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PubMed1976

Cyclic condensed metaphosphates and linear polyphosphates in brown and red algae.

The occurrence of linear condensed polyphosphates and cyclic condensed metaphosphates was studied by means of pulse-labeling with 32P-orthophosphate (3--5h) in a number of Phaeophyceae species: Pylaiella litoralis, Ilea fascia, Ectocarpus siliculosus and also Rhodophyceae species: Ceramiumdeslongchampsii, C. rubrum, Rhodomela confervoides, Porphyridium purpureum and P. aerugineum. Two-dimensional cellulose thin layer chromatography revealed that in all species studied 32P-radioactivity was generally present in all oligopolyphosphates containing 2 to 7 phosphate residues, in cyclic metaphosphates (tri-, tetra-, penta- and hexametaphosphates) and in high-molecular-weight condensed phosphates which remained at the starting point. Among the low-molecular-weight condensed inorganic phosphates the trimetaphosphate had a significantly higher specific activity than the other oligophosphates which were separated on the chromatography plates as measured by the direct scanning with a Geiger-Muller counter. The phosphate uptake strongly depends on the internal pool of reserve phosphates of the algae cells. The 32P-orthophosphate incorporation of the cells is low and sluggish when growning in a synthetic medium or in sea water. Accordingly 32P appeared preferentially in the low-molecular-weight fractions of condensed phosphates since the storage phosphates were not yet used. After previous incubation in a P-free culture medium of the algae the 32P was rather rapidly incorporated and was found mostly in the high-molecular-weight condensed phosphates. During MAK-chromatography the high-molecular-weight fractions were eluted together with the nucleic acids (tRNA and DNA) while most of the low-molecular-weight fractions left the column immediately on elution.

Eukaryota↗
PubMedDate not supplied

Isolation and characterization of a fucoidan-degrading marine bacterium.

Fucoidan, a mixture of sulfated fucose-containing polysaccharides, was prepared from the algal bodies of Cladosiphon okamuranus (class Phaeophyceae, order Chordariales, family Chordariaceae) with a yield of 2.0% of the wet weight of the alga. To obtain enzymes that digest the fucoidan, we screened bacteria in the guy contents of the sea cucumber Stichopus japonicus for their ability to decrease the fucoidan in their culture media, and successfully isolated one bacterial strain that could decrease it. The bacterial strain was gram-negative and possessed menaquinone 7 as the predominant respiratory quinone, and the GC content of its genomic DNA was 52%. The results of the phylogenetic analysis of its 16S ribosomal DNA sequence indicated that the bacterial strain was a member of the division "Verrucomicrobia." However, as the bacterial strain is phylogenetically and phenotypically distinct from verrucomicrobial species described previously, the strain was assumed to be a new member of the division "Verrucomicrobia." When the bacterial strain was cultivated in an algal fucoidan-containing medium, the strain decreased fucoidan from C. okamuranus (44%), Nemacystus decipiens (19%), Laminaria japonica (31%), Kjellmaniella crassifolia (23%), sporophyl of Undaria pinnatifida (22%), Fucus vesiculosus (42%), and Ascophyllum nodosum (61%).

Animals↗
PubMed2004

Isolation and characterization of a fucoidan-degrading marine bacterial strain and its fucoidanase.

A marine bacterial strain that degraded fucoidan from Kjellmaniella crassifolia (class Phaeophyceae, order Laminariales, family Laminariaceae) was isolated in our laboratory. The strain was gram-negative, ubiquinone 8 was the predominant respiratory quinone, and the GC-content of its genomic DNA was 36%. The cells of the strain were rod-shaped (2.0 microm long x 1.0 microm wide), and each cell was motile by means of one polar flagellum. Phylogenetic analysis of its 16S ribosomal DNA sequence indicated that it was a member of the family Alteromonadaceae. It produced a type of extracellular fucoidanase, an endosulfated fucan-digesting enzyme. The enzyme was purified with 3500-fold purity at 12.0% yield. Optimum conditions for the enzyme reaction were approximately pH 6.5 to 8.0 and temperature 30 degrees to 35 degrees C. The enzyme was activated by calcium ions, and maximum activity was observed in the presence of greater than 30 mM calcium ion.

Alteromonadaceae↗
PubMed2006

Homologs of the sexually induced gene 1 (sig1) product constitute the stramenopile mastigonemes.

The tripartite tubular mastigoneme on the anterior flagellum is a morphological feature that characterizes the stramenopiles. Mastigonemes are significant and potentially informative structures not only from the viewpoint of systematics, but also of cell biology. Nevertheless, few biochemical studies have been reported on stramenopile mastigonemes. The flagella of Scytosiphon lomentaria (Phaeophyceae) were successfully isolated and analyzed using SDS-PAGE followed by protein sequencing. The partial amino acid sequence of one flagellar protein (115kDa) showed high similarity with the sexually induced gene 1 (sig1) product of centric diatoms. A polyclonal antibody against the 115-kDa protein reacted not only to the shaft of mastigonemes in Scytosiphon lomentaria, but also another distinctly different stramenopile flagellate, Sulcochrysis biplastida (Dictyochophyceae). Therefore, we propose that the 115-kDa protein (i.e. Sig1 homologs) is a constituent of the tubular shaft of the mastigoneme.

Amino Acid Sequence↗
PubMed2000

Purification and characterisation of vanadium haloperoxidases from the brown alga Pelvetia canaliculata.

Two enzymes characterised as iodoperoxidases (PcI and PcII), with vanadium-dependent activity, have been purified from the brown alga Pelvetia canaliculata (L.) Decne et Thur. (Fucaceae, Phaeophyceae), collected in the Northern Portuguese coast, at Viana do Castelo. The relative molecular masses were 166 kDa for PcI and 416 kDa for PcII, as determined by gel filtration. SDS-PAGE shows that PcI has just one band corresponding to a subunit of 66 kDa, while PcII shows four bands (66, 72, 157 and 280 kDa). The following kinetic parameters have been determined from a steady-state analysis of the oxidation of iodide by H2O2: PcI, pHopt = 6.0, KM(I-) = 2.1 mM, KM(H2O2) = 110 microM, Ki(I-) = 127 mM; and PcII, pHopt = 6.5, KM(I-) = 2.4 mM, KM(H2O2) = 20 microM and Ki(I-) = 69 mM. These iodoperoxidases are thermostable, as also observed for vanadium bromo- and chloroperoxidases.

Catalytic Domain↗
PubMed2002

Oligoalginate recognition and oxidative burst play a key role in natural and induced resistance of sporophytes of laminariales.

Forty-five species of brown algae (Phaeophyceae) were surveyed for their capacity to respond by an oxidative burst to challenges with alginate oligosaccharides. Intertidal frondose brown algae (Fucales) constitutively released high quantities of peroxide. The capacity to recognize oligoguluronates and to react with an oxidative burst was confined to alginate-rich taxa with complex thallus morphology, epitomized by the sporophytes of Laminariales. When kelp sporophytes were impaired in their capacity to perform an oxidative burst by the NAD(P)H oxidase inhibitor diphenylene iodonium, they were readily degraded by their bacterial epiflora. Thus, in these algae, the oxidative response is an essential element of natural resistance. We also report on the establishment of a well-defined experimental system for investigations on kelp immunity, with Laminaria digitata as the host and its phaeophycean endophyte, Laminariocolax tomentosoides, as the pathogen. We found that an alginate-triggered oxidative burst significantly induces resistance in Laminaria digitata against infection. From these findings we infer that oligoalginate signals are important cues in the interaction between laminarialean kelps and potential pathogens.

Adaptation, Physiological↗
PubMed1985

Biosynthesis of algal pheromones. A model study with the composite Senecio isatideus.

Several cyclic and alicyclic C11 hydrocarbons have been shown to act as gamete releasing and/or attracting pheromones during sexual reproduction of brown algae (Phaeophyceae). The same compounds are also found in the essential oils of various plants, of which the occurrence of the cycloheptadiene-pheromone ectocarpene in Senecio isatideus (Compositae) is noteworthy. Administration of [3H]dodeca-3,6,9-trienoic acid to cuttings of this plant leads to incorporation of radioactivity into ectocarpene. Double-bond-deuterated nona-3,6-dienoic acid is converted to fucoserratene, the pheromone of several Fucales, which is certainly not present among the hydrocarbons of Senecio. This proves that the pool of medium-chain, multiply unsaturated fatty acids includes precursors of all types of highly unsaturated hydrocarbons. Appropriately labelled (deuterium markers) fatty acid homologues were synthesized and applied to Senecio plantlets to unravel the mechanistic aspects. The results strongly suggest radical initiation of the pheromone biosynthesis by abstraction of a single hydrogen from a 1,4-pentadienyl segment of the fatty acid followed by oxidation to the corresponding cation. This causes fragmentation of the reactive intermediate into an olefine and carbon dioxide by neighbouring-group participation of the flanking double bonds. A tentative biosynthetic scheme is deduced from the experimental results which also sets the stereochemistry of the algal pheromones into a uniform mechanistic concept.

Animals↗
PubMed1975

Polysaccharide production and the possible occurrence of GDP-D-mannose dehydrogenase in Azotobacter vinelandii.

During the growth of Azotobacter vinelandii in batch culture in Burk's 2% glucose medium supplemented with 50 mg EDTA per litre, water-insoluble capsular polysaccaride material accumulated in cultures prior to the appearance of water-soluble polysaccharide in the culture medium. On isolation, hydrolysis and chromatography, both these polysaccharides were observed to be composed of carbohydrate monomers having the same chromatographic mobilities as glucose, rhamnos, guluronic acid and mannuronic acid. The activity of GDP-D-mannose dehydrogenase recorded in crude cell-free extracts from Azotobacter vinelandii, when these polysaccharides were produced, may indicate a close similarity between the biosynthetic pathway of alginate synthesis in marine Phaeophyceae and this soil microorganism.

Alcohol Oxidoreductases↗
PubMed2002

A marine strain of flavobacteriaceae utilizes brown seaweed fucoidan.

Fucoidan, a mixture of sulfated fucose-containing polysaccharides, was prepared from Kjellmaniella crassifolia (class Phaeophyceae, order Laminariales, family Laminariaceae) with a yield of about 3.8% dry weight. To isolate enzymes that degrade fucoidan, we first screened marine bacteria for their ability to utilize fucoidan, and isolated one strain of Flavobacteriaceae from seawater that could do this. Phylogenetic analysis of the 16S ribosomal DNA sequence suggested that this strain appeared to belong to a new genus, and was tentatively named Fucobacter marina. The strain utilized L-fucose (17%), D-mannose (91%), D-galactose (46%), and D-glucuronic acid (66%) in the fucoidan from K. crassifolia. The strain partially utilized fucoidan from 2 other seaweeds that belong to the order Laminariales, Undaria pinnatifida (10%) and Lessonia nigrescens (48%).

Journal Article↗
PubMed1994

Structure and biosynthesis of marine algal oxylipins.

Diverse marine life, including algae, sponges, molluscs, corals, tunicates, and bacteria, have been found to possess a variety of structurally unique oxylipins. The algae are the best characterized of these organisms for their oxylipins, which have now been described from more than 30 species representing the three major groups of macrophytic algae (Rhodophyta = reds, Chlorophyta = greens, and Phaeophyceae = browns). A number of recent studies have sought to understand the biosynthetic origin and mechanistic chemistry which leads to the formation of these unique marine substances. In general, the red algae metabolize C20 acids via 12-lipoxygenase-initiated pathways, green algae metabolize C18 acids at C-9 and C-13, and brown algae metabolize both C18 and C20 acids, principally by lipoxygenases with n-6 specificity. This review updates the records of new oxylipins from marine algae and describes thoughts on their biogenesis as well as specific experiments aimed at probing these hypotheses.

Cyclopentanes↗
PubMed1996

Effects of soluble sodium alginate on cholesterol excretion and glucose tolerance in rats.

We studied the effects of a natural sodium alginate (isolated from Laminaria angustata Kjellman var. longissima Miyabe, Phaeophyceae) (average molecular weight: 2700 kDa; AG-270) and three water-soluble low-molecular weight sodium alginates (average molecular weights, 10, 50 and 100 kDa; AG-1, AG 5, and AG-10, respectively) on cholesterol excretion and glucose tolerance in rats. AG-270, AG-5 and AG-10 enhanced cholesterol excretion into faeces. AG-270 and AG-10 inhibited blood glucose and insulin levels from rising 30 min after glucose administration. AG-5 inhibited the blood glucose level from rising 30 and 60 min after glucose administration, without affecting blood insulin levels. AG-1 had no effect on cholesterol excretion or glucose tolerance. These findings suggest that the effects of the natural sodium alginate and AG-5 and AG-10 on cholesterol excretion and glucose tolerance may be due to the inhibition of cholesterol and glucose absorption from the small intestine by the gelling of the free alginic acid converted in the stomach. These experimental results indicate that the low-molecular weight sodium alginates, AG-5 and AG-10, should be useful as dietary fibers for the prevention of obesity, hypercholesterolemia, and diabetes.

Administration, Oral↗
PubMed2004

Morphological convergence characterizes the evolution of Xanthophyceae (Heterokontophyta): evidence from nuclear SSU rDNA and plastidial rbcL genes.

Xanthophyceae are a group of heterokontophyte algae. Few molecular studies have investigated the evolutionary history and phylogenetic relationships of this class. We sequenced the nuclear-encoded SSU rDNA and chloroplast-encoded rbcL genes of several xanthophycean species from different orders, families, and genera. Neither SSU rDNA nor rbcL genes show intraspecific sequence variation and are good diagnostic markers for characterization of problematic species. New sequences, combined with those previously available, were used to create different multiple alignments. Analyses included sequences from 26 species of Xanthophyceae plus three Phaeothamniophyceae and two Phaeophyceae taxa used as outgroups. Phylogenetic analyses were performed according to Bayesian inference, maximum likelihood, and maximum parsimony methods. We explored effects produced on the phylogenetic outcomes by both taxon sampling as well as selected genes. Congruent results were obtained from analyses performed on single gene multiple alignments as well as on a data set including both SSU rDNA and rbcL sequences. Trees obtained in this study show that several currently recognized xanthophycean taxa do not form monophyletic groups. The order Mischococcales is paraphyletic, while Tribonematales and Botrydiales are polyphyletic even if evidence for the second order is not conclusive. Botrydiales and Vaucheriales, both including siphonous taxa, do not form a clade. The families Botrydiopsidaceae, Botryochloridaceae, and Pleurochloridaceae as well as the genera Botrydiopsis and Chlorellidium are polyphyletic. The Centritractaceae and the genus Bumilleriopsis also appear to be polyphyletic but their monophyly cannot be completely rejected with current evidence. Our results support morphological convergence at any taxonomic rank in the evolution of the Xanthophyceae. Finally, our phylogenetic analyses exclude an origin of the Xanthophyceae from a Vaucheria-like ancestor and favor a single early origin of the coccoid cell form.

Base Composition↗
PubMed2003

Use of FTIR, FT-Raman and 13C-NMR spectroscopy for identification of some seaweed phycocolloids.

Many seaweeds produce phycocolloids, stored in the cell wall. Members of the Rhodophyceae produce polysaccharides the main components of which are galactose (galactans)-agar and carrageenan. In addition, alginic acid is extracted from members of the Phaeophyceae. This is a binary polyuronide made up of mannuronic acid and guluronic acid. The wide uses of these phycocolloids are based on their gelling, viscosifying and emulsifying properties, which generate an increasing commercial and scientific interest. In this work, the FTIR and FT-RAMAN spectra of carrageenan and agar, obtained by alkaline extraction from different seaweeds (e.g. Mastocarpus stellatus, Chondrus crispus, Calliblepharis jubata, Chondracanthus acicularis, Chondracanthus teedei and Gracilaria gracilis), were recorded in order to identify the type of phycocolloid produced. The spectra of commercial carrageenan, alginic acid and agar samples (SIGMA and TAAB laboratories) were used as references. Special emphasis was given to the 500-1500 cm(-1) region, which presents several vibrational modes, sensitive to the type of polysaccharide and to the type of glycosidic linkage. The FT-Raman spectra present a higher resolution than FTIR spectra, this allowing the identification of a larger number of characteristic bands. In some cases, phycocolloids can be identified by FT-Raman spectroscopy alone.

Agar↗
PubMed2003

Characterization of mannuronan C-5-epimerase genes from the brown alga Laminaria digitata.

Alginate is an industrially important polysaccharide obtained commercially by harvesting brown algae. The final step in alginate biosynthesis, the epimerization of beta-1,4-d-mannuronic acid to alpha-1,4-l-guluronic acid, a structural change that controls the physicochemical properties of the alginate, is catalyzed by the enzyme mannuronan C-5-epimerase. Six different cDNAs with homology to bacterial mannuronan C-5-epimerases were isolated from the brown alga Laminaria digitata (Phaeophyceae). Hydrophobic cluster analysis indicated that the proteins encoded by the L. digitata sequences have important structural similarities to the bacterial mannuronan C-5-epimerases, including conservation of the catalytic site. The expression of the C-5-epimerase genes was examined by northern-blot analysis and reverse transcriptase-polymerase chain reaction in L. digitata throughout a year. Expression was also monitored in protoplast cultures by northern and western blot, reverse transcriptase-polymerase chain reaction, and activity measurements. From both the structural comparisons and the expression pattern, it appears that the cDNAs isolated from L. digitata encode functional mannuronan C-5-epimerases. The phylogenetic relationships of the bacterial and brown algal enzymes and the inferences on the origin of alginate biosynthetic machinery are discussed.

Amino Acid Sequence↗
PubMed1995

Amplification of DNA polymerase gene fragments from viruses infecting microalgae.

Nested PCR with three highly degenerate primers was used for amplification and identification of DNA polymerase (pol) genes from viruses which infect three genera of microalgae. Group-specific primers (AVS1 and AVS2) were designed on the basis of inferred amino acid sequences unique to the DNA pol genes of viruses (PBCV-1 and NY-2A) that infect an endosymbiotic Chlorella-like alga (Chlorophyceae) and a virus (MpV-SP1) which infects the photosynthetic flagellate Micromonas pusilla (Prasinophyceae). In addition, a nested primer (POL) was designed on the basis of the highly conserved amino acid sequence YGDTDS found in most B-family (alpha-like) DNA pol genes. These primers were used to amplify DNA from the three viruses, PBCV-1, NY-2A, and MpV-SP1, for which the primers were designed, as well as eight clonal isolates of genetically distinct viruses which infect M. pusilla and others which infect Chrysochromulina spp. (Prymnesiophyceae), suggesting that these are a group of related viruses. In contrast, no product resulted from using DNA from viruses which infect the marine brown algae Ectocarpus siliculosis and Feldmannia sp. (Phaeophyceae), suggesting that these viruses may not be closely related to those that infect microalgae. These primers were also used to amplify DNA from natural virus communities. Our results indicate that nested PCR, even under low-stringency conditions, can be used as a rapid method to verify the presence in seawater of a group of related viruses which infect microalgae. Sequence analysis of these fragments should provide information on the genetic diversity and potentially the phyletic relationships among these viruses.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acid Sequence↗
PubMed1979

The route of entry of cytoplasmically synthesized proteins into chloroplasts of algae possessing chloroplast ER.

In 8 classes of algae, namely the Cryptophyceae, Raphidophyceae, Haptophyceae, Chrysophyceae, Bacillariophyceae, Xanthophyceae, Eustigmatophyceae and Phaeophyceae, the chloroplasts, in addition to being surrounded by a double-membraned chloroplast envelope, are also enclosed by a cisterna of endoplasmic reticulum called the chloroplast ER. Often this ER cisterna is continuous with the outher membrane of the nuclear envelope in such a manner that the nuclear envelope forms a part of the ER sac enclosing the chloroplast. In all these classes of algae except the Cryptophyceae, a regular network of tubules and vesicles, named the periplastidal reticulum, is present at a specific location between the chloroplast envelope and the chloroplast ER. In the Cryptophyceae, scattered vesicles are found between the chloroplast envelope and the chloroplast ER. Ribosomes which have been shown to be arranged to polysomes are found on the outer membrane of the chloroplast ER. It is proposed that nuclear-coded proteins which are destined for the chloroplast are synthesized on these polysomes, passing during synthesis into the lumen of the ER cisterna. Vesicles containing these proteins then pinch off the chloroplast ER and form the periplastidal reticulum. Vesicles containing these proteins then pinch off the chloroplast ER and form the periplastidal reticulum. Vesicles then fuse with the outer membrane of the chloroplast envelope thereby delivering their contents to the lumen of the chloroplast envelope. Proteins then cross the inner membrane of the chloroplast envelope in an as yet unknown manner. Experimental evidence for this hypothesis comes from studies on Ochromonas danica using chloramphenicol and spectinomycin, which inhibit protein synthesis on plastid ribosomes, and cycloheximide, which inhibits protein synthesis on cytoplasmic ribosomes. In cells of Ochromonas exposed to chloramphenicol or spectinomycin, the periplastidal reticulum proliferates markedly becoming several layers thick. Presumably this build up of periplastidal reticulum occurs because the transport of cytoplasmically synthesized plastid proteins is slowed down when protein synthesis in the chloroplast is inhibited. Conversely, when cells of Ochromonas are treated with cycloheximide, there is a reduction in the amount of periplastidal reticulum presumably because there are no cytoplasmically synthesized proteins to be transported into the chloroplast.

Biological Transport↗
PubMed2000

[Substance isolated from the kelp rhizoid identified as L-tryptophan shows high inhibition of breast cancer].

In general, the root of a seaweed is poorly developed as compared with its thallus and is called the rhizoid or holdfast. In Laminaria, belonging to Phaeophyceae, although the thallus is used for food, the rhizoid is considered an unuseful natural resource. We attempted to detect anti-breast cancer substances from that resource. As a result, a substance having a weak absorptivity to aluminium oxide and Sephadex G-25 was found. According to analysis of the FAB-MS spectra and 1H NMR spectra, the substance was identified as tryptophan, an amino acid. Finally, it was concluded by a chiral column-HPLC method that the tryptophan was the L-form.

Antineoplastic Agents↗
PubMed1964

STUDIES ON INHIBITION OF INTESTINAL ABSORPTION OF RADIOACTIVE STRONTIUM. I. PREVENTION OF ABSORPTION FROM LIGATED INTESTINAL SEGMENTS.

A method is reported which permits selective suppression of absorption of radioactive strontium from ingested food material, permitting the calcium to be available to the body. Studies were carried out in vivo by injection of Sr(89) and Ca(45) in the presence of inert carrier into ligated intestinal segments in rats, and the amount of absorption was measured by standard monitoring techniques. The pattern of absorption of both ions is very similar but the rate of absorption is different. It was found that the polyelectrolyte, sodium alginate, obtained from brown algae (Phaeophyceae), injected simultaneously with radiostrontium effectively reduces the absortion of Sr(89) from all segments of the intestine by as much as 50-80% of the control values. No significant reduction in absorption of Ca(45) was observed in equivalent concentrations. The reduction in blood levels of Sr(89) and in bone uptake corresponded to the absorption pattern. The difference in the effect on strontium and calcium absorption may be due to differences in the binding capacity of sodium alginate from the two metal ions under the conditions present in vivo.

Absorption↗