sp. strain SO-36 for low-temperature conditions, next-generation sequencing analyses were conducted. Complete genome sequence of the strain revealed that it has one main chromosome of approximately 6.8 Mbp with 4 plasmids, including 6855 coding sequences, 48 tRNA genes, 4 copies of rRNA operons, and 5 CRISPR regions. Putative genes for EPS biosynthesis were found to be conserved in Nostocaceae regardless of their habitat. These results provide basic information to understand the adaptation mechanisms at low temperatures, and the strain can be a model organism to analyze adaptation to extreme environments.
Sujet(s)
Nostoc , Adaptation physiologique , Régions antarctiques , Protéines bactériennes/génétique , Fixation de l'azote , Nostoc/génétique , Analyse de séquence d'ADN , Espace extracellulaire/métabolismeRÉSUMÉ
Basidiomycetes-X, of which Japanese vernacular name is Echigoshirayukidake, is a local speciality mushroom found and cultivated in Japan that has been distributed as a precious cuisine material or as a functional food with medicinal properties. Antioxidant activity-guided isolation of major ingredients in Basidiomycetes-X revealed the presence of ergosterol, trans-10,cis-12-octadecadienoic acid (a conjugated linolenic acid, 10(E),12(Z)-CLA) and 2,3-dihydro-3,5-dihydroxy-6-methyl4Hpyran-4-one (DDMP). Approximately 21% of the 2,2-diphenyl-1-(2,4,6-trinitrophenyl)hydrazino radical (DPPH) scavenging activities in the methanolic extract were related to 10(E),12(Z)-CLA, while approximately 6.2% of the activity was related to ergosterol. DDMP was present in both methanolic and water extracts, and the activity related to DDMP was conspicuously detected in water extracts. Moreover, uridine and adenosine were identified as major components of Basidiomycetes-X. The ingredients identified in Basidiomycetes-X are expected to be involved in biological functions observed in this mushroom, which is an attractive functional food resource.
Sujet(s)
Agaricales , Antioxydants , Ergostérol , Piégeurs de radicaux libres/composition chimique , EauRÉSUMÉ
The terrestrial cyanobacterium Nostoc commune is an anhydrobiotic organism with extreme longevity. Recovery of photosynthesis by rehydration was examined using our laboratory stocks of dry N. commune thalli after long-term storage in a desiccated state. In the samples stored at room temperature for over 8 years, photosynthetic oxygen evolution was barely detectable, whereas oxygen consumption was recovered. There was an exceptional case in which photosynthetic oxygen evolution recovered after 8 years of storage at room temperature. Both photosynthetic oxygen evolution and respiratory oxygen consumption were recovered in dry thalli stored at -20°C for over 15 years. Consistent with the recovery of photosynthetic oxygen evolution, Fv/Fm was detected in the samples stored at -20°C at levels similar to those of freshly collected N. commune colonies. Carotenoids, scytonemin and chlorophyll a appeared to be intact in the dry thalli stored at -20°C, but ß-carotene was not detected in the samples stored at room temperature. α-Tocopherol was intact in the samples stored at -20°C but was degraded in the samples stored at room temperature. These results suggest that dry thalli of N. commune are capable of sustaining biological activities for a long time, although they are gradually damaged when stored at room temperature.
Sujet(s)
Nostoc commune , Nostoc , Chlorophylle A/métabolisme , Photosynthèse , Oxygène/métabolismeRÉSUMÉ
The terrestrial cyanobacterium Nostoc commune has a cosmopolitan distribution. It is edible, and dry thalli are sold as a food in China under the name of Di Pi Cai. The pigment composition and the genotypes were characterized to identify the cyanobacterium Di Pi Cai from China as N. commune. Myxol glycosides and ketocarotenoids were detected, as expected in Nostoc sp., but ß-carotene and hydroxylated carotenoids were not detected. Nostoc-756, mycosporine-2-(4-deoxygadusoyl-ornitine), was found to be a main mycosporine-like amino acid, which indicates that Di Pi Cai belongs to the N. commune chemotype C. However, the 16S rRNA gene and the petH gene encoding ferredoxin-NADP+ oxidoreductase of Di Pi Cai did not exactly match those of genotype C found in Japan. These results suggest the unique molecular genetic features of Di Pi Cai and the global diversity of N. commune.
Sujet(s)
Nostoc commune , Nostoc , Acides aminés , Antioxydants , Nostoc/génétique , Nostoc commune/génétique , ARN ribosomique 16S/génétiqueRÉSUMÉ
Three pyrrole alkaloid derivatives were isolated from the edible mushroom Basidiomycetes-X (Echigoshirayukidake) by water extraction followed by ethyl acetate fractionation. The chemical structures determined by MS and NMR were 4-[2-formyl-5-(hydroxymethyl)-1H-pyrrol-1-yl] butanoic acid (compound I), 4-[2-formyl-5-(hydroxymethyl)-1H-pyrrol-1-yl] butanamide (compound II), and 5-(hydroxymethyl)-1H-pyrrole-2-carboxaldehyde (compound III). Compound I was found to be the major component, followed by compound II, and compound III was the minor component. The dry powder of Basidiomycetes-X contained approximately 825 µg g-1 compound I and 484 µg g-1 compound II. Compound II was found to be a novel pyrrole aldehyde homologue not previously reported and thus is a specific component of this mushroom.
Sujet(s)
Alcaloïdes/composition chimique , Basidiomycota/composition chimique , Mélanges complexes/composition chimique , Compléments alimentaires/analyse , Pyrroles/composition chimique , Acétates/composition chimique , Aldéhydes/composition chimique , Alcaloïdes/isolement et purification , Mélanges complexes/isolement et purification , Cuivre/composition chimique , Piégeurs de radicaux libres/composition chimique , Fer/composition chimique , Spectroscopie par résonance magnétique , Spectrométrie de masse , Pyrroles/isolement et purification , Solvants/composition chimiqueRÉSUMÉ
The aquatic cyanobacterium Nostoc verrucosum forms macroscopic colonies in streams, and its appearance is superficially similar to that of the terrestrial cyanobacterium Nostoc commune. N. verrucosum is sensitive to desiccation, unlike N. commune, although these Nostoc cyanobacterial species share physiological features, including massive extracellular polysaccharide production and trehalose accumulation capability. In this study, water-soluble sunscreen pigments of mycosporine-like amino acids (MAAs) were characterized in N. verrucosum, and the mysABCD genes responsible for MAA biosynthesis in N. verrucosum and N. commune were compared. N. verrucosum produced porphyra-334 and shinorine, with porphyra-334 accounting for >90% of the total MAAs. Interestingly, porphyra-334 is an atypical cyanobacteial MAA, whereas shinorine is known as a common and dominant MAA in cyanobacteria. Porphyra-334 from N. verrucosum showed little or no radical scavenging activity in vitro, although the glycosylated derivatives of porphyra-334 from N. commune are potent radical scavengers. The presence of the mysABCD gene cluster in N. commune strain KU002 (genotype A) supported its porphyra-334 producing capability via the Nostoc-type mechanism, although the genotype A of N. commune mainly produces the arabinose-bound porphyra-334. The mysABC gene cluster was conserved in N. verrucosum, but the mysD gene was not included in the cluster. These results suggest that the mysABCD gene products are involved in the biosynthesis of porphyra-334 commonly in these Nostoc species, and that the genotype A of N. commune additionally acquired the glycosylation of porphyra-334.
Sujet(s)
Cyclohexanones , Cyclohexylamines , Glycine/analogues et dérivés , Nostoc/composition chimique , Cyclohexanones/métabolisme , Cyclohexylamines/métabolisme , Glycine/biosynthèse , Glycine/génétique , Glycine/métabolisme , Glycosylation , Famille multigénique/génétique , Nostoc/génétique , Produits antisolaires/composition chimiqueRÉSUMÉ
The terrestrial cyanobacterium Nostoc commune forms macroscopic colonies in its natural habitats, and these colonies consist of both cellular filaments and massive extracellular matrixes. In this study, the biochemical features of the extracellular matrix components were investigated. Naturally growing N. commune was tolerant to desiccation, and produced massive extracellular polysaccharides that contained both neutral sugars and glucuronic acid as constituent monosaccharides. The extracellular polysaccharide contents and desiccation tolerance were compared in laboratory culture strains of Nostoc species. The laboratory culture of N. commune strain KU002 was sensitive to desiccation and produced smaller amounts of extracellular polysaccharides, unlike the field-isolated naturally growing colonies. Nostoc punctiforme strain M-15, which is genetically closed to N. commune, was able to tolerate desiccation, although the other Nostoc strains were desiccation-sensitive. A laboratory culture strain of the aquatic cyanobacterium Nostoc sphaericum produced massive extracellular polysaccharides but was sensitive to desiccation, suggesting that extracellular matrix production is not enough to make this strain tolerant to desiccation. WspA (water stress protein) and SodF (superoxide dismutase) were found to be characteristic protein components of the extracellular matrix of N. commune. Because the WspA proteins were heterogeneous, the wspA genes were highly diverse among the different genotypes of N. commune, although the sodF gene was rather conservative. The heterogeneity of the WspA proteins suggests their complex roles in the environmental adaptation mechanism in N. commune.
Sujet(s)
Adaptation physiologique/génétique , Protéines bactériennes/génétique , Dessiccation , Matrice extracellulaire/composition chimique , Protéines du choc thermique/génétique , Nostoc commune/composition chimique , Polyosides bactériens/composition chimique , Protéines bactériennes/composition chimique , Matrice extracellulaire/enzymologie , Matrice extracellulaire/métabolisme , Génotype , Protéines du choc thermique/composition chimique , Oses/métabolisme , Nostoc commune/enzymologie , Microbiologie du sol , Superoxide dismutase/composition chimique , Superoxide dismutase/génétiqueRÉSUMÉ
A UV-absorbing compound was purified and identified as a novel glycosylated mycosporine-like amino acid (MAA), 13-O-ß-galactosyl-porphyra-334 (ß-Gal-P334) from the edible cyanobacterium Nostoc sphaericum, known as "ge xian mi" in China and "cushuro" in Peru. Occurrence of the hexosylated derivative of shinorine (hexosyl-shinorine) was also supported by LC-MS/MS analysis. ß-Gal-P334 accounted for about 86.5% of total MAA in N. sphaericum, followed by hexosyl-shinorine (13.2%) and porphyra-334 (0.2%). ß-Gal-P334 had an absorption maximum at 334nm and molecular absorption coefficient was 46,700 at 334nm. Protection activity of ß-Gal-P334 from UVB and UVA+8-methoxypsoralen induced cell damage on human keratinocytes (HaCaT) was assayed in comparison with other MAA (porphyra-334, shinorine, palythine and mycosporine-glycine). The UVB protection activity was highest in mycosporine-glycine, followed by palythine, ß-Gal-P334, porphyra-334 and shinorine in order. ß-Gal-P334 had highest protection activity from UVA+8-methoxypsoralen induced cell damage followed by porphyra-334, shinorine, mycosporine-glycine and palythine. We also found an antioxidant (radical-scavenging) activity of ß-Gal-P334 by colorimetric and ESR methods. From these findings, ß-Gal-P334 was suggested to play important roles in stress tolerant mechanisms such as UV and oxidative stress in N. sphaericum as a major MAA. We also consider that the newly identified MAA, ß-Gal-P334 has a potential for use as an ingredient of cosmetics and toiletries.
Sujet(s)
Cyclohexanones/composition chimique , Glycine/analogues et dérivés , Nostoc/composition chimique , Stress oxydatif/effets des médicaments et des substances chimiques , Agents protecteurs/pharmacologie , Rayons ultraviolets , Acides aminés/composition chimique , Antioxydants/composition chimique , Antioxydants/isolement et purification , Antioxydants/pharmacologie , Lignée cellulaire , Survie cellulaire/effets des médicaments et des substances chimiques , Survie cellulaire/effets des radiations , Chromatographie en phase liquide à haute performance , Cyclohexanones/isolement et purification , Cyclohexanones/pharmacologie , Cyclohexylamines/composition chimique , Glycine/composition chimique , Glycine/isolement et purification , Glycine/pharmacologie , Glycosylation , Humains , Kératinocytes/cytologie , Kératinocytes/effets des médicaments et des substances chimiques , Kératinocytes/effets des radiations , Conformation moléculaire , Nostoc/métabolisme , Stress oxydatif/effets des radiations , Agents protecteurs/composition chimique , Agents protecteurs/isolement et purification , Spectrométrie de masse en tandemRÉSUMÉ
Mycosporine-like amino acids (MAAs) are UV-absorbing pigments, and structurally unique glycosylated MAAs are found in the terrestrial cyanobacterium Nostoc commune. In this study, we examined two genotypes of N.commune colonies with different water extract UV-absorption spectra. We found structurally distinct MAAs in each genotype. The water extract from genotype A showed a UV-absorbing spectrum with an absorption maximum at 335nm. The extract contained the following compounds: 7-O-(ß-arabinopyranosyl)-porphyra-334 (478Da), pentose-bound shinorine (464Da), hexose-bound porphyra-334 (508Da) and porphyra-334 (346Da). The water extract from genotype B showed a characteristic UV-absorbing spectrum with double absorption maxima at 312 and 340nm. The extract contained hybrid MAAs (1050Da and 880Da) with two distinct chromophores of 3-aminocyclohexen-1-one and 1,3-diaminocyclohexen linked to 2-O-(ß-xylopyranosyl)-ß-galactopyranoside. A novel 273-Da MAA with an absorption maximum at 310nm was also identified in genotype B. The MAA consisted of a 3-aminocyclohexen-1-one linked to a γ-aminobutyric acid chain. These MAAs had potent radical scavenging activities in vitro and the results confirmed that the MAAs have multiple roles as a UV protectant and an antioxidant relevant to anhydrobiosis in N. commune. The two genotypes of N. commune exclusively produced their own characteristic glycosylated MAAs, which supports that MAA composition could be a chemotaxonomic marker for the classification of N. commune.
Sujet(s)
Cyclohexanols/analyse , Nostoc commune/métabolisme , Antioxydants/métabolisme , Chlorophylle/analyse , Chlorophylle A , Chromatographie en phase liquide à haute performance , Cyclohexanols/isolement et purification , Cyclohexanols/métabolisme , Cyclohexanones/analyse , Électrophorèse capillaire , Génotype , Glycine/analogues et dérivés , Glycine/analyse , Glycosylation , Masse moléculaire , Nostoc commune/génétique , ARN ribosomique 16S/génétique , ARN ribosomique 16S/métabolisme , Spectrométrie de masse MALDI , Spectrophotométrie UVRÉSUMÉ
Mycosporine-like amino acids (MAAs) are water-soluble molecules that absorb UV-A and UV-B radiation and disperse the energy as heat. MAAs show great diversity in their molecular structures, which exhibit a range of molecular weights spanning 188 to 1050 Daltons. MAAs are utilized in a wide variety of organisms including prokaryotes and eukaryotic micro-organisms that inhabit aquatic, terrestrial, and marine environments. These features suggest that MAAs are stable and fundamental molecules that allow these organisms to live under UV irradiation. MAAs are thought to have been greatly important to ancient forms of life on Earth, functioning as a primary sunscreen to reduce short-wavelength light. Structurally different MAAs might have been developed in MAA-producing organisms during their environmental adaptation. Harmful irradiation directly damages biomolecules, including lipids, proteins and DNA, and induces oxidative stress through radical-propagating processes. Thus, MAAs are expected to play an additional role in the antioxidant system. This review focuses on MAAs with radical scavenging activities. To cover all the reported MAAs known thus far, we surveyed the CAS database and have summarized the structures and the chemical and physical properties of these MAAs, including their antioxidant activities.
RÉSUMÉ
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.
Sujet(s)
Cyclohexanols/composition chimique , Cyclohexanones/composition chimique , Glycine/analogues et dérivés , Nostoc commune/composition chimique , Thréonine/composition chimique , Acides aminés/composition chimique , Acides aminés/pharmacologie , Antioxydants/composition chimique , Antioxydants/pharmacologie , Cyanobactéries/composition chimique , Cyclohexanols/pharmacologie , Cyclohexanones/pharmacologie , Glycine/composition chimique , Glycine/pharmacologie , Glycosylation , Produits antisolaires/composition chimique , Produits antisolaires/pharmacologie , Thréonine/pharmacologie , Rayons ultraviolets , Eau/composition chimiqueRÉSUMÉ
Cyanobacteria have two types of sunscreen pigments, scytonemin and mycosporine-like amino acids (MAAs). These secondary metabolites are thought to play multiple roles against several environmental stresses such as UV radiation and desiccation. Not only the large molar absorption coefficients of these sunscreen pigments, but also their antioxidative properties may be necessary for the protection of biological molecules against the oxidative damages induced by UV radiation. The antioxidant activity and vitrification property of these pigments are thought to be requisite for the desiccation and rehydration processes in anhydrobiotes. In this review, the multiple roles of photosynthetic pigments and sunscreen pigments on stress resistance, especially from the viewpoint of their structures, biosynthetic pathway, and in vitro studies of their antioxidant activity, will be discussed.
RÉSUMÉ
Scytonemin is a 544-Da hydrophobic pigment that can absorb UV-A radiation. It is present in cyanobacterial sheaths and is thought to function as a UV protectant. In this study, scytonemin was purified from the terrestrial cyanobacterium Nostoc commune, and its radical-scavenging activity was characterized. The purified scytonemin quenched an organic radical in vitro and accounted for up to 10% of the total activity of an ethanol extract of N. commune. These results suggest that the extracellular UV-absorbing pigment scytonemin has multiple roles, functioning as a UV sunscreen and an antioxidant relevant to anhydrobiosis in N. commune.
Sujet(s)
Piégeurs de radicaux libres/métabolisme , Indoles/métabolisme , Nostoc commune/métabolisme , Phénols/métabolisme , Pigments biologiques , Rayons ultraviolets , Absorption , Cyanobactéries/métabolisme , Cyanobactéries/effets des radiations , Piégeurs de radicaux libres/composition chimique , Indoles/composition chimique , Nostoc commune/effets des radiations , Phénols/composition chimique , Produits antisolaires/composition chimique , Produits antisolaires/métabolismeRÉSUMÉ
The phylogeny of the terrestrial cyanobacterium Nostoc commune and its neighboring Nostoc species was studied using molecular genetic and chemotaxonomic approaches. At least eight genotypes of N. commune were characterized by the differences among 16S rRNA gene sequences and the petH gene encoding ferredoxin-NADP⺠oxidoreductase and by random amplified polymorphic DNA analysis. The genotypes of N. commune were distributed in Japan without regional specificity. The nrtP gene encoding NrtP-type nitrate/nitrite permease was widely distributed in the genus Nostoc, suggesting that the occurrence of the nrtP gene can be one of the characteristic features that separate cyanobacteria into two groups. The wspA gene encoding a 36-kDa water stress protein was only found in N. commune and Nostoc verrucosum, suggesting that these Nostoc species that form massive colonies with extracellular polysaccharides can be exclusively characterized by the occurrence of the wspA gene. Fifteen species of Nostoc and Anabaena were investigated by comparing their carotenoid composition. Three groups with distinct patterns of carotenoids were related to the phylogenic tree constructed on the basis of 16S rRNA sequences. Nostoc commune and Nostoc punctiforme were clustered in one monophyletic group and characterized by the occurrence of nostoxanthin, canthaxanthin, and myxol glycosides.
Sujet(s)
Anabaena/génétique , Nostoc commune/génétique , Anabaena/métabolisme , Anabaena/pathogénicité , Séquence nucléotidique , Canthaxanthine/métabolisme , Caroténoïdes/métabolisme , Gènes d'ARN ribosomique , Variation génétique , Japon , Données de séquences moléculaires , Nitrates/métabolisme , Nitrites/métabolisme , Nostoc commune/métabolisme , Nostoc commune/pathogénicité , Phylogenèse , ARN ribosomique 16S , Xanthophylles/métabolismeRÉSUMÉ
Mycosporine-like amino acids (MAAs) are UV absorbing pigments, and structurally distinct MAAs have been identified in taxonomically diverse organisms. Two novel MAAs were purified from the cyanobacterium Nostoc commune, and their chemical structures were characterized. An MAA with an absorption maximum at 335 nm was identified as a pentose-bound porphyra-334 derivative with a molecular mass of 478 Da. Another identified MAA had double absorption maxima at 312 and 340 nm and a molecular mass of 1,050 Da. Its unique structure consisted of two distinct chromophores of 3-aminocyclohexen-1-one and 1,3-diaminocyclohexen and two pentose and hexose sugars. These MAAs had radical scavenging activity in vitro; the 1050-Da MAA contributed approximately 27% of the total radical scavenging activities in a water extract of N. commune. These results suggest that these glycosylated MAAs have multiple roles as a UV protectant and an antioxidant relevant to anhydrobiosis in N. commune.
Sujet(s)
Acides aminés/composition chimique , Piégeurs de radicaux libres/composition chimique , Nostoc commune/métabolisme , Acides aminés/isolement et purification , Chromatographie en phase liquide à haute performance , Chromatographie en phase inverse , Cyclohexanones/composition chimique , Piégeurs de radicaux libres/isolement et purification , Glycine/analogues et dérivés , Glycine/composition chimique , Glycosylation , Spectrométrie de masse MALDI , Spectrophotométrie UVRÉSUMÉ
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.
Sujet(s)
Dessiccation , Matrice extracellulaire/métabolisme , Nostoc/métabolisme , Tréhalose/métabolisme , Protéines bactériennes/isolement et purification , Caroténoïdes/analyse , Chlorophylle/analyse , Chlorophylle A , Congélation , Gènes bactériens , Nostoc commune/métabolisme , Photosynthèse , Stress physiologique , Acides uroniques/analyse , Eau/physiologie , Microbiologie de l'eauRÉSUMÉ
We investigated the biochemical properties of the enzymes involved in trehalose metabolism in the cyanobacterium Nostoc punctiforme strain IAM M-15 to elucidate the mechanism of trehalose accumulation in response to desiccation and salt stress. There was no detectable trehalose in fully hydrated N. punctiforme cells; however, these cells accumulated trehalose upon desiccation. Moreover, NaCl treatment also induced trehalose accumulation. The three genes for trehalose metabolism, treZ (maltooligosyltrehalose trehalohydrolase, Mth), treY (maltooligosyltrehalose synthase, Mts), and treH (trehalase), were found as a gene cluster, and the mRNAs for these genes were detectable at similar levels during desiccation. Trehalase of N. punctiforme was heterologously expressed in E. coli cells in an active form with a molecular mass of 52 kDa. Trehalase activity was strongly inhibited in the presence of 10 mM NaCl while trehalose synthesis activity remained active in the presence of salt. These data suggest that the rate of trehalose production exceeds that of trehalose hydrolysis under water-stress conditions characterized by increased cellular solute concentrations. In the proposed mechanism, control of trehalase plays an important role in trehalose accumulation in terrestrial cyanobacteria under conditions of extreme desiccation.
Sujet(s)
Régulation de l'expression des gènes bactériens/physiologie , Nostoc/enzymologie , alpha, alpha-Trehalase/métabolisme , Tréhalose/métabolisme , Déshydratation/enzymologie , Déshydratation/génétique , Régulation de l'expression des gènes bactériens/effets des médicaments et des substances chimiques , Gènes bactériens/génétique , Nostoc/effets des médicaments et des substances chimiques , Nostoc/métabolisme , ARN messager/métabolisme , Protéines recombinantes/métabolisme , RT-PCR , Chlorure de sodium/pharmacologie , Stress physiologique/physiologie , Saccharose/métabolisme , alpha, alpha-Trehalase/génétique , Tréhalose/génétiqueRÉSUMÉ
The cyanobacterium Nostoc commune is adapted to the terrestrial environment and forms a visible colony in which the cells are embedded in extracellular polysaccharides (EPSs), which play a crucial role in the extreme desiccation tolerance of this organism. When natural colonies were immersed in water, degradation of the colonies occurred within 2 days and N. commune cells were released into the water. The activities that hydrolyze glycoside bonds in various N. commune fractions were examined using artificial nitrophenyl-linked sugars as substrates. A beta-D-glucosidase purified from the water-soluble fraction was resistant to 20 min of boiling. The beta-D-glucosidase, with a molecular mass of 20 kDa, was identified as a cyanobacterial fasciclin protein based on its N-terminal amino-acid sequence. The 36-kDa major protein in the water-soluble fraction was purified, and the N-terminal amino-acid sequence of the protein was found to be identical to that of the water-stress protein (WspA) of N. commune. This WspA protein also showed heat-resistant beta-D-galactosidase activity. The fasciclin protein and WspA in the extracellular matrix may play a role in the hydrolysis of the EPSs surrounding the cells, possibly as an aid in the dispersal of cells, thus expanding the colonies of this cyanobacterium.
Sujet(s)
Stabilité enzymatique , Matrice extracellulaire/enzymologie , Glycosidases , Température élevée , Nostoc commune/enzymologie , Séquence d'acides aminés , Protéines bactériennes/composition chimique , Protéines bactériennes/isolement et purification , Protéines bactériennes/métabolisme , Matrice extracellulaire/composition chimique , Galactosidases/composition chimique , Galactosidases/isolement et purification , Galactosidases/métabolisme , Glucosidases/composition chimique , Glucosidases/isolement et purification , Glucosidases/métabolisme , Glycosidases/composition chimique , Glycosidases/isolement et purification , Glycosidases/métabolisme , Polyosides bactériens/composition chimique , Spécificité du substratRÉSUMÉ
Ferredoxin-NADP(+) oxidoreductase (FNR) catalyzing the terminal step of the linear photosynthetic electron transport was purified from the cyanobacterium Spirulina platensis and the red alga Cyanidium caldarium. FNR of Spirulina consisted of three domains (CpcD-like domain, FAD-binding domain, and NADP(+)-binding domain) with a molecular mass of 46 kDa and was localized in either phycobilisomes or thylakoid membranes. The membrane-bound FNR with 46 kDa was solublized by NaCl and the solublized FNR had an apparent molecular mass of 90 kDa. FNR of Cyanidium consisted of two domains (FAD-binding domain and NADP(+)-binding domain) with a molecular mass of 33 kDa. In Cyanidium, FNR was found on thylakoid membranes, but there was no FNR on phycobilisomes. The membrane-bound FNR of Cyanidium was not solublized by NaCl, suggesting the enzyme is tightly bound in the membrane. Although both cyanobacteria and red algae are photoautotrophic organisms bearing phycobilisomes as light harvesting complexes, FNR localization and membrane-binding characteristics were different. These results suggest that FNR binding to phycobilisomes is not characteristic for all phycobilisome retaining oxygenic photosynthetic organisms, and that the rhodoplast of red algae had possibly originated from a cyanobacterium ancestor, whose FNR lacked the CpcD-like domain.