[Polar components of Black sea seaweed Colpomenia peregrina (Sauv.) Hamel]. / Poliarnye komonenty chernomorskoi buroi vodorosli Colpomenia peregrina (Sauv.) Hamel.

GC-MS of trimethylsilyl derivatives of the compounds present in the butanolic extract of biomass of brown seaweed Colpomenia peregrina from the Black Sea aided in identification of 24 components, including aliphatic hydroxy and keto and aromatic acids, glycerol, mannitol, floridoside, and monosaccharides. The polysaccharide composition of the biomass was also studied, with high sodium alginate and laminaran contents and a comparatively low level of fucoidan being revealed. The polysaccharides were isolated from the biomass by fractional extraction and purified by precipitation or ion exchange chromatography. The structures of alginic acid and laminaran were deduced from 13C NMR spectra and confirmed, in the case of laminaran, by methylation analysis. The sodium alginate was shown to contain more guluronic (G) than mannuronic acid (M) residues, the M/G ratio being 0.48. Laminaran was demonstrated to be a beta-glucan with 1-->3 linkages in its backbone and 1-->6 linkages in its branching points, which is characteristic of brown algae. Fucoidan turned out to be a complex heteropolysaccharide containing, in addition to fucose and sulfate, other neutral monosaccharides and uronic acids. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2004, vol. 30, no. 2; see also http://www.maik.ru

[Polar constituents of the ascidia Botryllus schlosseri]. / Poliarnye komponenty astsidii Botryllus schlosseri.

Eighteen compounds were identified by GC-MS of their trimethylsilyl derivatives in n-butanolic extract from the biomass of Botryllus schlosseri. Three of them, 5-oxoproline, 5-hydroxyhydantoin, and kinurenic acid, were found in marine invertebrates for the first time. In addition to cellulose, the biomass was also shown to contain complex water-soluble sulfated polysaccharides. These were extracted and fractionated, and sulfate content and monosaccharide composition were determined in the fractions; fucose, xylose, galactose, mannose, glucose, glucosamine, galactosamine, and uronic acids were found. Unlike several other tunicate species, Botryllus schlosseri does not seem to contain any simple galactan sulfate.

Characterization of novel methyl-branched chain fatty acids from a halophilic Bacillus species.

The 4-methylated fatty acids 4,9-dimethyldecanoic, 4,11-dimethyldodecanoic, 4,10-dimethyldodecanoic, and 4,13-dimethyltetradecanoic acid as well as the 2-methylated fatty acids 2,13-dimethyltetradecanoic and 2,12-dimethyltetradecanoic acid were identified for the first time in nature in the halophilic bacterium Bacillus sp. isolated from the salt pans of Burgas in Bulgaria. The principal fatty acids in this bacterium were a series of iso-anteiso fatty acids with chain lengths between C11 and C19, but an interesting series of linear alkylbenzene fatty acids with chain lengths between C10 and C14, such as 12-phenyldodecanoic acid, were also identified. The novel 4-methylated fatty acids were characterized using a combination of GC-MS and chemical transformations such as N-acylpyrrolidide derivatization. The 2-methylated fatty acids were also identified by GC-MS and gas chromatographic coelution with synthetic samples. The novel methyl-branched fatty acids probably originated from the selective incorporation of methylmalonyl-CoA by one of the fatty acid-synthesizing enzymes of the bacterium.

Development in a primary cell culture of the marine sponge Ircinia muscarum and analysis of the polar compounds.

We have established a primary cell culture of the marine demosponge Ircinia muscarum. The culture was started from a cell suspension obtained by a combination of mechanical chemical means. Microbial contamination was controlled by the use of a pool of antibiotics. Optical density, rather than hemocytometer count, is suggested to monitor the cellular growth. Analysis of the chemical composition of I. muscarum cells revealed absence of sterols, showing that the cells were unable to biosynthesize sterols. When the medium was supplemented with cholesterol an increase of about 70% in the number of cells was observed. These results suggest that the classic mammalian nutrient medium was not satisfactory for I. muscarum cell growth, and sterols were needed to satisfy the membrane requirements.

Chemical composition and biological activities of the Black Sea algae Polysiphonia denudata (Dillw.) Kutz. and Polysiphonia denudata f. fragilis (Sperk) Woronich.

The two investigated algae had almost identical sterol composition, but there were significant differences in the composition of the polar components and especially in the composition of the volatiles. P. denudata f. fragilis extracts possessed a stronger biological activity (antibacterial, antifungal and toxicity against Artemia salina). Despite the minute morphological differences between the two algae, we recommend P. denudata f. fragilis to be regarded as P. denudata subsp. fragilis.

Chemical composition of the lipophylic extract from the tunicate Botryllus schlosseri.

Eighteen sterols were identified in Botryllus schlosseri, stanols being the main sterols. The sterol composition is in agreement with our recent paper on taxonomic separation of tunicates into three groups. Again we found in invertebrates of the Black Sea sterols with a (22Z)-double bond. This confirms the presence of such sterols in nature. The composition of the phospholipids appeared to be complex, and twelve groups of them were identified in Botryllus schlosseri. The main representatives of phospholipids appeared to be phosphatidylcholine and phosphatidylethanolamine. Only four volatile compounds were identified, which is unusual for marine invertebrates.

Two novel iso-branched octadecenoic acids from a Micrococcus species.

The novel fatty acids 16-methyl-6(Z)-heptadecenoic acid and 16-methyl-8(Z)-heptadecenoic acid were identified for the first time in nature in a species of the bacterium Micrococcus isolated from Lake Pomorie in Bulgaria. The principal fatty acids in this bacterium were a series of iso-anteiso fatty acids with chain lengths between C(14) and C(24), while the most interesting series of monounsaturated fatty acids was a family of Delta(6) fatty acids with chain lengths between C(14) and C(17). The novel compounds were characterized using a combination of GC-MS and chemical transformations, such as dimethyl disulfide derivatization and catalytic hydrogenation. The results established for the first time a bacterial origin for some of these Delta(6) fatty acids.

Metabolites from a marine bacterium Pseudomonas/Alteromonas, associated with the sponge Dysidea fragilis.

The lipophylic extract from a Black Sea bacterium, associated with the sponge Dysidea fragilis, was investigated. Saturated hydrocarbons and fatty acids of the lipids were identified. The concentrations of the polycyclic compounds appeared to be negligible. The main components appeared to be phosphatidyl ethanolamine, followed by phosphatidyl serine. The first was investigated by FAB mass spectrometry and a series of molecular species partially identified.

Antiinfluenza virus effect of extracts from marine algae and invertebrates.

Sixty products, derived from marine organisms, typical of the Bulgarian Black Sea coast, were examined for inhibitory activity on the reproduction of influenza viruses in tissue cultures. The antiviral effect was investigated by the reduction of virus infectivity. Using representative strains of influenza virus it was shown that apparently the inhibitory effect was strain-specific. The most effective products were further studied in fertile hen's eggs and in experimental influenza infection in white mice.

Unusual lipid composition of a Bacillus sp. isolated from Lake Pomorie in Bulgaria.

The lipid composition of a Bacillus sp., isolated from Lake Pomorie in Bulgaria, was unusual and consisted of 26 different fatty acids between C12 and C26, with anteiso C15-C17 saturated fatty acids predominating. The furan fatty acid, 10,13-epoxy-11-methyloctadeca-10,12-dienoic acid, was also identified, a new finding for this genus. The hydrocarbons consisted of 30 different monounsaturated hydrocarbons, between C25 and C30, with the iso-iso, iso-anteiso, anteiso-anteiso, iso-normal, and anteiso-normal methyl branching for odd-numbered chains, and the iso-iso, iso-anteiso, iso-normal, and anteiso-normal methyl branching for even-numbered chains. The double bond positions in these hydrocarbons were determined by dimethyl disulfide derivatization followed by GC-MS, and the double-bond cis configuration was confirmed by infrared spectroscopy. Some previously unknown hydrocarbons in bacteria, such as (Z)-3,21-dimethyl-9-tricosene, (Z)-3,21-dimethyl-10-tricosene, (Z)-2,24-dimethyl-11-pentacosene, and (Z)-2,25-dimethyl-13-hexacosene were identified. Sterols were detected and were based on the sitosterol nucleus.

Novel di-, tri-, and tetraenoic fatty acids with bis-methylene-interrupted double-bond systems from the sponge Haliclona cinerea.

Unusual fatty acids, with up to 34 carbon atoms and containing one or two bis-methylene-interrupted double-bond systems, have been identified in the sponge Haliclona cinerea from the Black Sea. These include the dienes-5,9-16:2, 7,11-18:2, 9,13-20:2, 13,17-24:2, 15,19-26:2, 17,21-28:2, 19,23-30:2 and 21,25-32:2; trienes-5,9,23-30:3, 5,9,24-31:3, 5,9,25-32:3, and 5,9,27-34:3; and the tetraenes-5,9,19,23-30:4, 5,9,21,25-32:4, and 5,9,23,27-34:4. In addition, 5,9,13-eicosatrienoic acid was present. Many of these do not appear to have been described before, and only 5,9-16:2 and 5,9,23-30:3 are found often in sponges. They were identified by using silver-ion high-performance liquid chromatography to simplify the complex mixture of fatty acids for subsequent analysis by gas chromatography-mass spectrometry as picolinyl ester derivatives. Deuteration with Wilkinson's catalyst in homogeneous solution confirmed the structures. We speculate that the di- and tetraenoic fatty acids arise by chain elongation of 5,9-hexadecadienoic acid, also a major component of the lipids, followed by further insertion of double bonds in the 5 and 9 positions. The trienes may be formed from 9-hexadecenoic acid by similar mechanisms.

The fatty acid composition of a Vibrio alginolyticus associated with the alga Cladophora coelothrix. Identification of the novel 9-methyl-10-hexadecenoic acid.

The fatty acid composition of a new strain of Vibrio alginolyticus, found in the alga Cladophora coelothrix, was studied. Among 38 different fatty acids, a new fatty acid, 9-methyl-10-hexadecenoic acid and the unusual 11-methyl-12-octadecenoic acid, were identified. Linear alkylbenzene fatty acids, such as 10-phenyldecanoic acid, 12-phenyldodecanoic acid and 14-phenyltetradecanoic acid, were also found in V. alginolyticus. The alga contained 43% saturated fatty acids, and 28% C16-C20 polyunsaturated fatty acids of the n-3 and n-6 families.

Comparison of the fatty acids of the tunicate Botryllus schlosseri from the Black Sea with two associated bacterial strains.

The fatty acid composition of the tunicate Botryllus schlosseri and of two bacterial strains found within the tunicate, namely Vibrio parahaemolyticus and of an associated but previously unreported gram positive cocci were studied. The polyunsaturated fatty acids 6,9,12-octadecatrienoic acid, 5,8,11,14,17-eicosapentaenoic acid, and 4,7,10,13,16,19-docosahexaenoic acid were particularly abundant in B. schlosseri and were not detected in the two bacterial strains found in the tunicate. The iso/anteiso pair, 13-methyltetradecanoic acid and 12-methyltetradecanoic acid, were the principal fatty acids in the gram positive cocci, and the 9- and 11-hexadecenoic acids were particularly abundant in V. parahaemolyticus. The diunsaturated fatty acid 9,12-octadecadienoic acid was also shown to be present in V. parahaemolyticus. The fatty acid composition of a third bacterial strain, characterized as either a Pseudomonas or an Alteromonas species, and shown to be present only in the sea water from the Black Sea and not in B. schlosseri, is also reported. This is the first investigation on fatty acids from Black Sea bacteria.

An unusual fatty acid composition for a fresh-water mussel, Unio tumidus, from Bulgaria.

A combination of hplc in the silver ion mode and gc-ms of picolinyl ester derivatives was used to identify the fatty acids in a fresh-water mussel, Unio tumidus, from the Danube in Bulgaria. A number of novel fatty acids were found, including 14-methylpentadec-6-enoic and 17-methyloctadec-8-enoic acids. Eicos-7-enoic acid was a major component.

Comparative investigation of phospholipids and fatty acids of freshwater molluscs from the Volga river basin.

1. Four Gastropoda species and two Bivalvia species from the Volga river basin were examined. 2. Distribution of phospholipids in the molluscs was studied by qualitative and quantitative micro thin-layer chromatography. 3. Major phospholipid classes, phosphatidylethanolamine and phosphatidylcholine, were found to contain plasmalogens. 4. One mollusc species notably contained 67 fatty acids including 25 saturated (both iso and anteiso), 24 monoenoic, five dienoic, four trienoic and eight polyenoic compounds identified by capillary gas chromatography; fatty acid contents in the other studied species were considerably lower. 5. Relatively high concentrations of nonmethylene-interrupted fatty acids were detected in certain examined species.