To find the relation between the antioxidant activity and the chemical structure, several ω, ω'-bis-(3, 4-digydroxyphenyl) alkanes have been prepared⁽³⁾. In this work the activity of these compounds was examined through the Acitve Oxygen Method using fresh lard as the substrate. The results of experiments were shown in Table 1, and were summarized as follows:
(1) The activities of most of the ω, ω'-bis-(3, 4-dihydroxyphenyl) alkanes tested were ap-proximately equal or superior to that of NDGA.
(2) Among these compounds α, δ-bis-(3, 4-dihydroxyphenyl) butane was most effective and an increase or decrease of the carbon chain between the two phenyl groups led the activity to lower potency.
(3) It, is noteworthy to point out that the introduction of methyl groups into the carbon chain between the two phenyl groups of ω, ω'-bis-(3, 4-dihydroxyphenyl) butane lowered its antioxidant activity.
The authors wish to express their appreciation to Professor Dr. Y. SUMIKI for his guidance and advice.

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It had been demonstrated⁽¹⁾ that some ω, ω'-bis-(3, 4-dihydroxyphenyl) alkanes, particularly butane, pentane and hexane, have remarkable antioxidant properties. In relation to this fact several 4-alkyl catechols, shown in Table 1, were prepared and their activities were examined to seek further light upon the relationship. between the antioxidant property and chemical structure. To compare their activities, Active Oxygen Method was applied using fresh lard as the substrate. The results of the experiments were shown in Table 1 and were summarized as follows.
(1) Introduction of an alkyl group into the 4-position of the catechol nucleus made a great enhancement of activity.
(2) Among the 4-alkyl catechols, ethylcatechol is most active, propylcatechol the next and activity of the other 4-alky catechols decreased with the increase of the length of the side chain.
(3) It was noteworthy to point out that the structures of 4-ethyl-- and propylcatechols cor-responded to one half of those of ω, ω'-bis-(3, 4-dihydroxypenyl) butane and hexane respecti-vely.
(4) Introduction of an acyl group into the 4-position of the nucleus reduced the antioxdant activity of catechol.
(5) 4-Butyl and amylcatechols had a violent vesicant property and an increase or decrease of the length of the side chain leads the activity to lower potency.
The authors wish to express their appreciation to Professor Dr. Y. SUMIKI for his guidance and advice.

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It was described in our 3 rd paper that in the butyl-acetonic fermentation by Clostridium aceictutylicum 243, the precursors of biosynthetic riboflavin included in the soybean flake were contained in both the acidic and basic amino acid groups of its hydrolyzate.
According to the fermentation tests in this report with the pure amino acids instead of the hydrolyzate, it was confirmed that both glutamic acid (or aspartic acid) and arginine were the precursors of riboflavin-biosynthesis and phytin was not the precursor but was effective to remove the Fe ion which had an inhibiting action for synthesis of riboflavin.

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About the glycerol-oxidation by Bact. succinicum SAKAGUCHI et TADA, the following results were obtained by an experiment using dried cells or extract fronr ground cells.
1. Inorganic phosphate was required for glycerol-oxidation and this phosphorylation was de-monstrated as the so-called “oxidative phosphorylation”
2. The first metabolite of the phosphorylation was α-glycerophosphoric acid.
3. While a small amount of dioxyacetone was obtained as an other product of glycerol-oxi-dation.
4. Mainly, glycerol was oxidized to carbon dioxide by the TCA-cycle.

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(1) The effect of low oxygen tension on the inhibition of sweet potato β-amylase caused by ascorbic acid plus Cu⁺⁺ and the inhibition by the same inhibitor in its different stage of autoxidation were studied.
(2) Under low oxygen tension, the inhibition by Cu⁺ survived longer than under aerobic condition and the inhibition of the irreversible type was less serious. (Fig. 1 and Table 1).
(3) The intersection of curves, which showed the course of inhibition, as it was illustrated in Figures 5 to 7 in Part 1 of this series, was the effect of condition when the oxygen tension was rate-determining for the autoxidation of ascorbic acid.
(4) Under the present experimental condition (pH 5.6 and 2_??_3°), the inhibition by Cu⁺ survived for several hours even after the complete disappearance of titratable ascorbic acid. (Curve 4 in Fig. 2 and Curve 2 in Fig. 3)
(5) The probable cause of the inhibition of the irreversible type was an oxidation of -SH group of enzyme. From the data presented in Figures 2 and 3, it might be concluded that another mechanism of oxidation must be present in addition to the oxidation by the autoxidation product of ascorbic acid (probably by H₂O₂).

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Fluorometric determination of riboflavin in milk products by electronic photofluorometer was studied.
The conditions for photolysis.of riboflavin in milk products were examined(Table 1_??_7), and both from quantities of riboflavin added to deproteinized solution of milk products and from readings measured after photolysis, regression line was obtained by the application of methods of least squares (Table 8).
Obtained regression coefficients in different kinds of milk products were shown in Table 9.
Mean regression coefficient and estimated error with 5% of significant level obtained from measured samples of 188 were found to be 171.800 and 3.59. So the relation between reading in photofluorometer _??_ and content of riboflavin Y was shown by the next equation.
_??_=Y/171.800=0.582072Y
From the results of experiments, our method of determination of riboflavin in milk products by electronic photofluorometer is presented in this paper, and it was found to be well available for its determination.
Contents of riboflavin in some Japanese milk products were also determined (Table 10).

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In the previous papers^(1)(2)(3), it was demonstrated that the compounds, shown in Table 1, possessed remarkable antioxidant activities against the oxidative deterioration of lard. In the present paper, the effectiveness of these compounds for stabilizing vitamin A in fish liver oil was evaluated from the standpoint of the practical application. The antioxidants to be tested were incorporated into fish liver oil containing a definite quantity of vitamine A in concentrations of 0.05 and 0.10% and were stored at 24°. At regular intervals, vitamin A destruction was determined by the use of Beckman spectrophotometer. The results of experiments were shown in Table 1 and were summerized as follows.
(1) Though the ω-(3, 4-dihydroxyphenyl) butyric and caproic esters had a remarkable atitioxidant activity for the deterioration of lard, they were proved to be less effective for sta-bilizing vitamin A in fish liver oil.
(2) The activities of most of the ω, ω'-bis-(3, 4-dihydroxyphenyl) alkanes were approxi-mately equal or superior to that of NDGA. Among the compounds tested, α, δ-bis-(3, 4-dihydroxyphenyl) butane was most effective and α, ε-bis-(3, 4-dihydroxyphenyl) pentane and a, ζ-bis-(3, 4-dihydroxyphenyl) hexane ranked to the next.
(3) Among the 4-alkyl catechols, ethylcatechol was most effective and the activities of other two alkyl catechols decreased with an increase of the side chain.
The authors wish to express their appreciation to Professor Dr. Y. SUMIKI for his guidance and advice.

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In the previous papers^(1)(2)(3) it was reported that several ω-(3, 4-dihydroxyphenyl) alkanoic esters and ω, ω'-bis-(3, 4-dihydroxyphenyl) alkanes were prepared and were found to have marked antioxydant activities.
In the present paper, therefore, synthesis of ω-(2, 5-dihydroxyphenyl) alkanoic esters and ω, ω'-bis-(2, 5-dihydroxyphenyl) alkanes, the isomers of above compounds, were undertaken to search more effective antioxydants.
1) The synthesis of methyl γ-(2, 5-dihydroxyphenyl) butyrate (V): β-(2, 5-Dimethoxyphenyl) propionic acid (I) prepared by the treatment of hydroquinone dimethyl ether with succinic anhydride in the presence of AlCl₃, was demethylated with HI to give β-(2, 5-dihydroxybenzoyl) propionic acid (IV). This was then reduced to γ-(2, 5-dihydroxyphenyl) butyric acid (III) by the CLEMENSEN reduction and was then esterified.
2) The synthesis of methyl ε-(2, 5-dihydroxyphenyl) caproate (XIII): Hydroquinone dimethyl ether was condensed with polymeric adipic anhydride in the presence of AlCl₃ to give δ-(2, 5-dimethoxybenzoyl) valeric acid (VIII). This acid was reduced by the CLEMENSEN reduction to give ε-(2, 5-dimethoxyphenyl) caproic acid (X). The demethylation of (X) with HI gave ε-(2, 5-dihydroxyphenyl) caproic acid (XI), which was esterified in the usual way.
3) The synthesis of 1, 6-bis-(2, 5-dihydroxyphenyl) hexane (XIV): Two moles of hydroquinone dimethyl ether were condensed with one mole of adipyl chloride in the presence of AlCl₃ to give 1, 4-bis-(2, 5-dimethoxybenzoyl) butane (IX), which was reduced by WOLF-KISNER method to 1, 6-bis-(2, 5-dimethoxyphenyl) hexane (XIII) and was then demethylated.
4) The synthesis of 1, 10-bis-(2, 5-dihydroxyphenyl) decane (XVII): This compound was prepared in 'the same way as in (XIV), using sebacoyl chloride and hydroquinone dimethylether as starting materials.
The antioxydant activities of these compounds will be discussed in the following paper.
The authors wish to express their appreciation to Professor Dr. Y. SUMIKI for his guidance and advice.

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The carbohydrates in the tea leaves were divided into eight fractions. The component sugars of each fraction have been investigated by two-dimensional multiple development paper chro-matography. The following sugars were detected as the components: glucose, fructose, sucrose and two unknown oligosaccharides as free sugars ; glucose, rhamnose, galactose, and arabinose as the component of glycosides; glucose, galactose, arabinose, xylose, mannose, rhamnose, galacturonic acid and four unknown uronic acids as the component of water-soluble polysaccharides; ribose and desoxyribose as the component of nucleic acids;, and glucose, galactose, arabinose, xylose, fructose, rhamnose, glucronic acid and another unknown uronic acid as the component of hemicelluloses.

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The Japanese rice, which belongs to Oryza sativa L. japonica, and the rice which belongs mainly to Oryza sativa L. indica showed the same contents of moisture, ash, crude fat, crude protein, and crude carbohydrate. The starches of the Burmese, Siamese, and Californian rites showed greater affinity for iodine and they produced the aqueous paste with greater specific viscosity than the Japanese rice starch. The starch of Rikuu No. 132 (produced mainly in the northern Japan) probably contains the amylopectin which had the long terminal glucose chain.

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The Brabender's amylograph was used for comparing the viscocity of the non-glutinous and glutinous Japanese rices, which mainly belong to Oryza sativa L. japonica, and the Burmese, Siamese, and other rices which belong to Oryza sativa L. indica. The glutinous rice had lower gelatinating temperature than the non-glutinous rice. Generally speaking, the rice of indica type showed the highest Brabender Unit and the glutinous Japanese rice showed the lowest. It is assumed from these results that the amylose content of the starch separated from the rice of japonica type or/and the terminal glucose chain of amylopectin of the former starch is longer than that of the latter.

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In the alkali fusion of dihydroagroclavine (II), a small amount of methylamine has been obtained, but the formation of such a substance as naphthalene derivative could not be confirmed. On the other hands, oxalic acid and an acid, C₁₄H₁₁O₀N, have been isolated from the nitric acid oxidation of agroclavine (I). Titration of the latter acid has shown it to be dibasic and it has been found to contain an N-methyl group. On distillation with soda-lime, this acid has been found to yield β-methylquinoline (IV) together with methylamine and an acid (mp. 216_??_24°). It appears probable, therefore, that the dibasic acid is a quinoline derivative and possibly an 1, 3-dimethylquinoline betaine dicarboxylic acid, for which (III) is a possible formula. Moreover, it has been found that there is no difference between pH values of agroclavine and dihydroagroclavine hydrochloride solutions. The fact makes it probable that the easily reducible double bond in agroclavine must be located at position between carbon atoms s 8 and 9. These findings will support the proposed formula (I) for agroclavine.

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1. Partially purified preparation of the saccharogenic amylase of Aspergillus oryzae was isolated from Takadiastase. The principle of the method: Takadiastase solution is saturated completely with ammonium sulfate ahd the dialyzed supernatant is attacked with 1_??_2% HgCl₂ at 30° for one or two days. Other enzymes are destroyed completely and only the saccharogenic enzyme remains.
2. The followings are some properties observed on the preparation:
(1) The saccharogenic activity is always associated with maltase activity and these are not separated each other by any methods so far tested.
(2) Optimum pH: 4.2_??_5.2 (Fig. 1, Part 2).
(3) Optimum temperature: 50_??_55° (Fig. 2, Part 2).
(4) Stable between pH 3.8_??_6.5, most stable at pH 5.2 (Table 1, Part 2). It is destroyed completely at that pH within 10 minutes at 70° Calcium ions appear to have an unfavourable influence upon the stability. (Table 2, Part 2)
(5) All of the following substances have no inhibitive or activating actions: HgCl₂, CuSO₄, Pb(CH₃COO)₂, FeSO₄, AgNO₃, ZnSO₄, MgSO₄, MnSO₄, CaCl₂, BaCl₂, NaCl, NaF, KCl, KCN, phenol, streptomycin, chloromycetin, phenylhydrazine.
(6) The enzyme is not precipitated by trichloroacetic acid, phosphomolybdic acid, phosphotungstic acid, picric acid and tannic acid.
(7) 70_??_80% of its activity remains in the supernatant when the enzyme solution is saturated with ammonium sulfate. (Table 3, Part 2)
(8) Molecular weight: 7500_??_8000 by Northrop-Anson's diffusion method.
(9) Electrophoresis shows that the preparation yet contains two components (Photo 1, Part 1), perhaps the one may be a polysaccharide composed of glucose and xylose and the other may be a polypeptide containing tryptophan, arginine, isoleucine, valine, glutamic acid, lysine and one more that is not yet certained.
(10) Maximum absorption is at 278 mμ by Beckman Spectrophotometer. (Fig. 3, Part 2)
3. As it is shown above, the enzyme is not prepared in a pure state yet. However, these properties suggest us that the enzyme may be a very different one from other enzymes so far reported on its action and chemical compositions.
Further purification is following now.

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The constituents of the free fatty acids of the cuttle-fish oil, obtained from the dark oil by the method reported in previous paper, are as follows.
1) The free fatty acids contain about 31.1% solid acids and about 68.9% liquid acids.
2) Acids of the saturated saries are C₁₄H₂₈O₂, C₁₆H₃₂O₂, C₁₈H₃₆O₂, C₂₀H₄₀O₂ and C₂₂H₄₄O₂.
3) Acids of the oleic series are C₁₆H₃₀O₂, C₁₈H₃₄O₂, C₂₀H₃₅O₂ and C₂₂H₄₂O₂.
4) Acids of the polyunsaturated series are C₁₈H₃₀O₂, C₁₈H₂₆O₂, C₂₀H₃₂O₂, C₂₂H₃₆O₂ and C₂₂H₃₄O₂.

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The determination of fat in foods can be performed by the saponification method. The procedure of this method is as follows:
After the addition of 10ml. of 33% KOH solution and 40ml. of 96% EtOH to W g. of sample, the mixture is boiled for 30 min., then cooled, and the soap formed by the saponification is decomposed with 17 ml. of 25% HCI, and the free fatty acids are then extracted with 50 ml. of petroleum naphtha, and 25 ml. of this naphtha solution are titrated with a standard NaOH solution. When T is the volume of N/10 NaOH used for the titration, the fat content (%) can be calculated by the following formula:
T×284×2×1.04×100/W×10.000=5.907T/W For the determination in samples with high a starch content, KOH used for the saponification must be dissolved in EtOH.

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Molasses was successfully and continuously fermented in the laboratory plant with the cycle time of 24 hours and controlled pH for extended period without any apparent yeast degradation.
The fermentors consisted of two ten-liter glass jar fermentor in series. The mash was agitated mechanically and removed continuously by overflow system. When the yeast level, about 120×10⁶ cells per ml., was maintained in the secondary fermentor, the fermentation cycle (24 hours) was neary equal to that of batch fermentation which was inoculated with a small quantity of yeast and agitated mechanically. At the yeast level, about 190×10⁶, cellss per ml., the fermentation cycle was reduced to 16 hours.

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(1) A procedure for studying 542 strains of Aspergilli is described in this paper. Two types of proteolytic enzyme systems are obtained.
(2) Five hundred strains of Asp. orysae type are tested. Thirty-three strains produce, sufficient proteinase activity to warrant further study.
(3) Individual mold strains varied in their ability to attack various substrates, and these dissimilarities suggest differences in the specificity and composition of the proteolytic enzyme systems elaborated by the various strains.

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Sulfate formation in eystine-media(its nitrogen source was L-cystine only) by Aspergillus molds were studied. Addition of other nitrogen sources (amino-acids, ammonium salts, nitrate, urea) diminished its formation. From cysteic acid, taurine, dithiodilactic acid, dithiodiglycol, and thioglycolic acid, sulfate formation could not be recognized. Only mercaptopyruvic acid was the precursor of sulfate.
The data accumulated lead us to postulate tentatively the following pathways of cystine metabolism.
(-SCH₂CHNH₂COOH)₂→(-SCH₂COCOOH)₂→HO₂SCH₂COCOOH→SO₄''.

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The extract of green soybeans had the action of the higher fatty acid dehydrogenase. This dehydrogenase took only the following fatty acids as the substrate: myristic, palmitic, stearic, oleic, linoleic, and alpha-hydroxy, 12-hydroxy, 9, 10-dihydroxy, and 9, 10, 12-trihydroxystearic, acids, and the optimum pH of this enzyme was 6.8 and the optimum temperature was between 35 and 40°.
By the use of 2, 3, 5-triphenyltetrazolium chloride, as the hydrogen acceptor and the indicator, it was observed that the enzyme reaction proceeded linearly with time.

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A new crystalline glucofructan obtained from the water or 80% alcohol extract of the bulb of Lytoris radiata HERB. was confirmed to be a nonasaccharide consisted of one part of gluco-se and eight parts of fructose and was named “Lycorisin” by the authors.
Experiments on the intermediate hydrolysates and methylation of glucofructan showed that fructose residues linked through C¹ and C² and that glucose was Fan end group linked with fructose as the form of sucrose, and so the above-mentioned constitutional formula was assumed.

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On industrial acetone-butanol fermentation of Blackstrap molasses, authors had experienced three types of abnormal fermentation. Among them the B type (Fig. 1) was the one of the extremly prolonged acidity peak and was very sluggish, resulting poor yields of solvents.
To overcome this sluggishness, experiments were carried out to search out some symbiotic organisms among various aerobic bacteria and yeasts for several strains of Cl. acetobulylicum.
Among tested organisms (Table 1) Torula utilis showed an outstanding effectiveness for a rapid completion of the fermentation and the solvents yields were much increased. (Table 2).
The effective factor was thermolabile and was filtrable. A strong invertase activity was demonstrated in the filtrate and a close relation between high yield of solvents and degree of inversion of the molasses medium was revealed. (Table 5, Fig. 3).
Thus the effective factor of T. utilrs against sluggishness was positively ascribable to the invertase activity of the yeast.
The sluggish factor of solvents fermentation of molasses has hitherto been unknown. However the above experiments in this paper suggested that some inhibiting factors of the invertase activity of Cl. acetobutylicum which were contained in molasses should be the principal cause of sluggishness.

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1. In CuSO₄-NaOH-Na₃ citrate mixture, the moler ratio Cu/citrate of the Cu-citrate complex is 1:1 in strong alkaline solutions, while both complexes with molar ratios 1:1 and 1:2 coexist in less alkaline solutions.
2. In CuSO₄-Ca(OH)₂-glucose mixture, a Cu-glucose complex is formed, in which the molar ratio Cu/glucose is 1:1.
3. In the acidity, the activity of Cu⁺⁺ in CuSO₄-Na₃ citrate mixture is exceedingly smaller than that in CuSO₄-glucose mixture.
4. The following assumption as to the action of Bordeaux mixture has been suggested: under a strong alkaline condition as exists at the plant surface sprayed with the Bordeaux mixture, Cu forms a complex with glucose or similar substances, and goes into solution. As soon as the complex thus formded is absorbed in the cell, intracellular pH being between 5_??_7 it decomposes to liberate Cu⁺⁺. It is pressumed that the Cu⁺⁺ is biologically active.

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