The fat-globule membrane protein of cow's milk has been isolated, and its. electrophoretic properties studied.
The cream was washed by diluting with nine volumes of 0.95% NaCl solution and reseparated after each dilution by means of a DeLaval cream separator. The fat-globule membrane protein was obtained after four washings.
The fat-globule membrane protein was compared with casein, whey-protein and nonwahsed cream-protein on the basis of their electrophoretic properties.
Generally, 2 to 3 components were observed in the electrophoretic mobility pattern of the fat-globule membrane protein.
No positive identification of these components has been attempted from existing electro-phoretic evidence, but the fat-globule membrane protein was provisionally classified as either whey-protein (other than, β-lactoglobuline) or a protein with a whey-protein like nature.

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The adsorption of Ca⁴⁵-caseinate and whey-protein on the surface of milk fat has been studied.
Pure butter fat was emulsified in each of the following two media resulting in a fat content of 3.5%: (1) a sol containing Ca⁴⁵-caseinate (pH 6.6, casein content 2.5%), (2) acid casein whey (neutralized to pH 6.6).
The emulsion was separated by means of a DeLeval cream separator, and the cream obtained was washed by diluting it with 9 volume of 0.95% NaCl solution and reseparated after each dilution.
Results are as follows,
(1) Ca⁴⁵-caseinate was not detected on the surface of fat-globule after two washings.
(2) The paper electrophoretic diagrams showed that β-lactoglobulin was washed out after two washings, but the other whey-protein components were still adsorbed on the surface of fatglobule after four washings.
From these experiments it-is apparent that the fat-globule membrane protein of normal cow's milk does not contain Ca-caseinate.
It, therefore, seems probable that whey-protein components are contained in the fat-globule membrane protein of normal cow's milk.

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Numerous protozoaexist in the ruminant, belonging to ciliata and are moving in an active state. In order to cultivate the ciliata artificially, several experiments were undertaken and results obtained are as follows.
The rumen ciliata (Infusoria) was obtained through a permanent fistula from goat's rumen with its content and concentrated by gentle centrifugation after straining of the content throu adouble surgical gauze.
Species of Entodinium were the most among the rumen ciliata of the goat, so we have treated these species mainly, as the subject of cultivation.
(1) Infusoria can be alive in vitro for 100_??_120 hrs. in the rumen liquor requiring the coexistence of bacteria.
(2) The rumen liquor can be replaced dy the hay extract medium, which is prepared by extracting the hay with RINGER's solution and then incubated with the bacteria of goat's rumen for 3 days.
(3) There is one necessary factor for the cultivation of Iufusoria in the rumen liquor and this is produced in the hay extract medium by the rumen bacteria after about 3 days incubation.
(4) The effectiveness of the factor may be almost !entirely destroyed by heating at 60° for 10 minutes.
(5) The survival time of Infusoria varies with the species of bacteria in the culture medium.

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It was found that hydroquinone-formaline resin (Hq-F) and its oxidative product, quinone-formaline resin (Q-F) adsorp mercaptanes, cysteine and similar compounds selectively.
As an application of this facts we examined the exclusion of the cocoon-like odor caused by mercaptane in brewed products, such as Sake and beer.
By measuring the adsorption isotherm in both systems of Hq-F, Q-F and aqueous or alcoholic solutions of ethyl-mercaptane, we have shown that a Hq-F has larger capacity than Q-F in adsorption of mercaptane.
The adsorption velocity of the resins was explained by SAKURADA's equation referring heterogenous reaction velocity, and the rate determining step was dependent of the diffusion velocity into them.
The desorption of ethyl-mercaptane from the resins was carried out by steam distillation, and the resins were thus regenerated. Under a N₂ gas flow the adsorped mercaptanes were almost entirely recovered.

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Activated fuller's earth adsorbed the pigment of rape oil well and the adsorbed pigment was extracted with 3% sodium hydroxide. The pigment was precipitated the extracted alkali solution being acidulated to pH 4.3 with hydrochloric acid.
The purified pigment contained 0.17g % nitrogen and showed a biuret reaction, but did not give a ninhydrin reaction.

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The carbohydrates in black tea were divided into fourteen fractions (see Figure 1). Free sugars and the component sugars of each fraction have been investigated mainly by the method of paper chromatography and the following results were obtained (see Table 2).
Fructose, glucose and arabinose were detected as free monosaccharides, and sucrose, maltose, raffinose and stachyose as free oligosaccharides.
Inositol and two kinds of unknown substances as free sugar-alcohols were also detected.
R ha m nose, glucose, galactose, fructose, xylose and galacturonic acid were detected as the components of glycosides.
Arabinose, galactose, galacturonic acid, rha m nose and glucose were detected as the com-ponents of hot 50% ethanol-soluble polysaccharides.
Galactose, galacturonic acid, arabinose, rhamnose, ribose (?), desoxyribose (?) and glucuronic acid were detected as the components of cold water-soluble polysaccharides, and besides this arabinose, galactose, galacturonic acid, desoxyribose, ribose, glucuronic acid, rham nose and mannuronic acid as the components of hot water-soluble polysacharides.
Arabinose, galactose, galacturonic acid and small amounts of glucuronic acid and mannu-ronic acid were detected as the components of 0.5% ammonium oxalate-soluble polysaccharides.
Xylose, glucose, mannose, ribose, galactose, desoxyribose, mannuronic acid and rhamnose were detected as the components of hemicellulose.
α-Cellulose and a small amount of lignin were also detected.

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It was confirmed, by the application of the cutting method of paper chromatograms, that fructose, glucose, arabinose, sucrose, maltose, raffinose, stachyose, inositol and two unkown sugar-alcohols exist freely in black tea (see Figures 1, 2 and Table 1).
Moreover, the method of fractional quantitative analysis of these substances in black tea was established, and their quantitative distribution were studied (see Table 1).

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The carbohydrate composition of hydrol, obtained from sweet potato starch was analyzed and the sugar component of hydrol were fractionated by carbon column chromatography. Glucose, levoglucosan, scphorose, sakebiose, maltose, cellobiose, isomaltose and gentiobiose were isolated and identified by means of their crystalline sugars or acetic derivatives. Further trehalose, kojibiose and laminaribiose were identified by a paper chromatography and sugar-borate paper ionophoresis.

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It was demonstrated in this paper, by the use of paper partition chromatography that the acetone preparation of Schizosaccharoniyces Pombe No. 2 is capable of synthesizing sakébiose, kojibiose and isomaltose from glucose. These synthesized oligosaccharides were isolated by carbon column chromatography and identified as their crystalline acetic derivatives.

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Since the beginning of this century, many investigators have shown that fresh raw milk has a definite germicidal action which can be destroyed at the boiling point or by heating at 80° for 2 minutes. The above action was specific and dependent upon both the cow and the species of bacteria employed. And a substance that showed germicidal action was called “lactenin” by JONES and SIMMS in 1930.
The test organisms used here in this experiment consisted of 12 strains and 1 human type strain of Streptococcus pyogenes, cultured in serum broth. The germicidal action of milk was estimated by its inhibitory effect on acid production of these organisms using the pH response as the criterion. One of these strains “Yugo” (animal type) was found to be most suitable for the pH response assay, and the optimum condition for the experiment was obtained by using the 4th generation of the culture.
A considerable variation among the cow's milk in its germicidal activity was found. In general, however, the most significant manifestation of the action could be obtained after the heating of milk at 58° for 20 min. while, its action was destroyed by heating over 70° for 20 minutes. On the other hand, the fresh raw milk showed that the acid production of lactic acid bacteaia surviving in the milk would overcome the germicidal action of the milk above mentioned, whose dose-response curve would not be apparent.

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By means of ultraviolet irradiation, followed by LEDERBERG's replica method, two auxotrophic mutants were induced from one laboratory strain which seemed to belong to the Streptomyees griseoftavus group. One of them, strain No. 261, requires isoleucine and the other strain No. 305, cystine or cysteine. In both cases, the amount of the amino acid required to attain about. half maximal growth was determined to be 10_??_50mg/l. Though the dense inocula (10⁴ of No. 261 or 10⁸ of No. 305, spores per plate, ) provoked the growth of strain even on a minimal medium presumably by a leakage like phenomenon, the colonies appearing on minimal medium were auxotrophic and so far, after repeated subcultures, the strains have been kept stable no reversion being observed.

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(1) The partial purification of the active factors in Mikuni peptone and Bacto yeast extract, which produce the phenolsulphatase of Enterobacteriaceae, was studied. The fractions of MB-9 and YB, which have an activity ten times stronger than that of the original substance, have been obtained by the following method. The Mikuni peptone and Bacto yeast extracts were extracted with 95% aqueous solution of ethanol, and these extracts were adsorbed with active charcoal, and then eluted with an aqueous solution of phenol and acetic acid. In the case of the Mikuni peptone this MB fraction was separated by paper chromatography.
(2) A pyridine-water (4:1) solution was used as a developing solvent. Chromatography of the MB and YB fractions with this solvent revealed that almost all the active substances are to be at R_F 0.8_??_0.9 band in the former, and 0.6_??_0.9 in the latter.

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The constituents of Saké are systematically separated to 4 fractions corresponding to volatile substances, nitrogen compounds, acids, and sugars, by the use of a flash evaporator and ionexchangers. Each fraction is quantitatively recovered. The components of these fractions will be described in the next papers.

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The acid fraction adsorbed to the anion-exchanger in the systematical separation of Saké, has been studied by silica gel column chromatography, and paper chro natography.
Succinic, lactic, malic, citric, fumaric, pyruvic, and pyroglutamic acids were identified and determined.

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The presence of naturally occurring inhibitors for protease of Aspergillus oryzae was noted and some of its properties were also described. Generally speaking, the inhibiting substances are widely distributed in cereals and beans, whereby most root crops, fruits and vegetables do not contain the inhibitor except in the case of potato. Of the 23 natural products tested, barley, hull-less barley, rye, kidney bean, broad bean, and potato were found to be most inhibitive. No remarkable difference of inhibitor content could be found among the varieties of each natural product. The potato inhibitor is distributed in all parts of the tuber. In contrast to this, the inhibitor of hull-less barley is only found in the endosperm and not in the embryo. Germination did not cause any change on the nature of the inhibitors, while bacterial growth caused a complete loss of inhibiting power.

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Various studies upon the enzymic action on starch have been carried out and several specific hydrolytic products were identified. But when acids were used in the place of enzyme, most of the end product was found to be d-glucose.
This study was undertaken to clarify whether the selective cleavage of starch would occur by means of mineral and organic acids or their esters, as catalysers resulting the production of a certain length of α-1, 4-glucosidic linkage from amylose and hydrolysis on only the branching point of the α-1, 6-glucosidic linkage of amylopectin.
The present paper deals with the hydrolysis of amylose and amylopectin by hydrochloric, sulfric, phosphoric and glutaric acids. The reuslts are as follows:
1) It is impossible to find specific property in the individual catalysers, namely hydrochloric, sulfric, phosphoric and glutaric acids, under the following given condition. (1) substrate concentration, 1.0%, (2) catalyser concentration, pH 2.1_??_2.3 and 3.1_??_3.7 and (3) reaction time 52 hours under the condition of shaking in a boiling water bath. The substrates used were amylose (potato) and amylopectin. (potato).
2) Recombination reaction of the glucose did not occur in the above-conditions (sulfric acid).
3) It is noteworthy to have found that there was no significant difference between the two substrates, amylose and amylopectin, in the course of hydrolysis.
4) Difference in the production of monose, biose and triose between the catalysers used was not considerable.
5) In regard to kinetics of the hydrolysis, it will be better to consider this acid-catalysed reac tion as a “first order reaction” on the average.
The rate of hydrolysis of biose and triose will be published in the next paper.

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The WILLGERODT reaction has been applied to the synthesis of glutaric acid from levulinic acid in two methods; i) Levulinic acid was heated with sulfur and ammonium polysulfide in an autoclave, and the yield of glutaric acid was 18% of the theoretical amount, ii) levulinic acid and methyl levulinate was heated under reflux with sulfur morpholine. The yield of glutaric acid was 18%o from free acid (ca. 1% of succinic acid wrs obtained as the by-product) and 20% from methyl levulinate. This is one of the few examples which have ever been applied to the aliphatic compounds.

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(1) The authors carried out experiments on the effect of Mn and Co ions in a 1 mM concentration upon the splitting of glycyl-L-leucine by the dialyzed intestinal peptidase of pigs and remarkable variations, whether negative or positive, were found according to the methods of preparation of enzyme extracts.
(2) Negative effects (inhibitions) were obtained in the cases of using either a glycerine excract subjected to dialysis or a water extract subjected to dialysis and precipitated by acetone.
(3) Positive effects (activations) were obtained in the cases of using either a water extract subjected to dialysis after precipitation by acetone or a water extract subjected to dialysis after fractional precipitations by acetone and (NH₄)₂SO₄.
In cases (2) and (3), the dialysis was carried out at 1° in a refrigerator under reduced pressure using a collodion sack, following S. P. L. SÖRENSEN.
(4) A survey of these remarkable variations is given in Table 1.
(5) With regard to the explanation of the reason why these remarkable variations, whether negative or. positive, occur, the authors have reserved to express their opinions, since mechanism on the activation of the metals upon the splitting of the peptide has not yet been fully clarified.
(6) It should also be noted here that such remarkable variations of the effect are not to be found in the cases of splitting glycylglycine and DL-leucylglycine. (cf Table 3)

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White amorphous Spyrogyra tannin was prepared from fresh Spyrogyra arcta by the use of acetone. This tannin gave 1 mole of glucose and 5.3 moles of gallic acid when hydrolyzed with tannase. (Table 1). Methyl gallate and gallic acid were detected on the paper chromatogram by methanolysis; then 3, 4, 5-trimethylgallic acid, a small quantity of 4, 5-dimethylgallic acid, and monomethylglucose were obtained by the hydrolysis of the methylated tannin with sulphuric acid.
It has, therefore, been proved that Spyrogyra tannin is pentagalloyl-D-glucose similar to Turkish gallotannin, and for example, assumes the chemical structure of I as its main component. It contains moreover, a small quantity of galloylglucose, possesing two or more m-digalloyl groips.

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Concerning the process of biological synthesis of riboflavin, the author picked up barbituric acid, alloxan, uric acid, o-xylidine, dimethylbenzimidazole and dimethylphenvlendiamine as presumable intermediates, and the effect of these substances on riboflavin synthesis was tested following a method which was divised by the author. But it was not able to obtain the positive results on these substances.

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(1) It was found that the static cell suspension of acetone-butanol fermentation bacteria could not synthesize riboflavin even on a synthetic medium which was favorable for riboflavin formation under the growth of bacteria.
(2) It was found that certain fluorescent substances other than riboflavin, FMN and FAD were formed in a small amount in the culture medium, and these substances were extracted and isolated from 12l of the culture medium by the method of paper chromatography. And the R_F values in several solvents were determined and the absorption spectra of these substances (VB, YB and V) were taken. On these results certain discussions were carried out.

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Reaction of pentaacetylglucosamine and p-toluidine gave rise to formation of p-toluidine-N-glucosaminide tetraacete, a cyclic acetyl derivative of the N-glucosaminide. From the α- and, β-pentaacetylglucosamines, both isomers of the tetraacetate were obtained. These cyclic tetra-acetates gave upon acetylation in a mixture of pyridine and acetic anhydride the open-chain pentaacetyl derivative of the N-glucosaminide, indicating that the conditions of usual acetyla-tion rupture the lactol ring of the diamino sugars to form open-chain compounds.

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In the course of preparation of gentiobiose from N-(D-glucosyl) p-nitroaniline as a starting material, detritylation of N-(2, 3, 4-triacetyl-6-trityl-D-glucosyl) p-nitroaniline to N-(2, 3, 4-triacetyl-D-glucosyl) p-nitroaniline is required. Usual methods of detritylation gave low yields of the product because of lability of the N-glucosidic bond in acidic media. A new detritylation method has been described on the basis of the SWAIN's theory of concerted reaction, in which the tritylated compounds is treated in methanol containing a small amount of concentrated hydrochloric acid. By this procedure, N-(2, 3, 4-triacetyl-D-glucosyl) p-nitro-aniline was obtained in 27% yield. The method is open for wide application.

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