Hydroxyurea in an appropriate concentration inhibited DNA synthesis in Escherichia coli HF4701 (HCR-) and C (HCR+) without preventing the synthesis of protein and RNA. The drug also inhibited the formation of mature infectious particles of bacteriophage øX174 in E. coli HF4701 without preventing the synthesis of viral protein. The DNA synthesis and the phage formation were recovered upon removal of hydroxyurea.

View full abstract

The structure of fluorescein chromophore was studied with infrared and nuclear magnetic resonance spectrometries. Fluorescein chromophore formed acid salt with strong acid and this salt was decomposed with water.
Fluorescein derivatives were allowed to crystallize into trifluoroacetic acid salts. The new compounds, trifluoroacetic acid salts of fluorescein thiohydantoin (FTH)^* amino acids, were synthesized and they were studied with ultraviolet, visible, fluorescence and infrared spectrometries, as well as optical rotatory dispersion and nuclear magnetic resonance.
The trifluoroacetic acid salts of FTH-amino acids were superior to trifluoroacetic acid-free form as the standard materials of N-terminal analysis.

View full abstract

Inhibitory conditions of the tryptic hydrolyzing activity of benzoyl-DL-arginine p-nitro-anilide were investigated. Trypsin was inhibited probably according to its absorption on a colloidal dispersion which was formed in the reaction mixture containing zinc chelate of o-phenanthroline, the substrate, and phosphate of the buffer constituent over pH 6. Although zinc showed the same effect as its chelate compound did in the higher conc. of the substrate, the presence of more than 2 moles of o-phenanthroline to 1 mole of zinc prevented the formation of the colloidal dispersion.
It was also found that skeleton and side chains of o-phenanthroline and its derivatives were influential factors to show inhibitory effects on the hydrolase activity of trypsin.

View full abstract

The fermentative production of uridine diphosphate N-acetylglucosamine (UDPAG) from 5'-UMP and glucosamine by dried cells of baker's yeast was studied. UDPAG was found to accumulate in a reaction system containing 5'-UMP, glucosamine, fructose, in-organic phosphate and magnesium ions with dried baker's yeast as an enzyme source. UDPAG was separated from the reaction mixture by means of anion exchange column chromatography and was identified by several biochemical methods.
The reaction conditions for the fermentative production of UDPAG were examined. The yield of UDPAG was about 40_??_66% based on 5'-UMP added when fermentation con-ditions were optimized. The concentration of glucosamine and potassium phosphate buffer, and pH as well as the water content of dried cells greatly affected the formation of UDPAG.
The distribution of UDPAG forming activity among other yeasts was studied. Relatively strong activities were found in some Debaryomyces species by which 33_??_40% of 5'-UMP was converted to UDPAG.

View full abstract

The NADP analog and NAD diphosphate were tested for the coenzyme or inhibiting activity toward various dehydrogenases. These NAD derivatives showed little or no ac-tivity of as coenzymes for most of dehydrogenases tested. Only glyceraldehyde 3-phosphate dehydrogenase reduced the NADP analog under the high concentration of enzyme system. These NAD derivatives showed no inhibiting effect toward the reduction or oxidation of pyridine coenzymes.

View full abstract

The enzyme activity to synthesize pyridoxine glucoside was demonstrated in intact cells and cell extracts of genera, Sarcina and Micrococcus. The isolated and identified strain, Micrococcus sp. No. 431 was found to have high activity of this enzyme in its cell extract. The enzyme activity reached to a maximum after 20 hr of cultivation.
The enzyme which synthesized pyridoxine glucoside via transglucosidation from sucrose to pyridoxine was purified from Micrococcus sp. No. 431 by means of ammonium sulfate fractionation, DEAE-Sephadex, hydroxylapatite and Sephadex G-100 column chromatogra-phies. The enzyme was purified about 354-fold and confirmed to be homogenous in poly-acrylamide-gel electrophoresis and ultracentrifugation.

View full abstract

Pectolytic enzyme formation by whole cells of Erwinia aroideae was remarkably stimulated when nalidixic acid, mitomycin C or bleomycin, which are all known as inhibitors of deoxyribonucleic acid synthesis, was added to a culture medium. The enzyme formation required the presence of casein acid hydrolysate in the medium and was inhibited by chloramphenicol, indicating the involvement of de novo protein synthesis. The formation of pectolytic enzyme under such conditions occurred concomitant with or before cell lysis, suggesting the induction of bacteriophages. In fact, the enzyme formation was also induced by ultraviolet ray irradiation and the bacteriophage-like particles were observed in the cell lysate induced by mitomycin C.

View full abstract

Experiments were conducted for 4 or 8 weeks to examine the nutritive value of succinic acid and di-ethyl succinate as energy source for poultry. At the dietary level of 5% or lower, response of the chicks to succinic acid corresponded exactly with its caloric value, i. e. 2.99kcal/g and no more, suggesting that succinic acid is hopeful as energy source if the price be reasonable economically.
Succinic acid at the dietary level higher than 5% and di-ethyl succinate at 5% level retarded the growth of chicks significantly, mainly due to poor appetite.

View full abstract

The viscosity change of myosin A concentrated solution with or without other com-ponents was measured as the incubation time elapsed at 30°C.
The viscosity of myosin A solution increased, but that of F-actin solution did not. The shear stress at 0.04 sec^-1 was not increased to 1.0 dyne/cm² in the former, but in the latter was below 0.5 dyne/cm².
The viscosity of myosin B solution increased slightly, but that of native tropomyosin-free myosin B solution decreased remarkably. In both the shear stress at 0.04 sec^-1 was greater than or equal to 15 dynes/cm².
The speed of the viscosity increase in the presence of 3mM pyrophosphate and 3mM MgCl₂ was higher in concentrated solution of myosin B than in that of native tropomysin-free myosin B. The shear stress at 0.04 sec^-1 after 6 hr at 30°C was 11.5 and 8.2 dynes/cm², respectively.
The effect of native tropomyosin and actin on the viscosity change was discussed.

View full abstract

L-Asparaginase [EC 3.5.1.1], antitumor enzyme, was purified to a crystalline form from the cell free extract of Escherichia coli A-1-3 KY3598, by ethanol fractionation and chromato-graphies on DEAE cellulose and CM Sephadex. The crystalline enzyme was homogeneous by the criteria of ultracentrifugation: s₂₀, w was 7.87S.
The molecular weight was estimated to be 141, 000 by the short column method. The pI of the enzyme protein was 4.75 according to isoelectric electrofocusing.
Amino acid analysis revealed the absence of cysteine or cystine residues in the molecule.
The enzyme exhibited optimal activity between pH 6 and 8. It was stable in the pH range 5.5_??_9.0.
The enzyme activity was cleared very slowly in the plasma of dog. Intravenous administration of the enzyme caused a complete regression of the Gardner lymphoma implanted in the C3H mice.

View full abstract

The effectiveness of N-methyl-N'-nitro-N-nitrosoguanidine (NG) in inducing mutations in Streptomyces cacaoi was demonstrated with two systems: reversion to prototrophy of an isoleucine-requiring mutant and the induction of forward mutation of a prototrophic strain. An optimal condition for yielding a high frequency mutation was set as follows: Treat-ment of spores with 1mg/ml of NG in 0.016M phosphate buffer, pH 6, at 42°C for 60min. In Streptmyces cacaoi, the mutagenesis depended highly on the temperature of NG treatment; elevation of the treatment temperature resulted higher frequency of mutation. Only a little dependency on temperature was shown in Escherichia coli, but the dependency was also observed when the Streptomyces was treated with ethyl methanesulfonate. The significance of these results for considering mechanism of NG action was discussed.

View full abstract

1. The nutritional value of tempeh in comparison with that of unfermented soybeans was studied in rat feeding experiment. It was observed that the PER value of tempeh (fresh or stored) was not significantly different from that of the unfermented soybeans (fresh or stored).
2. The peroxide value of the oil of stored tempeh powder was only 10% of that of stored soybean powder. Red blood cells of rats receiving tempeh were less than 20% in hemolysis by dialuric acid whereas hemolysis was 100% for the rats receiving the un-fermented soybeans. Serum tocopherol was 0.14±0.05mg/dl in the former group, but 0.07±0.05mg/dl in the latter. Liver TBA values were 0.20±0.05 O. D./g and 0.65±0.13 O. D./g in the tempeh and the unfermented soybeans group, respectively.
3. Subsitution of whole egg for tempeh to supply 30% of protein in the diet improved the quality of the protein as measured by the protein efficiency ratio. An equal improve-ment was accomplished by supplementation of tempeh with lysine, methionine, and threo-nine in such amounts as the level of these amino acids equal to that in the tempeh-egg diet.

View full abstract

Determination of small amounts of polybasic and hydroxy acids in biological materials by gas-liquid chromatography was investigated. Extraction of free acids from their aque-ous salt solution was conducted with methanol by treating the solution with Dowex 50 (H⁺). The free acids obtained were esterified with diazomethane in the presence of 2, 2-dimethoxypropane. The ester solution was concentrated under reduced pressure with an aspirator at room temperature. Finally, the concentrate, after mixing with an internal standard ester, was injected into a gas-liquid chromatograph with a hydrogen flame ioni-zation detector. Succinic, adipic, malic, mesaconic, citric, mevalonic, and 3-hydroxy-3-methylglutaric acids were found to be determinable by the present method with a high accuracy.

View full abstract

The number and reactivity of the sulfhydryl (SH) groups of potato phosphorylase [EC 2. 4. 1. 1] were investigated, in comparison to those groups in muscle phosphorylase b. Potato phosphorylase was found to contain 17.2 moles of total half-cystine residues per 200000g of protein; all of which seemed to exist in the form of the SH group. Reactivity of SH groups in the native state with various SH reagents was low. Inactivation with p-chloromercuribenzoate and iodoacetamide proceeded gradually. No inactivation was observed with 5, 5'-dithiobis-(2-nitrobenzoic acid), N-ethylmaleimide and iodoacetic acid. The enzyme apparently did not dissociate into subunits upon reacting with SH reagents. The difference in quarternary structure may be directly related to differing control mechanisms in potato and rabbit muscle phosphorylases, although their other properties are surprisingly similar.

View full abstract

The physicochemical properties and chemical constituents of the blue protein from rice bran were investigated. The blue protein was a copper-containing glycoprotein, the molecular weight of which was found to be 18, 300 Daltons by the sedimentation equilibrium method assuming the partial specific volume 0.72cm³g^-1. The hexose and pentose contents were 5.49 and 4.01g per 100g protein respectively. The copper content was 0.38%, which corresponded to 1.09 atoms per one molecule of the protein. The electron spin resonance spectrum showed that the copper was in a cupric state. The standard oxidation-reduction potential of the copper was found to be +275 mV at 20°C and at pH 7.39. The visible and near infrared absorption maxima were found at 450, 600 and 890mμ, and the 450mμ band was optically active in the optical rotatory dispersion exhibiting a large Cotton effect.

View full abstract

By continuing flavor analysis of green tea from a previous paper, ¹⁾ further twenty seven compounds were newly identified. These compounds are limonene, α-cubebene, α-copaene, caryophyllene, α-humulene, α- and γ-muurolene, β-sesquiphellandrene, δ-cadinene, cala-menene, cubenol, α-cadinol, α-terpineol, n-heptanol, n-nonanol, furfurylalcohol, n-nonanal, N-ethylformylpyrrole, pyrrylmethylketone, 6-methyl-trans-3, 5-heptadien-2-one, 2′, 2″-dihydro-α-ionone, 6, 10, 14-trimethyl-2-pentadecanone, cis-3-hexenylcaproate, cis-3-hexenylbenzoate, α-terpinylacetate, coumarin and diphenylamine.
Relative quantities of known compounds in intermediate- and high-boiling fraction were determined.

View full abstract

The synthesis of (±)-epilupinine¹⁾ from trans-1-cyanoquinolizidines (IVa) and (IVb), the intermediates of the (±)-lamprolobine synthesis is described.

View full abstract