In the course of study on the utilization of methyl-substituents of mono-cyclic aromatic hydrocarbons by Pseudomonas aeruginosa S668B2, some organic acids and phenolic compounds were found to be produced in culture broth.
Strain S668B2 was capable of producing ultraviolet absorbing and fluorescent substances from m-xylene. These substances were isolated in the form of crystal and identified as 3-methyl salicylic acid and m-toluic acid.
Strain S668B2 also produced ultraviolet absorbing and fluorescent substances from pseudocumene (1, 2, 4-trimethyl benzene). These substances were isolated in the crystalline form and identified as 3, 4-dimethyl benzoic acid and 3, 4-dimethyl phenol.
Strain S668B'' did not attack o-xylene. Under the similar conditions Pseudomonas desmolytica S449B3, which produced a large amount of cumic acid from p-cymene, did not oxidize o-xylene, but grew on p-xylene, m-xylene and 1, 2, 4-trimethyl benzene.
None out of 364 soil samples gave microorganisms which utilize o-xylene as a sole carbon source.

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Lipases produced by _{Mucor lipolyticus} Aac-0102 were separated into three different fractions (F-1, F-2 and F-3) by CM-Sephadex column chromatography.
Molecular weights of them were estimated to be higher in the order of F-1, F-2, and F-3 by gel filtration on a Sephadex G-100 column. F-1 and F-2 could hydrolyze water soluble substrate such as Tweens. F-3 showed a strong hydrolytic activity toward triglycerides but the activity toward Tweens was almost negligible.
Sodium lauryl sulfate inhibited the olive oil hydrolyzing activity of F-3. However, Tween hydrolyzing activity of F-2 was not affected with it. These results suggested that they are different with each other with respect to their molecular weights, substrate specificities and sensitivities to some inhibitors.

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Preincubation with spermine, of λ, T7 and P465 phages which were sensitive to oxidized spermine, resulted in a decrease of their susceptibility to the action of oxidized spermine. Phages resistant to oxidized spermine such as T4 and φX174 became susceptible to this agent after dialysis.
The mechanism of phagocidal action of oxidized spermine was examined with ³²P-labelled λ phage. Oxidized spermine interfered neither with the absorption of λ phage, nor with the injection of its DNA into the host cells. The injected DNA, however, did not lead to the formation of mature phage.
The interaction of oxidized spermine with the DNA of phages T4 and T7 was investigated by thermal denaturation studies. DNA treated with oxidized spermine showed the same Tm as untreated DNA. However, the treated DNA was decreased in its hyperchromicity and was renatured to a great extent, even after rapid cooling. These facts areexplained by the formation of cross-links which prevents the separation of complementary DNA strands.

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The incorporation of ¹⁴C of acetate-l-¹⁴C into the lipids of the liver and carcass, and the changes in the concentrations of nucleotides and citric acid in the liver were studied in the rats fed individual nutrients; starch, casein or corn oil. And the metabolism of citric acid-l, 5-¹⁴C was also investigated after the feeding of nutrients. Lipogenesis in the liver and carcass was more markedly stimulated with starch than with casein or corn oil. In the liver of rats fed starch, the concentration of ATP doubled and that of citric acid was one-half of those with casein or corn oil, respectively. And the conversion of citric acid to carbon dioxide and lipids was stimulated with starch.

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Dietary protein-carbohydrate ratio affected the change in rat hepatic threonine de-hydratase (TDH) activity: 7% casein-85.5% carbohydrate, 0.ll; 20% casein-72.5% carbohydrate, 7.07; 40% casein-52.5% carbohydrate, 26.71; 90% casein-2.5% carbohydrate, 63.71 μ mole α-ketobutyric acid/min/g liver. The variation of TDH activity thus covering a wide range exceeded that due to the regulation by gluco-corticoid alone observed previously by us in the case of disorder in dietary amino acid balance. Alloxan-diabetic rats showed enormously high TDH activity. Insulin was effective in lowering the TDH activity which had been elevated by feeding 20% casein diet. Insulin also had a suppressing effect on the elevation of TDH activity by feeding higher protein diet. The function of adrenals measured by adrenal glucose-6-phosphate dehydrogenase activity did not show so wide variation throughout these studies. Effects of adrenalectomy, alloxan treatment and/or cortisone treatment on TDH activity were as follows: adrenalectomy, 2.33; adrenalectomy-alloxan, 3.92; adrenalectomy-cortisone, 10.98; adrenalectomy-cortisone-alloxan, 32.57 μ mole α-ketobutyric acid/min/g liver. These results indicate that the depression of insulin secretion is independent of the increase in TDH activity, but gives rise to the increase in the TDH activity through the lowering of counteraction of insulin against gluco-corticoid. Actinomycin S₃ treatment on rats disclosed that the change in TDH activity based on insulin related to de novo synthesis of this enzyme via transcriptional step.

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Methods for differential determination of 3-ketosucrose and 3-ketoglucose were established. For determination of 3-ketosucrose, alkaline treatment with 0.1N NaOH was found to be most effective. In this method, 3-ketosucrose gave a characteristic absorption spectrum with a molar extinction coefficient of 6.5×10³M-1cm^-1 at 340mμ, while 3-ketoglucose did not show a significant absorption spectrum within a range from 300 to 400mμ.
By mixing with 0.2M phosphate buffer, pH 7.0, 3-ketoglucose gave a characteristic absorption spectrum with a molar extinction coefficient of 3.8×10³M^-1cm^-1 at 310mμ, while 3-ketosucrose showed little absorbance.
From the reduction rate of 2, 6-dichloroindophenol with 3-ketoglucose, the ketosugar was determined. 3-Ketosucrose was not able to reduce the reagent at all.
The methods established here were not affected by fructose.

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A seed medium and a fermentation medium for nucleotide fermentations such as 5'IMP, 5'GMP (plus GDP and GTP) and 5'AMP (plus ADP and ATP) with Brevibacterirm ammoniagenes ATCC 6872 were entirely chemically defined, with the use of a mixture of five amino acids.
As a result, the presence of Zn²⁺, Fe²⁺ and Ca²⁺ in addition to Mn²⁺ was found to be essential for the nucleotide fermentations. In particular, Zn²⁺ levels as well as Mn²⁺ affected nucleotide productions remarkably. Various fermentations proceeded favorably only when suboptimum levels of manganese (20_??_30μg/liter) and zinc (100_??_200μg/liter) were simultaneously present. This effect of trace metals was attributed to the fact that the excretion of R5P, a precursor of nucleotides, and those enzymes catalyzing reactions synthesizing nucleotides from R5P, ATP and purine bases were greatly stimulated by trace metals in cooperation with two vitamins, Ca-pantothenate and thiamine, and presumably high concentrations of phosphate and magnesium.
Furthermore, it was revealed that some metals were able to control the amounts of nucleotides accumulated when they were added to the broth during fermentation. For example, Hg²⁺ and Ag⁺ could increase the amounts of 5'GMP or 5'AMP, and decrease those of GTP and ATP.

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Growth responses of Brevibacterium ammoniagenes ATCC 6872, capable of accumulating purine nucleotides, were investigated by the use of completely defined media.
(1) Casamino acids required for its growth could be replaced by a mixture of L-histidine, L-homoserine, glycine, D, L-alanine and L-lysine. Acompletely defined medium for mucleotide productions was thus established by the use of this mixture.
(2) High levels of phosphate inhibited growth markedly, and this inhibition was overcome by the simultaneous addition 1) of hign levels of Mg²⁺ and 2) of Mn²⁺, 3) pantothenate and 4) thiamine. Ca²⁺ had also a stimulatory effect on the growth. Therefore, a clear growth response to Mn²⁺ levels and the requirement of the two vitamins for growth emerged only under the conditions of high phophate and magnesium salts. These 4 factors were found entirely the same as factors essential for nucleotide accumulations by Br. ammoniagenes.

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The flavor concentrate obtained by the extraction of “Katsuobushi” of bonito (Katsuwonus pelamis) with 80% ethanol and by the subsequent steam distillation of the extract was fractionated by the usual method and the resulting neutral, non-carbonyl oxygenated fraction was investigated by gas chromatography. The following components were tentatively identified: 2-pentanol and 2-methyl-l-heptanol as free alcohols, and 4 alcohos of n- and isobutanol, n-pentanol and n-dodecanoll and 9 carboxylic acids of propanoic, n-butanoic, n-pentanoic, n-octanoic, n-nonanoic n-decanoic, n-dodecanoic, n-tetradecanoic and n-hexadecanoic acid as the constituents of esters. A constituent alcohol existing in the largest amount was isolated by gas chromatography and identified as 2-methyl-l-heptanol by elemental analysis, NMR, IR, and MS. A constituent acid existing in large amount was also isolated and investigated similarly, and the structure was partially estimated. 2-Methyl-l-heptanol holds a fresh woody aroma and seems to have a major effect on “Katsuobushi” flavor.

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In order to know the substrate specificity in a hydrocarbon utilizing bacterium, the following materials were examined: n-alkanes, n-alkenes, monohydric alcohols, aldehydes, monobasic carboxylic acids, dihydric alcohols and dibasic carboxylic acids.
It was found that dibasic carboxylic acids were well utilized, and a great deal of L-glutamic acid was accumulated from them. Then suberic acid, which is C₈ dibasic carboxylic acid, was compared with n-dodecane in the effects of thiamine, penicillin, C/N ratio and substrate concentration on L-glutamic acid accumulation and cell growth.

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The kinetics of hydrolysis of acetyl-DL-methionine in DEAE-cellulose-aminoacylase (EC 3. 5. 1. 14) column and DEAE-Sephadex-aminoacylase column was studied.
The rate of hydrolysis of substrate was shown to be dependent on the flow rate and independent to the dimension of the enzyme column. The rate of hydrolysis of the substrate was equal in cases of down-waard flow and of up-ward flow. The deteriorated aminoacylase columns by long period operation were reactivated by the recharge of aminoacylase to them. The continuous enzyme reaction using an aminoacylase column was superior to the batch enzyme reaction using native aminoacylase.

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The enzymatic properties of the water-insoluble aminoacylase prepared by linking mold aminoacylase (EC 3. 5. 1. 14) to DEAE-Sephadex were studied and compared with those of the native aminoacylase.
Optimum pH values for hydrolysis of several substrates by the DEAE-Sephadex-amino-acylase complex (DSA-complex) shifted about 0.5_??_1.5 pH units more to the acid side than those by the native enzyme. On the effects of metal ions and inhibitors, substrate specificity, optical specificity and kinetic constants, no marked difference was observed between the native enzyme and the DSA-complex. Heat stability, optimum temperature and resistance towards proteases were increased by conversion from the native form to the insoluble enzyme. It was also observed that the DSA-complex was activated by urea.

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Orally administered 3, 4-dimethylphenyl N-methylcarbamate labelled with carbon-14 at 4-CH₃ was easily absorbed from the gastrointestinal tract of male Wistar rats and distributed into the tissues. Elimination of the radioactivity was rapid and essentially complete; namely during 48hr approximately 92% and 5% of the total radioactivity were excreted respectively into urine and feces. The content of the intact carbamate compound in the urine was less than 0.5%. Major degradation products were identified as 3-methyl-4-car-boxyphenyl N-methylcarbamate, its N-hydroxymethyl analog and its component phenol. Much less amount of direct hydrolysis product of the original carbamate, 3, 4-dimethyl phenol and its conjugated forms was demonstrated. 3, 4-Dimethylphenyl N-methylcarbamate is presumed to undergo biodegradation through oxidative pathways.

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Of five strains of yeast tested, three which produced 5-hydroxy-4-ketohexanoic acid (HKH) were also found to form a new keto acid, when grown on medium containing ethanol as the sole carbon source. The new product was isolated in pure form and the structure was investigated by elementary analysis, infrared spectral analysis, NMR analysis and determination of its degradation product.
The compound was shown to be 5-acetoxy-4-ketohexanoic acid. 5-Acetoxy-4-keto-hexanoic acid (AKH) was synthesized and found to be identical with the isolated product.

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An alternative route to dl-prostaglandin-B₁ using the Grignard reaction of 2-(6'-tert-butyloxycarbonylhexyl)-3-methoxy-2-cyclopenten-1-one (XII) with 3-tetrahydropyranyloxy-1-octyne was developed.
An easy synthesis of dihydrojasmone was also described.

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Rats fed a low protein diet containing choline in which casein supplemented with sulfur-containing amino acids is the protein source develop moderately fatty livers. Effects of some hormones on this type of fatty liver were investigated. The accumulation of fat in the liver was alleviated by injection of thyroxine. Adrenalectomy also prevented the induction of fatty liver but fat accumulated excessively in adrenalectomized rats by the injection with cortisol.

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