Changes in M:G ratios of extracted and residual alginate fractions on boiling with water the dried brown alga Kjellmaniella crassifolia (Laminariales,Phaeophyta)

Kjellmaniella crassifolia, the edible macro-brown alga in Japan contained nearly 27% of alginates of which nearly 7% was extractable from the fronds with boiling water for 6 h and the residual alginates in the frond were almost exhaustively extracted with a dilute alkali at 60 °C for 6 h. The alginates dissolved in all these extracts with both boiling water and dilute alkali were purified by fractionation with MgCl₂ and alcohol.The content of MM blocks in the boiling water-soluble alginate sample increased remarkably during heating for 6 h while that of GG blocks from the same sample decreased. In contrast, MM blocks in the alkali-soluble alginate sample decreased during 6 h heating while GG blocks continued to increase. Since the amounts of MG blocks showed slight fluctation, the M:G ratio of alginates extracted with boiling water increased towards the end of extraction whereas the reverse is true for the alkali-soluble alginates.     

Direct determination of alginate content in brown algae by near infra-red (NIR) spectroscopy

The methods used to quantify total alginate in brown algal tissue are time-consuming and may also be misleading, so faster and simpler methods for measuring alginate content would be beneficial in a variety of applications. This study reports on the use of near infra-red (NIR) analysis to monitor the alginate content of Laminaria hyperborea stipe during biodegradation. NIR reflectance spectra were recorded for 78 different freeze-dried samples of its stipe. The samples were collected during several biological degradation experiments and the total alginate content varied from 2.2 to 40.8% Na-alginate (w/w), determined by established methods based on ion exchange. Data analysis was performed using multivariate calibration methods in order to relate the spectral data to the alginate content. PLS2 analysis revealed some dependence on material type, probably reflecting differences in polyphenol content. In the end, a PLS1 model with 9 components was selected. The calculated model was validated both with internal data and with an external test set. Internal full cross validation explained 96.6% of the variance in alginate content. The external validation showed that the PLS1 model was able to predict the alginate concentration with a root mean square prediction accuracy of 2.1%. This revised version was published online in June 2006 with corrections to the Cover Date.     

Sugar constituents of fucose-containing polysaccharides from various Japanese brown algae

Sugar constituents of the fucose-containing polysaccharides (FCPs) from 21 species of brown algae were analyzed. FCPs were extracted with hot water (100 °C, 4 h), separated by precipitation with 20% (v:v) ethanol in the presence of 0.05 M MgCl₂ to remove contaminating soluble alginate, and purified by DEAE-Sephadex column chromatography. The samples were hydrolyzed with HCI, and neutral sugar and uronic acid were separated by anion exchange chromatography. Their amounts were determined by gas-liquid chromatography. The neutral sugars in the FCPs from Ishige okamurae, Laminaria ochotensis, Myelophycus simplex, Padina arborescens and Sargassum thunbergii all contained arabinose, fucose, galactose, glucose, mannose, rhamnose and xylose residues. The FCPs from Ishige okamurae, Padina arborescens, Sargassum hemiphyllum, S. patents and S. sagamianum contained the four uronic acids, galacturonic acid, glucuronic acid, guluronic acid and mannuronic acid.     

Association between exudates of brown algae and polychlorinated biphenyls

Exudates from the brown algaeCaepidium antarcticum andDesmarestia sp. were investigated for their ability to associate with hydrophobic pollutants such as polychlorinated biphenyls (PCB s). The percentage of PCB associated with algal exudates ranged from 79% for decachlorobiphenyl to 23% for the pentachlorobiphenyl congener No. 95. Exudates from the tested brown algae may therefore alter the bioavailability of PCBs in natural or artificial ecosystems.     

Growth and size distribution of some large brown algae in Ohori,east coast of Korea

About fifty individuals of four dominant large brown algae were collected by random sampling at monthly intervals using SCUBA. Size frequencies of Sargassum confusum and S. horneri were concentrated mainly in the small size classes during the initial growth phase, but were more or less evenly distributed in most size classes during the fast growing phase. Undaria pinnatifida and Costaria costata showed the same tendency as Sargassum spp., that frequency distribution became even when the plants reached their maximum growth. The kelp species recorded their maximum lengths in March, whereas Sargassum spp. showed their peaks in early summer. The relationships between the length and weight in different growth phases appeared to be dependent on the phenology of each species.     

Extraction, isolation and cadmium binding of alginate from Sargassum spp.

Sargassum brown algal species have recently shown promise for use in flow-through column systems that rely on a passive ion-exchange mechanism for the remediation of toxic heavy metals such as Pd, Cd, and Zn from contaminated waters. To elucidate the metal binding mechanism and optimise this so-called biosorption process, detailed information on the biochemistry of the raw biomass and the alginate in particular is essential. This study focuses on the detailed characterisation (e.g., percentage of yield, block co-polymer structure) of the various fractions of material isolated from S. fluitans and S. oligocystum following a (i) standard neutral, (ii) alkaline (NaOH) and (iii) high-temperature alkaline alginate (80 °C; Na₂CO₃) extraction. Results indicate that the alginate yield was independent of the temperature or the extraction method employed (21.1 to 22.8% and 18.9 to 20.5% yields for S. fluitans and S. oligocystum, respectively). Furthermore, ¹H-nuclear magnetic resonance (NMR) analyses revealed that the alginates isolated by the three methods displayed nearly identical doublet -L-guluronic acid frequencies (F _GG; between 0.55 to 0.58 for both S. fluitans and S. oligocystum). Cadmium binding experiments (pH 4.5) further demonstrated that the three alginate extracts have similar metal binding capacities (uptake ranging from 1.59 to 1.81 mmol Cd/gram). The implementation of the high-temperature alkaline extraction procedure resulted in the isolation of a new acid-soluble fraction (ASF), capable of binding cadmium at pH 4.5, which cannot be isolated by the standard neutral extraction protocol. A preliminary characterisation of this ASF revealed the presence of minor quantities of proteins and sulphated polysaccharides, as well as traces of alginate and possibly other low-molecular weight uronic acid-containing polymers.     

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 m long × 1.0 m 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° to 35°C. The enzyme was activated by calcium ions, and maximum activity was observed in the presence of greater than 30 mM calcium ion.     

Bacterial communities of some brown and red algae from Peter the Great Bay, the Sea of Japan

The structure of microbial communities of brown algae, red algae, and of the red alga Gracilaria verrucosa, healthy and affected with thallus rot, were comparatively investigated; 61 strains of heterotrophic bacteria were isolated and characterized. Most of them were identified to the genus level, some Vibrio spp., to the species level according to their phenotypic properties and the fatty acid composition of cellular lipids. The composition of the microflora of two species of brown algae was different. In Chordaria flagelliphormis, Pseudomonas spp. prevailed, and in Desmarestia viridis, Bacillus spp. The composition of the microflora of two red algae, G. verrucosa and Camphylaephora hyphaeoides, differed mainly in the ratio of prevailing groups of bacteria. The most abundant were bacteria of the CFB cluster and pseudoalteromonads. In addition, the following bacteria were found on the surface of the algae: Sulfitobacter spp., Halomonas spp., Acinetobacter sp., Planococcus sp., Arthrobacter sp., and Agromyces sp. From tissues of the affected G. verrucosa, only vibrios were isolated, both agarolytic and nonagarolytic. The existence of specific bacterial communities characteristic of different species of algae is suggested and the relation of Vibrio sp. to the pathological process in the tissues of G. verrucosa is supposed.     

Local and chemical distribution of phlorotannins in brown algae

The local and chemical distribution of phlorotannins among the Japanese Laminariaceae, Eisenia bicyclis, Ecklonia cava and Ecklonia kurome, was investigated. As a result of light microscopy observations with vanillin-HCl staining, phlorotannins were found to be accumulated within the vegetative cells of the outer cortical layer of the thalli, regardless of the species, stage of growth or organ. Crude phlorotannins comprised about 3.0% of the algal powder for each of the algae. High-performance liquid chromatography (HPLC) showed that the phlorotannins of E. bicyclis were composed of phloroglucinol (0.9%), phloroglucinol tetramer (4.4%), eckol (7.5%), phlorofucofuroeckol A (21.9%), dieckol (23.4%), and 8,8'-bieckol (24.6%), plus some other unknown phenolic compounds (17.3%). The composition of the phlorotannins differed little among the Laminariaceae, except for a significantly larger amount of the tetramer, MW 478, in E. bicyclis.     

Host-virus interactions in marine brown algae

Ectocarpus-like marine brown algae are frequently parasitized by polyhedric DNA viruses. Infected hosts have been studied in unialgal and axenic cultures, and the present state of knowledge is summarized in regard to stage-specific virus expression, discharge and survival time of virus particles, infection mechanism, association with host's nuclear genome, passage of the virus genome through mitosis and meiosis of the host, suppression of symptoms and spontaneous recovery of infected plants, host specificity and intergeneric transmission, vitality of infected plants, pandemic occurrence of virus infections, molecular data on Ectocarpus and Feldmannia viruses, and algal DNA-viruses as potential vectors for gene transfer. A scheme for the nomenclature of brown algal viruses is proposed.     

Extracellular secretion of phenolic substances from living brown algae

The chemical structures of ultraviolet (UV)- absorbing substances secreted from the healthy living brown algae, Eisenia bicyclis and Ecklonia kurome, were demonstrated. The living activity of algal cells was critically examined using a confocal laser-scanning microscope after incorporation of fluorescein diacetate (FDA) into the cells. Using thin-layer chromatography (TLC), reversed-phase three-dimensional high-performance liquid chromatography (RP-3D-HPLC) and gas chromatography-mass spectrometry (GC-MS), it was found that the UV-absorbing substances (λ_max 265–270 nm) secreted from the living brown algae mainly consisted of the three monomeric bromophenols, 2,4-dibromophenol, 2,4,6-tribromophenol and dibromo-iodophenol, but not phloroglucinol or phlorotannins. The other minor compounds detected in the secretions were as follows: benzothiazole, fatty acids (14:1, 16:0 and 18:0 acids), franesol, 3-hydroxy-2,4,4-trimethylpentyl 2-methylpropanoate and squalene. Exudation of phloroglucinol and phlorotannins was ascertained to take place only after the cell death of these algae. These results indicate that, whilst the algae are alive, polymeric phlorotannins are strictly kept within the algal body, and only monomeric bromophenols are secreted into the seawater medium.     

High rhenium enrichment in brown algae: a biological sink of rhenium in the sea?

Twenty-one species of seaweed from the California coast were analyzed for rhenium. For the first time, high enrichment (thousandfolds) of rhenium relative to seawater was found in brown algae, but not in green or red algae. Brown algae was suggested as a biological sink of rhenium in the sea and the analogous behavior of technetium to rhenium was found in marine algae. Preliminary incubation experiments with a common brown alga (Pelvetia fastigiata) showed that algal surface is not a major accumulating locus of rhenium.     

The plastome of a brown alga,Dictyota dichotoma

Summary Plastids of the brown algaDictyota dichotoma contain a single homogeneous DNA species which bands at a buoyant density of 1.693 g/cm³ in neutral CsCl equilibrium density gradients. The corresponding nuclear DNA has a density of 1.715 g/cm³. The molecular size of the plastid DNA is 123 kbp as calculated by both electron microscopy of spread intact circular molecules and gel electrophoresis following single and double digestions with various restriction enzymes. A restriction map has been constructed using the endonucleases Sal I, Bam HI, and Bgl II which cleave theDictyota plastome into 6, 12, and 17 fragments, respectively. No large repeated regions, as found in chlorophycean andEuglena plastid DNAs, were detected.Dictyota dichotoma is the first member from the chlorophyll c-line of the algal pedigree for which a physical map of plastid DNA has been established. Dedicated to Professor Dr. W. Stubbe on the occasion of his 65th birthday.     

Metal concentrations in Sargassum algae from coastal waters of Nha Trang Bay (South China Sea)

The concentrations of metals, viz., Fe, Mn, Cu, Zn, Cd, Pb, and Ni, were investigated in brown algae of the genus Sargassum collected in the coastal waters of Nha Trang Bay (South China Sea). Metal concentrations in algae growing in the zone of influence of the city of Nha Trang were higher than those in macrophytes collected to the south and north of the city. Similar levels of Cu and Fe concentrations were observed in macrophytes from the urbanized coastal areas of Nha Trang Bay and in algae from the Peter the Great Bay (Sea of Japan). This is suggestive of comparable metal inputs from the anthropogenic sources of the cities of Nha Trang and Vladivostok (Peter the Great Bay). However, the concentrations of Pb, Ni, and Cd in algae from the coastal waters of Nha Trang Bay were lower than in macrophytes from the inshore zone of Vladivostok. The high level of pollution of sea water with Zn compounds near Nha Trang was local. The metal levels in macrophytes from background areas reflect the geochemical features of the environment. Sargassum algae from Nha Trang Bay had lower concentrations of Zn, Cd, and Ni and higher concentrations of Mn than algae from Peter the Great Bay. The background concentrations of Cu, Pb, and Fe in macrophytes from both bays were virtually the same.