Isolation and characterization of β-d-glucan,heteropolysaccharide, and trehalose components of the basidiomycetous lichen Cora pavonia

Cora pavonia, a lichen having a basidiomycetous mycobiont, is rich in protein (36%), of which 10% is tyrosine. α,α-Trehalose is present and was isolated in 4.4% yield. The lichen was found to contain polysaccharides typical of basidiomycetes and different from those of ascomycetes and ascomycetous lichens. Isolated and characterized were a β-d-glucan and a heteropolysaccharide containing l-rhamnose, l-fucose, d-xylose, d-mannose, d-glucose, and d-galactose. The β-d-glucan was highly branched with 21% of nonreducing end-groups, contained 3-O-, 6-O-, and 3,6-di-O-substituted β-d-glucopyranosyl units, and had a main chain consisting of (1→3)- and (1→6)-links. The heteropolysaccharide component contained mainly mannose and xylose, having a mannose-containing nucleus and a main chain with preponderant (1→3)-linked α-d-mannopyranosyl residues. These were unsubstituted (10%), and 4-O- (10%) and 2,4-di-O-substituted (10%) with residues of β-d-xylopyranose. On methylation analysis of the heteropolysaccharide, a capillary column of DB-210 proved to be particularly useful for gas-liquid chromatographic resolution of partially O-methylated alditol acetates.     

Comparative studies on the polysaccharides of Cladonia alpestris (reindeer moss), Cladonia confusa,and Cladonia amaurocraea

The chemical structures of polysaccharide components of three species of the lichen genus Cladonia were compared. C. alpestris and C. confusa are similar in overall growth appearance despite different habitats, and each contains traces of water-insoluble nigeran. The residual lichens gave almost pure d-galacto-d-mannans isolated via insoluble Cu complexes formed with Fehling solution. They were not identical but structurally related having (1→6)-linked α-d-mannopyranosyl main-chains substituted in different patterns by β-d-galacto- and α-d-mannopyranosyl groups. Supernatant solutions of the Fehling-solution precipitation contained high proportions of β-d-galactofuranosyl residues. The polysaccharide of C. alpestris contained consecutive (1→2)-linked α-d-mannopyranosyl units substituted in the 6-position by β-d-galactofuranose, whereas that of C. confusa was a d-galactan with both pyranosyl and furanosyl forms. The d-glucan component of C. amaurocraea was isolated together with d-galacto-d-mannan as insoluble Cu complexes. The former was isolated in good yield and proved to be water-insoluble pustulan. The galactomannan had the same overall structure as those of C. alpestris and C. confusa, but showed differences according to the ¹³C-n.m.r. spectra.     

Composition of turpentine from Pinus edulis wood oleoresin

Monoterpenoids and sesquiterpenoid hydrocarbons of Pinus edulis wood oleoresin were analyzed by chromatographic and spectroscopic methods. Monoterpenoid hydrocarbons (20·3%) were composed mainly of α-pinene, with camphene, β-pinene, 3-carene, sabinene, myrcene, limonene, β-phellandrene, trans-ocimene and terpinolene in secondary to trace amounts. Oxygenated terpenoids (0.28%) contained bornyl acetate and verbenone as major constituents, and linalool, camphor, terpinene-4-ol, citronellyl acetate, borneol, neral, α-terpineol, citronellol, nerol, and geraniol in smaller amounts. Oleoresin contained 1·1% of acetogenins, composed mainly of ethyl caprylate. Sesquiterpenoid hydrocarbons were high (5·7%) in oleoresin) and were composed of germacrene D as a major constituent (36·6%), of γ-amorphene, α-copaene, and longifolene as secondary constituents (5–20%), and β-farnesene, α- and γ-murolenes, β₁-, γ-, δ-, and ε-cadinenes, α-amorphene, δ-guaiene, sibirene, α-cubebene, β-copaene, β-ylangene, sativene, cyclosativene, β-bourbonene, α- and γ-humulenes, caryophyllene, α-longipinene and longicyclene in smaller amounts. Composition of P. edulis and of P. monophylla turpentines was found to be similar, with percentage of ethyl caprylate being the best distinguishing criterion.     

Über das auftreten α-alkyl-substituierter äpfelsäuren und β-hydroxy-β-alkyl-substituierter dicarbon- und tricarbonsäurederivate als normale stoffwechsel-produkte

Occurrence of α-alkyl-substituted malic acids, and β-hydroxy-β-alkyl-substituted dicarboxylic and tricarboxylic acid derivatives in normal urineUrine contains a number of α-hydroxy acids so far unknown to occur in biological liquids. Besides the already as urine constituent known methylmalic acid, also the ethyl, isopropyl and butyl derivatives of malic acid were found. Further metabolites in urine are a β-propyl-substituted β-hydroxyglutaric acid, a β-hydroxy-β-[methyl-carbomethoxy]-adipinic acid and two isomeric α-methylcitric acids.     

O-Methylated mannogalactan from the microalga Coccomyxa mucigena, symbiotic partner of the lichenized fungus Peltigera aphthosa

A structural study of the carbohydrates from Coccomyxa mucigena, the symbiotic algal partner of the lichenized fungus Peltigera aphthosa, was carried out. It produced an O-methylated mannogalactan, with a (1 → 6)-linked β-galactopyranose main-chain partially substituted at O-3 by β-Galp, 3-OMe-α-Manp or α-Manp units. There were no similarities with polysaccharides previously found in the lichen thallus of P. aphthosa. Moreover, the influence of lichenization in polysaccharide production by symbiotic microalgae and the nature of the photobiont in carbohydrate production in lichen symbiosis are also discussed.     

Evidence for novel linkages in a glycoprotein involving beta-hydroxyphenylalanine and beta-hydroxytyrosine.

Cutinase, an extracellular enzyme from Fusarium solani f. pisi contains about 4% covalently attached carbohydrates. Treatment of the enzyme with alkali resulted in β-elimination and generation of dehydroamino acids absorbing at 241 nm. NaB³H₄ treatment in 0.1 N KOH followed by hydrolysis of the labeled protein gave rise to tritiated alanine, α-aminobutyric acid, phenylalanine, and tyrosine. Chemical and enzymatic degradation of the labeled phenylalanine showed that this amino acid was a 1:1 mixture of D- and L-stereo-isomers and that³H was equally distributed between the α- and β-positions. Therefore it is concluded that this glycoprotein contained 0-glycosidic linkages not only at serine and threonine residues but also at β-hydroxyphenylalanine and β-hydroxytyrosine; the latter two have not been found heretofore.     

Amino acid sequences of allophycocyanin α- and β-subunits isolated from Anabaena cylindrica: Presence of an unknown derivative of aspartic acid in the β-subunit

Allophycocyanin (APC) α- and β-subunits were isolated from Anabaena cylindrica and their amino acid sequences were determined. The α- and β-subunits consisted of 160 and 161 amino acid residues, respectively, and lacked tryptophan. The β-subunit contained an unknown derivative of aspartic acid. The sequences were compared with those of other allophycocyanins.     

Lipids of the tropical seagrass Thallassia hemprichii

The component sterols, alcohols, hydrocarbons, monocarboxylic, α,ω-dicarboxylic and α- and ω-hydroxy acids from the leaves and roots of the tropical seagrass Thallassia hemprichii are reported. The leaves contained significant concentrations of cholest-5-en-3β-ol, a sterol not normally detected in either higher plants or seagrasses. The lower abundance of polyunsaturated fatty acids found in both the leaves and roots compared to other seagrass species may be a result of the warmer waters from which this species was collected. Solvent-extractable, long-chain (> C₂₂)α,ω-diacids, α- and ω-hydroxy and monocarboxylic acids were also isolated from the leaves. The distribution pattern of these lipids should enable these components along with other distinctive components to be used as chemical markers for this seagrass.     

The final structure of robinin and biorobin and their total synthesis

Robinin has been proved by total synthesis and by methylation analysis using GC-MS to be kaempferol-3- O-(6-O-α-l-rhamnopyranosyl -β-d-galactopyranoside)-7-O-α-l-rhamnopyranoside and not, as claimed by Maksjutina et al., a mixture of 4 glycosides containing the 7-O-rhamnosyl moiety in the α- and β-furanoside as well as in the α- and β-pyranoside forms. The assumption that the 3-O-rhamnosido -galactose moiety contained furanoid rings was also disproved.     

Phospholipid and acid composition of Nocardia and nocardoid bacteria as criteria of classification

Phospholipid and acid composition of 5 strains of ‘true’ Nocardia and 4 strains of nocardoid bacteria have been studied. A great homogeneity was found in all the Nocardia species: phospholipids consist of cardiolipin, phosphatidyl ethanolamine, phosphatidylinositol and phosphatidylinositol mannoside. Streptomyces (Nocardia) mediterranei did not contain phosphatidylinositol and Oerskovia (Nocardia) turbata had no phosphatidyl ethanolamine. The fatty acid composition of these phospholipids was determined and was found different in Nocardia and nocardoid species. Nocardia were rich in straight chain fatty acids and tuberculostearic acid while the phospholipids of nocardoid bacteria contained greater amounts of branched fatty acids. The fatty acids from acetone soluble lipids consisted of hydroxy and non-hydroxy compounds. Hydroxy acids were found in Nocardia which contained nocardic acids: high MW β-hydroxy α-branched acids and in S. mediterranei which contained β-hydroxy acids with 15–17 carbon atoms. Non-hydroxy acids were essentially palmitic and tuberculostearic acids in Nocardia species while S. mediterranei and O. turbata contained great amounts of iso acids from C₁₄ to C₁₇. Phospholipid and acid composition are discussed as criteria of taxonomic classification of Nocardia and related Actinomycetes.     

Studies on extracellular and intracellular purified amylases from a thermophilic Bacillus stearothermophilus

Extracellular and intracellular amylases have been purified from a thermophilic Bacillus stearothermophilus and further studies have been made with the purified enzyme. The molecular weights for extra- and intracellular α- and β-amylases were found to be 47 000, 58 000, 39 000 and 67 000, respectively. α-Amylase (1,4-α-d-glucan glucanohydrolase, EC 3.2.1.1) and glucoamylase (1,4-α-d-glucan glucohydrolase, EC 3.2.1.3) were glycoproteins, whereas β-amylase (1,4-α-d-glucan maltohydrolase, EC 3.2.1.2) had little or no carbohydrate moiety. Extracellular FI (α-amylase), FIII (glucoamylase), FIV and FV (α-amylase) had carbohydrate moieties of 14.4, 27.0, 11.0 and 12.5%, respectively, whereas intracellular amylases FI (α-amylase), FII (β-amylase) and FIII (α-amylase) contained 15.2, 0.8 and 13.4% carbohydrate, respectively. The amino acid profile of the amylase protein digest showed a total number of 16 amino acids with aspartic acid showing the highest value followed by glutamic acid and leucine plus isoleucine. Compared to other thermostable amylases, proline and histidine contents were low. Both α- and β- amylase had the - SH group at their active site, which was essential for enzyme activity. EDTA and parachloromercuribenzoate exhibited dose dependent non-competitive inhibition of enzyme activity indicating the involvement of a divalent cation and the - SH group for activity.     

The phenyl - and -L-idopyranosiduronic acids and some other aryl glycopyranosiduronic acids

Condensation of 1,2,3,4,6-penta-O-acetyl-á-l-idopyranose (1) with phenol yielded phenyl 2,3,4,6-tetra-O-acetyl-α- (2) and α-l-idopyranoside (4). Deacetylation of 2 and 4 afforded phenyl α and β-l-idopyranosides (3 and 5), respectively, the structures of which were verified by periodate oxidation studies. A platinum-catalyzed oxidation of 3 and 5 produced the amorphous phenyl α- and β-L-idopyranosiduronic acids (9 and 11), respectively, which were isolated as the crystalline cyclohexylammonium salts. Phenyl β- and α-d-glucopyranosiduronic acids are apparent minor byproducts of the catalytic oxidations, resulting from an inversion at C-5. _p-Nitrophenyl α-d-mannopyranosiduronic acid and _p-nitrophenyl α- and β-d-galactopyranosiduronic acids are also described.     

Chromatographic methods for the determination of alpha- and beta-5-phospho-D-ribose-alpha-1-pyrophosphate pools in bacteria.

Different methods for the determination of 5-phospho-d-ribose-α-1-pyrophosphate (PRPP) are described. PRPP from ³²P-labeled microorganisms is determined directly either alone or together with the ribo- or deoxyribonucleosidetriphosphates in thin-layer chromatographic systems, using chemicals of high purity. As little as 0.1 pmol of both the α- and β-anomeric forms can be detected. α-PRPP is also determined by a new enzymatical radioactive microassay; the lower limit of detection is 1 pmol. An anomerization from α- to β-PRPP was found to take place during the application of the PRPP onto polyethyleneimine cellulose plates if formic acid or cell extract or both is added to α-PRPP. If the α-PRPP is purified from the cell extract before application, no anomerization will take place. It is also found that PRPP is stable in dilute organic acids. During accumulation of PRPP in purine-starved S. typhimurium, 10% of the PRPP was found as β-PRPP in vivo.     

Bacillus subtilis SSE4 produces subtulene A, a new lipopeptide antibiotic possessing an unusual C15 unsaturated β-amino acid

Subtulene A, a new cyclic lipopeptide, was isolated from the culture broth of Bacillus subtilis SSE4. This antibiotic compound contained the seven common α-amino acids, l-Asn-1, d-Tyr-2, d-Asn-3, l-Gln-4, l-Pro-5, d-Asn-6, l-Ser-7 and the unique β-amino acid-8 present in the iturin family. 1D and 2D NMR, as well as MS analyses, identified the β-amino acid as 3-amino-13-methyltetradec-8-enoic acid, an Iso C15 long chain β-amino acid. B. subtilis SSE4 was also found to produce iturin A. B. subtilis SSE4 culture filtrate exhibited both antifungal and antibacterial activities.     

STRUCTURAL STUDIES ON THE GALACTOMANNANS OF LICHENS OF THE GENUSCLADONIA

Galactomannans were isolated fromCladonia signata,C. furcata,C. imperialis, andC. clathratavia successive alkaline extraction and precipitation with Fehling solution and Cetavlon. They were investigated using¹³C-NMR spectroscopy, methylation analysis, and Smith degradation, and their specific rotations and monosaccharide compositions determined. As with galactomannans of otherCladoniaspecies, they contained (1→6)-linked main chains of α-mannopyranose, which were non-substituted (structure4in Fig. 2), mono-substituted at O-2 with α-mannopyranose (structure6) or α-galactopyranose (structure1), O-4 with β-galactopyranose (structure2), and disubstituted at O-2 and O-4 with α-mannopyranosyl and β-galactopyranosyl units, respectively (structure5). Disubstitution was present to a greater extent in the galactomannans ofC. clathrataandC. imperialisthan in those ofC. signataandC. furcata. In the case of the galactomannans ofC. furcata,C. clathrata, andC. imperialis, substitution also occurred at O-2 withO-β-galactofuranosyl-(1→6)-O-α-mannopyranosyl units (structure7). As observed in previous investigations, the C-1 portion of the¹³C-NMR of mannose-containing polysaccharides is typical of the lichen species. However, those of galactomannans ofC. imperialisandC. clathrataare almost identical and, although other chemical data showed many structures in common, some differences were evident.     

Main regulatory element (MRE) of the <Emphasis Type="Italic">Danio rerio</Emphasis> α/β-globin gene domain exerts enhancer activity toward the promoters of the embryonic-larval and adult globin genes

In warm-blooded vertebrates, the α- and β-globin genes are organized in domains of different types and are regulated in different fashion. In cold-blooded vertebrates and, in particular, the tropical fish Danio rerio, the α- and β-globin genes form two gene clusters. A major D. rerio globin gene cluster is in chromosome 3 and includes the α- and β-globin genes of embryonic-larval and adult types. The region upstream of the cluster contains c16orf35, harbors the main regulatory element (MRE) of the α-globin gene domain in warm-blooded vertebrates. In this study, transient transfection of erythroid cells with genetic constructs containing a reporter gene under the control of potential regulatory elements of the domain was performed to characterize the promoters of the embryonic-larval and adult α- and β-globin genes of the major cluster. Also, in the 5th intron of c16orf35 in Danio rerio was detected a functional analog of the warm-blooded vertebrate MRE. This enhancer stimulated activity of the promoters of both adult and embryonic-larval α- and β-globin genes.     

L-Galactosaccharinic acids: new saccharinic acids formed by calcium hydroxide treatment of L-sorbose

The new saccharinic acids, α- and β-L-galactosaccharinic acids (2-C-methyl-L-xylonic and 2-C-methyl-L-lyxonic acids), have been synthesized. The same acids were isolated, as their lactones, from the syrup obtained by treatment of L-sorbose with calcium hydroxide. The acids were produced in comparable amounts, and the factors that might regulate the stereoselectivity of the benzylic acid type of rearrangement are discussed.     

Characterization of surface n-alkanes and fatty acids of the epiphytic lichen Xanthoria parietina, its photobiont a green alga Trebouxia sp., and its mycobiont, from the Jerusalem hills.

Surface alkanes and fatty acids from the thalli of the lichen Xanthoria parietina, its photobiont Trebouxia sp., and its mycobiont were analysed by GC-MS. The green alga Trebouxia sp. synthesized mainly unsaturated fatty acids such as (Z,Z,Z)-9,12,15-18 : 3 (Z,Z)-9,12-18 : 2 and (Z)-9-18 : 1, and light alkanes C8-C15 (up to 83% of total n-alkanes). However, the mycobiont contained mainly saturated fatty acids such as hexadecanoic (16 : 0) and octadecanoic acid (18 : 0), and also very long-chain n-alkanes C22-C34. Dehydroabietic acid was found in both lichen and mycobiont. The occurrence of different amounts of n-alkanes and fatty acids in the photobionts and mycobionts of X. parietina was shown for the first time. Lichens collected from different locations in the Jerusalem hills contained n-alkanes ranging in concentration from 187 to 211 mg x (g dry wt)-1; n-alkane concentrations in the photobiont and mycobiont were 17-24 and 215-262 mg x (g dry wt)-1, respectively.     

Teichoic acids of three type strains of the Bacillus subtilis group, Bacillus mojavensis VKM B-2650, Bacillus amyloliquefaciens subsp. amyloliquefaciens VKM B-2582, and Bacillus sonorensis VKM B-2652

Cell walls of three type strains of the Bacillus subtilis group, Bacillus mojavensis VKM B-2650, Bacillus amyloliquefaciens subsp. amyloliquefaciens VKM B-2582, and Bacillus sonorensis VKM B-2652, are characterized by the individual set of teichoic acids. All strains contained 1,3-poly(glycerol phosphates), unsubstituted, acylated with D-alanine, and glycosylated. The latter differ in the nature of the monosaccharide residue. Teichoic acids of B. mojavensis VKM B-2650^T and B. amyloliquefaciens subsp. amyloliquefaciens VKM B-2582^T contained α-glucopyranose, while those of B. sonorensis VKM B-2652^T contained β-glucopyranose and N-acetyl-α-D-glucosamine. Moreover, cell walls of B. mojavensis VKM B-2650^T contained a teichoic acid of poly(glycosylglycerol phosphate) nature with the following structure of the repeating unit: -4)-α-D-α-D-GlcpNAc-(1 → 3)]-Glcp-(1 → 2)-sn-Gro-(3-P-. The type strains have been characterized according to the composition of cell wall sugars and polyols. Application of teichoic acids (set and structure) as chemotaxonomic characteristics is discussed for six type strains of the Bacillus subtilis group. Polymer structures were determined by chemical and NMR spectroscopic techniques.     

Depsidones and fatty acids of Parmelia stygia

One Russian sample of the lichen Parmelia stygia has yielded fumarprotocetraric acid, while a second contained the same compound accompanied by colensoinic, lobaric, caperatic and norcaperatic acids.