The Oligosaccharides Produced from Sucrose by Dematium pullulans

A tetrasaccharide produced from sucrose by Dematium pullulans was obtained in a crystalline state, and its properties and physical constants were studied. By elementary analysis, molecular weight, tetrazolium test, the Raybin test, aldose value, paper chromatographic examination of partial acid hydrolyzate and enzymic hydrolyzate, periodate oxidation, and molar optical rotation, the crystalline compound was identified as β-fructofuranosyl-1-kestose trihydrate.     

Evidence for hemocyanin formation in the compound eye of Squilla mantis (Crustacea,Stomatopoda)

Summary In the retina and in the subretinal space of the compound eye of Squilla mantis a special kind of pigment cell is present. The crystalline inclusions of this cell have been identified as hemocyanin, as determined (i) by the dimensional congruence of the crystalline substructure with the dimensions of isolated, purified hemocyanin, and (ii) by the immunofluorescence reaction using anti-hemicyanin antibodies. The ultrastructure of these cells, their location and the presence of crystalline bodies in their cytoplasm suggest that they are sites of hemocyanin synthesis and homologous to the cyanocytes or cyanoblasts of Limulus.Supported by grant No. 3,012-0.76 of the Swiss National Science Foundation     

Isolation of (+)-abscisin II from dormant aerial tubers of Dioscorea batatas

Summary The occurrence of at least three growth inhibitors was demonstrated in the dormant aerial tubers of Dioscorea. One of the inhibitors was isolated in crystalline form and identified as (+)-abscisin II by the infrared and ultraviolet spectra as well as by the optical rotatory dispersion.     

Flavonoid Pigments in Rice Leaves and the Isolation of Glucotricin

The distribution of flavonoid pigments in the leaves of indica and japonica type rice plants was studied by paper chromatographic and electrophoretic techniques. A fluorescent flavoniod compound prominent in the leaves of indica was isolated in a crystalline form and identified as glucotricin (tricin-7-β-glucoside), the structure of which was characterized by Kuwatsuka in 1962.     

Studies on Production of Biotin by Microorganisms

In a preceding paper we reported that Rhodotorula flava 194 effectively converted biotin to biotinamide. In a present paper the metabolism of desthiobiotin by R. flava 194 was studied under the same condition as in the conversion of biotin to biotinamide. Two desthiobiotin derivatives (Vitamer I and II) were isolated. Vitamer II (crystalline) was identified as bisnordesthiobiotin and Vitamer I was chromatographically determined as desthiobiotinamide.     

Biochemistry of Filamentous Fungi. XIII. Metabolites of Helicobasidium mompa Tanaka

Five crystalline compounds, two colored and three colorless, have been isolated from the mycelium of Helicobasidium mompa Tanaka, a root-infecting plant pathogen. One of the colorless crystals was identified as D-arabitol. Chemical studies of the other substances are going on.     

Isolation of zeatin riboside from the chicory root

Summary From 250 kg of fresh chicory roots about 2 mg of a crystalline cytokinin were obtained. This substance was identified as ribosylzeatin (trans isomer). From the procedure employed it seems unlikely that the isolated cytokinin comes from the degradation of tRNAs; rather, it may constitute a separate pool of cytokinins.     

Characterization and Cytopathology of Hibiscus Latent Ringspot Virus Isolated from Kenaf (Hibiscus cannabinus L.) in Italy

Hibiscus latent ringspot virus (HLRV) was isolated from kenaf plants (Hibiscus cannabinus L.) in Italy. and identified as III. RV by its host range, purification, electron microscopy and serology. Ultrastructural studies showed that infected plants contained membranous cytoplasmic inclusion bodies. characteristic of nepoviruses, and sometimes also crystalline aggregates.     

Anthraquinone production by a fungus causing black heartwood stain in yellow cedar

Summary A yellow crystalline fungal metabolite was obtained from cultures of sterile, darkly pigmented fungi which had been isolated from the heartwood of yellow cedar. This chemical was identified as 1-hydroxy-3-methylanthraquinone and showed negligible toxicity against a range of wood-destroying Basidiomycetes and moulds. It also greatly reduced conidial formation inT. virgatum.     

α-Aminoisobutyric Acid as the Constituent Amino Acid of Protein

α-Aminoisobutyric acid is the only tertiary amino acid which is reported to occur in the proteins. Nevertheless, this amino acid has not been yet isolated from the proteins. Recently we succeeded in isolating this amino acid as white prismy crystalline substance from both acid and pepsin hydrolysate of horse hind leg muscle proteins, and this crystal was identified to be α-amino-isobutyric acid by elementary analysis, properties of this derivates, etc.     

p-Hydroxybenzoic Acid, a Growth Regulator, Isolated from Woody Cuttings of Ribes rubrum

Woody cuttings of Ribes rubrum, an easy plant to root, were extracted with methanol. One fraction of this extract was both water and ether soluble; a part of it was only water soluble. From the water and ether soluble fraction a crystalline compound was isolated which was fully identified with p-hydroxybenzoic acid (PHB) by m.p., mixed m.p., infrared spectrum, ultraviolet absorption in neutral and alkaline methanol and chemical analyses. For Avena coleoptiles straight growth test, p-hydroxyhenzoic acid was found to have a significant growth promoting activity in the range 20–100 μg/ml. and it showed strong growth inhibition at concentrations higher than 200 μg/ml. From the same extract fraction a phenolic ester was isolated by paper chromatography which showed a weak growth inhibiting activity. This compound was identified as an ester of p-hydroxybenzoic acid by micro-scale reactions. From the extract soluble only in water further amounts of crystalline p-hydroxybenzoic acid were obtained by alkaline hydrolysis. This PHB may have arisen from the breakdown of pelargonidin glucosides.     

Regulators of Cell Division in Plant Tissues

Procedures were devised for purification of the cytokinins in the milk of mature coconuts (Cocos nucifera fruits). One cytokinin isolated in crystalline form was unequivocally identified as 9-β-D-ribofuranosylzeatin. This compound appeared to account for a large proportion of the cytokinin activity in n-butanol extracts of coconut milk. The activity which was not extracted by n-butanol was largely due to unidentified compounds which could be partially purified by adsorption onto and elution from charcoal.     

Studies on the Hypertrophic Disease of Cherry (Genus Prunus), So-called “Witch’s Broom” Caused by Taphrina wiesneri

Metabolites of Taphrina wiesneri (Rath.) Mix. were examined. Brassicasterol, stearic acid, and p-hydroxyphenylacetic acid were isolated in crystalline form. p-Hydroxybenzoic acid and vanillic acid were identified by paper chromatography and UV measurement. Palmitic acid was identified by gas-chromatography. The fungus produced usually these compounds on any one of four kinds of medium used. p-Hydroxyphenylacetic acid promoted germination of rape seeds at the concentration of 20 ppm in water and showed inhibition at 250 ppm.

Phenolic acids and their related compounds in Japanese flowering cherry leaves infected by Taphrina wiesneri were examined. In the acidic and neutral extracts of infected cherry leaves (I), eighteen compounds positive to diazotized sulfanilic acid and two fluorescent compounds were detected by paper chromatography. Of these compounds, coumarin, 3, 4-dihydrocoumarin, melilotic acid, o- and p-coumaric acids, p-hydroxybenzoic melilotic acid, ferulic acid and caffeic acid were identified. Melilotic acid and coumarin were obtained in crystalline form. The amount of melilotic acid in I was higher than that in healthy leaves independent of sample source, although increased with the growth of cherry leaves.     

Urocanic Acid,Produced by Sporogenous Bacteria

On the screening of microorganisms which accumulate ultra violet light absorbing substances, some sporogenous bacteria were selected as powerful strains for production of a crystalline substance having a maximum absorption at 267.5 m_μ (pH 6.0 in water). These strains were microbiologically examined and named B. subtilis var. thermophilus. Submerged fermentation was carried out for 48 hrs at 37°C in glucose bouillon medium and the isolation of substance was performed by absorption to active carbon and elution with ammonia water.

On the basis of chemical studies of this crystal, it was identified as urocanic acid.     

Studies on dl-Alanine Formation by Thermophilic Bacteria

During the course of the screening of thermophilic microorganisms, several strains were found to accumulate amino-acids. Of those strains that were isolated from feces source with Bennett medium, one strain was found to produce a large amount of an amino acid. This amino acid was isolated in crystalline form and identified as dl-alanine by IR absorption spectrum, specific rotatory power and elementary analysis. The taxonomic studies were carried out and this strain was identified as Bacillus coagulans. The strain B. coagulans B₉-17 produced dl-alanine as much as 1000 mg/l after 24 hours at 50°C in shake culture.

The yield of dl-alanine was increased up to 16.5 g/l with some improvements.     

Synthesis of Oligosaccharides by Growing Culture of Leuconostoc mesenteroides

When Leuconostoc mesenteroides NRRL B-1299 was grown on a medium containing sucrose and lactose, a trisaccharide named ‘Lactsucrose’ (O-β-d-galactopyranosyl- (l→4) -O-α-d-glucopyranosyl- (l→2) -β-d-fructofuranoside) was produced. This sugar was obtained as a crystalline acetyl derivative and its structure was identified by the methods of paper chromatography, paper ionophoresis, hydrolysis with acid or enzymes and periodate oxidation.     

New delphinidin glycosides isolated fromVinca major flowers

Four kinds of delphinidin glycoside were found in the bluish-purple flowers ofVinca major. Among them, two glycosides were isolated in a crystalline state by application of Sephadex and Avicel columns. The names of vincanin A and vincanin B were tentatively assigned to those new glycosides. They were respectively identified as delphinidin 3-robinobiosido-5-rhamnoside and delphinidin 3-robinobioside on the basis of chemical, spectral and paper chromatographic analyses. The identity of two other glycosides could not be fully confirmed because of the smallness of the amount.     

(Z)-3-Hexenyl-β-d-glucopyranoside in Fresh Tea Leaves as a Precursor of Green Odor

The glycoside fraction from fresh tea leaves was acetylated and separated by silica gel column chromatography.

A crystalline product was identified as (Z)-3-hexenyl-(tetra-O-acetyl)-β-d-glucopyranoside from spectrometric data which were identical with those of an authentic synthesized sample in all respects.

There are two possible processes for the formation of the greenish odor of plant materials, these being a biosynthetic process from the lipid and enzymatic hydrolysis of (Z)-3-hexenyl-β-d-glucoside.     

Haemoglobin producing cells in freshwater snails

Summary One of the cell types occurring in the connective tissue of gastropods and bivalves is thepore cell. Cells of this type have a characteristic morphology in that their plasmamembrane has many invaginations bridged by cytoplasmic tongues. In the well developed granular endoplasmic reticulum and in these invaginations fine granular material, sometimes arranged in crystalline patterns, is present. InLymnaea stagnalis this material has been identified as haemocyanin on a morphological basis. Histochemical and ultrastructural results on pore cells of 2 snail species(Biomphalaria glabrata, Planorbarius corneus) having haemoglobin in their blood indicate that the pore cells in these species synthesize haemoglobin. Possible other functions of pore cells are briefly discussed.     

Cloning and expression of theClostridium thermocellum celS gene inEscherichia coli

Clostridium thermocellum ATCC 27405 produces an extremely complicated multi-component cellulase aggregate (cellulosome) highly active on crystalline cellulose. From the cellulosome, two subunits, CelS (or S _s ;M _r = 82 000) and CelL (or S _l , CipA;M _r = 250 000), have been identified as essential for crystalline cellulose degradation [Wu et al. (1988) Biochemistry 27:1703]. We have determined the DNA sequence of thecelS gene from four cloned DNA fragments encompassing this gene [Wang et al. (1993) J Bacteriol 175:1293]. To express the entirecelS gene inEscherichia coli, thecelS structural gene was amplified by the polymerase chain reaction (PCR) employing the PCR primers corresponding to sequences flanking the desired gene. This PCR product (2.1 x 10³ bases; 2.1 kb) was cloned into anE. coli expression vector pRSET B. Subsequent expression of the cloned gene resulted in a fusion protein (rCelS;M _r = 86 000) as inclusion bodies. The rCelS protein was recognized specifically by an anti-CelS antiserum in a Western blot analysis. The inclusion bodies were purified and solubilized in 5m urea. The refolded rCelS produced very little reducing sugar from carboxymethylcellulose. However, it showed a higher activity on the crystalline cellulose (Avicel) and an even higher activity on phosphoricacid-swollen Avicel. These results indicate that the CelS is an exoglucanase.