The following facts were found on the constituents of fatty acids of cuttle fish oil.
1. Fatty acids of cuttle fish oil is composed of about 22% saturated fatty acids, about 33% monoethenoid acids and about 45% higher unsaturated, acids.
2. Saturated fatty acids consist of 13.6% palmitic, 4.8% stearic, 3.6% myristic and small amonut of arachidic, lauric, capric acid.
3. Monoethenoid acids consist of 14.3% oleic, 9.8% gadoleic, 5.8% zoomaric and small amount of docosenic, tetradecenoic acid.
4. Higher unsaturated acids consist of 20% eicosatetraenoic, 6.2% docosapentaenoic, 4.4% docosahexaenoic, 3.5% docosatetraenoic, 2.0% tetracosapentaenoic and small amount of eicosapentaenoic, octadecatetraenoic, hexadecatrienoic acid.
5. The facts are noticeable that the amount of C₂₀ acids are that largest and docosa-hexaenoic acid is present.

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In the search for amylase-producing bacteria, a powerful strain was isolated from a herbal plant after screening a large number of bacteria from various sources. According to its fundamental characteristics, this bacterium was found to belong to the group of Bacillus subtilis, but to differ from the authentic B. subtilis in the lack of ability of forming acids from xylose, arabinose, mannit, glycerine, dextrine, salicine and starch. The most salient feature of this strain lies in its enormous amylolytic activity displayed both in surface and submerged cultures.
The optimum conditions for the amylase formation by this bacterium are: 35_??_37°, pH 6.0_??_7.2, and incubation for 3_??_5 days in the surface culture. The starch-dextrinizing acti-vity was higher in acidic media such as about pH 6.0, while the starch-liquefying activity was increased in alkaline media such as about pH 8.0. The ratio of dextrinizing and lique-fying activities was found to be variable in normal cultural conditions, indicating that the two phenomena are brought about by different enzyme-systems.
In view of its peculiar characteristics, this bacterium was concluded to be a variety of Bacillus subtilis and was named Bacillus subtilis var. amyloliquefacus.

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By using amylase-producing bacteria (Bacillus subtilis var. Amyloliquefacus) as the test organism, various cultural conditions favourable for the amylase formation were investigated in the media prepared from cereal wastes.
The media prepared from the cereal wastes, especially from defatted soybean flour, partially digested by Bacillus natto were shown to be favorable for amylase production as compared with those prepared by alkaline extraction from respective cereal wastes.
On preparing the partially digested soybean medium (designated as “natto medium”) optimum time for the digestion exhibited by B. natto was revealed to be 3 days and the most economical concentration of soybean flour to be 10 per cent. No more remarkable improvement in the amylolytic activity was observed with the addition of other carbon and nitrogen sources into the medium.
Amylolytic activity as influenced by the C:N ratio of the media (synthetic and natto media) was also investigated. It was found in this experiment that the C:N ratio favourable for the amylase formation lies between 5:1_??_8:1 (synthetic medium) and 3:1_??_5:1. (natto medium) In this connexion natto medium thus prepared is considered to be one of the most favourable media for amylase production, the C:N ratio of which was found indeed to be 3.8:1.

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In tea flavones, very water-soluble glycosides were isolated by use of chromatopile. thousand pieces of circular filter paper were used for this purpose and fifty pieces contain-ing water-soluble pigments were inserted in this pile and developed with an organic solvent (Fig. 3).
By this method, six flavonoid pigments were isolated, i.e., kaempferol-3-rhamnoglucoside, rutin, kaempferol-3-rhamnodiglucoside, quercetin-3-rhamnodigluco side, kaempferol-triglucoside, and quercetin-triglucoside (Table 2).
These pigments were unchangeable during the black-tea fermentation, very water-soluble, and abundant in tea leaves. So the yellow color of green and black tea is mainly due to these pigments.

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(1) The complex nature of the inhibition of sweet potato β-amylase caused by ascorbic acid plus Cu⁺⁺ was demonstrated by testing the degree of reactivation at different stages with cysteine. (Fig. 3 and Fig. 4).
(2) As MAPSON suggested in the case of urease, -SH group of enzyme reacts rapidly with Cu⁺ formed by ascorbic acid and results in inhibition which is completely reversed by excessive cysteine. The effect of cysteine concentration on the degree of reactivation shows that -SH group of enzyme competes with cysteine for Cu⁺. (Fig. 2).
(3) While inhibition by Cu⁺ diminishes as ascorbic acid decreases, another type of inhibition proceeds, which is not reversed by cysteine. Thus the observed inhibition is the sum of these two components. (Fig. 3 and Fig. 4)
(4) The mechanism of the latter (not reversed by cysteine) is not yet determined, but there may be a change occurring in -SH group of enzyme intramolecularly.
(5) This inhibition may be of no importance as the regulatory mechanism of β-amylase action in vivo. But, it is rather interesting that the enzyme acts as a kind of tracer indicating the mechanism of autoxidation of ascorbic acid in the presence of Cu⁺⁺ as a catalyst.

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(1) Free L-tyrosine was the propigment involved in darkening of white potatoes and it was crystallized from the tubers by means of extraction with ethanol, precipitaion with Hg(NO₃)₂ and liberation with H₂S.
(2) On treatment with 3 kg of the fresh tissues, L-tyrosine, L-asparagine, and L-glutamine were obtained in the yieleds of 0.75g, 4.5g and 0.175g, respectively.
(3) Chlorogenic acid and probably a related compound also were propigments, and the acid was separated by the method same in principle with that of RUDKIN and was identified from the resulting solution by means of paper-chromatogaphy, absorption spectra curve (max. 325mμ) of the water extract of the spot, and the color reaction.
(4) The buffered (pH 4_??_5) solution of the crude tyrosinase preparation obtained from the potatoes was effective in detecting the enzymatic discoloration by adding to isolated crystalline subtances or by spraying to the separated spots on paper-chromatograms.

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(1) Melanic pigments were formed by exposure to sunlight from L-tyrosine solutions. As intermediate of the pigmentation, 3, 4-dihydroxyphenylalanine (DOPA) was isolated in crystelline form by buffered solutions and identified by microanalysis, melting point, and other characters.
(2) The detection of the separated DOPA and tyrosine on paper chromatogram was made successfully by spraying the FOLIN's phenol reagent and then exposing to ammoniacal vapor.
(3) The formation of DOPA by the photoxidation of tyrosine was maximal at pH 4_??_5, but the color development was greatest at pH 6. The mechanism of the phenomenon was explained.
(4) DOPA was relatively stable for the oxidation at lower pH values than 5 and was little destroyed in a room. The oxidation proceeded rapidly at pH 6 or higher with melanization.
(5) Thiourea was preventive for tyrosine-oxidation, but uneffective for DOPA-oxidation.

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Pantethine was synthesized from methyl pantothenate and cystamine by heating at 100_??_140° for several hours with or without solvent. Attempts to complete purification were unsuccessfull, which included solvent precipitation, countercurrent distributions, ionexchange, and regeneration from mercuric mercaptide. The synthetic pantethine was converted into coenzyme A by pigeon or mouse liver enzyme in the presence of A. T. P.

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A new and improved method is described for synthesis of cystamine hydrochloride. β-Bromoethylamine hydrobromide [II]. was treated with equimolecular amount of thiourea in ethanol to give β-aminoethylisothiouronium bromide [III] quantitatively. The bromide was subjected to alkaline hydrolysis, the reaction mixture was neutralized with sulfuric acid, oxidised with iodine, and the cystamine hydroiodide was converted to hydrochloride thus
cystamine hydrochloride was obtained as white crystals. Overall yield was more than 60 per cent from monoethanolamine [I].

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In the hope of getting better antiseptics from the results of antimicrobial spectra, an experiment was undertaken to examine the antibacterial, antifungal and antiyeasty activities of several 3-acetyl-4-hydroxy-coumarin derivatives, having some positive and negative substituents in their benzene nuclei, the syntheses of which had been described in the previous paper⁽¹⁾. The activities of these compounds were shown in Tables 1_??_5, using some species of molds, yeasts and bacteria as the testing microörganisms.
The following conclusion might be postulated from the results.
(1) 3-Acetyl-4-hydroxy-7-acetamino-8-bromo-coumarin was found to be most effective, having a broad antimicrobial spectrum against molds, yeasts and bacteria.
(2) 3-Acety-4-hydroxy- and 3-acetyl-4, 5-dihydroxy-coumarins did not inhibit the growth of bacteria even at a concentration of 5×10^-3 but completely inhibited the growth of molds at 8×10^-5.
(3) The bromo derivative seemed to be unsuitable for the antifungal food preservative, since it did not inhibit at molds at a concetration of 5×10^-4 and was rather insoluble to the acidic side than the alkaline, in spite its high antibacterial activity.
(4) The antibiotic activity increased in the following orders, i.e. AcNH>OH, H>Br against molds, AcNH>Br>OH>H against bacteria and AcNH>Br, H>OH against Bac. saprogenus Sake, respectively, as the influence of substituents, in so far as the tested compounds concerned.
A discussion on the toxity of these compounds will be published in the following paper.
The author wishes to dedicate his great appreciation to Prof. Dr. Y. SUMIKI for his kind direction and valuable suggestions. The advices of many researchers of Sumiki Laboratory, University. of Tokyo, is gratefully acknowledged.

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It was seen that the sound part adjacent to the injured part of black-rotten swepotato turned green in the presence of air, when it was dipped in NaHCO₃ solution, as well as the cortev and the vessel of sound sweet potato. This chromogens were proved to be several caffeic acid esters such as chlorogenic acid.
2. This coloration is based on the oxidation of polyphenols in sweet potato by polyphenol oxidase and partially on the autoxidation of polyphenols; the noticeable coloration in the parts described in 1 may be explained by the accumlation of polyphenols and increase of polyphenol oxidase activity in them.
3. The presence of any one of NH
, amino acids and peptides was found to be indispensable for the coloration.
4. The coloration was considered to occur through the following process; the quinone substances, produced as the result of the oxidation of caffeic acid esters by polyphenol oxidase, oxidized the amino acids, peptides or proteins in the alkaline solution, to liberate NH₃. The reduced substances from quinones were condensed with the, quinorie substances or further with NH₃ to give the green pigment in the similar process to ninhydrin reaction. Thus the reaction was assumed as one of STRECKER reaction.
5. Amino acids and primary amines (perhaps, also, peptides and proteins) are able to be detected by this reaction. The sample to be tested is heated at 50° with methyl caffeate in the Na₂CO₃-NaCO₃ solution (pH 9.1), if the solution turns green, one or more of the compound are present in the sample.

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1. An experiment was done on the coloration of 33 different kinds of synthetic and natural polyphenolic substances oxidized by polyphenol oxidase and Purpureo Salz under the slightly alkaline condition. Caffeic acid esters, such as chlorogenic acid and methyl caffeate, and caffeyl amide turned into green, but caffeic acid, dihydrocaffeic acid and its esters, protoctechuic acid and its esters, 3, 4-dioxycinnamoyl ketone and caffeyl chloride etc. turned into brown or reddish brown.
2. The polyphenols, which into green, must have a sidechain of 3 or higher odd number of carbon atoms at the para position of a phenol ring. This chain is forming a conjugated double union together with a benzene ring, and it links, at the terminal, with a radical which has +E effect of a certain intensity.
3. The identification of each polyphenolic substance which in existing in many parts of a plant would be carried out by the following method
(A) A sample was dipped in NaHC0₃ solution and heated. Or the exracting solution of the sample was mixed and heated with the solutions of NaHC0₃ and polyphenol oxidase or Purpureo Salz.
(B) By the change of color or the solution, the kind of the substance could be identified. When it turned into green, caffeic acid ester would be present; when it was reddish brown, catechin or caffeic acid; and when it was brown, tannin.

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1. It was shown that an increase in the volume of respiration occured in the black-rotten sweet potato was caused by a polyphenol oxidase system as well as a certain cytochrome oxidase system which were activated by the infection of a fungus, Ceralostomella fimbriata.
2. It was found that there was a tendency that these two enzymes were inactivated during the oxidation of polyphenolic compounds contained in the enzyme solutions.
3. When the root tissues of sweet potato were affected by Ceralostomella fimbria, the function of the protoplasm of cells penetrated by the mycellium, was destroyed and the accumulated polyphenolic substances were oxidized by the activated polyphenol oxidase and peroxidase, to corresponding quinones irreversibly. The respiratory enzymes in the penetrating mycelliumn were assumed to be inactivated during such an oxidation on polyphenols. The assumption will be an explanation of the evidence that polyphenolic substances in sweet potato prevent penetration of the fungus.

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1. Proteins from fresh meat of twenty six of marine fish, fresh-water fish, crustacea and mollusca were prepared and their methionine and threonine contents were determined.
The results showed that meat of aquatic animals are the good sources of these two amino acids.
2. Methionine contents in dehydrated proteins of marin and fresh-water fish, cuttle fish, octopus, sea-cucumber etc. are 3_??_4.8%, except 6.3% of tunny; 2_??_3% in crustaceas and less than 2% in shells.
3. Threonine contents in dehydrated proteins are 3_??_4% in all kinds of fish; about 3% in cuttle fish, octopus, lobster, crub and less in sea-cucumber and shells.
4. Fish meat contains more methionine and threonine than mollusca and crustacea meat and there is almost no difference in their contents according to the kinds of fishes.

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In the previous paper on the growth of etiolated pea epicotyl, section it has been shown that the effect of gibberellin (GIB) is markedly dependent on the endogenous growth of the section. Since GIB is inactive in Avena curvature test, it is felt desirable to obtain the evidence as to whether or not GIB promotes the growth of coleoptile sections. This paper, therefore, reports the experiments with influence of GIB, comparing with that of indoleacetic acid (IAA), on the coleoptile sections isolated from wheat and oat.
GIB had no effect in promoting the growth of wheat coleoptile sections under any experi-mental conditions as in Tables 1_??_3 and Fig. 1.
GIB was, however, found to be slightly active in increasing the growth of the upper sec-tions of oat coleoptile, when these were cut from about 2.5cm. long plants of age 60 hours or less, but inactive in the other parts of the coleoptile as well as in the sections from the older plants (Table 4, Fig. 2). The effect of the optimal concentration of GIB is of the same order of magnitude as that of the lower concentration of IAA (0.02γ_??_0.05γ/cc.).
In Avena curvature test GIB is inactive, because the upper part of the coleoptile, GIB is effective, has been removed in carrying out this test.
Wheat coleoptiles are about ten times lower than oat in sensitivity to IAA, as measured by straight growth. This will be a tentative explanation for the reason why GIB has no effect on the straight growth of wheat coleoptile sections.

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The author collected about 3, 500 samples of soils, cereals and many kinds of agricultural and aquatic products from many districts in Japan, and isolated 47 different strains of Aspergilli, which were characterized giving violet-black colonies, and carried out morphological and taxonomical studies on them together with 2 strains of Asp. japonicus SAITO and I strain of Asp, violaceo-fuscus GASPERINI NRRL 360.
One new species, Asp. acideatus (6 strains), two new varieties, Asp. japonicus var. viridiflavus (5 strains) and Asp. japonicus var. atrofuscus (1 strain), were isolated among them along with 35 strains of Asp. japonicus.
According to THOM and RAPER⁽¹⁾, Asp. japonicus together with Asp. luchuensis and Asp. violacca-fuscus were classified into the group of Asp. nigcr or the black Aspergilli, which were divided into two main subgroups by the arrangements of sterigmata, single or double series. According to INUI'S original descriptions⁽²⁾, Asp. luchucnsis produces heads with single sterig-mata exclusively.
K. SAKAGUCHI and the present author⁽³⁾ have, however; pointed out that 3 strains of Asp. luchuensis in the collection of Institute for Fermentation (in Osaka) and Department of Agricultural Chemistry, Univ. of Tokyo, are all provided with double sterigmata predominantly and single or mixed sterigmata partly or very rarely. Also Asp. violaceo-fuscus occasionally has double sterigmata as observed on NRRL 360 which the author received from Dr. K. B. RAPER in Oct. 1948. All the strains of Asp. japonicus and 47 strains in this experiment had, however, sterigmata in one series.
As it was shown in the electron micrographs (Fig: 1 to 4), the surfaces of the conidial wall of Asp. japonicus and these 47 strains were provided with densely aculeate processes, while those of Asp. luclzuensis and Asp. violacea-fuscus were rough and somewhat roughened.
Colonies of Asp. japonicus and these 47 strains were characterized by their violet-black heads, but those of Asp. luchuensis are brownish olive and of Asp. violaceo-fuscus NRRL 360 are light purple drab.
It is, therefore, conspicuous characteristics of Asp. jcponicus and these 47 strains that they produce exclusively violet-black conidial heads with single sterigmata and conidia which possess aculeate processes over the surface.
From the points mentioned above it seems to be unsuitable to sum up Asp. japonicus and these 47 strains in Asp. luchuensis series together with Asp. luchuensis and Asp. violaceo-fuscus, and it is not appropriate to put these “violet black Aspergilli” in Asp. niger group with Asp. carbonarius series and Asp. niger series. This is why the author has proposed here to classify them in the new “Asp. japonicus group” The above relations are shown in the following Key.
A Key for the classification of the violet black Aspergilli
I. Sterigmata in two series.
A. Conidia more than 5μ in diameter…………………………………A. carbonarius group.
B. Conidia mostly less than 5μ in diameter.
1. Conidial wall distinctly echinulate when mature………A. Niger group.
2. Conidial wall smooth, rough or rarely echinulate………Kuro-Koji mold group.
II. Sterigmata in one series, conidial walls densely aculeate………A. japonicus group.
A. Conidia strongly elliptical 4.0_??_6.5 ×2.8_??_3.2μ, vesicle 50_??_80μ, conidiophores long 3_??_4mm……………A. acuteatus IIZUKA sp. nov…(6 strains)

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[in Japanese]

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By the method of two-dimensional paper chromatography of the polyphenolic substances in apple fruit, three spots were detected, i.e., L-epi-catechin which was epimerized to L-cate-chin by heating, D-catechin which gave a small spot and was epimerized to D-epi-catechin, and chlorogenic acid which was hydrolized to caffeic acid (Fig. 1).
In these, L-epi-catechin and chlorogenic acid were isolated in crystalline form from fresh apple fruit (Kokko). These were expectedly oxidized by polyphenol oxidase of apple fruit (Fig. 2).
Catechins were estimated by nephelometry (Fig. 4), and chlorogenic acid was estimated by cdlorimetry (Fig. 4), and the relation was studied between the contents of these polyphenols and the quality of apple fruit, but no evidence was obtained (Table 2).

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On the pilot plant of continuous fermentation of sulfite waste liquor (S. W. L.), Yeast Reuse Method was compared with Suspension Method which had been put into operation in Japan, and ADAMS Method which utilized the optimum phase of yeast generation.
1) ADAMS Method was successful by feed of 13 hours cycle time (v. z. total fermentation times was 23 hours), but was not suitable for the fermentation of S. W. L. even for the purpose of shortening the fermentation times, because the fermentation times was not different from that of one-batch fermentation, being attended with the serious consequences to require abundant nutriments and to lose the yeast in vain.
2) Yeast Reuse Method was better both in the fermentation recovery and fermentation times than Suspension Method.
3) In case of Reuse continuous fermentation, it had a good influence on the yeast propagation and fermentation to active yeast by passing through the medium of molasses.

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