1-Naphthoic acid derivatives substituted with chlorine, bromine, methyl or nitro groups at various positions of the ring were assayed for their activities in the pea straight-growth and the callus formation tests. Their growth activities are discussed from two points of view, i.e. the spatial structure hypothesis of Veldstra and the ortho reaction hypothesis advanced for benzoic acid derivatives proposed by Hansch and Muir. It appears that a combination of these two views are appropriate for the present authors' results.

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The author has isolated an obligate halophilic yeast from old soy-mash and named it Torulopsis halonitratophila nov. sp. It is of particular interest that though this yeast shows obligate halophilic property at 30° it exhibits facultative halophilism when tested at a lower temperature such as 20° This observation suggests that cultural temperature is one of the most important factors for halophilism.

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D-Glucosaminic acid has recently been found to be an oxidized product of D-glucosamine formed by Ps. fluorescens. It has been revealed that many strains of oxidative bacteria can oxidize glucosamine. The formation of glucosaminic acid has been recognized among a large number of strains of Pseudomonas, Acetobacter and Gluconobacter, by means of paper chromatography. Furthermore, one of these strains, A. melanogenum Beijerinck, oxidized glucosamine to glucosaminic acid with the theoretical consumption of oxygen as Ps. fluorescens liquefaciens. Glucosaminic acid was proved by isolation and identification by means of using resting cells.
The experiment of growth shows that Ps. fluorescens liq. could not secure any energy by means of the oxidation of glucosamine.

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Aiming at the enhancement of total lethal effect of radiations, chemical agent actions on irradiated cells were investigated. It was found that the sensitivity of irradiated cells to NaCl was markedly increased by radiation. In other words, the over-all lethal effect of radiations on microorganisms was enhanced by the pre- or post-treatment with NaCl. Some microbiological aspects of radiation enhancement of NaCl sensitivity were described in this paper, connecting with the possible mechanism. The fact that this phenomenon of radiation enhancement was observed in the case of the cells which gave the one-hit type survival curve suggests that radiation damage responsible for this phenomenon is not restricted to biological targets. Further discussion and experimental evidences will be presented in the subsequent paper.

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(1) Cell-free enzyme preparation which catalyzes an oxidative breakdown of gentisic acid with rupturing of benzene ring was obtained from m-hydroxybenzoic acid induced cells of Pseudomonas oxalis S-5. This new benzene ring splitting enzyme was named “Gentisic acid oxidase”
(2) Comparisons of various methods to obtain a cell free enzyme preparation were made. The enzyme was partially purified, and ferrous ion was proved necessary, for the activity of this enzyme. Further, some other properties of the gentisic acid oxidase were also investigated.
(3) From the various evidences obtained by manometrical experiments and chemical reaction of the product of this enzyme, the oxidative pathway of gentisic acid by Ps. ovalis was r presumed to be as follows.

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1. A highly purified gentisic acid oxidase preparation free from protocatechuic acid oxidase was obtained by the repeated ammonium sulfate precipitations. The specific activity increased up to twenty-fold. This enzyme preparation is free from oxidases of other aromatic compounds, as maleylpyruvate isomerase and fumarylpyruvate hydrolyase.
2. The activity of this enzyme preparation was remarkably inhibited by metal reagents such as α, α'-bipyridyl, ethylenediaminetetraacetic acid, ο-phenanthroline and nitroso-R-salt, and the inhibition was reversed by the addition of ferrous ions, and reactivation increased by the further addition of reducing agents, but not by the reducing agent such as ascorbic acid alone.
3. Enzyme and ferrous ion combination was proved to be necessary for the activity of the gentisic acid oxidase as the results of experiments using several combinations of iron-free enzyme, substrate and ferrous ions.
4. The loss of activity of iron-free enzyme could be recovered by the addition of ferrous ion. Other metalic ions and reducing agents tested were not effective.
5. Participation of SH-radical in the enzymesubstrate combination was presumed from the inhibition pattern by SH-reagents such as p-chloromercury-benzoate.
The authors wish to express their sincere thanks to Honorary Prof. K. Sakaguchi for his continued guidance throughout this research.

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1) One of the presumed intermediates of the gentisic acid oxidation was obtained in a crystalline form using dialyzed crude gentisic acid oxidase, free from fumarylpyruvic acid hydro-lyase, from the cells of Pseudomonas ovalis S-5.
2) From the results obtained, the crystalline intermdediate was identified as fumarylpyruvic acid, which was splitted by crude extracts into pyruvic and fumaric acids.
3) Maleylpyruvic acid, the initial product of gentisic acid oxidation, was recognized spectro-photometrically in the reaction mixture using purified enzyme. And the pathway of gentisic acid oxidative metabolism was established.
4) Colorimentric determination of fumarylpyruvic acid is also described.
The authors wish to express their sincere thanks to Honorary Prof. K. Sakaguchi for his continued guidance throughout this research.

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Many phosphates containing vinyl radical such as Phosdrin¹⁾ were prepared in order to study the correlation between structure and toxicity. From the results obtained it seems certain that in regard of toxicity, it is the carboalcoxy radical rather than the vinyl radical that is necessary and furthermore the toxicity is connected with the atom or the radical linked with the vinyl radical. As a results of these studies, some new toxic phosphates were synthesized.

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Several years ago, one of the authors, M. Yamada has found that the fermentation of straightchain amino acid like alanine or α-amino-n-butyric acid by yeasts gives rise to iso-butyl alcohol or active amyl alcohol in contrary to F. Ehrlich's scheme. From this respect we examined the fermentation of dl-threonine which one of the hydroxylic amino acids, and obtained an active amyl alcohol as in the case of α-amino-n-butyric acid and it was found that threonine seems to be reduced to the corresponding straight-chain amino acid.

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The activity of tyrosinase, isolated from potatoes is suppressed in the presence of redox dyes and the composed dehydrogenase systems. The oxygen uptake of potato tuber increases, but after being cut into slices the dehydrogenase activity of tuber decreases in the lapse of time. This increase in oxygen uptake is caused by tyrosinase. Accordingly, tyrosinase does not seem to play the role as a terminal oxidase in the electron transferring systems.

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The combustion-zone temperature of cigarettes made from each individual tobacco leaf of several varieties, was measured as follows: Turkish-846°C, Bright-844°C, Japanese Native “Nambu” -832°C, Japanese native “Matsukawa”, “Daruma” and Burley-812°C. Within the range of 1.0 and 1.2g for Turkish and Bright, and 0.7 and 0.9g for the others, it was found that throughout the varieties, thickness of the cigarette has no affect on the combustion-zone temperatures. The amount of nicotine contained in the smoke (Ymg) per one gram of the consumed cigarette is mainly affected by the contents of nicotine in the cut leaves (X%), and the relation between both values of each variety of tobacco leaves is shown as follows: Y=1.7X.

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1. Glucosyl transfer from p-nitrophenol-α-D-glucoside to several alkyl- and arylalcohols was observed with crystalline mold maltase. α-Ethylglucoside was isolated as a transglucosidation product, indicating the evidence of such action.
2. No reaction of α-glucoside synthesis from free glucose and alcohol could be observed with mold maltase.
3. By using glucose, fructose or mannose as glucosyl acceptors, the glucosyl residue of p-nitrophenol-α-D-glucoside was transferred to these acceptor sugars. Paperchromatography showed the formation of isomaltose from glucose, three unknown oligosaccharides from fructose and one unidentified oligosaccharide from mannose, respectively.

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Isomaltose, dextrantri-, tetra-, penta- and hexasaccharides, panose, and 4-α-[dextrantri-, tetra-, penta-, hexa- and heptaosyl]-D-glucose were isolated as transglucosidation products formed in the reaction of crystalline mold maltases with maltose.

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When the sample in the Fink's method ot staining yeast cells with methylene blue was taken from the yeast suspension after determining the fermenting activity, the measure-ment was carried out more easily and exactly. Next the existence of the positive correlation (r=0.839) was confirmed between the ratio of yeast cells negative to methylene blue staining (MBN) and Qco₂ (20';_??_40';). So the change of dry yeast during storage was examined con-cerning MBN and three quotients of ferment-ing power (Qco₂ (20';_??_40';), ΔQco₂ Qco₂ (max)). Besides, the interrelation of these quotients was discussed in connection with the influence of the storage temperature.
1) The relation between moisture content and MBN retention of dry yeast in the storage at various temperatures (0°_??_5°, 20°, 30°, 37°C) was similar to that reported formerly about Qco₂ (20';_??_40';). The lesser the moisture content of dry yeast, the smaller was the value of MBN but the value retained during the period of storage was larger. Besides, the lesser the moisture content, the lesser was the influence of the storage temperature. But such a tendency in the storage was seen more clearly in Qco₂ (20';_??_40';) than in MBN.
2) In the storage at 30°C, the relation between the retention ratios of MBN and above three quotients was examined. In the initial period of storage, Qco₂ (20';_??_40';) decreased and the induction period appeared. Afterwards, the decrease of ΔQco₂ (the increase of the induc-tion period) went on more rapidly than that of Qco₂ (20';_??_40';). Of the three quotients Qco₂ (max) decreased most slowly. MBN decreased more slowly than Qco₂ (max), and had more connection with Qco₂ (max) than with Qco₂ (20';_??_40';) or ΔQco₂, in the three quotients.
3) The decrease of MBN was slowest also in the storage at 0°C. Qco₂ (20';40';) also was almost constant in the case of dry yeast of a moderate moisture content, but only AQco₂ was clearly inconstant.
4) In the samples stored at 30°C after the storage at 0°C, the decrease of MBN and Qco₂ (20';_??_40';) was more remarkable than in those stored at 30°C soon after drying. On the contrary, both MBN and Qco₂ (20';_??_40';) decreased only a little when the same samples were stored at 0°C after the storage at 30°C.

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A number of organisms were screened to test activity of accumulating aspartate from fumarate and ammonium salts and, Pseudomonas fluorescens 6009-2 and mutant Ki-1023 derived from Escherichia coli K-12 by ultraviolet radiation were chosen as the suitable strains.
By employing these strains various cultural conditions suitable for the production of L-aspartic acid were investigated and, the results obtained are as follows:
a) Ammonium fumarate is the most favorable as ammonium salts.
b) On the molar ratio of fumarate and ammonium ion in the medium, the ratio 1:1.5_??_2.0 gives the best result.
c) On the effect of pH, culture without pH adjustment starting from initial pH 7.0, is preferable for Pseudomonas fluorescens than in case of pH adjustment during incubation, while on Escherichia coli the addition of calcium carbonate results in better accumulation of aspartate.
d) The addition of glucose causes formation of amino acids other than aspartic acid and lowers the yield of aspartate.
e) On the concentration of fumarate in the medium, 5.% gives the best result.
f) Of the various organic nitrogen compounds in the medium, peptone is most favorable.
g) Of the wild strain and mutants of Escherichia coli K-12, Ki-1023 is ascertained to be the most suitable strain.
h) Incubation at 37°C accelerates formation of aspartate than incubation at 30°C, though there is no difference of the attained conversion rate.
i) On the influence of shaking, partially stationary culture, that is, after shaking at initial day stationary culture is done, enables better result than stationary or shaking culture throughout the period of incubation.
Under the optimum conditions revealed by the above experiments, practical formation of aspartate was carried out, and formation over 95% conversion rate from consumed fumarate was attained by both strains. The formed aspartate was readily isolated from the culture broth and identified as L-aspartic acid. This method is considered to be an advantageous procedure for the production of L-aspartic acid.

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The pentose isomerase of heterofermenters was found to have specific action on the substrate. Only L-arabinose was isomerized by L-arabinose isomerase and no inhibitory effect was demonstrated by the addition of D-isomer, and xylose isomerase acted on D-xylose, and L-xylose did not inhibit such activity. No ketose from glucose, mannose, galactose, L-xylose, D-arabinose, D-ribose and mannitol was produced by these heterofermenters. The equilibrium constants for D-xylose- and L-arabinose isomerases were 20_??_28% and 13_??_16% respectively, and this equilibrium was shifted to ketopentose in the presence of borate. Michaelis constants were found to be 2×10^-2M for D-xylose isomerase and 5_??_10×10^-2M for L-arabinose isomerase.

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Six strains of homofermenters produced L-arabinose isomerase in only the L-arabinose-medium, and with these bacterial cells which were obtained from the medium containing glucose, fructose or mannose, any activity of this enzyme was not pointed out. Moreover the L-arabinose isomerase of six strains of heterofermenters and of Pediococcus lindneri, and also D-xylose isomerase of six strains of heterofermenters and of L. xylosus are produced even in glucose, fructose, mannose, galactose or pentose-media. The amounts of D-xylose isomerase of L. fermentum varied according to the kind of carbon source of the growing culture. This xylose isomerase is produced abundantly in the cells grown on mannose, L-arabinose and D-xylose-media. Glucose, galactose, maltose, sucrose, and gluconate do not show any stimulating effect on the production of xylose isomerase.

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Preparations of glucan obtained from baker's yeast and sclerotia of Sclerotinia Libertiana were found to be completely hydrolysed by enzymes of the Sclerotinia fungus. Some differences in the molecular structure of the glucans were found upon examination of the modes of degradation by the successive action of Rhizopus- and Sclerotinia enzyme preparations of which the former had only a partial hydrolytic effect.
The dissolution of glucan in intact cells of yeast, that could be estimated from the rate of autolysis of the cells, was proved to be insignificant on the action of glucanase alone in the Sclerotinia enzyme solution. The combined action of glucanase with lipolytic enzyme in the fungus enzyme solution are shown to promote the solubilization of intact yeasts and sclerotium cells.

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