Inositol deficiency of Sehizosaccharomyces pombe did not induce significant change of contents of various cellular components except for phospholipids and inositol. The most remarkable decrease in inositol content by the deficiency occurred in the mitochondrial fraction. Electronmicroscopic observation of the inositol-deficient cells of Sch. pombe showed no remarkable thickening of cell wall as occurred in the inositol-exacting mutant of Saccharomyces cerevisfae Strain A-21-20.
Marked loss of fermentative activity under the aerobic condition was caused from inositol deficiency without significant change of activities of respiration and anaerobic fermentation. This seemed to indicate elevated regulatory control of the fermentative activity by oxygen in the inositol-deficient Sch. pombe.
Phosphorylative activities of intact cells and the isolated mitochondria coupled with oxidation was also remarkably suppressed by inositol deficiency.

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Effect of feeding millet (Sorghum vulgarie) at 5, 10 and 15 per cent protein levels respectively for a period of six weeks to rats on their liver DNA, RNA and proteins of liver, its subcellular fractions and plasma has been studied, and results compared with rats fed casein at 10 per cent level. Both liver DNA and RNA of rats fed millet at 5 per cent protein level were significantly increased. Liver proteins (mg/100g body weight) of rats fed millet at 5 and 10 per cent protein level were significantly increased and plasma proteins decreased. Incorporation of leucine-1-¹⁴C into both liver and plasma proteins of rats fed millet was significantly higher than the control.

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A peptidoglutaminase activity in microorganisms was detected using carbobenzoxy-L-glutamine or tertiary-amyloxycarbonyl-L-glutaminyl-L-proline as substrate. By screening, an organism which produces a relatively large amount of peptidoglutaminase was isolated from soil. The organism was identified as Bacillus circulans. The highest enzyme formation by the bacterium occurred during stationary growth phase in the basal medium containing lactose (0.5%) and polypepton (1%).

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The synthesis of L-tyrosine or 3, 4-dihydroxyphenyl-L-alanine (L-dopa) from pyruvate, ammonia and phenol or pyrocatechol was studied with intact cells of Erwinia herbicola ATCC 21434 containing high tyrosine phenol lyase activity. By elemental analyses and determination of optical activity, the tyrosine or dopa synthesized was confirmed to be entirely of L-form. Maximum amount of L-tyrosine (60.5g/liter) or L-dopa (58.5g/liter) was formed using this enzymatic method by feeding sodium pyruvate, and phenol or pyrocatechol. However, large amounts of by-products were formed in the L-dopa synthetic reaction mixture. By-products were proved to be formed from L-dopa and pyruvate by a nonenzymic reaction. pH and the temperature of reaction had intensive effects on the formation of by-products. A simple method using a boric acid-pyrocatechol complex was devised, as the feeding procedure of substrates was complicated.

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Studies on absorption, translocation and metabolism of 3-allyloxy-1, 2-benzisothiazole 1, 1-dioxide (Oryzemate), a new rice blast controlling agent, in rice plants were undertaken using the ³⁵S-labeled preparation. Oryzemate was administrated to the plants by liquid application. Uptake of labeled compound into rice plants was demonstrated by autoradiography and quantitative ³⁵S-analyses. Preferential accumulation of radioactive compounds in the leaves was observed. The metabolites in the plants were identified as allyl o-sulfamoylbenzoate, saccharin and N-o-glucopyranosylsaccharin by cochromatography and cocrystallization with synthesized authentic compounds. Most of ³⁵S-labeled compounds accumulated in the plants were saccharin and N-D-glucopyranosylsaccharin. A small amount of allyl o-sulfamoylbenzoate was detected and Oryzemate was detectable in trace quantities at all harvest.

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Pseudomonas O-3 strain which was isolated from soil can grow on polyvinyl alcohol (PVA) as a sole carbon source. When 0.5 per cent of PVA (500, 1500 or 2000) was employed as the carbon source in the culture medium, PVA was almost completely lost from the culture fluid after a week and the concentration of total organic carbon measured by a TOC analyzer decreased from the initial value of about 2700ppm to 250_??_300ppm after 7_??_10 days culture. This bacterium was found to produce and secrete an inducible enzyme which degrade PVA. The way by which this enzyme degrades PVA was examined and the results were obtained which suggested that PVA was broken down oxidatively in a way of endowise splitting. However, the mechanism of PVA degradation has not been clarified yet. The optimum pH and temperature for enzyme activity were examined and they were 7.5_??_8.5 and 35_??_45°C, respectively. Morphological and biological characteristics of this bacterium were examined and it was similar to a strain of Pseudoinonas boreopolis.

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An aminopeptidase from Aspergillus orvzae 460 was purified from the rivanol precipitable fraction. The partially purified enzyme was not homogeneous in disc electrophoresis, although symmetric profiles were obtained for enzyme protein and activity in Sephadex gel filtration. Its optimum pH is at pH 8.5 for L-leucyl-β-naphthylamide. The enzyme activity was inhibited by metal chelating agents and S-S dissociating agents, but not inhibited by SH reagents. The molecular weight of the enzyme was estimated to be about 26, 500 by gel filtration. The enzyme was named leucine aminopeptidase I of Asp. oryzae 460, since it preferentially hydrolyzed oligopeptides that possess leucine as the amino terminal amino acid.

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Aminopeptidase II from Aspergillus oryzae 460 was purified from the rivanol precipitable fraction. The purified enzyme was homogeneous in polyacrylamide gel disc electrophoresis. Its optimum pH is at pH 8.0 for L-leucyl-glycyl-glycine and L-leucyl-β-naphthylamide. The enzyme was inhibited by metal chelating agents and S-S dissociating agents, but not inhibited by SH reagents. The molecular weight of the enzyme was estimated to be about 61, 000 by gel filtration. The enzyme was most active toward oligopeptides of amino terminal leucine, indicating that it was a kind of leucine aminopeptidase. Thus, the enzyme was proposed to be called leucine aminopeptidase II of Asp. oryzae 460.

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Leucine aminopeptidase III from Aspergillus oryzae 460 was purified from the rivanol non-precipitable fraction. The purified enzyme was homogeneous in polyacrylamide gel disc electrophoresis. Its optimum pH is at pH 8.0 for L-leucyl-glycyl-glycine. The enzyme was inhibited by SH reagents. The molecular weight of the enzyme was estimated to be about 56, 000 by gel filtration.

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A κ-casein-like fraction was prepared from human whole casein by gel filtration with Sephadex G-150 and Biogel A-150m. The fraction was calcium-insensitive and its solution became turbid by rennin. In polyacrylamide gel electrophoretic (PAE) analysis, the fraction gave 11_??_12 bands after reduction with 2-mercaptoethanol. It appeared to exist as a disulfide-bound complex of many components. The occurrence of human para-κ-caseins from the fraction after rennin treatment was confirmed by PAE. When the reduced, alkylated human κ-casein-like fraction was chromatographed by DEAE-cellulose, several fractions were obtained. After rennin treatment, they formed either a para-SCM-κ-casein band moving toward the cathode on PAE pattern or the one which moved toward the anode. These results suggest that two para-κ-caseins were formed from human whole casein or human κ-casein-like fraction by the action of rennin.

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Osmophilic Aspergillus responsible for spoilage of rice, corn, milo and wheat have been isolated and identified. Fifteen strains were classified as members of the Aspergillus glaucus group, and were subdivided into A. ruber, A. repens, A. mangini, A. chevalieri and A. tnontevidensis. Nine strains were classified as members of the A. restrictus group, and were subdivided into A. gracilis, A. vitricolae and A. casiellus. The other 7 strains were classified as A. versicolor in the A. versicolor group, A. sulphureus in the A. oehraceus group, and A. niveus in the A, flavipes group.
All of dose-survival curves obtained with the conidia of 10 strains showed the sigmoidal type having the D₁₀ values between 18 and 30 krad. The survival curves obtained with the ascospores of A. glaucus group also showed the sigmoidal type having the D₁₀ values of 54 krad. Radio-sensitivity of the dry conidia was similar to that of the dry ascospores, having D₁₀ values between 50 to 58 krad.

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The maximum yield of lytic enzyme was obtained from shake flask cultures of Streptomyces globisporus 1829 which were grown at 30°C for 48 hr in a medium containing 2% dextrin, 0.5% soybean meal, 0.2% polypeptone, 0.5% Na₂HPO₄•12H₂O, 0.1% KH₂PO₄, 0.1% to MgSO₄•7H₂O, 1.0% NaCl and 0, 02% CaCl₂, pH 7.5. The activity of successively transferred substrains of St. globisporus 1829 gradually decreased. However, a mutant strain obtained by ultra-violet irradiation has been shown not to have lost any lytic activity for 2 years. The enzyme exhibited maximum activity at 60°C in the pH range of 6 to 6.5 and was lytic against the intact cells of Streptococci, Lactobacilli and Bacilli but inert against the intact cells of Staphylococcus aureus and Micrococcus Iysodeikticus.

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A new method for the preparation of whole egg protein was presented and its qualification as a standard reference for evaluating dietary protein was made in the course of this experiment: e. g., Purity, 96.5% and Biological value, 99.9. The amino acid composition was almost identical with that shown in the table of FAO (1970).
Using this whole egg protein, casein and yeast, the nutritive values of dietary proteins in the adult rats at the normal level of body protein and in those after 8 day's protein depletion were compared. It was found that the evaluation after protein depletion had a tendency to overestimate the nutritive value of proteins rather making the difference little between each other. The significance of the relative evaluation by use of the purified whole egg protein was discussed with special regard to the actual retention of body nitrogen, i.e., the net nitrogen retention.

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A viridicatin derivative having anti-auxin action, i.e. 3-(4-phenylcarbostyriloxy) acetic acid (V-OCH₂COOH) was found to increase the formation of both IAA (indole-3-acetic acid)oxidase and -synthetase in rice and pea seedlings. With the IAA synthetase, the activity on indolepyruvic acid was markedly increased. V-OCH₂COOH stimulated the induction of IAA oxidase in the excised segments from pea epicotyl, but did not IAA synthetase. The effect of V-OCH₂COOH on the former was inhibited by cycloheximide. Activity of the IAA oxidase extracted from pea epicotyl and dialyzed was also stimulated by V-OCH₂COOH in the presence of a cofactor such as 2, 4-dichlorophenol. Effect of IAA per se on enzyme regulation was tested in parallel and discussed.

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Peptidoglutaminase-I and II that catalyzed the hydrolysis of the γ-amide of peptidebound glutamine, were purified from the cell-free extracts of Bacillus circulans by streptomycin sulfate precipitation, ammonium sulfate fractionation, DEAE-Sephadex, Sephadex G-200, QAE-Sephadex, hydroxylapatite-cellulose column chromatography, and finally preparative polyacrylamide gel disc electrophoresis. The purification steps resultd in a 714-fold increase in specific activity for peptidoglutaminase-I and in a 223-fold for peptidoglutaminase-II over the original extracts. The both enzymes were homogeneous in disc electrophoresis in polyacrylamide gel, immunoelectrophoresis in agar gel, and sedimentation analysis. Using gel filtration, the molecular weights of peptidoglutaminases I and II were estimated to be 90, 000 and 125, 000. However, during the purification steps, the both enzymes were observed to cause the dissociation and aggregation reaction which did not so much affect on their enzyme activities.

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It was recognized that Geotrichurn candidum Link which was selected as the efficient lipase producer formed lipase only in the presence of substrate or its relating compounds such as oils or fatty acids in a cultivation medium. From the experimental results obtained by the cultivation of the microorganism and also by using of washed cells, it seemed that lipase was formed inducibly. It is likely that the produced lipase is localized around the cell wall and membrane and it is released from the cells after a certain period from the inducible synthesis.

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To study the mechanism of fatty infiltration in the liver due to added sulfur-containing amino acids to low casein diet, the effect of sulfur-containing amino acids and glutathione (GSH) on the incorporation of acetate-1-¹⁴C into lipid fractions were studied in liver slices from rats fed on 8% casein diet (Basal diet) with or without added methionine (Met).
The liver acetyl Co A carboxylase activities of rats on basal diet with or without added Met were similar.
Addition of Met, cystine or cysteine to the incubation medium had little effect on lipogenesis of slices. On addition of GSH to liver slices from rats fed on basal diet, lipid formation increased appreciably. On the other hand, addition of GSH to liver slices from rats fed on Met supplemented diet showed no accelerative effect on lipogenesis.
Addition of GSH to the incubation medium of liver slices from rats fed on basal diet tended to reduce the incorporation of acetate into the phospholipid fraction and to increase into the fatty acid fraction of liver slices.
The content of liver GSH was lower in rats on basal diet than in those on Met supplemented diet. The higher GSH level in rats on Met supplemented diet may be one factor causing fatty infiltration in the liver of these animals.

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For the purpose of the fermentative production of flavin-adenine dinucleotide (FAD), the authors attempted to select the microorganisms which could produce FAD in culture medium from flavin mononucleotide and adenylic acid for adenine, and discovered that bacteria and actinomycetales could accumulate FAD more or less, but yeasts and moulds could not. Among the microorganisms used in the experiments, the strains belonging to the genera Sarcina and Brevibacterium accumulated relatively large amounts of FAD. Especially, Sarcina lutea and Sarcina aurantiaca seemed to be the most favorable microorganisms for the fermentative production of FAD. By using Sarcina lutea the investigation of the culture conditions for the production of FAD was carried out.

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Greater production of pantothenic acid 4'-phosphate and pantetheine 4'-phosphate by a microorganism were described. The incubation of pantothenic acid and adenosine 5'-triphosphate with resting cells of Brevibacterium mnmoniagenes IFO 12071 gave pantothenic acid 4'-phosphate in a high yield. Cultivation of the organism with pantothenic acid and 5'-adenylic acid also gave pantothenic acid 4'-phosphate in a high yield. In a similar fashion pantetheine 4'-phosphate was readily obtained in a good yield. The products were identified chemically and enzymatically.

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Cellular fatty acid compositions of Candida tropicalis pK 233 and Candida lipolytica NRRL Y-6795 and the time-course changes during yeast growth were studied using individual n-alkanes of various chain lengths (from C₁₁ to C₁₈) and a mixture of n-alkanes (C₁₀-C₁₃) as a sole carbon source. Observed relationships of the chain-length of n-alkane substrate to time-course changes and final patterns of the fatty acid compositions of these yeasts, especially those of the cells grown on odd-carbon alkanes, indicated that “intact incorporation mechanism, ” that is, accumulation of the fatty acid having the same chain-length as that of the alkane substrate used was predominant in the yeasts cultivated on a longer alkane such as n-heptadecane and n-octadecane. On the other hand, “chain elongation pathway” and “de novo synthesis pathway” following β-oxidation of substrate were simultaneously operative in the cells growing on a relatively shorter alkane such as undecane and dodecane.

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Ion exchange resins have been used to separate soy sauce into three fractions of distinctly different composition: a cation fraction, a neutral fraction and an anion fraction. Almost all of the constituents responsible for browning were recovered in these three fractions.
Storage experiments show that when the three fractions were stored separately, only the cation fraction darkened considerably. When they were combined and stored, the color of the mixture increased at nearly the same rate as that of the original soy sauce. Neutral sugars are important constituents of the neutral fraction with respect to browning. The browning rate of a sugar-amino acid mixture (simulated soy sauce), was about 10% of soy sauce. The effect of the anion fraction (mainly caused by organic acids) and the ashed cation fraction on the over-all browning of soy sauce is calculated to be 10_??_12% and 20%, respectively.
The sum of the contribution rate of the anion fraction, the neutral fraction, the amino acids and the ashed cation fraction in the browning of soy sauce was concluded to be approximately 40%. Compounds responcible for residual part of 60% should be considered to exist in the cation fraction. It was suggested that such compounds have strong reducing power and O₂-uptaking ability.

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To establish a novel process for the economical production of citric acid from n-paraffins by yeast, attempts were made to obtain some mutant strains capable of producing citric acid in higher yield without (+)-isocitric acid.
From among the mutant strains derived from Candida lipolvtica ATCC 20114, which produced citric acid and (+)-isocitric acid in the ratio of about 60:40 from n-paraffins, a citrate non-utilizing mutant strain, K-20, and a fluoroacetate-sensitive mutant strain, S-22, were selected on the basis of high citric acid and low (+)-isocitric acid productivity.
The mutant strain S-22 showed extremely poor growth in a medium containing sodium citrate as the sole carbon source and extremely high sensitivity to fluoroacetate. The production ratio of citric acid and (+)-isocitric acid by the mutant strain was changed to 97:3, and the yield of the citric acid from n-paraffins, charged to the fermentation medium, reached 145% (w/w).

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Fluoroacetate-sensitive mutant strains, K-20 and S-22, of Candida lipolytica could not grow or could only slightly grow on agar media containing di- or tricarboxylic acid involved in the TCA-cycle as the sole source of carbon. Relative activities of aconitate hydratase in the cells of the mutant strains, K-20 and S-22, were approximately 1/10 and 1/100, against that of the parent strain, respectively. This facts support the statement that the mutant strains were extremely sensitive to monofluoroacetate.
The aconitate hydratase activities of these mutant strains and the parent strain corresponded well to the citric to (+)-isocitric acid ratio in the final fermented broths.

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From nutmeg, a seed of Myristica fragrans Houtt., were isolated thirteen phenylpropanoids. Among them, seven compounds have been hitherto unknown. Most of the compounds isolated here revealed growth inhibitory activity to silkworm larvae, Bombyx mori L.

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The reaction of rotenone, which has the 5' β-isopropenyl grouping, with boron tribromide in dichloromethane gives the 1', 5'-seco-5'-bromo compound having the opened E-ring. When treated with sodium bicarbonate in aqueous acetone, the compound closes the E-ring to form two products having the 5'-isopropenyl grouping in the α- and β-configurations. By this cycle, rotenone (5'β-rotenone) gives 5' α-epirotenone as well as rotenone, while d-epirotenone (5' β-epirotenone) gives 5 'α-rotenone (the antipode of natural rotenone) as well as d-epirotenone.

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The interconversion between variotin and ketovariotin is described.

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The structure of a minor component in the oxidation products of crude natural variotin oxidized with manganese dioxide is elucidated. The synthesis of the investigated compound, N-(6-oxo-2, 4-trans, trans-heptadienoyl) 2-pyrrolidonide, and the reduction of the substance are also described.

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The total synthesis of dl-variotin is described.

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Antimycinone A₃, which is a neutral fragment of mild alkaline hydrolysate of antimycin A₃, and its stereoisomers were synthesized stereoselectively from methyl trans-2-n-butylpent-3-enoate or methyl cis-2-n-butylpent-3-enoate, and natural antimycinone A₃ was proved to possess Hα-Hβ and Hβ-Hγ trans configuration.

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