In the course of investigation of the metabolism of p-xylene, we detected p-cresol, 4-methylcatechol and p-hydroxybenzoic acid from culture broth.
Pseudomonas aeruginosa S668B2 was found to utilize benzoic acid, p-toluic acid, m-toluic
acid and 3, 4-dimethyl benzoic acid as a carbon source.
The strain S668B2 was shown to produce p-cresol from p-toluic acid, 3-methyl salicylic acid from m-toluic acid and 3, 4-dimethyl phenol from 3, 4-dimethyl benzoic acid.
The strain S668B2 also produced p-hydroxy benzoic acid from p-cresol.
Pathways of the metabolism of m-xylene and pseudocumene were speculated.

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The oxidation of aromatic compounds by Pseudomonas aeruginosa S668B2 was an induced
phenomenon.
p-Xylene grown cells oxidized p-methylbenzyl alcohol, p-toluic acid, p-cresol, p-hydroxy-benzyl alcohol, p-hydroxybenzaldehyde, p-hydroxybenzoic acid and protocatechuic acid with-out lag.
The strain S668B2 also oxidized benzyl alcohol, benzaldehyde and benzoic acid.
The strain did not oxidize, gentisic acid.
The formation of p-toluic acid from p-methyl benzylalcohol and p-methyl benzaldehyde, and p-hydroxybenzoic acid from p-cresol, p-hydroxybenzyl alcohol and p-hydroxybenz-aldehyde were investigated.
The pathway for the metabolism of p-xylene was intensively studied.

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Screening was carried out to obtain microorganisms which produce organic acids from n-paraffins. Three strains of yeast like microorganisms showed strong ability to produce fumaric acid. One of them turned out to be a new species and was named Candida hydrocarbofumarica n. sp.
Shaking culture of this microorganism was carried out in a medium containing 8% (v/v) n-paraffins, mineral salts (NH₄Cl 0.3%, KH₂PO₄ 0.05%, K₂HPO₄ 0.05%, MgSO₄•7H₂O 0.05%, FeSO₄.•7H₂O 0.001%, MnSO₄•4H₂O 0.0002%, ZnSO₄•7H₂O 0.0005%, CaCl₂ 0.001%) 0.02% yeast extract and 4% CaCO₃. We found that the yield of fumaric acid reached as high as 65% of n-paraffin added after 7 days cultivation.

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L-Arabinose ketol isomerase (EC 5.3.1.4) was extracted from cells of Candida utilis which were grown in sulfite pulp mill waste.
The enzyme was partially purified 135-fold by means of fractionation with ammonium sulfate, on DEAE-cellulose and DEAE-Sephadex A-50 column chromatographies and gel filtration on Sephadex G-200.
Optimal pH and temperature for the activity of the enzyme were 7.0 and 60°C, respectively. The enzyme reaction was activated by Co²⁺ and Mn²⁺, and was inhibited by Na-pyrophosphate.
The Michaelis-Menten's constant for L-arabinose was 1.07 × 10^-1M, and the thermodynamic quantities, ΔH, ΔG (35°C), ΔS (35°C) and E were 8914 cal/mole, -1397 cal/mole, 33.5 cal/deg. mole and 9588 cal/mole, respectively.

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Miniaturized bioassay technique, mini-test, was developed to estimate biologically available energy of a sample of only 5_??_50g. Chicks were previously starved to keep body size minimum, then given either standard diets of three energy levels or test diets containing 5% of the sample for 6 days. Available energy of the sample was estimated on the standard curve showing linear relationship between dietary energy level and response of the chicks on the standard diets.
Reliability of the estimate by this mini-test was discussed based on linearity of the standard curve, 95% confidence interval of the estimate and available energy of 2 known carbohydrates estimated by this mini-test.

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Growth rate, efficiencies of utilization of dietary energy and protein, and carcass fat level of the chicks decreased linearly with the increase of the intensity of restriction of either feed or energy alone, while the levels of carcass components other than fat were almost kept constant regardless of the restriction. The data indicated that chicks have ability to tolerate severer restriction than feeding moderately lipogenic diet at the level of 40% of full-feeding. It was confirmed that responses of chicks can be predicted based on the dietary composition and feeding conditions, using equations presented in this and previous papers.

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Studies were conducted on the influence of Kitazin analogues on the incorporation of ¹⁴C-glucosamine into the mycelial cell wall fraction of Pyricularia oryzae. Compounds of thiolates and phosphates, both having in vitro inhibitory activities toward the mycelialgrowth, inhibited the incorporation, whereas those of thionates and dithioates, either having no fungitoxicity, did not inhibit the incorporation. Mycelia of P. oryzae treated withKitazin-P (S-benzyl 0, 0'-diisopropyl phosphrothioate; IBP) accumulated about twice as much an amino sugar derivative as untreated ones. Mycelia treated with thiono or dithio analogues, which have no fungitoxicity, showed no accumulation. The accumulated substance gave a identical spot with authentic UDP-N-acetyl glucosamine on paper chromatograms developed with four solvent systems.

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The activity of histidase (L-histidine ammonia-lyase EC 4. 3. 1. 3) of rat liver was inhibited by cystine competitively with the substrate histidine. Histidase was inactivated by preincubation with cystine, but the inactivated enzyme was reactivated by the addition of glutathione to the reaction mixture. Cysteine, at a lower level, partly reactivated the enzyme inactivated by cystine, but, at a higher level, did not reactivate it. Cysteine itself acted inhibitory on the enzyme. Glutathione did not reactivate histidase activity inhibited by cysteine. No other amino acid affected histidase activity at the level of 2mM. Apparent Km of rat liver histidase was l.8×lO^-3M.
Neither inhibitory nor activating effect on histidase activity was observed by the addition of liver homogenate obtained from rats fed a histidine imbalanced diet to that from those fed a basal diet and vice versa.

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Rice embryo peroxidase 556 was purified to the extent as indicated by the absorbance ratio, RZ greater than 4.0. The enzyme was found to be major basic component among isoenzymes of rice embryo. The preparation was homogeneous as examined by sedimentation analysis, and the sedimentation coefficient, ^s°_{20, w}, was 3.76S. The prosthetic group of the enzyme was identified as protohematin and its content was 1.36%. The minimum molecular weight was calculated to be 46, 700. From the typical spectra of ligand-enzyme compounds, peroxidase 556 was found to react with carbon monoxide, cyanide, fluoride, and azide. However, at neutral pH, neither fluoride nor azide reacted with the enzyme. The high affinity of the enzyme to ammonia was one of the most remarkable characteristics of the enzyme. The hydrogen peroxide compounds I and II have been observed in the enzymic reaction, and therefore rice embryo peroxidase 556 is also concluded to follow the common reaction mechanism of plant peroxidases. Overall results show the close resemblance of rice embryo peroxidase 556 with wheat germ peroxidase 556 and hemoprotein 550.

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The 2, 3-Dihydro-1H-pyrrolo [1, 2-a] indole ring system was synthesised by the condensation reaction of toluquinone and 2-cyanomethylenepyrrolidine.

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The cold-insoluble fraction of soy protein was hydrolyzed with pepsin under usual conditions and the hydrolyzate was dialyzed. The nondialyzable part was treated with aqueous ethanol to obtain a 50% ethanol-soluble and 90% ethanol-insoluble fraction. From this fraction, a bitter tetracosapeptide was separated with the aid of column chromatography using Sephadex G-10 and DEAE Sephadex A-25, thin layer chromatography using silica gel G and cellulose powder, and paper electrophoresis. The partial structure of the bitter tetracosapeptide was proposed as follows: H•Phe-(Arg, Asp₂, Glu₂, Gly, Ile, Leu, Lys₂, Pro, Ser, Thr)-Trp-(Ala, Arg, Asp, Gly, Val)-Gln-Tyr-Phe-Leu•OH. Pepsin hydrolyzed this peptide to lessen its bitterness, whereas trypsin was ineffective in lessening the bitterness. Either carboxypetidase A or aspergillus acid carboxypeptidase was effective in debittering. N-Bromosuccinimide treatment on this peptide produced a nonapeptide bearing the intact C-terminal structure, which was found to be still bitter. However, further degradation of the bitter nonapeptide with the carboxypeptidase A or the aspergillus acid carboxypeptidase from its C-terminal side lenssened the bitterness. These results indicate that the bitterness of the bitter tetracosapeptide relates intimately to the hydrophobic C-terminal sequence, -Tyr-Phe-Leu•OH. Experimental results concerning chemical modification on this tetracosapeptide and also concerning some synthesized model oligopeptides supported the above conclusion.

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The site of action of antimycin A is known to lie between cytochrome b and c in the respiratory chain of mammalian cells. But in general, bacteria, even those which have cytochromes similar to those of mammalian cells such as Bacillus subtilis, are naturally resistant to this antibiotic.
The mechanism of this natural resistance is studied using a strain of B. subtilis. Succinoxidase activity of the intact cells of this bacterium showed very low sensitivity to the antibiotic, but on disruption of the cells, the sensitivity increased 7.5 times. Moreover, the activity of the intact cells could be sensitized by treatment with cationic detergent. In addition to the permeability barrier suggested by the above results, it was found that the electron transport system of this bacterium contained antimycin A insensitive by-path.

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A method was given for determination of alkaloids in culture filtrates using alkaloid reagents, e. g., modified Dragendorff's and Mayer's reagents, in conjunction with several pharmacological tests such as Magnus method employing the isolated guinea pig intestine and the influence on blood pressure etc. In a screening of about 1250 strains of actinomycetes, 8 strains yielded basic materials that gave positive reaction with both of the above reagents. Among them, HCl extract from the culture filtrate of Streptomyces strain No. FFD-101 caused a transient drop of blood pressure of rabbit. Furthermore one fraction of HCl extracts from the culture filtrate of Streptomyces strain 227×1 caused a continous rise of blood pressure.

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In the course of screening research on alkaloids among metabolites of microorganisms, Streptomyces strain No. FFD-101 which was isolated from a soil sample collected in Bankok of Thailand, was found to produce a new alkaloid, nigrifactin both in its culture filtrate and mycelium. This alkaloid possessed strong inhibitory activity against histamine and caused a transient fall of blood pressure. From the results of taxonomical study on the strain No. FFD-101, it was found that this is a variant of Streptomyces nigrifaciens, to which the name, Streptomyces nigrifaciens var. FFD-101 was proposed. Examination of the alkaloid productivity with media in various compositions indicated that the medium containing glycerol 3% as carbon source and casamino acid 0.35% as nitrogen source was most suitable for the production.

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Fusamarin, C₁₈H₂₄O₄, a metabolite of Fusarium sp., has been shown to be ((-)-(R)-(3)-trans-pentenyl-(3)-5-n-butyl-6, 8-dihydroxy-3, 4-dihydroisocoumarin) (1) on the basis of chemical and spectroscopic evidences.

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Histidase (L-histidine ammonia-lyase EC 4.3.1.3) activity of thyroidectomized rats was higher than that of intact animals. The levels of protein bound iodine (PBI) in plasma of rats fed a basal diet were higher than those of an imbalanced diet group under ad libitum condition, while if the food intake of a basal diet group was restricted to that of the imbalanced diet group, the levels of PBI of a paired fed group were practically the same as those of the imbalanced diet ad libitum group. Histidase activity of paired fed rats was twice as high as that of ad libitum fed animals, and was about 60% of that of the imbalanced ones. Thyroxine did not affect histidase activity in vitro.
Neither sex difference nor castration affected the activity of histidase of young rats fed the imbalanced diet.

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Rice embryo γ₁., globulin, composed of ten polypeptide chains, was found to dissociate into the subunits in the presence of urea by monitoring sedimentation and gel electrophoresis. No disulfide bond appeared to participate in the binding between subunits. In connection with this dissociation, the environments of tyrosine side chains varied from hydrophobic to hydrophilic ones, which resulted in the blue shift of the ultraviolet absorption bands and gave a difference spectrum having peaks at 280.5 and 288.0 mμ. Simultaneously the difference spectrum gave a vibrational bands due to phenylalanine, but no bands could be observed due to tryptophan. These results suggested the participation of hydrophobic bonds in the interaction between subunits. p-Chloromercuribenzoic acid also affected on the dissociation into subunits in the presence of urea, so that the sulfhydryl groups should participate in the association of the subunits.

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The structure of aeruginoic acid which was isolated from the culture medium of P. aeruginosa was detremined and confirmed synthetically.

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β-BTC (3, 4/5, 6), ¹⁾ γ-BTC (3, 4, 6/5), and ε-BTC (3, 4, 5/6) were synthesized from α-BTC (3, 6/4, 5) by stepwise routes.

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In DMSO (dimethyl sulfoxide) and some other polar solvents thermal isomerization of α-, β-, γ-, δ-, and ε-BTC²⁾ was studied. The time course of interconversion was by studied means of gas-chromatographic analysis. At 100°C more than 100 hr were required to reach an equilibrium, where α-BTC was an almost exclusive component. NaI had a catalytic effect in the reaction. The CS₂-AICl₃ system caused the isomerization, in some cases very rapidly, at room temperature.
Before reaching equilibrium in the thermal isomerization, convenient chances could be found to isolate δ-BTC from β, and γ-isomer, and γ-BTC from β-BTC. Possible mechanisms and intermediates are discussed.

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