Photo-oxygenation of α-ionone was studied to clarify the relationship between the maturity of aroma and photo-oxygenative change of α-ionone. α-Ionone was converted to oxygenated derivatives which were identified as 2, 3-epoxy-β-ionone, 3, 4-epoxy-α-ionone, 4-keto-β-ionone (trans- and cis-form), 5-keto-α-ionone and 3, 4-dihydroxy-α-ionone.

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The acid protcase produced from Rhodotorula glutinis K-24 was purified and crystallized by precipitation with ammonium sulfate, fractional precipitation with acetone, gel-filtration with Sephadex G-100, and pH adjustment. The recovery of activity was about 40 per cent, and about 80_??_100mg of third crystallized enzyme was obtained from one liter of broth. The purified enzyme was chromatographically homogeneous and confirmed to be monodispersive by physicochemical criteria. The enzyme was most active at pH values between 2.0 and 2.5 toward casein, and was stable at pH values from 2.5 to 6.5 on twenty hour incubation at 37°C. Comparing with several proteases obtained from various sources, the specific activity toward casein was potent and was 1.7 times that of pepsin. The enzyme was inactivated by pepsin inhibitor (S-PI) produeecl from Streptomyces naniwaensis, wheares it was not inactivated by SLS, divalent metal ions and sulfhydryl reagents.

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Some physicochemical properties, amino acid composition, and substrate specificity of the acid protease of Rhodotorula glutinis K-24 were determined. The molecular weight was estimated to be about 30, 000 by the sedimentation equilibrium method. This value also coincided with the data obtained from Andrews's method.
The isoelectric point was pH 4.5, and the amino terminal amino acid was identified to be alanine. The enzyme contained 14.5% of nitrogen and was composed of 285 residues of amino acid. Substrate specificity toward synthetic peptides was similar to that of pepsin, but its activity was considerably weak.
The enzyme was inactivated by diazoacetyl glvcine ethylester, p-bromophenacyl bromide, et al., which attacked the active center of pepsin.

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Chymotryptic peptides of CGMMV^* protein were separated by chromatography on a AG50W-X2 column. Tryptic peptides obtained before and chymotryptie peptides which contain basic amino acids were aligned to yield a partial amino acid sequence (residue 41 to 129 in the corresponding TMV protein) of the protein. Amino acid sequences of coat proteins of TMV group were compared and CGMMV protein showed some variations even in such sections as 87 to 94 or 113 to 122, which is identical in the other coat proteins. Distributions of hydrophobicity in CGMMV, TMV, and HR proteins were found to be very similar each other in spite of fairly large difference in amino acid sequences. This suggests that the specific distribution of hyclrophobicity in the primary structure would be significant for specific conformation of the protein which construct the rod-shaped virus particle.

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When azelaic acid was used as _??_ sole carbon source on the growth of Micrococcus sp. which was isolated from soil, intact cells of the organism catalyzed the enzymie condensation of fatty acids with hydrosylamine. Some of the characteristics of fatty acid hydroxamate formation were investigated.
The enzyme activity was tested with azelaic acid compared to other fatty acids. Azelylhydroxamate formation was activated with the addition of reduced glutathione or 2-mercaptoethanol. The reaction was inhibited by p-chloromercuribenzoate (PCMB), ethylene diamine tetraacetate (EDTA), NaF and benzoate.

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When Micrococcus sp. which was isolated from soil assimilated azelaic acid as a sole carbon source, cell-free extract of the organism catalyzed enzymic fatty acid hydroxamate formation. The enzyme was effective only for mono-carboxylic acid, but not for cli-carboxylic acids such as azelaic acid. The activity was high with higher fatty acid such as oleic acid. Some of the properties of higher fatty acid hydroxamate formation were investigated.
Olelylhydroxamate was formed with the high concentration of hydroxylamine. The reaction was inhibited by PCMB, but recovered by the addition of SH-compounds (such as cysteine).
On the other hand, when methylacetate was used as a sole carbon source and cell-free extract of Micrococcus sp. hydrolyzed several fatty acid esters. The fatty acid hydroxamate degradation by esterolysis are also discussed.

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The gamma-radiolysis of methionine in 10^-3_M aqueous solution was studied. From the yield vs. dose plots, the following values were obtained: G(-Methionine)=3.0, G(Methionire sulfoxide)=0.30, G(α-Aminobutyric acid)=0.46, G(3-Methylthio propylamine)=0.59, G(Methional)=0.08, G(Carboxylic acid)=0.24, G(Mercaptan+Disulfide)=0.61, G(Ammonia) =1.48, and G(Carbon dioxide)=1.45. From these results, it was concluded that major reaction was deamination and decarboxylation by _??_OH radical attack.
Addition of methanol as the second solute at concentration above 4×10^-3_M could not lower G(Methionine sulfoxide), suggesting the reaction of methionine with secondary radical produced from methanol.
Existence of similar reaction was suggested in the radiolysis of methionine in aqueous pork extract.

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In this study, the mechanism of the extracellular accumulation of L-glutamic acid by the glycerol auxotroph was partially clarified. Whenever Corynebacterium alkanolyticum GL-21 (glycerol auxotroph) accumulated a large amount of L-glutamic acid in the fermentation broth, the content of its cellular phospholipids was not more than 50% of that of C. alkanolyticum No. 314 (prototroph).
Moreover, biotin, oleic acid or thiamine had no influence on the cellular phospholipid content of the auxotroph.
Under limited supply of glycerol, the efflux of L-glutamic acid in the auxotroph was extremely enhanced, but its enzyme activities participating in L-glutamic acid biosynthesis remained at the same level as those of the prototroph.
From the results, it is considered that the regulation of phospholipid content gave rise to the destruction of the permeability barrier to L-glutamic acid in the cell membrane.

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A novel process for the microbial production of L-glutamic acid on an industrial scale was successfully established by using a glycerol auxotroph.
The most suitable carbon source for producing L-glutamic acid was n-paraffins (C₁₃-C₁₅). The production of L-glutamic acid was not affected by a large amount of biotin or oleic acid in the absence of penicillin, and occurred maximally at the glycerol concentration of 0.02% at pH 6.6. The most effective temperature was 28°C.
Under optimal conditions in a 200 liter fermentor, the mutant produced 72g/liter of L-glutamic acid. On the other hand, the parent produced 53g/liter of L-glutarnie acid in the presence of penicillin.
It is believed that the low productivity of L-glutamic acid by the parent strain was mainly due to the occurrence of the marked decrease in the viable cell counts at the later phase of the fermentation caused by the action of penicillin added.

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Ciliatine (2-aminoethylphosphonic acid) (76mg) was isolated from 72g of lipids of the oyster with _??_ combination of ion exchange chromatographic techniques and was identified from the results of elementary analysis, infrared spectrum, and chromatographic behaviors. The phosphonic acid was also detected in hydrolysates of a chloroform-methanol insoluble fraction of the oyster. It has been demonstrated that the oyster contains high concentration of ciliatine.

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The constituents of steam volatile aroma, which were responsible for topnote of Ceylon tea aroma, were identified. A total of 57 compounds were identified, of which 10 (terpinolene, n-nonanal, trans-2-pentenal, trans-3-octen-2-one, 6-methyl-3, 5-heptadien-2-one, n-nonanol, cis-3-hexenylbutyrate, cis-3-hexenylcaproate, α-terpinylacetate and nerylacetate) had not previously been reported as associated with aroma of black tea. Approximate composition of topnote aroma from Ceylon flavory tea was also determined.

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Lipogenesis in the fatty liver of rat which was induced by feeding an amino acidimbalanced diet containing 8% casein supplemented with 0.3% DL-methionine has been investigated by measuring the incorporation of acetate-1-¹⁴C into various lipid fractions during in vitro incubation of liver slices.
In the incorporation of acetate-1-¹⁴C into the total lipid per one g of the slices, no significant difference for the imbalance group was observed. However, the total radioactivity of liver lipid per 100g of the body and the incorporation into triglyceride in the lipids were significantly higher in the imbalance group than in the control group. Conversion of acetate-1-¹⁴C to CO₂ was not impaired in the imbalance group.
It is evident from these results that the induction of this type of fatty liver is due mainly to the synthesis of triglyceride.

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Phosphatidyl ethanolamine and lecithin dispersed in water rapidly absorbed oxygen. The autoxidation was promoted with copper and inhibited with EDTA. Phosphatidyl ethanolamine was more sensitive both to copper and EDTA than lecithin. Their peroxide decomposition under the deaerated condition was promoted with copper and inhibited with EDTA. The copper-catalyzed O₂ uptake of phosphatidyl ethanolamine was lowered, when its peroxide was decreased. Thus, the mechanism of promoting the oxidation with copper may be explained as catalytic supply of initiator radicals from the peroxides by copper.

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Strain MR-12 which was derived from Candida cloacae M-1 as a mutant unable to assimilate n-alkane showed marked increase in dicarboxylic acid (DC) productivity from n-alkane.
Resting cells of strain MR-12 produced 42.7g/liter of n-tetradecane 1, 14-dicarboxylic acid (DC-16) from n-hexadecane (n-C₁₆) after 72hr' incubation. DC degradation activities of strain M-1 and MR-12 were found to be markedly reduced and their activities against DC-16 decreased to 40% and 10% of that of the parent strain, respectively.
Strain M-1 and MR-12 produced DC from the various oxidized derivatives of n-alkane such as alcohol, diol, aldehyde, fatty acid and methyl- or ethylester of fatty acid other than n-alkane.
The carbon balance in n-C₁₆ oxidation seas determined by using resting cells of strain MR-12 and about 6% of utilized carbon was recovered as DC-16 and about 40% was recovered as CO₂.

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Proteolytic activity was found in the culture fluids of numerous psychrophilic bacteria isolated from terrestrial or marine samples. Among these organisms, a marine psychrophilic bacterium, Pseudomonas sp. No. 548, showed the highest proteolytic activity. This organism required salts of sea water for both growth and protease formation. The optimum temperature for the growth of this organism was 20°C. The formation of protease was the greatest at 5°C and decreased with increasing temperature. The extracellular protease system was fractionated into at least two components having proteolytic activities by chromatography with DEAE-cellulose. Increasing culture temperature tended to increase the activity ratio of Fraction I to Fraction II. Some cultural conditions for protease formation were investigated.

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Protease which was found in the culture fluid of Pseudomonas sp. No. 548 was fractionated into four components with protease activity by a two step chromatography using DEAE-cellulose. Each protease was further purified by gel filtration on Sephadex G-100 and/or G-75. The protease of Ia was obtained in crystalline form and was shown to be homogeneous by analysis with electrophoresis, while the other three enzymes were also highly purified. The enzymatic properties of the proteases were investigated. All of the four enzymes were inactivated by ethylene diamine tetraacetate. Proteases Ia, Ib, and IIb were inactivated by diisopropylfluorophosphate. The optimum activity of protease Ia was shown to be at pH 10.0, and that of the other enzymes were at pH 7.0 to 8.0. The proteases of Ia, Ib, and IIb were stabilized by calcium ion. The effect of temperature, pH, and metal ions on the activity of the enzyme were also investigated.

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Intact cells of Kluyvera citrophila KY 3641 produced enzymatically D(-)-α-aminobenzylpenicillin from 6-aminopenicillanic acid and phenylglycine derivatives. The optimum pH of the aeylase was 6.5. Among various phenylglycine derivatives examined as substrates, D-phenylglycine methylester HCl was the best compound giving the yields of about 10.7mg/ml of D(-)-α-aminobenzylpenicillin in the enzymic reaction mixture. The product was isolated in a crystalline form and identified as D(-)-α-aminobenzylpenicillin.

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The reaction of alanine with glyoxal was studied at both 50° and 100°C. Effects of reactant concentration, temperature, solvent, ionic strength and quinoline concentration on decarboxylation rate was investigated. From kinetic data, it was considered that decarboxylation took place through the unionized Schiff base. Experiments with reductones and 2, 6-dichlorophenol indophenol proved that generated reductone was one of the causative factors for oxygen absorption in the amino acid-dicarbonyl reaction.
Quantitative studies of reactants and products in the reaction showed that deamination reaction occurred to a small extent, and that the quantity of generated carbon dioxide was much smaller than that of decomposed alanine. Investigation with Ala-1-¹⁴C showed that about three ¹⁴C-products were produced. It was also concluded that in the degradation of alanine in the presence of glyoxal, decarboxylation, i.e. Strecker degradation, was not the main pathway at low temperature.

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L-Threonine production by strain BB-69, which was derived from Brevibacterium flavum No. 2247 as a α-amino-β-hydroxyvaleric acid resistant mutant and produced about 12g/liter of L-threonine, was reduced by the addition of L-lysine or L-methionine in the culture medium. Many of lysine auxotrophs but not methionine auxotrophs derived from strain B-2, which produced about 7g/liter of L-threonine, produced more L-threonine than the parental strain. Except only one methionine auxotroph (BBM-21), none of lysine and methionine auxotrophs derived from BB-69 produced more L-threonine than the parental strain. Homoserine dehydrogenase of crude extract from strain B-2 was inhibited by L-threonine more strongly than that from BB-69. Strain BBM-21, a methionine auxotroph derived from BB-69, produced about 18g/liter of L-threonine, 50% more than BB-69, while accumulation of homoserine decreased remarkably as compared with BB-69. L-Threonine production by BBM-21 was increased by the addition of L-homoserine, a precursor of L-threonine, while that by BB-69 was not. No difference was found among BBM-21, BB-69 and No. 2247 in the degree of inhibition of homoserine kinase by L-threonine. L-Threonine production by revertants of BBM-21, that is, mutants which could grow without methionine, were all lower than that of BBM-21. Correlation between L-threonine production and methionine or lysine auxotrophy was discussed.

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Mass spectra of the δ-lactones of the following 5-hydroxy-2-enoic acids were determined: 5-hydroxyhex-2-enoic acid (I), 5-hydroxyoct-2-enoic acid (II), 5-hydroxydec-2-enoic acid (III), 5-hydroxydodec-2-enoic acid (IV), 5-hydroxy-8-methylnon-2-enoic acid (V), 5-hydroxy-6-ethyloct-2-enoic acid (VI), 5-hydroxy-5, 6, 6-trimethylhept-2-enoic acid (VII), and 5-hydroxy-5-methylnon-2-enoic acid (VIII). The following modes of fragmentation are consistent with observed m/e values, metastable peaks, and established modes of breakdown in compounds containing similar atomic groupings: -l. Loss of side chain, resulting in ions at m/e 97 for I-VI and at m/e 111 and 153 for VII and VIII (diagnostic peaks); 2. 1, 4-Rupture of the ring giving an ion at m/e 68 (diagnostic peak) which loses CO to give m/e 40; 3. Loss of CO from m/e 97 fragment to give m/e 69 which breaks down further to m/e 41→m/e 39; 4. 1, 4-Rupture of m/e 111 and m/e 153 fragments to give m/e 43 and 85, further breakdown of m/e 85→57→41→39; 5. Loss of H₂O from the molecular ion providing there is a hydrogen atom on C₅ and the side chain is at least 3 carbon atoms in length, further loss of H₂O when the side chain is equal to C5 or C7; 6. Loss of CO₂ from the molecular ion of I, IV-VIII; 7. Loss of CO from all molecular ions; 8. Loss of 2×28 from the molecular ions of III, IV, V, VI; 9. Loss of (18+28) from the molecular ion of III, IV, V, VI; 10. Loss of 60 from the molecular ion of II, III, IV, V, VI; 11. Formation of M+1 ion (169) of VII and VIII; 12. Formation of M+1 ion (143) of saturated_??_-octalactone and loss of H₂O from this M+1 ion.

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Polyoxins D, E, and F which possess 5-carboxyuracil as the nucleobase were reacted selectively with sodium bisulfite at pH 4.0 resulting in facile decarboxylation to afford corresponding 5, 6-dihydrouracil-6-sulfonates and uracil type polyoxins (polyoxins L, M, and K) in good yield. The former compounds were also converted to the latter almost quantitatively with mild alkali treatment. Biological activities of the transformed compounds were described.

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