Using the synthetic medium which was set up by the author and was favorable for ribofla-vin formation by acetone-butanol fermentation bacteria, the necessity of arginine and phytic acid and the substitutive abilities of aspartic acid and guanidine for asparagine and arginine were confirmed. In the next place the effect of eighteen kinds of salts on bacterial growth and ribo-flavin formation was examined and the salts were divided into the following six groups.
(1) Salts in this group do not affect both growth and riboflavin formation of bacteria. Li₂SO₄.7H₂O, MgSO₄•7H₂O, Sr(NO₃)₂ are included in this group. But in the case of Sr(NO₃)2 it causes the inhibition of growth of bacteria when it is employed at high concentration.
(2) Salts in this group depress the riboflavin formation slightly, but do not affect the, growth of bacteria. CaCl₂ is included in this group.
(3) Salts in this group cause acid fermentation, but do not inhibit riboflavin formation. CaCO₃ and BaCO₃ are included in this group.
(4) Salts in this group inhibit the formation of riboflavin strongly. FeSO₄•7H₂O, Na₃AsO₄•12H₂O, Co(NO₃)₂•6H₂O and NiSO₄•7H₂O are included in this group. FeS0₄•7H₂O do not affect the growth or fermentation of bacteria at all, but inhibits ribobavin formation perfectly. However, Na₃AsO₄•12H₂O, Co(NO₃)₂•6H₂O and NiSO₄•7H₂O inhibit the growth of bacteria to a certain extent.
(5) Salts in this group inhibit the fermentation. AgNO₃ and UO₂(C₂H₃O₂)₂ are included in this group. The ratio of glucose consumption was depressed into 1/3 by the addition of 10 mg/100cc of them, but the riboflavin formation was observed to some extent notwithstanding such poor glucose consumption. From this fact it is considered that these salts do not inhibit the riboflavin formation essentially,
(6) Salts in this group inhibit the growth of bacteria strongly. SeO₂, HgCl₂, Na₃Co(N0₂)₆•1/2H₂O and Pb(CH₃COO)₂ are included in this group.

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Using a synthetic medium which was set up by the author and was favorable for the formation of riboflavin by acetone-butanol fermentation bacteria, the author investigated on the inhibitory action of iron, and found several facts as follows:
(1) The minimum concentration of ferrous sulfate to cause perfect prevention of riboflavin formation is identical with that of ferric sulfate and it is 1.8×10^-5 mole of Fe⁺⁺ or Fe⁺⁺⁺
(2) Biotin, folic acid, PABA, Ca-pantothenate, B₁, B₆;, nicotinic acid, B₁₂, phytic acid and. CaC0₃ do not show any competitive action against the inhibitory action of iron.
3) The value of B/A ratio (the ratio of butanol to acetone produced) on Fe-free synthetic medium is always high than that of Fe-added medium.
(4) For this result it is concluded that the change of the value of B/A ratio is caused by the direct action of iron and not by the indirect action of riboflavin formed.
(5) The evolution of CO₂ is not affected, but the evolution of H2 is slightly increased by the addition of ferrous sulfate.
(6) When iron is added to the fermenting culture, the aditional formation of riboflavin is stopped immediately and perfectly at the time of the addition of iron.
(7) It is concluded that the inhibitory action of iron on riboflavin formation may be attributed to the inhibition on the action of the riboflavin forming enzyme system of acetone-butanol fermentation bacteria.

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Investigating the inhibitory action of Na-arsenate upon riboflavin formation by acetone-butanol fermentation bacteria, it was found that Na-arsenate inhibited the fermentation along with riboflavin formation when it was used in high concentration. But in its low concentration, riboflavin formation was depressed markedly. And it was concluded that the inhibitory action of arsenate upon riboflavin formation would be attributed to the inhibition on the action of the riboflavin-synthesizing enzyme system as in the case of iron.

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Of the nitrogen metabolism in rice-plants, it is too well known to introduce that many con-tributions to these problems have already been published in Japan. But, on the contrary, few papers appeared from the view point of the recent science related to the protein metabolism.
Therefore, the present authors have attempted further chromatographic studies on protein metabolism in rice plants prepared with the water culture by KASUGAI's method. At first, free amino acids were separated from various parts of 'rice-plants at different stages from seedtime to harvest, and the residual parts were susequently treated with several kinds of solution such as cold water, 10% NaCl, 75% ethanol and 0.2% NaOH. Each fraction was decomposed with 25% hydrolytic acid and the composition of the hydrolyzates thus obtained was tested by the ordinary chromatographic process. Results may be briefly summarized shown in Table 1_??_7.
From the results of experiments, the varieties and contents of amino acids in protein meta-bolism of rice plants was confirmed at every stages from seedtime to harvest.

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A new method to is devised. It applies the ion exchanger. determine the distribution of complex chromium ion in chrome tanning liquor.
This method was applied to the analysis of K₂Cr₂O₇ solution which was treated with reducing agents, such as gluco3e, methanol, glycerine, ethyleneglycol, Na₂S₂O₃, and SO₂ gas. The following is the result.
(1) Chrome liquors have special charge distributions of complexes in accordance with the kind of reducing agent employed.
(2) The amount of individual chromium complex was changed immediately after the reduc-tion, but it became constant with the progress of aging for approximately one week.
(3) Ordinary aged chrome liquors may be classified into two groups in regard to the type of cationic chromium complexes present in the solutions. One is mainly consisted of [Cr III complex]¹⁺ [Cr III complex]³⁺ and the other [Cr III complex]³⁺ alone.
(4) The liquors prepared from K₂Cr₂O₇ by adding glucose or methanol and then H₂SO₄ were found to belong to the former group.

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The increase in particle size of chromium complexes with the degree of alkali additions was determined by Styrene Type Ion Exchangers having diffleent cross linkings. The following facts were recognized.
1) There are two groups, one may increase their molecules somewhat homogeneously and the other rather heterogeneously.
(2) We presume that the solutions of [Cr III complex]¹⁺ or the liquors containing [Cr III complexe]¹⁺ as a chief constituent, increase their particle size heterogeneously.

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The previous paper dealt with isolating a phenolic compound, C₁₅H₁₄O₃ m. p. 188_??_189 [α]²²_D-20, from cow's urine from spring to autumn, while this paper has to do with the determination of the chemical structure of that phenolic compound.
Resorcin and p-oxybenzoic acid were obtained by alkali-fusion of the phenolic compound at 300_??_320°. Dimethylether, C₁₇H₁₈O₃, m. p. 88_??_89, [α]²²_D-17.4, was oxidized at room temperature to anisic acid by the excess of CrO₃ and to a ketone, C₁₇H₁₆O₄, m. p. 121_??_122°., by 3.2 moles of Cr0₃. Diacetate, C₁₉H₁₈O₅ m. p. 120_??_122° [α]²²_D-19.3, was also oxidized to a ketone, C₁₉H₁₆O₆, m. p. 155_??_156°. These ketone groups were proved to be conjugated with the phenyl groups by UV-absorption spectrometry.
This phenolic compound seemed to be 7-oxy-3 (4'-oxyphenyl) chroman and the latter was synthesized by Marrian's method. The identity of the two compounds was shown by the mixed melting point and infra-red absorption spectra. This phenolic compound is identical with “Equol” isolated from mare's urine by Marrian et al.

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1. The leucineless and histidineless mutant strains of Asp. sojae; which, had been known to be comparatively stable, were irradiated with ultraviolet rays, and the following double auxotroph is mutants were isolated.
From 1OS his* strain; reduced nitrate (11 strains), a rgininelessr⁽¹⁾, and. methionineless⁽¹⁾.
From 14S leu**strain; reduced nitrate⁽²³⁾, arginnineless⁽¹⁾, histidineless⁽¹⁾, lysineless⁽¹⁾, methionineless⁽²⁾, nicotinic-acidless⁽¹⁾, and adenine or hypoxanthineless⁽¹⁾-(Table 2).
2. As to the genetic markers such as colony type and nutrient requirement, 10 double-less-Mutant strains excepting reduced nitrates, were proved to be more stable than single-less mutant strains.
3. Allelism-tests were carried out in every combination of 3, arginineless strains, 3 methio. nineless strains, and 4 histidineless strains, by prototrophic heterocaryon method. with the results that the strains belonging to the two formers were observed to be non-allelic each other and the latter (histidineless mutants) were allelic.
4, Prototrophic heterocaryon-tests between 3 tester strains originating from 10S his and 6 auxotrophs from 14S leu were performed. The frequencies of prototrophic heterocaryon forma-tion were found to be 0.5_??_8.9%, indicating that there is little difference between single-less (0.4_??_4.8%) and these double-less mutants.
5. The heterocaryons formed between wild type tester strains and mutant strains were pro-ved to have the character of wild type in their phenotypes such as colour of colony, sporulation, and growth factor etc.
Therefore, it may be reasonable to say that wild type characters are dominant to mutant ones so far as the above mentioned characters concerned.
* (Sporogenous type, histidineless) ** (Sporogenous type, leucineless)

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L-Glutamic acid content of mangolds which were fertilized with ammonium sulfate or urea were determined by enzymic procedure⁽⁶⁾. The content of free L-glutamic acid were present 0.02_??_0.04%, and the total L-glutamic acid 0.11_??_0.31% of fesh roots.

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1. Comparative studies were carried out on fermentable sugar production in secondary phase of starch hydrolysis with six mold enzyme solution (mold bran extracts of Aspergillus oryzae, Asp. usamii, Asp. awamori, Rhizopus tonkinensis, and α-amylase added extracts of Asp. usamii and Rhiz. tonkinensis), in the presence and absence of yeast.
2. It was concluded that the initial velocity of glucose formation from starch, referred here as “Glucose-Direct” activity⁽⁸⁾ (G. D.) represents the amount of saccharogenic amylase (Table 1).
3. Generally speaking, as has been described by PAN et all ^{(17, 18, 22)}, transglucosidation of mal-tose into unfermentable oligosaccharides plays an important role upon the rate of fermentable sugar formation from starch with the solution of those mold enzymes. Without yeast, the effect of the action continues to appear up to the secondary phase of hydrolysis. In the presence of yeast, the effect disappeares rather fast on account of the reconversion of the presynthesized unfermentable oligosaccharides into fermentable sugars by the rapid yeast removal of maltose and the reversibility of transglucosidase. This makes the difference between the two curves, T_a (total fermentable sugars produced in the absence of yeast) arid Tp, (total fermentable sugars produced in the presence of yeast). It however, is not always as vivid as in Fig. 2., because the degree of the transglu-cosidation reaction is different in each enzyme solution, due to their different transglucosidase activity and their different value of D/G. D. (ratio of α-amylase to saccharogenic amylase).
4. It was concluded that the “Glucose-Direct” activity shows close correlation to the velocity of starch attenuation in the presence of yeast.

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The author selected 198 strong acid-producing strains (1.0_??_1.4N in mash, including physiological acidic nitrogen-source) from 1, 928 strains of black Aspergilli, most of which were isolated from samples collected in almost whole districts of Japan, and using, these strains studied on -citric acid fermentation which have been applied to industrial production of citric acid from 1937 to 1948. The author carried out mycological studies on the representative strong strains and according to the SAKAGUHI's key for the classification of the “Kuro-Koji” mold⁽¹⁾, found that most of them (Group I) belonged to Aspergillus saitoi SAKAGUCHI, II7_UKA et YAMAZAKI and a few of them (Group II) to Aspergillus saitoi var. R-16 SAKAGUCHI, IIZUKA, et YAMAZAKI.
Characteristics are as fsllows:
1. Shape of the organs: same as black species of SAKAGUCHI's “Kuro-Koji” mold; especially the surfaces of the conidial wall are provided densely with fine aculeate processes, which are same as A. saitoi and A. usamii; colour of colonies are Fuscous Black XLVI-Blackish Brown (3) XLV (sometimes more brownish by group 11); sclerotia (+) (Group I), (-) (Group II); perithecia (-).
2. Dimension of the organs; see Table 1.
3. Abundant growth and good sporulation on Koji-agar, potato slice, bread, cooked rice, PFEFFER-oryzanin-agar and CZAPEK-agar. (On some media by Group II colonies are somewhat brownish.
4. Thermal death point: 55°, 30 min.
5. Optimum temperature 35° for growth, 30° for sporulation, time for sporulation 21 hrs.
6. Optimum pH for growth: 2.6_??_5.0 (Group 1), 2.6_??_4.4 (Group II).
7. Gelatin liquefaction PFEFFER-oryzanin gelatin (+); tap-water gelatin (+) (Group I), (-) (Group II).
8. Coagulation and digestion of milk (+).
9. Fermentation and assimilation of carbohydrates: see Table 2.
10. Assimilation of N-materials: see Table 3; nitrite is not assimilated.
11. Acid-producing power: very strong.
12. Saccharifying power: strong.

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The products formed by irradiating of aqueous solution of tryptophan with sunlight in the presence of air were investigated.
(1). Kynurenine was isolated as kynurenine sulfate monohydrate and confirmed by elemental analysis, paper chromatography and ultraviolet absorption spectrum. L- and DL-kynurenine were derived from L- and DL-tryptophan respectively.
(2). The red and the blue crystalline substances which were produced by mild heating of some intermediates of the photochemical degradation, were also isolated by column chromatography, recrystallization and sublimation procedure.
These pigments were identified the former as indigo red and the latter as indigo with each authentic samples for analyses, ultraviolet and infrared absorption spectra. (See Fig. 1. 2. 3)
(3). The possible degradation course of tryptophan by the irradiation was proposed.

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The two oligosaccharides previously reported as unknown in the free-sugar fraction of tea leaves and seeds were investigated.
One (b-sugar) was orcin reaction positive, and its R_F value of paper chromatogram, when developed with n-butanol-acetic acid-water (4:1:1), was similar to that of raffinose but with n-butanol-pyridine-water (3:2:1.5), the R_F value was definitely smaller. Its hydrolyzate developed three spots of galactose, glucose and fructose on the chromatogram, but the amount of galactose was larger than that of raffinose.
The other sugar (a-sugar) was also orcin reaction positive, and gave similar R_F value to that of stachyose in two developing solvents while the hydrolyzate showed four spots of galactose, glucose, fructose and xylose.

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Effects of sulfur-containing compounds, such as n-butylthiolsulfinic acid ester (B. S. A.), thio-cyanoacetic acid isopropyl ester (SCN-P), thiocyanoacetic acid n-butyl ester (SCN-B) and thio-cyanoacetic acid isoamyl ester (SCN-A) upon the respiration of both E. coli and Sacch. cerevisiae were observed.
The inhibiting effects of all drugs upon E. coli were 54_??_63%, and to Sacch. cerevisiae 85_??_97%. To establish from what cause the decrease of 0₂ uptake of microbes occurred, the number of active microbes under the same condition of respiratory experiments were counted. B. S. A. combined with the essential SH-radicals of microbes under 20 minutes reaction and inhibited their growth 100%, while SCN-compounds inhibited 30_??_40% under 90 minutes reaction.
In view of the above results it is concluded that the decrease of 0₂ uptake of resting cell is due to the inhibition of the essential stages in metabolism, which destroys the normal respiration of microbes.

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We investigated whether the direct spectroanalysis could be used for the determination of the quality of alcohol, and found that the reliability of the result of the analysis was about equal or higher than the sensorial test that had been the only method of final determination of the quality of alcohol.
1. We concluded that the following two methods were fit for above purposes, one was the direct measuring of 100-% transmittance of alcohol at near-ultraviolet region by using water as the standard (Awa.-method), the other was the direct measuring of absorbance at the same region comparing with standard alcohol which was, refined as purely as possible (Aalc.-method).
2. The absorption curve of the standard alcohol by Awa.-method between 2550Å (0%) and 2100Å (36%) showed an almost perfect parabola. Various alcohol tested by Awa.-method showed -the different types of curves which were plotted above this curve by influences of those impuriti-es. So it was found that we could evaluate the quality of alcohol absolutely or relatively by the form and the value of the peak of the curve and that the results were almost parallel to that of the sensorial test.
3. The curve for high class alcohol took place in zone of 2600 A (<5%)_??_2100Å (<55%) by Awa.-method, but the curves obtained by Aalc.-method showed larger differences between each other. Therefore if we could easily have more purified standard alcohol, Aale.-method may be more suitable for the detection of alcohol impurities.

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1) Seasonal variation of the results in the micro determination of carbon and hydrogen was discussed statistically from the results of the test analyses.
2) The micro-balances used had an accuracy of σw_??_2λ, sucrose of 2_??_4mg was used chiefly as the test material, and its contents of carbon and hydrogen were determined after the method of PREGL with some modifications using an automatic controlled combustion apparatus⁽³⁾
3) In 406 analyses (during 2.5 years), C% found showed a good consisteney with that calculated, namely mean error=+0.025% and σc=0.15%. No special care for the air conditioning was taken, yet no seasonal variation of the data was observed (Table 1 ). As somewhat higher results (ca. 0.2%) were expected in summer by the diffusion of the humidity into the absorption tube, it was supposed that the sample might absorb humidity from the air while it was being weighed.
4) In 108 analyses (during 1 year), H% found showed a good consistency with that calcu-lated on the average (mean error=-0.03%), but the fluctuation of the data (OH=0.19%) was somewhat larger than that of C%. Besides, H% found was high in summer and low in winter. These facts might be explained as follows.
a) The amount of water introduced into the reaction tube while the platinum boat was taken out and in it might be canceled, on the average, by the amount of water temporarily taken hold by PbO₂.
b) But the amount of the former was high in summer and low in winter, and that of the latter might not be constant in every determination.

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We have cultured Aspergillus sp. in molasses solution contained 1% carbohydrate under several agitation-aeration conditions of laboratory fermentor (Part 1, Fig. 3:turbine type) and examined the growth curves by the theory of aerobic fermentation. The results are as follows
1. Maximum oxygen supply is proportional to K_dP_a (c. f. Fig. 1 ). Growth rate of log phase is independent on K_dP_a but the maximum cell concentration of log phase (Cmc) depends upon K_dP_a.
2. By K_dP_a-Cmc diagram, we can decide the necessary and sufficient oxygen supply for the culture. Also, the oxygen supply is determined by changing K_dP_a during culture and by exa-mining growth curve.
3. K_d-Cmc diagram in conditions that P_a is equal shows the characteristics of oxygen diffusion coefficient through mycelia.
4. Shaking culture is different in character from submerged culture in fermentor. The difference is considered partly due to mechanical strike of fermentor and also due to surface culture-like character of shaking culture, but deciding factor could not be found. So we may say that it is dangerous to consider shaking culture as basal experiment for fermentor culture.
5. Usually cultures, by which K_d is const., are performed, but we consider that cultures, by which K_dP_a is changed corresponding to cell concentration and Pm=const.>Pmc, is maintained, , are more economical.

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By means of dynamometer (Fig. 4) and by using the sulfite oxidation method, agitation-aera-tion conditions of three geometrically similar fermentors (Fig. 1) were studied. The results are as follows:
1. Without agitation, the expression of air rate by average superficial linear air velocity, Vs, is generalized one. Differences of Kd by form and size of fermentor are slight, if this expression is used. (Fig. 3)
2. With agitation, the equation,
K_d=kVs^0.67p,
(where, k:a constant, p:agitation horsepower per unit volume)
is applicable to any geometrically similar fermentor of different size. (Fig. 8, 9)
3. Decrease of agitation horsepower by aeration (Fig. 6) could not be expressed in genera-lized form. But this is important when the above equation is applied.
4, To design a fermentor not only the above equation is necessary, but also power number and decrease of agitation horsepower must be known.

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1) In the genus Streptomyces, there is a group of morphologicaly readily distinguishable groupspecies, that is Streptomyces reticuli group.
2) A 3724 strain, one of the typical reticuli group, is observed its micromorphological characteristics.
3) The author suggests that Streptomyces reticuli group should be classified by its morphological charactesistics.

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1) A new species of Streptomyces, Streptomyces albireticuli, which produces three antibiotics, Eurocidin, Enteromycin, Carbomycin, has been described.
2) And their taxonomic relationships are studies.

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1) A new antibiotic, Eurocidin was extracted from the mycelium of Streptomyces albireticuli.
2) Eurocidin is active against fungi, yeast and Trichomonas vaginalis.

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1. For the purpose to clarify the chemical properties of impurities in the protein preparation of red clover, acid insoluble humin obtained by acid hydrolysis of protein preparation was degradated by alkali fusion, and dihydroxyphenyl mono-carboxylic acid and oxalic acid were found in the degradation products.
2. If the impurities were polysaccharide-like substances -mucilage etc. -as J. W. H. LUGG⁽²⁾ assumed, phonolic substances would have not been found. As oxalic acid is obtained from polysaccharides by alkali fusion and also from lignins, the presense of oxalic acid in the degrada-tion products do not prove that the impurities are polysaccharides. Also, polysaccharides were not detected in the protein preparations as described in Part I.
From these facts it may be more reasonable to assume that the impurities are the phenolic substances rather than the polysaccharides.

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