Quantitative studies on yeast 5'-nucleotidase are presented.
Km values for purine 5'-nucleotides were generally smaller than those for pyrimidine 5'-nucleotides and, among purine series, Km value for 5'-AMP was the smallest, while their V values were almost same.
The enzyme activity was inhibited in the competitive type by bases, nucleosides, 3'-or 2'-nucleotides, and NMN and in the mixed type by NAD and NADP.
Base-, ribose-, 3'- or 5'-phosphate moiety of nucleoside and nucleotide had some effects on binding with enzyme; especially the structure of base moiety characterizes the Km or Ki value.
The enzyme activity was accelerated by Ni⁺⁺ or Co⁺⁺, which increases V value but never affects Km value.
The relationship between the structure of substrate and its affinity towards enzyme is discussed.

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A method for obtaining a highly purified preparation of yeast uricase was developed. The procedure included extraction of uricase from the uricase-induced yeast cells, fractionation with ammonium sulfate and chromatography on a DEAE-cellulose column. The purified yeast uricase was shown to be ultracentrifugally homogeneous. The enzyme acted best at pH 8.5 and was stable in a range from pH 7.0 to 11.0 and at temperatures lower than 40°C. The Michaelis constant for urate was calculated to be 5.88×10^-6M at pH 8.5, borate buffer. Activity and stability of the enzyme, however, were found to be significantly affected by the kind of buffer used. The enzyme was sensitive to heavy metal ions such as mercuric and cupric ions, but the sensitivity was influenced by the kind of buffer used.

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The first effluent of essential oil of black tea in the gas chromatographic analysis is important for characterizing black tea flavor. The isolation and identification of main unknown components were conducted by means of gas chromatography, and also by infrared spectra and ultraviolet spectra. Ethylacetate, ethylalcohol and β-myrcene were identified as the increasing compounds during fermentation. cis-β-Ocimene and trans-β-ocimene were also identified only in completely manufactured black tea. These three terpenoid hydrocarbons are newly found constituents of essential oil of black tea and are supposed to contribute considerably to the black tea flavor.

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The nutritional values of nonessential amino acids as the nitrogen source in the crystalline amino acid diet for the chick growth were examined. The nitrogen of the nonessential amino acids in the basal diet for chick was substituted for a nonessential amino acid to be tested on the nitrogen base. The experimental methods were the same as in the evaluation of the nutritional value of D-amino acids previously reported. Nonessential amino acids were classified into four groups.
A. Very useful nitrogen source: Glutamic acid, Aspartic acid
B. Useful nitrogen source: Alanine, Diammonium citrate
C. Insufficient nitrogen source: Glycine, Proline
D. Harmful for chick growth: Serine
At the end of experiment chicks were killed and the concentration of free amino acids in the serum were measured. The concentration of glycine and serine in the serum increased when glycine was tested, but that of serine in the serum only increased when serine was tested. This result suggested the pathway from glycine to serine was fast and the opposite one was very slow.

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The water-extracted proteins, C and D fractions prepared from defatted soybean meals were fractionated by a method of gel filtration with Sephadex G-200, resulting in higher purification of the C and D components. The dissociated subunits of the C and D components were seen as bands B and B' on the starch-gel electrophoretical pattern of system without urea. By the starch-gel electrophoresis in system with urea, the subunits of C component were mainly corresponding to the bands 7, 8 and 9, and those of the D component mainly to the band 10. Those subunits were fractionated by column chromatography on DEAE-cellulose contained urea.

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The nature and the origin of major polyphenols in Hoji-cha were elucidated from the investigation on the effects of heating on individual flavanols by using paperchromatographic technique. Furthermore, the direction of thermal transformation of each flavanol, and the quantitative changes of major product and original flavanol were also shown. The heating of flavanol solution resulted in epimerization, polymerization and decomposition, though the aspect of change was different with the kind of flavanol and thermal condition.

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A specific inhibitor to fluoroacetanilidase was searched in order to block the first step of the fluoroacetanilides poisoning to mammals.
Fluoroacetanilidase prepared from chicken liver was inhibited by organophosphates, phosphorothioates, phosphorodithioates, phosphites, and phosphonates. Above all, the inhibition by TPP, TBT, TPPi, Dipterex, and DDVP were extremely remarkable. Their concentrations for 50% inhibition were about 10^-7M or 10^-8M, and the inhibition by TPP was non-competitive to the substrate.
These inhibitors were slightly or moderately toxic to mammals even when administered intraperitoneally.

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The effects of the fluoroacetanilidase inhibitors on the fluoroaceto p-bromoanilide (FBA) poisoning to mammals and insects were studied. Triphenyl phosphate (TPP) and tri-n-butyl phosphorothioate (TBT) were mainly used as the inhibitors.
1) In the case of the oral and cutaneous toxicities to mice and rats, TPP was effective to reduce the toxicities of FBA only when applied previously to FBA.
2) In the cases of the inhaled toxicity to mice and the combined inhaled-cutaneous toxicity to mice and rats, the effects of TPP and TBT were clearly found even when applied simultaneously to FBA.
3) When TPP or TBT was administered to mice by the combined inhaled-cutaneous application, the fluoroacetanilidase activities in the liver and kidney were rapidly decreased, but recovered within 4 to 6 days after the_??_plication.
4) The insecticidal activities of FBA to use fly, Comstock mealybug, two-spotted ider mite, and carmine mite were not afcted at all by either the previous or simulneous applications of inhibitors.
5) The properties of the fluoroacetanilidase in Mealy plum aphid seemed to be considerably different from those in worm-blooded animals. Especially, the former was not inhibited by TPP even at the concentration of 10^-4M.

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Protoplast-bursting factor (P. B. factor) has a little antibacterial activity and is capable of inhibiting the growth of Bacillus megaterium.
The cell suspensions required P. B. factor and Mg⁺⁺ for the oxidation of glucose-6-phosphate but did not require them for that of glucose.
Leakage of various cellular components into the surrounding menstruum occured when the cell suspension was subjected to treatment with P. B. factor. These materials were identified as protein, deoxyribonucleic acid, ribonucleic acid, and amino acids.
Under an electron microscope, the cytoplasm of the cells treated with P. B. factor was apparently less dense than the control, which seemed to suggest that the cytoplasm had leaked out of the inside of the cell through the membrane by the treatment of P. B. factor.

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It is well known that biotin has a marked effect on L-glutamic acid fermentation.
The authors have intended to find strains which are independent of the amounts of biotin in the culture medium. As a result, oleic acid-requiring mutants were obtained from a strain of Brevibacterium thiogenitalis which is an auxotroph for biotin. The growth of the mutant was remarkably stimulated by Tween 20, 40, 60, Ca ions and a small amount of corn steep liquor. And also, the mutant was found to have lost its requirement for biotin and showed growth response only to oleic acid or unsaturated fatty acids.

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The effect of biotin, oleic acid and other unsaturated fatty acids on the production of L-glutamic acid was investigated by using an oleic acid-requiring mutant of Brevibacterium thiogenitalis No. 653. The results described in the present paper showed that the oleic acid-requiring mutant D-248 produced a large amount of L-glutamic acid in the excess biotin-containing media, and that oleic acid seemed to be completely replaced by other unsaturated fatty acids such as palmitoleic acid and linoleic acid.

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Strefitococcus cremoris was cultivated for 7 days at 30°C in sterilized skim milk or in the sterilized 10% solution of dry skim milk. This skim milk culture was divided into precipitate and supernatant by centrifugation. The absorbancy at 280mμ of the supernatant prepared from the skim milk culture of S. cremoris was higher than that of the control supernatant.
Casein prepared from the skim milk culture of S. cremoris was less hydrolyzed by rennet than control casein at pH 7.0.
According to the free boundary electrophoretic analysis of the treated casein in M/10 veronal buffer of pH 8.5 containing urea, a-casein seemed to be hydrolyzed by S. cremoris but β-casein did with more difficulty.

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A synthetic sequence for the conversion of 2-methoxycarbonyl-methyl-3-methoxycarbonyl-4-methylindanone to (±)-7-deoxyepiallo-gibberic acid methyl ester norketone is described.

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A synthetic sequence for conversion of m-methoxy benzaldehyde to 2-methoxycarbonylmethyl-3-methoxycarbonyl-5-methoxy indanone and its reaction with methyl vinyl ketone are described.

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In the course of investigation of aromatic hydrocarbon-utilizing bacteria, a certain strain S668B2 was found to produce a large amount of p-toluic acid from p-xylene.
Strain S668B2 was shown to be capable of producing an ultraviolet-absorbing substance from p-xylene. This substance was isolated in the form crystal and identified to p-toluic acid by several observations.
From the results of taxonomic studies on S668B2, it was identified to be Psuedomonas species.
The optimal medium composition and culture condition were studied in order to increase p-toluic acid production. The addition of CaCO₃ was favorable for the p-toluic acid production. The amount of p-toluic acid formation increased from 0.4g/l to 3.4g/1 after the investigation of optimal culture condition.

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By means of a modified Konigs-Knorr reaction using 2, 3, 4, 6-tetra-O-benzyl-D-glucopyranosyl chloride, all the possible isomers of D-glucopyranoside of trans-2-aminocyclohexanol were synthesized. The total yield of the a-anomers exceeded that of the corresponding βanomers.

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Treatment of methyl tri-O-acetyl-6-azido-6-deoxy-D-glucopyranoside (II) with benzyl chloride afforded the corresponding tri-O-benzyl derivative. Reduction of the azido group with zinc-acetic acid or sodium amalgam gave 6-amino derivative. N-Acetylation, hydrolysis of the 1-0-methyl group, acetylation and treatment with dry hydrogen chloride in dioxane afforded the title compound (VIII). Königs-Knorr reaction of the chloride with (+)- and (-)-2-aminocyclohexanol produced the corresponding glycosides, one of which was proved to have the a-configuration.

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An adenine-requiring mutant (KY7208) of Brevibacterium ammoniagenes ATTC 6872 was found to accumulate an appreciable quantity of IMP and hypoxanthine in the culture liquid.
Crystalline IMP was isolated from culture broth of KY7208 by the use of ion-exchange columns. The preparation obtained was definitely identified as 5'-IMP, based on the results on paperchromatography, UV and IR absorption spectra, and analyses of its hydrolysates.
Growth responses of this mutant were demonstrated to adenine and adenosine, but not to 5'-AMP, 3'-AMP and 5'-AMP.
Over 5 mg of IMP per ml of broth were produced by the organism in natural medium consisting of glucose, yeast extract, urea, high concentrations of phosphate and magnesim salts, and others. The chemical changes showed that hypoxanthine first accumulated in the earlier stage of fermentation, and IMP synthesis then took place with the disappearance of hypoxanthine in the later stage of fermentation.

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The lipase produced by a strain of Penicillium crustosum Thom was fractionated into three lipase components, I_??_III by DEAE cellulose column chromatography, and two of them, I and II were purified and obtained in crystalline form respectively, which proved homogeneous by electrophoresis and ultracentrifugal analysis. Lipase I was an ordinary lipase with molecular weight about 29000 hydrolyzing olive oil and tributyrin favourably in almost the same degree, while II, rather, a so-called tributyrinase with M. W. about 32000 hydrolyzing tributyrin more efficiently than olive oil. The site of the activity on olive oil in these lipase was generally sensitive to sodium desoxycholate, ethylenediamine-tetraacetate (EDTA), and p-chloromercuribenzoate (PCMB), and lipase I was converted to a lipase II by a treatment with these reagents. Also, partial degradation of I by proteinase (‘pronase’) yielded the enzyme fragment of type II. On the other hand, treatment of the enzymes with hydrogen peroxide or sodium borohydride caused the conversion of type II into I. From the observation of UV difference spectrum during incubation with sodium desoxycholate it was indicated that the situation of tryptophane residue in enzyme molecule may have a significance in the activity of lipase I on olive oil.

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Certain Streptomyces strains were found to accumulate an unknown substance in culture broth when the microorganisms were grown in the medium containing DL-methionine. The substance was isolated from the culture broth as hydrochloride and was identified as 3-methylthiopropylamine (MTPA), decarboxylated product of methionine, from its melting point, chemical composition, infrared spectrum, and other properties. Cultural conditions for MTPA formation in Streptomyces sp. K 37 were investigated. The yield of MTPA from L-methionine reached about 9000 with a culture medium containing corn steep liquor. Namely, 6.47mg of MTPA per millilitre of culture broth was produced from 10mg of L-methionine per millilitre of the growth medium. The transforming activity was found in the cells of the early culture period. MTPA-producing activity was induced by L-methionine in the medium. D-Methionine was not utilized as a substrate of the reaction with intact cells. Optimum pH for the reaction appeared to be 6.0_??_8.0.

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The time-course of changes in total amount of proteins of sarcoplasmic, myofibrillar and stromal fractions in muscle of the rats fed a protein-free diet for 8, 16, 24 and 32 days, together with the referential data of those changes in the rats fed a protein-free diet up to time of death and a 60% casein diet for 12 days was determined respectively. The results were as follows: (1) The sarcoplasmic and the myofibrillar fractions decreased much more than the stromal fraction in the earlier stages of protein depletion following the same pattern as seen in reserve proteins. (2) The sarcoplasmic fraction decreased slightly more than the myofibrillar fraction as early as 8 days of the depletion, but the relative proportion between these two fractions was thereafter almost the same as that of the standard diet group. (3) In rats fed a 60% casein diet, the sarcoplasmic fraction increased markedly than the others.

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