In order to produce microbial cell substances from petroleum, 83 strains of kerosene-utilizing yeasts, as a sole source of carbon, were isolated from 37 materials in contact with petroleum in the petroleum refinery. They could be distributed in either of 15 cultural groups with their colony appearances. Fifteen representative strains in 15 cultural groups were served for determination and identified with the following species: Candida tropicalis, 9 strains; C. guilliermondii, 2 strains; C. intermedia, 2 strains; C. pulcherrima, I strains; Torulopsis pinus, 1 strain.
In order to clarify what the ability of hydrocarbon utilization means biologically, 46 standard strains were served for test, of which the following 5 strains could utilize kerosene as a sole source of carbon: Candida albicans IAM 4888; C. arborea IAM 4147; C. lipolytica IAM 4947; C. tropicalis IAM 4862 and IAM 4924. Considering the result, the ability of utilizing kerosene would seem to characterize the genus, but it was not evident that it would characterize the species.
C. tropicalis Pk-233 gave the best cell yield among the above strains when kerosene was employed as a sole source of carbon and moreover, in the production of the cells of Pk-233, employing kerosene as u carbon material was compared with employing glucose.

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The alkaline proteinases of Gliocladium roseum (Link) Bainier were purified in crystal-line forms by procedures of alcoholic precipitation, fuller's earth- and acrinol-treatment, and isolated in two types. (Proteinases I and II). Both of these proteinases were homo-geneous on zone electrophoresis with polyacrylamide gel (Cyanogum 41), and had the optimal pH values of 11 (Proteinase I) and 10 (Proteinase II), and the optimal temperature of 45°C.
The enzymatic reaction of proteinase I was remarkably promoted by Fe⁺⁺ and Co⁺⁺, and that of proteinase II was promoted by Fe⁺⁺, Co⁺⁺ and Ca⁺⁺, and both proteinases were protected from heat-inactivation by Ca⁺⁺, Proteinase II was activated remarkably by Cl- under the existence of Fe⁺⁺, but proteinase I was unaffected by the anion.
The order of strength of proteolytic power of these proteinases and chymotrypsin on casein was as follows; proteinase I> proteinase II> chymotrypsin.

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The Amount of ethyl acetate-extractable phenolic compounds in brandies or whiskies of differed aging grade was determined. Extractable phenolic compounds under neutral condition were increased along with proceeding of aging process.
In comparison of chromatographic pattern obtained using silicic acid column, the first fraction from neutral extract was increased with aging. The fraction had a specific aroma.

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A study was made on the elaborate separation and identification of the fraction, boiling higher than monoterpene hydrocarbons, of cold-pressed peel oil of Citrus natsudaidai Hayata, by using fractional distillation under reduced pressure, adsorption and gas chro-matography, infrared, mass and proton magnetic resonance spectrometry. The following components were identified; in sesquiterpene hydrocarbon series: copaene, β-elemene, β-ylangene, caryophyllene, aromadendrene, farnesene, a-selinene, b-cadinene, calamenene, a-calacorene; and among oxygenated compounds: n-nonyl alcohol, n-decyl alcohol, linalool, α-terpineol, terpinen-4-ol, nerolidol, elemol, n-octyl aldehyde, n-decyl aldehyde, perillalde-hyde, carvone, n-octyl acetate, n-decyl acetate, geranyl acetate, perillyl acetate and acetic ester of p-methadiene-1, 8(10)-ol-9.

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A procedure for obtaining a highly purified preparation of the putrescine oxidase of Micrococcus rubens has been developed. The method involved fractionation with ammonium sulfate and separation on successive columns of DEAE-cellulose, DEAE-sephadex and hydroxylapatite. The enzyme was purified about 200-fold from the cell extract and the final preparation showed a symmetric peak in the ultracentrifugal analysis. The purified enzyme was yellow in color and showed absorption maxima at about 380 and 460mμ. The yellow color was lost by the substrate as well as by sodium dithionite, and was subsequently restored by oxygenation. The purified enzyme contained 12.2mμ moles of flavin adenine dinucleotide (FAD) per mg of protein. The enzyme was inhibited by p-chloromercuric benzoate.
The putrescine oxidase could oxidatively degrade spermidine into 1, 3-diaminopropane and γ-aminobutyraldehyde stoichiometrically, and no ammonia was liberated.

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A remarkable increase in polyphenol oxidase activity was observed in tea leaves after plucking. When the tea leaves treated with the specific inhibitors for the protein biosynthesis such as blasticidin S and puromycin A by the absorption through stem or by the vacuum infiltration, the increase in the enzyme activity was severely prevented. Therefore, it was thought that these antibiotics inhibited the activation of polyphenol oxidase. In addition, CM-cellulose column chromatogram of the enzyme protein of the withered leaves was different from that of the fresh leaves. It was also considered that new protein containing the enzyme activity developed in the withered leaves.
These results suggest that the synthesis of the enzyme protein occurred in tea leaves after plucking.

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The properties of yeast 5'-nucleotidase, one of NAD-metabolic system in yeast, were studied.
1) The enzyme has optimum pH at 5.8_??_6.1 for its activity and is most stable at pH 6. It is inactivated completely at 55°C for 6min, pH 7, but never at 40°C for 6min. 2) The enzyme hydrolyzes only 5'-nucleotides of guanine, adenine, hypoxanthine, uracil and cytosine, but never splits nicotinamide mononucleotide, thiamine monophosphate, ribose 5-monophosphate and flavin mononucleotide. 3) The enzyme seems to have specially high affinity for 5'-AMP. 4) The enzyme activity is accelerated by addition of Co⁺⁺ and Ni⁺⁺, but inhibited by Ag⁺, Cu⁺⁺, EDTA, I₂ and N-bromosuccimide. Mg⁺⁺, KCN, NaF and thiol reagents except p-chloromercuribenzoate have no effects. 5) Nucleosides have inhibitory effects, among which adenosine is most effective inhibitor. 6) The activity is reduced up to 30% by dialysis against 1mm EDTA solution, and the reduced activity is completely reactivated by addition of Co⁺⁺ or Ni⁺⁺, but not by Mn⁺⁺ or Mg⁺⁺.

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The effects of solute and hydrochloric acid concentrations on optical rotation were studied using 20 naturally occuring amino acids.
There appeared to be no common factor among the amino acids as far as the inclina-tion of optical rotation was concerned. Lutz-Jirgenson's rule could be applied to few amino acids in the cationic form. Therefore, in the determination of the optical rotation, the concentration of the solute, nature of solvent and temperature must be rigorously controlled. The optical conditions of measurement and the specific rotation of 20 amino acids were recommended based on this work.

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A steep decrease in value of specific rotation was observed when concentration of sulfuric acid reached more than constant normality for each amino acid. Ultraviolet ab-sorption, nuclear magnetic resonance and Raman spectra showed that these decrease of specific rotation in concentrated sulfuric acid are attributed to the protonation of carboxyl group. The concentration of sulfuric acid in which amino acids showed maximum specific rotation are as follows.
Amino Acids Normality of sulfuric acids
L-Arginine 10 to 11
L-Aspartic acid 2 to 3
L-Isolcucine 20
L-Leucine 20
L-Phenylalanine 18
L-Valine 20

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The utilization of hydrocarbons by microorganisms was studied in many fields, but the production of biotin vitamers by hydrocarbon-utilizing bacteria has never been reported.
We have screened many hydrocarbon-utilizing bacteria which produce biotin vitamers in the culture broth. The effects of cultural conditions on biotin vitamers production by strain 5-2, tentatively assigned to the genus Pseudomonas, were studied.
More than 98% of biotin vitamers produced from hydrocarbons by strain 5-2 was chromatographically determined as desthiobiotin. As nitrogen source, natural nutrients were more effective than inorganic nitrogen sources. The production of biotin vitamers was increased under the condition of good aeration. Exogenous pimelic or azelaic acid enhanced biotin vitamers production by strain 5-2.

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The production of biotin vitamers from n-alkanes, n-alkenes or glucose by an isolated bacterium, strain 5-2, tentatively assigned to the genus Pseudomonas, was investigated. Among these carbon sources, n-undecane was the most excellent for biotin vitamers production.
The biosynthetic pathway of biotin vitamers, especially desthiobiotin, from n-undecane was also studied. It was found by thin-layer and gas-liquid chromatographical methods that pimelic and azelaic acids were the main acid components in n-undecane culture.
This result, together with previously reported enhancement of biotin vitamers pro-duction by these acids, suggests that pimelic and azelaic acids may be the intermediates of biotin vitamers biosynthesis from n-undecane.

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A glucose isomerase which reversibly catalyzes the reaction between D-glucose and D-fructose was demonstrated in the cell-free extracts of a strain of Streptomyces sp. isolated from soil. The enzyme was produced when the strain was grown in the medium con-taining xylan or xylan-containing material such as wheat bran. A medium which consists of 3% of wheat bran, 2% of corn steep liquor and 0.024% of C_oCI₂•6H₂O is recommendable for the production of the glucose isomerase enzyme with the strain. With the enzyme, some conditions for the conversion of D-glucose to D-fructose were also studied. The method is very useful for the production of invert sugar from D-glucose and is now on the way to be applied to the practical use.

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The F-1 β-glucan from naked barley endosperm, a main component of water soluble β-glucan, has been subjected to degradation with the laminarinase from Bacillus circulans. This enzyme converts the F-1 β-glucan to a trisaccharide, 3-O-β-cellobiosyl-D-glucose*, and a tetrasaccharide, 3-O-β-cellotriosyl-D-glucose*, as the main products. These products, which constitute 74% of the polymer, have been identified by chemical methods. As the minor or trace components, laminaribiose, cellobiose, cellotriose** and two unidentified tetrasac-charides are detected. Overall data show that F-1 β-glucan mainly consists of two types of structural sequences; one is trimeric unit in which a single linkage alternates with two consecutive β-(1→4) linkage, and the other, α tetrameric unit in which a single β-(1→3) linkage alternates with three consecutive β-(1→4) linkages.
It is shown that this laminarinase from B. circulans hydrolyses the glucoside bond of the reducing side of 1, 3-linked β-D-glucopyranose residues.

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A crystalline alkaline protease was prepared from B. amylosacchariticus, which was isolated as a strain of saccharogenic α-amylase-producing Bacillus subtilis. The enzyme was most active at pH values between 10.3 and 10.7 towards casein and was stable at pH values from 6 to 11 on twenty hour incubation at 30°C. Calcium ions were effective to stabilize the enzyme especially at higher temperatures. The enzyme was markedly in-activated by DFP as well as protease inhibitor from potato and slightly by surface active agents, but not affected by sulfhydryl reagents and divalent metal ions except Hg⁺⁺. Hemoglobin was the best substrate for the enzyme and more than 20% of the peptide bonds were hydrolyzed. Of numerous synthetic peptides tested, only the two compounds, glycyl-L-prolyl-L-alanine and Cbz-L-tyrosyl-glycinamide, were found to be hydrolyzed. A cyclic peptide, gramicidin S, was split by the enzyme only at the peptide bond of -L-valyl-L-ornithyl-. Methyl n-butyrate and tributyrin were also good substrates for the alkaline protease obtained here.

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Investigation was carried out on demonstration of two substances constructing a pre-cursor system located at a late stage of streptomycin biosynthesis by Streptomyces griseus. One of them is thought to be a natural precursor of Streptomycin(L) and the other is suggested as an enzymatic substance(H) transforming L to streptomycin. Both substances had no antibiotic potency and H was inactivated at low pH. L was obtained from a cell-free supernatant (active supernatant) prepared from suspension of young mycelium of Streptomyces griseus in glucose solution. H was obtained not only from active supernatant but also from cell-free extract of the organism.
Two ways of isolation were established for L. Active supernatant was adsorbed on a CM-cellulose column equilibrated with 0.05 M Tris-maleate buffer (pH 8.0). Elution of this column with the same buffer as was used for equilibration gave L-containing fraction separated from streptomycin which was eluted with the buffer including 1% of sodium chloride. L was adsorbed also on active carbon in aqueous solution at neutral pH and liberated from it at acidic pH with 95% methyl alcohol. The former method was useful to separate L from streptomycin, and the latter one was so to concentrate L.
H was isolated by using a column chromatography on DEAE-cellulose. After adsorb-ing active supernatant or cell-free extract of organism on a column equilibrated previously with the same buffer as above, H was eluted with the buffer including 1% of sodium chloride. Cell-free extract of S. griseus was a better source of H supply than the active supernatant.

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The auxin activities of the homologs of racemic and enantiomeric α-alkylphenylacetic acids were estimated by pea straight growth test. The α-methyl, -ethyl and -propyl acids were moderately active whereas the longer and branched alkyl chain were found to make the molecule inactive. The more active enantiomers were shown to have the same con-figuration as the more active enantiomers in the other series of the optical active synthetic auxins.

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The auxin activities of the cyclic homologs of 1, 2, 3, 4-tetrahydro- and 3, 4-dihydro-l-naphthoic acids were determined by pea straight growth test. In the tetrahydro-acid series, it was observed that the alicyclic ring expansion from the 6-membered to the 7-membered made the molecule inactive. In the 3, 4-dihydro-acid series, on the other hand, the activity remained almost unchanged by such a structural change. Structure-activity relationships were discussed in terms of their molecular structures, in particular, the configuration of the carboxyl group.

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