An acidic polysaccharide was isolated from the culture broth of Aureobasidium sp. K-1. The polysaccharide ([α]²⁰_D-10°) was composed of D-glucose and sulfoacetic acid. The glucan moiety was hydrolyzed by exo-β-1, 3-glucanase to give glucose and gentiobiose (molar ratio, ca. 1:3). The methylated glucan yielded on acid hydrolysis 2, 3, 4, 6-tetra-O-methyl-, 2, 4, 6-tri-O-methyl- and 2, 4- di-O-methyl-D-glucose (molar ratio, 2.6:1.0:2.7) together with small amounts of 2, 3, 6-tri-O-methyl- D-glucose. Smith degradation of the glucan moiety yielded glucose, glycerol and erythritol (molar ratio, 4:3.1:trace). From these results, it was suggested that the glucan consists of a backbone of β-1, 3-linked glucose residues containing single branches of glucose residues joined by β-1, 6-linkages, roughly three out of every four glucose residues in the backbone.

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When mice were prevented from resting under the conditions of being forcibly fatigued by means ofa rotary cage, PCBs which accumulated in fat tissues were found to move towards the liver to be concentrated through so-called "fat mobilization" at a level about 10 times as great as that in mice kept under normal conditions. It was supposed that excretion of PCBsconcentrated in the liver might be promoted through fat mobilization. From the results of experiments on fat mobilization, active charcoal was suggested to considerably accelerate the excretion of PCBs accumulated even in fat tissues, although the main effect of the charcoal was considered to be the prevention of absorption of PCBs. Besides the effect of active charcoal, other effects have also been observed with some organic matters, including glutathione and unmetabolizable sugars or acids such as arabinose and the acid-hydrolysate of pectin.

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In a study on excretion and accumulation of PCBsin the animal body there arose the new concept of conjugate formation of PCBs with some organic substances, including glutathione, unmetabolizable sugars and acids such as arabinose and the acid-hydrolysate of pectin. PCBs conjugated probably with glutathione, including their intermediate metabolites, were found to becomethe same mercapto-chlorobiphenyl on alkali-hydrolysis, depending exclusively on the chemical structure of their original form. The same product as the above was also found to be excreted in feces of mouse or rat even as one of the partial metabolites from PCBs. The same phenomenon has also been recognized even with chlorobenzene thus supporting the possibility for conjugate formation ofPCBs with glutathione. A similar possibility was also seen for other organic matters mentionedabove.

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In in vivo and in vitro experiments using mice or rats, an N-acetyl-cysteine derivative, so-called mercapturic acid, was demonstrated with PCBs by gas chromatography-mass spectrometry with, use of the sample after methylation and/or acetylation of their metabolites. The derivative was suggested from its chromatographic pattern to be derived from the conjugate of PCBs with glutathione, although direct evidence has not been obtained as yet, owing to the extreme difficulty in purifying this conjugate on account of its characteristic of being soluble only in water, and to the limited capacity of the apparatus for mass spectrometric analysis. And yet, in the experiment using chlorobenzene an N-acetyl-cysteinylglycine derivative was found, whichcould further support the possibility of conjugate formation of PCBs, because the former was observed to behave quite similarly to the latter.
On the other hand, it was recognized in urine that these conjugates could further be metabolized into an acetic acid derivative via pyruvate without only mercapturic acid as an end metabolite.

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From the findings that cell fractions (membrane and cytoplasm) of Streptococcus lactis 527 hydrolysed phosphatidyl ethanolamine to monoglyceride, diglyceride and fatty acids without accumulating lysophosphatidyl ethanolamine, the main contribution of phospholipase C to the enzymatic degradation was presumed. Although the fractions have strong activity toward lysophosphatidyl ethanolamine, the contribution of lysophospholipase was ruled out. Phospholipase A showing activity at optimum pH 5 was considered to be bound to the cell membrane and was different from the enzyme showing activity at pH 9 in the influences of Ca²⁺ addition, cholate addition, and heating at the optimum pH. In bacteria lysis during lactic acid fermentation of milk, phospholipases C and A, and lipase in spheroplasts of S. lactis were presumed to mainly contribute to degradation of phosphatidyl ethanolamine in the cell membrane.

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Manyyeast strains which grew on palm oil were obtained from natural sources. Ayeast strain capable of assimilating crude palm oil effectively was isolated from field soil, and named Torulopsis Candida Y-128.
When the substrate was emulsified with a nonionic surfactant, yeast cell growth was promoted. In a shaking culture of this strain, corn steep liquor as a natural nutrient was good for cell growth, and ammoniumsulfate was moreeffective than any other nitrogen source. Theprotein content of dried cells was over 40% and amino acids of the yeast protein were well-balanced.

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Glucotriosyl asparagine 2, a proposed structure for the glycan part of nephritogenic glycopeptide 1 isolated from the glomerular basement-membrane of rat, was synthesized in regioand stereo-selective manners starting from a-D-glucopyranosyl azide 8. The key intermediate for this synthesis was designed to be glucotriosyl azide 21, based on retrosynthetic considerations.

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The concentrations of Fe, Mn, Zn, Cu, Pb, Ni, Cd and Hg in the muscle, liver and kidney of fifty nine striped dolphins captured during 1977- 1980 were determined by atomic absorption spectrophotometry. The levels of most the metals examined were higher in the liver than the other tissues, except for the highest renal Cd. The frequency distribution of the metal concentrations was mostly log-normal. For all of the organs, correlation of the metal concentrations to the body length, weight and age was positive for Fe, Pb, Ni, Cd and Hg, and negative for Mn, Zn and Cu. A higher correlation of the concentrations of Fe, Mn, Zn and Cu to the body length and weight than to the age indicates that the metabolic turnover is more important than age or exposure time in determining the levels of those metals. Whilst the concentration of Pb, Ni, Cd and Hg correlated closely with the age of dolphins, and this suggests that the age or exposure time is a dominant factor for accumulation of these metals.
The concentrations of Pb, Ni, Cd and Hg in the muscle, liver and kidney increased with age until 1 year, leveled off through 1 to 18 years by physical dilution with increased body weight, and thereafter steadily increased year by year, but in case of Hg such physical dilution effects were not significant. Relatively wide variation of the metal concentrations in the pup and 8-25 years females was probably due to the effects of suckling, and of parturition and lactation, respectively.

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Heating of 7S globulin alone did not cause precipitation, but addition of 11S globulin to the system did. Gel nitration and ion exchange chromatography, followed by electrophoresis of the precipitate and supernatant, suggested an interaction through disulfide bonding between the 7S and US globulin subunits. The basic and β subunits tended to be located in the precipitate by interaction through a secondary force on heating. On the other hand, the acidic and αα' subunits tended to be located in the supernatant by interaction through disulfide bonding on heating.
Using the predictive parameter of amino acids, each macroscopic parameter of the soybean globulin subunits was calculated on the basis of their aminoacid composition and a relationship with their behavior on heating is discussed.

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Six strains of radiation-resistant gram-positive cocci were isolated from sewage sludges and animal feeds in Japan after gamma-irradiation of more than 1.0 Mrad. All six strains were able to growon nutrient agar slants, and somestrains were also able to growon glutamate agar slants. Cells of the six strains were single or diplococci, and occasionally seen in tetrads, being spheres averaging from 0.8 to 1.0μm in diameter. The peptide subunit of cells of all the strains contained ornithine, and the predominant fatty acid component was a Cl_16:1. The GCcontent of the DNA of these strains ranged from 59 to 66 mol%thus indicating them as belonging to the genus Deinococcus Brooks and Murray 1981 which was previously called the "Micrococcus radiodurans" group. From the similar cultural characteristics and morphology, the six strains, TD1, TD3, TD9, T843, Fr 3 and Fr 7, were identified as D. proteolyticus. However, the predominant component of cellular fatty acids of strain T843 was similar to that of D. radiodurans.
The resistance to gamma-radiation of these new isolates was similar to that of D. radiodurans R₁, and D₁₀ values in phosphate buffer ranged from 0.10 to 0.25 Mrad, and the low oxygen enhancement effect caused by radiation was distinct from other kinds of bacteria.

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Two new m-cresol derivatives, 5-methyl-2-(2'-oxo-3'-butyl)-phenol (1) and 5-methyl-2-(3'- oxo-2'-pentyl)-phenol (2), together with a terpene alcohol with a tetrahydrofuran structure, 3- (5', 5'-dimethyltetrahydrofuran-2'-yl)-cw-2-buten-l-ol (3), were isolated and identified in peppermint oil.

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A methanol-utilizing phototrophic bacterium, strain M402, was isolated from surface water of an acidic hot spring. The isolated strain was identified as Rhodopseudomonas acidophila from its morphological and physiological characters. Profiles of the utilization of non-aromatic compounds as carbon sources by this strain were in good agreement with those of somestrains of R. acidophila reported by Pfennig [J. Bacteriol., 99, 597 (1969)]. However, strain M402 was found to be capable of utilizing vanillic acid, vanillin, vanillyl alcohol, ferulic acid, veratric acid, syringic acid, syringaldehyde and benzyl alcohol as carbon sources under anaerobic-light conditions. Although Pfennig did not refer to these abilities of his strains, these notable characters of strain M402seem to be additional new characters of R. acidophila.

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The distribution of glutathione peroxidase was investigated among microorganisms. Glutathione peroxidase was found in the cell-free extracts of molds belonging to the genus Mucor, and Mucor hiemalis showed the highest enzyme activity. The glutathione peroxidase production by M. hiemalis was almost completely associated with the mycelial growth. The addition of oxidants such as hydroperoxides and antioxidants such as glutathione and α-tocopherol did not significantly affect the enzyme production.

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Translation of total RNAprepared from cultured tobacco cells by the guanidine-HCl method in a cell-free reticulocyte lysate system results in the synthesis of a larger precursor of β-1, 3- glucanase (EC 3.2.1.39). This identification was based on immunoprecipitation with specific anti-β- 1, 3-glucanase antibody. Furthermore, sodium dodecyl sulfate polyacrylamide gel electrophoresis after limited proteolysis and two dimensional tryptic fingerprintings of β-1, 3-glucanase and its precursor show a high degree of similarity. The precursor is larger than the mature protein by 4, 000 daltons. β-1, 3-Glucanase mRNAwas partially purified by oligo(dT)-cellulose column chromatography and sucrose density gradient centrifugation. The sedimentation coefficient of the mRNAwas estimated to be about 17S.

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Heat evolution during the microbial degradation of glucose in soil was measured with a multiplex heat-conduction calorimeter in the presence of mercury, cadmium, selenium and iodoacetic acid as pollutants. Effects of the pollutant concentration on the degradation thermograms were compared and the results implied that each pollutant had a different type of inhibitory effect on the degradation process of glucose in soil. The dependence of peak times on the thermogramson the pollutant concentration was quantitatively analyzed with a simple mathematical model and possible parameters describing the inhibitory effect of each substance were presented.

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Brevibacterium flavum cells obtained from different growth phases were immobilized with kcarrageenan and the stability of the fumarase activity was investigated. The stability of fumarase activity of the immobilized preparation of cells of the stationary growth phase was highest. The highest stability of the immobilized cells seemed to be correlated to the high stability of fumarase activity in free cells of the stationary phase. High rigidity of the cell wall and membrane of B. flavum cells of the stationary phase and firm binding of fumarase protein to the cell membranewere suggested from several lines of evidence obtained on treatment of the cells wi h lysozyme and detergents or sonication of the cells. Electronmicrographsshowedthat the cells of the stationary phase retained the original shape after repeated batch reactions. Solubilized fumarase prepared from cells of the stationary phase showedthe highest stability. Experiments using the partially purified enzymestrongly suggested the existence of fumarase-stabilizing components in the cells.

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The effect of dietary nucleic acid constituents (purine and pyrimidine bases) on the metabolism of serum and urinary uric acid, allantoin, creatinine, urea nitrogen and urea was examinedin the rat. The results were compared with the morphological changes. It was found that guanine is readily converted to uric acid and allantoin, whereas adenine, due to its unique metabolism, is metabolized quite differently from guanine and pyrimidine bases. Furthermore, an increase of creatinine, urea nitrogen and urea in the serum as well as a reduction in their urine excretion was observed in rats fed on the adenine diet. Dietary adenine produced a nephrotoxic condition as reflected in the histopathological changes. That is, deposits of amorphous birefringent crystal (2, 8-dihydroxyadenine) with formation of foreign body granuloma were demonstrated in the tubular lumina and the interstitium of the kidney, and also in the urinary bladder and urine. However, there were not seen in the glomeruli and other organs. On the other hand, in the kidneys of rats fed on guanine, uracil, cytosine and yeast RNAdiets, these histopathological changes were not noticed.

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During the seed development of Nicotiana tabacum, appreciable accumulation of the soluble protein fraction started to occur at around the 6th day after anthesis and finally reached 12%on the basis of dry weight when seed maturation was accomplished. In the soluble fraction of mature seeds, four protein fractions were observed on analytical ultracentrifugation, and the protein having a sedimentation coefficient of 11.7S was the major one. The 11.7S protein was isolated and SDS polyacrylamide gel electrophoresis indicated that the protein consisted of at least five subunits with molecular weights of 49, 000, 31, 000, 29, 000, 21, 000 and 19, 000. The 11.7S protein was rich in glutamic acid or glutamine and arginine, and the presence of carbohydrate was confirmed.
During development, all of the five subunits started to appear during the period between the 12th and 15th day after anthesis.

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A simple in vitro assay method for erythropoietin, a plasma glycoprotein which stimulates erythroid differentiation, has been developed. In this method, a liver cell suspension from 13-day mouse fetus was planted in the wells of a flat-bottomed 96-well microtiter plate and incubated for culturing. The formation of erythroid colonies, which were identified with benzidine staining, was dependent on added erythropoietin. Since this method is inexpensive, sensitive, simple, and only requires a small sample, it could be adopted for assaying the erythropoietin levels in blood, urine, and other materials.

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Two β-D-glucan components have been obtained from mycelia of Neurospora crassa by fractionation with alkali. The component obtained by extraction with 4% NaOH had a unit structure of a linear β-1, 3-D-glucan consisting of about 86 glucosyl residues, two of which were substituted with single β-D-glucosyl residues at C6. The unit chain is linked in turn through two β- 1, 6-glucosidic linkages. The alkali insoluble component had a structure essentially similar to that of the alkali soluble component. But, the main chain of the unit structure of the latter was much shorter (ca. 20) and had both glucosyl and laminaribiosyl side chains.

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A new flavoprotein enzyme, L-glutamate oxidase, was purified to homogeneity from an aqueous extract of a wheat bran culture of Streptomyces sp. X-119-6. It showed absorption maxima at 273, 385 and 465nm and a shoulder around 490nm, and contained 2 mol of FAD per mol of enzyme. The enzyme had a molecular weight of approximately 140, 000 and consisted of three sizes of subunits with molecular weights of 44, 000, 16, 000 and 9, 000. Balance studies showed that 1 mol of L-glutamate was converted to 1 mol of α-ketoglutarate, ammonia and hydrogen peroxide with the consumption of 1 mol of oxygen. In addition to L-glutamate, L-aspartate was oxidized by the enzyme but only to an extent of 0.6% at pH 7.4; the Michaelis constants were as follows: 0.21 mM for L-glutamate and 29mM for L-aspartate. The isoelectric point was pH 6.2, and the enzyme activity was optimal between pH 7.0 and 8.0. When the enzyme was heated at pH 5.5 for 15 min, the remaining activity was 100% of the original activity level at 65°C, 87% at 75°C and 47% at 85°C.

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Quinoxalines derived from D-galactose with o-phenylenediamine (OPD) in acidic media under reflux were studied by using GLCand NMRmeasurements. Four quinoxaline derivatives were obtained from the reaction mixture, and were identical with those derived from D-glucose. The yields of 2-(D-lyxo-tetrahydroxybutyl)quinoxaline (GA-III), and the stereoisomeric derivative of GA-III, i.e., 2-(D-arabino-tetrahydroxybutyl)quinoxaline (ATBQ), were 13.2 and 5.3%, respectively. The ratio of GA-III to ATBQ derived from D-galactose was reciprocally coincident with that from D-glucose. Someproposals are made on the relationship between the isomerization of these sugars and the formation of quinoxaline derivatives.

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This study was undertaken to elucidate the toxic nature of dietary hydroperoxides in rats by the measurement of tissue chemiluminescence (CL). By the oral administration of methyl linoleate hydroperoxide (HPO) for two days, the tissue CL intensities and thiobarbituric acid (TBA) reactants were significantly increased in the liver, kidney, heart and lung, and the increases of both indices were inhibited by the supplemental administration of antioxidants, i.e. d-α-tocopherol (TOC) and riboflavin-tetrabutyrate (RTB). TOC was especially effective on the liver and kidney and RTB was effective on the liver, lung and heart. Hepatic glutathione peroxidase activity was evidently elevated at the seventh day of HPO-feeding, and this activation resulted in the decay of CL intensities and TBA reactants in rat organs. The spectrum of CL from liver homogenates of rats dosed with HPO for two days revealed five emission bands in the visible region and was similar to that for a singlet oxygen.
The results indicate that TOC and RTB act as protective agents against the tissue lipoperoxidation caused by dietary HPO in rats, and that liver glutathione peroxidase is effectively induced as one of the bio-antioxidative functions in rats against the toxic effect at an early stage of the HPO dosage.

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The purified inorganic pyrophosphatase from the leaf of Amarantus blitum shows absolute magnesiumrequirement for its enzymeactivity. For maximumenzymeactivity, the Mg: PP_i ratio was found to be 10:1 at the optimum pH of 9.0. This ratio varies with the shift of the pH. MgPP_i^2- was found to be the true substrate for the enzyme in the presence of the free Mg²⁺ ion. Some divalent metal ions and the fluoride anion severely inhibit the enzyme activity in the presence of Mg²⁺. The Michaelis constant (Km) and molecular weight of the enzyme were found to be 4.95 10^-6 m and 32, 860, respectively. This enzyme is strictly specific for inorganic pyrophosphate.

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