The phenolsulphatase of Aerobacter aerogenes was found to be formed in cell suspensions containing tyramine and casein hydrolyzate under aerobic conditions in a few hours.
Factors affecting phenolsulphatase synthesis in cell suspensions were studied.
The relationship between the rate of enzyme activity formed and the level of tyramine concentration in 10^-4M to 10^-3M is linear. The optimum concentration of tyramine for its utilization and phenolsulphatase induction is in both cases 5×10^-3M. The opt. concentration of casamino acids for phenolsulphatase induction is 0.2 per cent and the opt. pH is 7.2. Agents such as 2, 4-dinitrophenol, sodium azide, arsenate and potassium cyanide inhibit the induction of this enzyme. The external inducer of phenolsulphatase should be highly specific. On addition of tyramine, hydroxytyramine and noradrenalin are effective, although the effect of noradrenalin is very weak.

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Ilexol, a new triterpenoid alcohol isolated from the barks of several species of the Genus Ilex, has now been oxidized with chromium trioxide to yield a ketone, designated as ilexone of C₃₀H₄₈O, m. p. 239.5-240.5°, and [α]¹⁴D-68.74°, which gives oxime of m. p. 256.5-257.5°, 2, 4-dinitrophenyihydrazone of m.p. 260-261°, and positive Zimmermann test.
On reduction with sodium and ethanol as well ai with lithium aluminum hydride, it has been shown that ilexone is regenerated to yield an alcohol proved to be identical with natural ilexol.
On reduction according to the procedure of Huang-Minlon, ilexone has been shown to yield an oxygen-free compound, designated as ilexene of C₃₀H₅₀, and m. p. 241-242°.
It seems most likely that ilexol might be a triterpenoid alcohol of C₃₀H₄₉OH, an alcoholic hydroxyl group of which is secondary in nature, and moreover attached at C-3 in its molecule.

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We have been studying on the genesis of lignin and the changes of some chemical constituents contained in the gourd fruits which were taken in turn in certain periods after flowering.
The results obtained are as follows:
(1) Lignin was at first generated on the fifteenth-day after flowering, and since then increases rapidly and finally occupies the whole fruit. The distributions of peroxidase and phenoloxidase in the tissue of fruit agreed with the lignification, and, it is suggested that these enzymes have relations to the biosynthesis of lignin.
(2) Lignin is mainly synthesized at the tissue of the shell-part of fruit. The content of lignin was only 2.3% on the fifteenth-day but it reached to 32% in hardened fruit. The methoxyl content of lignin was only 2% on the fifteenth-day but it increased along with the increase of lignin reaching to a level of 12% on the fifteenth-day, and to 15.3% after drying. In addition to these changes of lignin, other components except lignin also showed characteristic changes.
(3) In order to investigate the increase of the methoxyl content of lignin further, lignin was oxidized with nitrobenzene in an alkaline medium. Among p-hydroxybenzaldehyde, vanillin and syringaldehyde in that oxidative mixture, the former was the richest component in the younger fruit, but the latter two, which were richer in their methoxyl content increased with the growth of fruit, and after ripening vanillin was the richest and p-hydroxybenzaldehyde the poorest component.

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In concern of the amylase system of blackkoji mold, the following facts are described:
(1) The dextrinogenic amylase fraction has an extremely weak action on raw starch, independent of the presence or absence of maltase activity, as compared with the saccharogenic amylase fraction.
(2) The saccharogenic and dextrinogenic amylase fractions, when used combined, interact each other, to promote digestion of raw starch up to about 3-times the sum of activity of each fraction.
(3) This accelerating-interaction effect solely depends upon the “debranching activity” of saccharogenic amylase fraction.
(4) Corn, wheat and glutinous rice starches in a raw state are able to be digested to completeness with this combined system.
(5) Glutinous rice starch shows peculiar easiness in digestion either by saccharogenic amylase fraction alone or by the interacting system combined with the dextrinogenic amylase fraction.

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The enzymatic resolution of acetyl-DL-methonine previously reported was studied further, in detail. As a result, it was found that metal ion plays and important role on the asymmetric hydrolysis of acyl-DL-methonines by the enzyme preparations of Aspergillus oryzae.

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The advantageous resolution procedure of DL-lysine previously reported, consisting of the enzymatic hydrolysis of ε-benzoyl-α-acetyl-DL-lysine with mold enzyme preparation was innovated. Susceptibility of ε-benzoyl-α-acetyl-DL-lysine towards various enzyme preparations such as kidney, pancreas extracts, renal acylase and carboxy-peptidase, was investigated. As a result, it was found that although the substrate was readily hydrolyzed by the mold enzyme preparations, the other enzyme preparations showed, almost no, if any, hydrolytic activity.

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The distribution of activity hydrolyzing acyl amino acids was investigated, so as to obtain an advantageous enzyme source for the resolution of amino acids. For this purpose, a number of molds such as Aspergillus, Penicillium, Rhizopus, Mucor, Abshidia, and Cunninghamella were cultivated on wheat bran and the extracts were tested in relation to their hydrolytic activity of acyl amino acids. Although almost all molds yielded acylase more or less, high activity was shown in cases of Aspergillus and Penicillium. As one. of the most suitable strains, Aspergillus oryzae No. 9 was chosen and the optimum cultural conditions for acylase formation was also inveitigated.

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With the mold acylase preparation, optimal pH and relative activity to the various acyl amino acids were investigated. As a result, a difference of optimal pH was observed between acetyl and chioroacetyl derivatives. A comparison of the susceptibility disclosed that mold preparation has either a wider or loose specificity than renal acylase. Namely, besides aliphatic amino acids, the acyl derivatives of basic amino acids and aromatic amino acids were readily hydrolyzed by the mold preparation. These results indicate that the mold acylase preparation can be used for the resolution of a great variety of amino acids. As an example of this, the resolution of acetyl-DL-tryptophan was presented.

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The “Paste-medium” technique, described here, has made the separate treatment of the mycelial-part and the substrate-part of koji materials, possible in the study of koji. The ground koji materials were mixed with water to be brought into the state of paste. The paste-medium, whose surface was flattened in a petri-dish and covered with a piece of nyloncloth, was sterilized in the usual way, in an autoclave. After inoculation and growth of the koji mold on the surface, the mycelial-part together with a piece of nylon, was stripped off, and the corresponding substrate-part was digged out of the paste-medium.
This new method enables us to investigate the effects of some cultural conditions on the growth of Asp. soyae KS.

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By employing the paste-medium technique, the effects of some conditions of medium on the protease production by Asp. soyae KS were investigated. The medium of initial pH of 7-8, initial moisture content of 50 per cent, high C/N ratio, and of rice bran, copra meal had each favorable influence on protease production. Protease which was produced in the paste culture was observed in the mycelium as well as in the medium, while in the case of liquid culture it was mostly secreted into the medium. The secreted protease in the pastemedium was found to difuse to the depth of 2.5mm under the surface of the mycelium.

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In the preparation of koji from various raw materials, it was recognized that the lower the cultural temperature, the more the three kinds of protease are produced by Asp. soyae KS. In the paste-culture, the amounts of production of mold protease both in the mycelium and in the medium were also more at lower cultural temperatures than at higher ones. The low temperature should be applied at a period which coincides with that of spore-production because at such period the mold protease production is observed to be maximum. The limit of the practical low temperature was about 12°C, which was also the limit to growth of the mold tested.

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