(1) The author examined survival rates, mutation rates and genetically fixed mutant types by X-ray radiation in Aspergillus sojace.
(2) Conidia of Aspergillus sojae were almost sterilized at 65000r and it will be seen that the mutation rate increased with dosage in a linear manner, the highest dosage, 65000r, resulting in a mutation rate of 6.24 per cent.
(3) It seems that in production of induced mutations soft X-ray radiation was proved less effective as compared with ultraviolet radiation, since sterilities and mutation rates of soft X-ray radiation has been found comparatively low in Aspergillus sojae.
(4) Mutant types by X-ray radiation were almost the same as those by ultraviolet radiation. In the present experiment a new “yeast type” which had not been obtained in the case of ultra-violet radiation was obtained, but no “heavy type” was obtained.
(5) It was observed that morphological changes occurred most frequently in especially “olive” and “light” types through X-ray induced mutants.

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We experimented what components of soybean oil gave a remarkable effect on penicillin yield when added in the shaking culture of P. chrysogenum as antifoam agents and obtained the following results.
(1) Oleic acid showed decrease in penicillin yield or no increase when added to the medium at the time of sterilization, but revealed a remarkable increase in potency when added during 24 to 48 fermentation hours in a relatively high percentage up to 0.5%.
(2) Glycerol showed no remarkable effect.
(3) Saturated fatty acids showed no remarkable effect but palmitic acid had a little unfavorable effect and stearic acid less than 0.1% showed the increase in potency. (Lauric acid worked as stimulant next to oleic acid).
(4) Oleic acid in 0.5% showed the same effect with or rather a little better than 0.5% of soybean oil and the use of oleic acid together with phenylacetic acid showed more remarkable results than the separate use of the two.
Judging from the above results, we are convinced that we have chosen a splendid formula of antifoam agents of natural fats and oils which give interesting results.

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We made comparison tests of effects of soybean oil and other various oils on penicillin yield in the shaking culture, and the results are shown below.
1. Addition of oils after 24 hours of fermentation:
A remarkable increase in potency was observed by adding soybean, rape seed, sesame oil, cotton seed, peanut, rice-bran, and olive oils, lard, and oleic acid.
2. Addition of oils after 48 hours of fermentation:
The addition of oils at this time resulted in some inhibition of mycelium growth and autolysis. In cases of rape seed oil, sesame oil, and lard, unfavorable effect was slight and the broth recovered its vigor and showed the increase in potency. In soybean oil, effect of addition time on penicillin yield was remarkable. On, the contrary, higher potency was obtained when rape-seed and sesame oil were added 48 hours after fermentation than 24 hours after fermentation. Addition of castor oil showed a bad result.
3. In the case of adding oils 24 hours after fermentation, remarkable bad results were obtained when phenylacetic acid was employed together. However, bad effects were slight in the case of adding machine oil, defoamer B, rape-seed oil, lard, or whale oil.
4. When vegetable oils including castor oil were used, pH value were low and no increase in potency was observed in these cases even though the propagation was sufficient. The pH value was low in the case of fish oils, but satisfactory increase in potency was not obtained except in the, case of whale oil. Oleic acid, machine oil, heptadecanol, and synthetic defoamer B revealed high pH values.

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(1) For the studies of antibiotic action, we sxnthesized several dehydroacylacetocarboxylic acids, among which dehydro-n-valeryl acetocarboxlic acid (m, p. 78_??_79°C), dehydro-isovalerylacetocarbo-itylic acid (m. p. 74°C) and dehydro-n-caproylacetocarboxyli_??_acid (M. P. 90_??_492°C) are new compounds.
(2) We also synthesized several dehydroacylacetic acids among which dehydro-n-valerylacetic acid (m. p. 44_??_45°C) dehydro-isovaleryladetic acid (b. p. I50_??_155°C/6mm.) end dehydro-n-caproyl acetid acid (m. p. 47_??_48°C) are new compounds.
(3) In the synthesis of dehydroacylacetocarboxylic acid from acetonedicarboxylic acid and acid chloride we obtained pure products in good yields by increasing the volume of sulfuric acid added. to acid chlorides or anhydrides as a catalyzer.

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(1) On studying antibiotic action of dehydroacetic acid (I) and dehydrobutyrylacetic acid (II) against several microorganisms, we found these compounds have antibiotic activitities on yeasts and molds such as Saccharomyces, Zygosaccharomyces, Pichia, Penieillium, Aspergillus and Rhizopus; but not so strong on Lactobacillus and Acetobacter.
(2) (I) inhibited the growth of those molds at 1:10, 000, and yeasts at 1:5, 000; (II) at 1:50, 000 and 1:30, 000, respectively.
(3) (II) showed stronger activity than p-oxybenzoic acid butyl ester in antibiotic test on the pellicle-forming yeast of soy sauce.

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(1) Several methods of preparing the food yeast from water extracts or acid-hydrolyzates of a brown seaweed (Eckeloria cava KJELLMAN) by using Candida sp. No. 1 are described.
(2) The yield was 8.56g. of dry yeast per 100g. of the seaweed in the water extract-culture.
(3) In the case of the hydrolyzate-culture under the meximum-saccharifying condition for the seaweed, the yields were 11.5 to 12.1g., and even in the partial hydrolysis 10.5 to 12.3g. of dry yeast could be obtained. In the acid-hydrolyzate, an amorphous deposit was produced during a longer storage and its removal, always delayed the growth of yeast, as it resulted in the removal of some phosphates.
(4) Some experiments on the production of yeast from a green seaweed(Ulva pertusa KJELLMAN) have been carried out. The result proved that it was not suitable for this. purpose.

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Two methods for the determination of lipoxidase activity were described: one was based upon the decolorization of crocin, a carotenoid pigment obtained from Gardenia jasminoides ELLIS, the other upon the manometric measurement of O₂ absorbed by the action of this enzyme. In the latter method, methyl linoleate emulsified by ultra-sonic waves was employed. This enzyme was inactivated at the early stage of its action, presumably, by the inhibiting action of the oxidized products.

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When flavones are chromatographied on paper, Rf values are affected not only by the distribution coefficient but also by the adsorption based on the filter paper (Table 3. ). When the ethyl acetateacetic acid-water system (50:2:50) is used as the developing solvent, the Rf values of spots of flavones are controled by those sugars, and when the n-butanol acetic acid-water system (4 1 :5) is used, the Rf values of spots of flavones containing the same sugar are controled bythose-OH and -CH₃ radicals. (Table 4.)(This journal. 24, 50 (1950)). So by these methods, the two dimen-sional paper chromatogram of flavones is obtained (Figure 1.).

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Paper chromatography of carboxylic acids by means of the hydroxamic acid method has been applied to determine the constitution of unsaturated fatty acids. Each of pure oleic, erucic, linoleic, and linolenic acids was subjected to ozonolysis, and the products of ozonolysis were treated with alkaline silver oxide suspension, in order to oxidize thoroughly the, carbonyl group of aldehydes and semialdehydes in the decomposition products to the carboxyl group. Thus obtained salts of mono-and dicarboxylic acids were esterified and then converted into their hydroxamic acid derivatives in the usual manner. [Cf. INOUYE and NODA This Journal, 23, 368 (1950); 24, 291 (1951).] These hydroxamic acids could be easily separated and identified on the one- and two-dimensional paper chromatograms.
Therefore, by using this paper chromatographic technique, the analysis of the ozonolysis products of unsaturated fatty acids, namely the determination of the constitution of these acids, can be carried out simply and minutely even from very small quantities of the original acids.

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The filter paper impregnated with silicic acid has been used to separate higher fatty acids which were inseparable on ordinary filter paper. Several solvent systems were employed for paper chromatography of hydroxamic acid derivatives of saturated fatty acids on the impregnated paper, and among those systems, n-butanol-benzene-water system gave the best results. The paper chromato-graphic separation of free saturated fatty acids was also tried on this paper using methanol as the stationary solvent and petroleum benzine as the mobile solvent, but it was less accurate than the separation by the hydroxamic acid method.
Although the use of the impregnated paper enlarged the range of separation of fatty acids or their hydroxamic acid derivatives, it was still incomplete or impossible by this method to separate such higher fatty acids having twelve or more carbon atoms from each other.

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The transformation of β-glucosides (acetates) into α-form has been studied by using SeCl₄ as a catalyser which is found by us to be effective under mild conditions.
β-Ethyl-, -methyl-, and -isopropyl-D-glucbside (acetate) were generally dissolved in purified chloroform, and SeCl₄, (3 or 4 moles ratio) was added. After refluxing for a given time(10_??_12 hours), the chloroform layer was shaken with ice water, and the mixture was filtered through an active carbon layer, and then the filtrate was concentrated to a thick syrup under a reduced pressure. After addition of a small amount of water to the syrup dissolved in alcohol, the 'solution was allowed to stand for several days, then the a-glucoside crystallised was filtered, and w4is, recrystallised from alcohol. The results obtained under various experimental conditions were described.
In order to distinguish the a-form, specific rotation and melting point were employed. The crystals obtained gave respectively no depression in melting points on admixture with the a-gluco. sides (acetates) prepared from β-glucosides by using SbCl₅, as catalyser.
The yields of α-methyl-, -ethyl-, and -isopropyl-D-glucoside (acetates) were 75, 45, and 60% respectively. It appears that SeCl₄, gives rise to the transformation of β-glucosides under a mild action.

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In the preceeding paper of this series, it has been reported that the function of 3 oxygen atoms in gibberic acid, C₁₉H₂₂O₃, was identified as being one carbonyl and one carboxyl. In this paper, the results of oxidation of gibberic acid by selenium dioxide are described which were attempted in order to acertain the position of the ketone group.
The oxidation does not take place by the usual method, but advances smoothly when the alcoholic solution of gibberic acid is heated at 144° during 3 hours in a sealed tube using six times of the theoretical amount of selenium dioxide. By this oxidation, gibberic acid gives a substance named “gibberdionic acid” Gibberdionic acid, crystallizing into yellowish plates, mp 188°, has an empirical formula, C₁₉H₂₀O₄, and shows an absorption spectrum of ultraviolet ray very similar to that of gibberic acid. This fact shows that there occurs no change in the mother nucleus of gibberic acid but oxidation occurs in one of the side chains introducing a new carbonyl group at the position of the next carbon atom of the ketone group which existed originally. Gibberdionic acid is negative against Schiff's reagent, the reaction of Tschugaef's diketone-Ni-dioxime, but -positive for the reaction of tolidine-acetate, showing the existence of an aldehyde group. Moreover, this acid gives the ppt. of cadmium-yellow and colourless crystals when recrystallized from alcohol containing water, showing just the same reactions of phenylglyoxal.
By treating gibberdionic acid with hydrogen peroxide in an alkaline solution, we obtain one mole of a hydroxy-dicarboxylic acid and one mole of carbon dioxide, but no aldehyde and volatile acid. This acid has been named “gibberdicarboxylic acid” and it crystallizes out in the form of colourless needles and decomposes at 145° We propose as the empirical formula of gibberdicarboxylic acid C_{17_??_18}H_{22_??_24}O₆, and its absorption spectrum of ultraviolet ray is observed to be very similar to that of gibberic acid. Therefore, gibberdionic acid must be an acid which contains a glyoxal, -CO-CHO.
From these results, it is ascertained that the position of ketone group in gibberic acid is located not in the mother nucleus but in one of side chains and, moreover, adjacent to the terminal methyl group of its side chain.

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It has been pointed out that the amino acids of Japanese green tea are essential components of its taste. Amino acids which were discovered previously in Japanese green tea were as follows asparagine⁽¹⁾, arginine⁽²⁾, theanine⁽³⁾, glutamic acid⁽⁴⁾, and aspartic acid⁽⁴⁾.
And it, has been supposed that not only these amino acids but also other amino acids would exist.
The author thought it was desirable to apply the technique of paper partition chromatography to the identfication of amino acids contained in Japanese green tea. In the the application of this technique to this subject, the author improved a few defects of LONGENECKEE and OTOZAI's developing apparatus and could identify the existence of glycine, alanine, valine, leucine, threonine, and glutathione.
Accordingly the author concluded that the delicacy of Japanese green tea is harmoious, combined taste of these amino acids, and that the most important role is played by glutamic acid, theanine, glycine, and alanine. It is necessary to establih.the reliable quantitative partition chromatography.

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1. The refined meal of glutinous Italian millet (TocHiot No.1) contains 1.75% of nitrogen of-the pure protein type in the dry matter and the majority of it is of the prolamin type.
2. A white prolamin preparation is separated from the defatted meal and its chemical properties are examined as follows:
a) Judging from color reactions, its content of tryptophan is doubtful, but the existence of trypto-phan is undeniable from the specific Liebermann's reaction.
b) It is the protein with high purity, containing 16.65% of nitrogen, 0.039% of phosphorus, 1.42% of sulfur, and 0.174% of ash in the dry matter.
c) The diamino acids content of it is determined by the R. J. Block's method, the dry matter contains 1.5% of arginine, 1.4% of histidine and 0.3% of lysine. Comparing with the result obtained by the Van Slyke's method, the diamino acids content by the former, is a little smaller for arginine and histidine, and remarkably smaller for lysine.

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(1) The auther has isolated an antibiotic substance as the quinine-salt. This antibiotic substance is produced by some strains of Aspergillus japonicus in modified Czapek solution and inhibits the pellicle forming yeast of “Shoyu”
(2) The molecular formula of this antibiotic substance is C₈H₁₀O₄. (M. W. 170)
(3) The toxicity for mouse is L. D. (50%) 0.3g/kg ; M. L. D. 0.2g/kg
(4) The maximum dilution of the antibiotic activity of its K salt (powder) against the pellicle forming yeast in Shoyu is 1:600000.

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