A detailed procedure for the separation of diastatic enzymes of molds was establi-shed, which was based on their electrophoretic behaviours on a paper strip. For the detection of enzymes, a bioautographical method was newly devised. Comparing the enzymatic images appeared on the paper strip, the authors classified mold enzyme systems into five types.

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The components of mold diastatic enzymes which were successfully separated from one another on a filter paper by the paper electrophoretic technique were extracted with water, and the resulting enzyme solutions were tested with starch and maltose as substrates. By determining the reaction products, the author attempted a specula-tion on the actual nature of each component enzyme.

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Oxidatie metabolism of D-glutamate by microorganism was investigated. It was found that several isolated strains of bacteria belonging to the genus Aerobacter could metabolize D-glutamate more rapidly than the L-form even when they were grown in the DL-glutamate medium. Pure L-glutamic acid could be successfully obtained from the DL-form by decomposing D-glutamic acid using one of these strains. From several experimental results, the occurrence of a specific oxidase responsible for the oxidation of D-glutamic acid was suggested.

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In order to examine the possibility of mass-cultivation of Chlorella for the pur-pose of fat production, the cultural con-ditions for fat accumulation were studied, Chlorella ellipsoidea being used. It was seen that Chlorella grew in the nitrogen- free medium, accumulating lipide in its cells, and that the accumulation degree was greatly affected by the initial algal concentration, the depth of the suspension and the incident light intensity. In order to obtain cells of a high lipide content, it was required that the initial density is low and the layer of the suspension thin. As for the light intensity, it was found that a stronger light has a stronger retarding effect on the nitrogen-free growth in its later stage than a weaker -light over 5-10 kilolux. This is explained to be due to more severe chlorosis caused by the stronger light. The presumptive yield of fat by mass-culture was roughly estimated as 4.9 ton per acre per year under favourable conditions.

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Changes of carbohydrate fractions and chlorophyll and carotenoid content in the cells during the nitrogen-free cultivation were studied. It was found that free reducing sugar increased and sucrose decreased remarkably with the progress of nitrogen deficiency in the cells. Solu-ble and insoluble polysaccharides were also found to increase in the earlier stage. It is considered that fat is not synthesized at the expense of carbohydrate in the early stage of fat accumulation, up to 50%, but that, in the later stage, not only the synthesis of carbohydrate is reduced, but the accumulated carbohydrate, the greater part of which is free reducing sugar, is converted to fat.
The chlorophyll content diminishes faster than the nitrogen content, especially at high light intensity. Carotenoid decreases as fast as or faster than chloro-phyll, and carotene disappears in the very early stage of N-deficiency.

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Magnesium as a controlling factor in the multiplication and the formation of cell material or chlorophyll has been in-vestigated at about 15 kilolux, being contrasted with the full nutrient or the nitrogen-free condition. In the full nutrient culture, the relative relation of these processes proceeded in a rate almost constant and the cells of about 3.5% chlorophyll content were produced in the same size at the same linear growth rate. The cell growth was continued even without taking magnesium, giving the cell size more than 3.5 times larger in volume than the normal cell, and the cell digestion occured in the later stage, of culture. From the sequence in which the activity of the respective processes de-creased, the level of magnesium require-ment was suggested. It is probable that the formation of chlorophyll takes place at the highest level of magnesium, in the cell, and the synthesis of other cell materials occurs at lower level than the cell division.

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Islanditoxi^{1, 2)} has been isolated from the cultured broth of Penicillium islandicum, and C₂₅H₃₃O₈N₅CL₂ was proposed as its molecular formula. The Presence of the peptide linkage in islanditoxin was ascertained and the general properties were des-cribed.

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Complete hydrolysis of islanditoxin, C₂₅H₃₃O₈N₅CI₂, with 12 N hydrochloric acid gave three ninhydrin-positive substances, termed fractions A, B and C, and small amounts of a humin-like material. Fractions A and B were identified as serine and α-aminobutyric acid, respectively. To fraction C the molecular formula C₉H₁₁O₂N was assigned and this seemed to be an amino acid containing a benzene ring. Mild hydrolysis of islanditoxin with 4.5 N hydrochloric acid has shown that 1 mole of is-landitoxin produces 2 moles of serine.

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It has been confirmed that gibberellin A is a mixture of three components, gibberellin A₁, gibberellin A₂ and gibberellic acid (namely gibberellin X), by treating their methyl ester through the chromatography on Al₂O₃ column. Attempts to separate them in free acid were made. The physical and chemical properties of each gibberellin as well as its physiological properties are described.

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Some different results from previous reports on acid degradation and catalytic reduction studies of the three components were obtained. From gibberellin A_l only gibberellin C and no gibberic acid were obtained. From gibberellic acid (namely gibberellin X and A₃) only gibberic acid and no gibberellin C were obtained. In addition to this, some experimental results of gibberellin A on animal tissues were also, presented.

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(1) In the present paper, further con-firmation was obtained by repeated in-tensive experiments with Co-porphyrin-derivatives freshly prepared from pure chlorophyll a and b by the column chromatographic method of Gattermann and Wieland. The effect on Euglena
(chlorophyll free) was always revealed to be positive.
(2) Moreover, vitamin B₁₂.-mass pro-ducing experiments with Co-porphyrin-derivative and 5, 6-dimethylbenzimidazole was attempted by Euglena gracilis strains in B₁₂-Free basal medium, and vitamin B₁₂ was isolated from both cells and broth by paper electrophoresis. The increase of a fair quantity of total vitamin B₁₂ was always seen in the cells and the broth of control strains.
(3) Therefore, the authors are inclined to propose the close implication of cobalt-porphyrin-derivative in the biosynthesis of vitamin B₁₂ by Euglena.

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