1. Saccharomyces cerevisiae growth in a medium containing radioactive sulfate produced radioactive sulfur-containing thiamine of high specific activity in small amounts.
2. Radioactive sulfur-containing thiamine, its phosphates, and its precursor were investigated on paper chromatograms. However, no intermediate of thiazole biosynthesis accumulated in detectable amounts in yeast cells.

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The production of thiamine per unit weight of the cells by the growing culture of Escherichia coli was increased by addition of aspartic acid and sulfur-containing amino acid to the growth medium. The presence of glucose in the growth medium depressed the production of thiamine but the presence of ribose was stimulative.

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Radioactive sulfur of S³⁵-labelled L-cystine and L-methionine added to a growing medium was incorporated into the thiazole of thiamine by the growing cultures of Escherichia coli.

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1. Escherichia coli 26-43 (thiazole-requiring mutant) did not response to sulfur-containing amino acids and thiazolidine-4-carboxylic acid which had been reported to be the precursor of thiazole.
2. α-Amino-β-(4-methylthiazol-5-yl)propionic acid was active to Escherichia coli 26-43 and considered to be the possible precursor of thiazole.
3. Investigations to obtain the accumulated compound by Escherichia coli 26-43 and to decide the immediate biochemical lesion of this mutant were carried out but had not been successful.

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1. As reported in the previous paper, the R_F value of a new flavin mononucleotide, FMN′, in a solvent system of phenol-n-butanol-water (16:3:10) was 0.27, which is higher than that of FMN, 0.18, and lower than that of 4′, 5′-cyclic monophosphate, 0.50. It was further confirmed that FMN′ migrated faster than FMN toward the anode in paper electrophoresis.
2. One mole of NaIO₄ was consumed per mole of FMN′.
3. The yellow oxidation product of FMN′ by NaIO₄ was chromatographycally different from that of FMN, and more stable than that in the course of oxidation.
4. FMN′ was more stable than FMN against the hydrolysis by 5′- or 3′-nucleotidase or by the cell-free extracts of Aspergillus oryzae.
5. It was suggested that the possible structure of FMN′ might be 3′-, 2′, 5′-cyclic or 2′, 3′-cyclic monophosphate.

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1. A method for the purification of D-erythrose using Dowex 1 borate column has been presented.
2. A product formed from D-erythrose by treatment with pyridine has been isolated by Dowex 1 borate column chromatography and characterized as D-erythrulose by preparation of the crystalline o-nitrophenylhydrazone.

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1. When D-erythrose was incubated with beef liver preparation, a distinct decrease in reducing power, measured by Willstätter-Schudel's method was observed.
2. The enzyme has been prepared in a soluble form from beef liver and purified about thirty fold.
3. The enzyme shows pH optima at 6.0 and 7.5. During the reaction little oxygen uptake, little carbon dioxide output and litle change of optical density at 340mμ were observed. The enzyme acts more effectively upon D-threose than upon D-erythrose, but has no activity on the following compounds: D-ribose, D-xylose, D-arabinose, D-lyxose, D-glucose, D-galactose, D-mannose.
4. From the reaction mixture containing D-erythrose as the substrate, erythritol and D-erythronic acid were detected by paper chromatography. When D-threose was used as the substrate, the reaction products were D-threitol and D-threonic acid.
5. By treatment with alumina Cγ gel, the partially purified preparation was fractionated into a supernatant component (A) and an adsorbable, sodium citrate-extractable component (B). A or B alone showed little or no activity. When these components were mixed, almost the full activity was recovered.
6. When B was mixed with catalytic amounts of NAD, the activity was largely regained. NAD could not be replaced by NADP. A as well as B is heat labile and non-dialyzable. Therefore, A seems to be the bound form of NAD-like substance.
7. On the basis of these results, it can be expected that dismutation of D-tetrose to the corresponding polyol and acid in the presence of NAD-like substance takes place by the action of liver enzyme.

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A colorimetric determination of orotic acid reported by Tsuji was examined in detail and improved in several points. Ascorbic acid gave better results than thioglycollate in the step of reduction. Satisfactory results were obtained when the improved method was applied to several multivitamin preparations. The mechanism of the color development was also described. It has been elucidated that bromination of orotic acid results in the production of 5, 5-dibromobarbituric acid and the brominated compound is reduced with ascorbic acid to barbituric acid which condenses with P-dimethylaminobenzaldehyde to yield an orange color, 5-(P-dimethylaminobenzylidene)-barbituric acid.

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When free pyrimidine base, uracil is administered to normal healthy animals, not only the decrease of RNA in the liver, but also the decrease of FAD in the liver and kidney due to the disturbance of riboflavin biosynthesis of intestinal microorganisms and of FAD formation in the liver and kidney resulted, and the animal growth was suppressed and the blood sugar level rose. As the incorporation of uracil into nucleotides took place in the regenerating liver, uracil is to be considered to be acting effectively in the case of liver damage. The authors examined and studied the action of uracil as a free pyrimidine base and also on its concern in the riboflavin metabolism.

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1. Addition tests of synthetic ascorbic acid to raw materials were performed according to the methods of Kadin, Mizutani and Miki. The former two methods showed extremely low recoveries, while the latter was not quite satisfactory for food analysis.
2. The cause of the fluctuations of the recovery of added ascorbic acid was studied using the norit-hydrazine method and by paper chromatography. The results obtained were corrected by the recovery curve plotted against each drying time, but the significant errors could not be avoided.
3. The effect of drying with air from a drier the spotted sample on the paper and of development on the oxidation of the vitamin was tested. No effect of hot or cold air was recognized for a short time, but the recovery was considerably low when a great amount of the solution was spotted.
4. The use of a vaccum high frequency induction heater for drying was proved to give practically accurate results.
5. The effect of various solvent systems on the prevention of the oxidation of the vitamin were tested and the reasonable results could be obtained by paper partition chromatography using a vacuum high frequency induction heater for drying and a mobile phase containing 10% metaphosphoric acid for development.

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1. The growth-promoting effect of ω-methyl pantothenic acid on Saccharomyces sake (strain No. 6) was studied and it was found that pantothenic acid can be replaced with β-alanine in Saccharomyces sake as in other yeasts. ω-Methyl pantoic acid, formed by the removal of β-alanine from ω-methyl pantothenic acid, has a slight inhibitory effect but it has a remarkable growth-promoting effect after addition of an equimolar amount of β-alanine. It supports the hypothesis that the β-alanine moiety contained in ω-methyl pantothenic acid can be reutilized for the biosynthesis of pantothenic acid in this yeast.
2. Biological activities of pantothenic acid analogues other than ω-methyl pantothenic acid on Saccharomyces sake were also studied. Bis(N-pantoyl-β-amino ethyl) disulfide and bis(N-methyl pantoyl-β-amino ethyl) disulfide were found to have an inhibitory effect. These analogues do not contain the β-alanine moiety in the molecules. Panthenol, which was formed by the replacement of the β-alanine moiety in pantothenic acid with ethanolamine, gave no effect on the growth of the yeast. These results support the possibility that the growth-promoting effect of ω-methyl pantothenic acid is due to the β-alanine moiety contained in the molecule.
3. Though ω-methyl pantethine contains a β-alaine moiety in the molecule, it showed an inhibitory effect on the yeast. It was also shown that pantethine can not replace pantothenic acid and that bis (N-β-alanyl-2-amino ethyl) disulfide, a moiety of pantethine or ω-methyl pantethine molecules, had only a slight promoting effect. β-Alanine contained in these molecules of the pantethine level cannot be utilized effectively. The reason is still unknown, but it seemed probable that it is related not only with the permeability of the cell membrane but also with the enzymatic system in the pathway of coenzyme A biosynthesis.
4. Biological activities of pantothenic acid and pantethine were also studied. The growth-promoting effect of pantothenic acid on Lactobacillus arabinosus 17-5 and Leuconostoc mesenteroides P-60 were found to be greater than that of pantethine. Lactobacillus arabinosus 17-5 showed exceptionally high specificity. On the other hand, pantethine was more effective than pantothenic acid on Lactobacillus helveticus IAM-1042 and Lactobacillus bulgaricus B₁. However, the growth of these bacteria in this basal medium was very poor, but the growth was improved by the addition of skim milk, so that it seemed likely that the medium is devoid of some nutrional factors necessary for these bacteria.

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A patient with pernicious anemia in relapse was treated with coenzyme B₁₂. The red blood cell count increased significantly following the single administration of 5μg of coenzyme B₁₂. The hematologic effect of coenzyme B₁₂ was compared with the effect of CN-B₁₂ according to the reported data and was found to be almost equivalent in each other on a molar basis. The discussions was extended to the clinical application of coenzyme B₁₂ in future.

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