Using a new determination method of tannin by means of colloid-titration with methy-glycol-chitosan, the reactions between various tannins and hide powder were investigated from the viewpoint of absorption isotherm. The absorption mechanism of tannins such as qebracho (SO2 treatment), chestnut, myrobalan, and lignosulfonic acid can be represented by LANGMUIR's equ-ation, but in the case of tannins such as mangrove and Acacia Mollissima it may rather follow FREUNDLICH's equation. Or the basis of various data of absorption isotherm, it seems reason-able to assume that absorption of the tannins of the former group can be partly ionic, whereas that of the later group non-ionic.

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This work was undertaken in order to quantitatively determine the selective absorption of the tanning materials from the solution containing both lignosulfonic acid and the vegetable tannin. For this purpose, a method to determine the vegetable tannin and lignosulfonic acid separately has been first established by colloid-titration. The absorption of the vegetable tannin by hide powder is disturbed by lingosulfonic acid, and this disturbance is remarkable, in case of the tannin having the ionic character. However, if the absorption of tannin by hide powder is non-ionic such as that of Acacia Mollissima, the selective absorption of lignosulfonic acid and tannin does not occur. This is due to the fact that the absorption active center of Acacia Mollissima is different from that of lignosulfonic acid.

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(1) Using a mechanical smoking de%, ice (smoking conditions: regular sized blended cigarettes, volume of smoke/1 puff-40cc., puff/30 sec.-once, length of time required to take each puff-2 sec.), the authors studied on the distribution of nicotine in cigarette-smoke and butt, and measured the burning velocity and smoke-flow resistance of cigarettes.
(2) The ratio of nicotine going into the main smoke vs. the nicotine in one cigarette incr-eased along with the length of the smoked portion, and was larger in soft cigarettes than in hard ones. When 3/4 of the entire length of the cigarette was smoked, the ratios in soft, medium and hard cigarettes were 21, 19 and 13% respectively.
(3) By “3/4-smoking” about 49% of nicotine in one soft cigarette escaped into side smoke or decomposed, while values in medium and hard cigarettes were 52 and 59%, respectively.
(4) The rate of the nicotine accumulated in butt was larger in short butts than in long ones, and higher in soft cigarettes than in hard ones. In smoking of 3/4 of initial length, the rate of increased nicotine in butt were: soft cigarettes 18%, medium 15%, and hard 14%. In “1/4-smok ing” the rates were the largest in the center of butts in both soft and medium cigarettes, while in the case of hard ones largest in the neighbouring portion of the glowing zone. In “2/4-smoking”, however, every cigarette showed the same trend having lower values in mouth side of butts.
(5) Advancing of the glowing zone produced by each puffing in soft, medium and hard cigarettes was. 3.8, 3.2 and 2. 5 mm respectively.
(6) Lighted cigarettes showed increased values of “smoke-flow resistance” different from the initial air-flow resistance through unsmoked cigarettes. The ratios of the maximum increase .of the resistance of soft, medium and hard cigarettes were approximately 70, 60 and 30%.

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In order to elucidate the fermentative process in the rumen, goats whose rumen has a permanent fistula with fistula plug have been raised soundly for a long period (Fig. 1).
In foreign country, the feeding of ruminant with a permanent fistula was hitherto reported by many workers. But our success of the raising should be the first one in Japan. The rumen gas was collected through the fistula plug under several conditions and subjected to analyses (CH₄ C0₂, 0₂ and H₂).
(1) The animal was fed on a measured quantity of a fodder composed of mixed concentr ate and prairie hay once a day and the rumen gas was collected at definite time. Its composi-tion and the ratio C0₂/CH₄, changed with the season. The pH of rumen liquor was always lower than 7 (Table 2).
(2) In animal fed on concentrates the C0₂ content of rumen gas increased, reaching maximun 1_??_2 hrs after feeding, then decreased slowly. Its content was higher in the morning than in the afternoon. The CH₄ content changed almost the same in manner as C0₂, but. a little irregularly.
The ratio of C0₂/CH₄ was not influenced by feeding, but' its value was increased from morning to afternoon (Table 4). When the animal was fed on grazing, the increasing tendency of the ratio of C0₂/CH₄ was the same as fed on concentrate, but its value was higher than the latter (Table 5).

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We studied on the synthesis of oligosaccharides by Shizo. Pombe from the sirup containing glucose, and 1, 4-α-linked glucosidic polymers. The washed cells of Schizo. Pombe were suspended in the sugar solution (Total sugar 14. 10, Reducing sugar 5. 64 per cent) and the mixture incubated for two hours at 30°. It was demonstrated by paper chromatography that Schizo. Pombe synthesized sakebiose (0.28%), kojibiose (0.44%), isomaltose(1.25%), panose (0.33%) and isomaltotriose (0.40%). The digest was fractionated by carbon column chromatography, and oligosaccharides obtained from each fraction were identified either as crystalline acetates or free sugar.
The new trisaccharide which probably consisted of two 1, 2-α-glucosidic linkages, herein designated as “Kojitriose” was isolated from the digest.

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Oxalic, succinic, gallic and pectic acid were isolated from the water extracts of dried tea leaves, and malic acid was proved by papaer partition chromatography.
It is a very interesting fact that these acids are abound in Gyokuro (shaded tea) in their free state.

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Myo-inositol was isolated from the water extracts of dried tea leaves. The method is as follows: neutral lead-acetate was added to the water extracts of dried tea, leaves, and after removing the salts the solution alkalized by dil. NaOH soln. in order to precipitate the lead-salts of myo-inositol and other glucosides.
The basic lead-salt was next decomposed by hydrogensulphide gas and evaporated in vacuo to a thin syrup, then absolute methanol was added.
The insoluble fraction, after the removal of methanol, was passed through the columns of ionexchange resin in order to remove cations and other acids.
The solution, after passing through the two columns, were then evaporated in vacuo to a thin syrup and recrystallised from dill ethanol and methanol-ether solvent.
The yield is about 0.01 wt. % of dried tea.

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This paper is concerned with the separation and determination of lysine from protein hydro-lysine, by means of paper ionophoresis.
In regard to the separation and the recovery of lysine, the following conditions were found suitable: current; 3. 5 mA/4cm (width), voltage; 300 volts; duration, 180 minutes; electro-lyte; 5/100M solution.
Under the present experimental 'condition, teh decomposition rate of lysine was estimated during ionophoresis, and data for the determination of lysine is given in Figures 3 and 5.

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Cl. felsineum var. sikokianum, a useful retting bacteria, produces an enormously large amoun.t of liquefying polygalacturonase and protopectinase, while the production of saccharifying polygalacturonase is low. When the culture media of this bacteria was passed through a column of Amberlite IRC-50 (H-form), liquefying PG and protopectinase were adsorbed on the resin, while saccharifying PG passed through the column (Table 1). The resin which adsorbed enzymes, was taken from the glass tube and washed with N/20 HC1. Protopectinase and colouring matter in the resin were eluted by HC1. After washing the resin with water, liquefying PG of the adsorbent was completely eluted by 0.5 ammonia soin. PG in the eluate was purified by the process as shown in Figure 2. The optimum pH value of this enzyme was found to be 6.0. When purified liquefying PG acted on the 0.5% pectic acid soln. at pH 6.0. 35°, a considerable decrease of viscosity was observed within several hours. The result will be shown in Table 5. When action of liquefying PG was continued for 10 days, until the rate of hydrolysis reached 90%, a small amounts of mono- and digalacturonic acids were produced. After 15 days' hydrolysis, 175cc of ethanol was added to 50 cc of the reaction media. A white ppt. was produced and when salts in the ppt. were removed by ion-exchange resins, 0.177 g of low polygalacturonic acid was obtained. The yield was found to be 78.7%.
Saccharifying PG was obtained from a crude enzyme soln. passed through ion-exchange resin. This PG did not act on pectic acid but reacted on low polygalacturonic acid produced from pectic acid by liquefying PG. The end products of the reaction media proved to be monoand digalocturonic acids (Table 6).
From the results mentioned above, it was found that the liquefying PG first acted on pectic acid, and saccharifying PG hydrolysed low polyuronide into mono- and digalacturonicacids.
The liquefying PG revealed a weak action on pectin.

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The influence of various inorganic actions and anions upon activity of Aspergillus-proteinase (Takadiastase) were investigated. Of the twenty cations tested, some ions of heavy metals, especially silver and mercuric ions, were found effective to cause inactivation of the enzyme. But the addition of other metals which have been known to affect the activity of peptidases, such as cobalt, manganese, and magnesium ions, did not increase the activity of this preparation. Of the twenty anions, ferrocyan and iodic acid ions seemed to be somewhat inhibitive toward this enzyme. It was found that the enzyme inactivated by the heavy metallic ions is reactivated by either hydrogen sulfide or thiosulphate. Despite of the enzyme characteristics suffering reversible inactivation heavy metals, it is difficult to assume whether the sulfhydryl groups are essential to the activity of this fungal proteinase, since specific reagents for the sulfhydryl groups, such as ferricyanide, iodoacetate, and p-chloromercuribenzoate had no significant influence on enzyme activity.

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An attempt was made to search for bacteriophages affecting lactic acid bacteria of various groups. Eighteen strains of phage were isolated; 7 were found to ba active against Streptococcus brevis, 6 against S. faecalis, 2 against Lactobacillus arabinosus, 2 against L. brevis and 1 against leuco nostoc mesenteroides.

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Temperature effects on the reproduction of a bacteriophage previously isolated from sakestarter were studied. The phage attacked its host, a strain of Leuconostoc naesenteroides, likely within the temperature range 5° to 30°, but differently at 38°. At 38°, some of the cells were abortively infected and the others could neither grow nor produce any phage. Though it was shown that the phage synthesis was really inhibited, the liberation of preformed virus particles occurred at that temperature. Thus the authors were able to draw a curve similar to so-called “intracellar growth curve” of viruses, by raising the incubation temperature from 30° to 38° at intervals during the normal latent period. Data were presented which indicate the considerable heat-lability of the infected cells. Some possible mechanisms regarding these phenomena are considered.

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We have isolated a crystal and obtained fairly good yield by melting glutamic acid at 200° in vacuum under 30 mm Hg.
It gave a new crystal after recrystallization from hot water, under slow operation.
From the properties as described in Table 1, the former crystal was concluded to be 3, 5, 8, 10-tetraketoperhydrodipyrrolo [a, d] pyrazine, and the latter, N-pyroglutamyl pyroglutamic acid-a substance previously unreported.

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1. A, brief procedure for the detection of ribonucleolytic enzymes on agar plate was devised.
2. With the presence of adenylic acid, guanylic acid, cytidylic acid, and uridylic acid in yeast ribonucleic acid hydrolysates obtained from a ribonucleodepolymerase preparation was recognized by means of paper electrophoresis, chromatography, and ultraviolet spectrophotometry. (This enzyme preparation, free from ribonucleophosphatase, was prepered from culture filtrates. of Aspergillus oryzae var. No. 13 with the aid of anion exchange resin, Amberlite IRA-400.)
As degradation products resulting from the action of the enzyme preparation, containing both ribonucleophosphatase and ribonucleodepolymerase, the following were obtained adenosine, guanosine, cytidine, uridine, and inorganic phosphate.

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The products formed by the action of Aspergillus oryzae ribonucleolytic enzyme system on various nucleotides, nucleosides, and purine or pyrimidine bases were studied by paper electro-phoresis, chromatography, and ultraviolet spectrophotometry.
It was then concluded that the ribonucleolytic enzyme system from Aspergillus oryzae should contain mononucleotidase acting on 3'-adenylic acid, 3'-guanylic acid, 3'-cytidylic acid, and 3'-uridylic acid, adenyl deaminase acting on 3'-adenylic acid and adenosine, and purine nucleoside hydrolase acting on inosine and guanosine, as well as ribonucleodepolymerase.
The principal pathways in the enzymatic degradation of yeast ribonucleic acid by Aspergillus oryzae var. No. 13 were shown (Fig. 31).

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The effect of medium pH on the inhibitory power of sorbic acid toward microörganisms was determined by the dilution method. The results are as follows:
1) Without distinction of the test strain, the medium pH did significantly influence the inhibitory power of free sorbic acid: i.e. in an acidic environment, below pH 4, sorbic acid prevented the growth of microorganisms even at the concentration of about 0.002 per cent. But proportional to the increase of pH value, the inhibitory action decreased, and at a neutral pH condition it disappeared (see Table 1 and Fig. 1).
On the contrary, the inhibitory powers of esters of sorbic acid was invariable in any pH condition, and maintained the same level of inhibitory powers at about 0.002 per cent (see Table 2 and Fig. 2).
2) It may be understood that such a behaviour of sorbic acid is due to selective permiability of cell membrane toward nondissociative molecules of this agent; i.e. sorbic acid dissociates in proportion to the medium pH respectively, and, in case of such molecules, the nondissociative molecules penetrate into the cell injuring cell metabolism, but the ionized molecules cannot permeate the cell membrane and thus do not have. influence upon the physiology of microbes. Therefore, the inhibitory power of sorbic acid should depend on the concentration of nondissociativemolecules, and not upon that of the ionized molecules, and as the former is decided as a function of pH, so the inhibitory power varies as a function of medium pH. It was also found for the same reason that esters, nondissociable compounds, indicate the constant maximum inhibitory powers.
3) This information is neteworthy in the application of sorbic acid to beverage etc.

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Streptomyces No. 310 is an antibiotic streptomyces which was isolated by the authors from the soil of Sayama-hill in1949. The antibiotic substance (No. 310 substance) produced by this organism crystallizes from acetone in rhombic form; d. p. 228_??_230°, elemental analysis (Found: C 51.49, H 5.06, N 4.94, Cl 13.15), antibacterial spectrum and toxicity of this substance show that No. 310 substance is identical with aureomycin monohydi-ochloride⁽³⁾⁽⁴⁾. After the taxonomic studies on Streptomyces No. 310, the authors recognized that Streptomyces No. 310 was different from Streptomyces aureofaciens; an aureomycin producing strain which was reported by B. M. DUGGAR in 1948⁽³⁾⁽⁵⁾. Streptomyces No 310 is easily differentiated from Streptomyces aureofaciens in the following points; the conidium of S. No. 310 is short rod to short, cylindrical in intact form, and when stained with PFEIFFER's method it is right prism to cube-shaped having slightly rounded corner, about 1.0 by 1.5 microns, on the other hand the conidium of S. aureofacieas is spheroidal to oval. (Table 4). On the potato plug S. No. 310 showes heavy growth, pinkish gray with purplish tinge, pale reddish brown soluble pigment formed, rugous and potato darkened after long time, while S. aureofaciens does not formed soluble pigment and not darkened potato. (Table 4). The culture of litmus milk; S. No. 310 shower yellowish gray collar, coagulation and peptonization with alkaline reaction, while S. aureofaciens showes yellow-brown above, neither significant pH change nor apparent peptonization in 15 days (Table 4). After the analytical comparisons of chemical changes in l0% skim-milk powder medium, S. No. 310 is different from S. aureofaciens in the surveys of total N and NH₃-N in centrifuged culture medium, redoxpotential and pH; EhmV of S. No. 310 broth in 6 days is +107.2 mV at pH 6.6, 37° while +163.2 mV at pH 6.2, 37°, in S. aureofaciens. On 12 th day S. aureofaciens showes higher potential than S. No. 310. pH of S. No. 310 broth on 17th day culture is pH 8.2 and that of S. aureofariens is pH 5.8. The observation on total N and NH₃-N of the two strains. is different as Fig. 3.
According to the T. G. PRIDHAM's method⁽⁷⁾, S. No. 310 cannot utilize xylose, arabinose and lactose, while S. aureofaciens utilizes these three carbohydrates.
In the description of BERGEY'S Manual of Determinative Bacteriology (1949), S. No. 310 is only somewhat related to S. flavovirens but it is differenciated from this species by the color of soluble pigment, and on the culture of gelatin stab; S. No. 310 does not liquefy gelatin (Table 3).
Hence, the authors designated this strain as Streptomyces sayamaensis n. sp. after the name of the place, whence this organism has been isolated.

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In previous papers, the authors described on the photolysis of tryptophan and its resulting products.
Afterwards, further experiments were attempted to confirm those products when oxidizable L- and DL-tryptophan were treated with peracetic acid and performic acid.
Among the oxidized products of L-tryptophan, L-kynurenine was isolated and identified as L-kynurenine sulfate monohydrate, and DL-kynurenine sulfate monohydrate was similary derived from DL-tryptophan.
Now L- and DL-kynurenine may be prepared easily by the use of peracetic acid without depending upon the biological method.

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The estimation of pantoic acid (PA) and its precursors contained in the enzymatic reaction mixture was studied. The results were as follows:
(1) α-Ketopantoic acid (KPA) was determined by means of the specific color reaction of 2, 4-dinitrophenylhydrazone of a-ketopantolactone (KPL-DNP), with ammonia water (cf. Part 2 No 1). The conversion of a-ketpoantolactone (KPL) to KPL-DNP was promoted by the addition of hydrochloric acid and especially ethanol. The condition of the quantitative formation of KPL-DNP and the standard curve for determination are shown in Table 2 and Figure 4, respec-tively.
(2) β, β-Dimethylpyruvic acid (DMPA) was determined by measuring the red color due to the alkalinized 2, 4-dinitrophenylhydrazone of DMPA (DMPA-DNP). The standard curve is shown in Figure 9. The steric structure of DMPA-DNP was assigned to the syn configuration from the infrared absorption spectra data and the difficulty of isomerization to an anti isomer may be attributed to effects of steric hindrance caused by the terminal methyl groups of DMPA-DNP molecule.
(3) The total amount of KPA and PA was determined by the color reaction of KPL and pantolactone (PL) using 2, 7-naphthalenediol reagent. The standard curves are shown in Figure 11.
(4) R_F values of pantothenic acid precursors and their related compounds are shown in Table 4.

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