Spheroplasts were disrupted with 0.2% Brij 58 and the separation of intact cells, spheroplasts, disrupted spheroplasts, fragmented membrane, and supernatant was performed on a linear 40_??_55% sucrose gradient. About half an amount of nucleic acid components was distributed in disrupted spheroplast fractions, while only a small amount of protein components was found in these fractions.
DNA polymerase in the fragmented membrane fraction incorporated ³H-TTP more rapidly than that in the supernatant fraction for the first 5 to 6 min, and then the incorporation rate decreased, while DNA polymerase in the supernatant fraction incorporated ³H-TTP linearly up to 20min when native DNA was used as a primer. The former required native DNA as a primer and showed little activity towards denatured DNA, while the latter incorporated ³H-TTP at a similar rate to both the primer DNA's.
DNA polymerase of the fragmented membrane fraction synthesized various sizes of DNA from short to a size of primer when native DNA was used as a primer, while when denatured DNA was used, products were only short. DNA polymerase of the supernatant fraction synthesized various sizes of DNA when both native and denatured DNA's were used as primers.

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The distribution of tyrosine phenol lyase activity in microorganisms was studied with intact cells in a synthetic reaction mixture containing L-serine and phenol or pyrocatechol. This activity was found in various bacteria, most of which belonged to the Enterobacteriaceae; especially to the genera Escherichia, Proteus and Erwinia. Cells of Erwinia herbicola ATCC 21434 were selected as a promising source of enzyme.
Intact cells of Erwinia herbicola ATCC 21434 prepared from a broth cultured for 24 hr contained markedly high enzymic activity and catalyzed the synthetic reaction of L-tyrosine or 3, 4-dihydroxyphenyl-L-alanine (L-dopa) from L-serine and phenol or pyrocatechol in significantly high yields.
Results of the isolation and identification of the products showed that the amino acid synthesized by this enzymatic method was identical with L-tyrosine or L-dopa.

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Tyrosine phenol lyase catalyzes a series of α, β-elimination, β-replacement and racemization reactions. These reactions were studied with intact cells of Erwinia herbicola ATCC 21434 containing tyrosine phenol lyase.
Various aromatic amino acids were synthesized from L-serine and phenol, pyrocatechol, resorcinol or pyrogallol by the replacement reaction using the intact cells. L (D)-Tyrosine, 3, 4-dihydroxyphenyl-L (D)-alanine (L (D)-dopa), L (D)-serine, L-cysteine, L-cystine and S-methyl-L-cysteine were degraded to pyruvate and ammonia by the elimination reaction. These amino acids could be used as substrate, together with phenol or pyrocatechol, to synthesize L-tyrosine or L-dopa via the replacement reaction by intact cells. L-Serine and D-serine were the best amino acid substrates for the synthesis of L-tyrosine or L-dopa. L-Tyrosine and L-dopa synthesized from D-serine and phenol or pyrocatechol were confirmed to be entirely L-form after isolation and identification of these products. The isomerization of D-tyrosine to L-tyrosine was also catalyzed by intact cells.
Thus, L-tyrosine or L-dopa could be synthesized from DL-serine and phenol or pyrocatechol by intact cells of Erwinia herbicola containing tyrosine phenol lyase.

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Some properties of sucrose-P synthetases obtained from various plant tissues, including sweet potato roots, potato tubers and leaves of barley, rape and ladino clover were studied. The specific enzyme activity of the sucrose-P synthetase from sweet potato roots was much lower than that of the sucrose synthetase of the other tissues. The enzyme activity decreased gradually as the roots developed. The optimum pH did not differ between enzyme preparations from sweet potato roots and barley leaves. Manganese chloride exhibited a marked stimulative effect on the sucrose-P synthetase from sweet potato roots and potato tubers, whereas it was inhibited the barley leaf enzyme.
Kinetic studies of sucrose-P synthetase showed that the behavior of the enzyme to the substrates did not differ in the enzyme sources examined. The substrate saturation curve of the enzyme with respect to fructose-6-P was sigmodal in shape, giving a straight line with a slope of 1.35_??_1.5 (n value) in a plot of the data using the empirical Hill equation. On the other hand, enzymes from all the various tissues exhibited a hyperbolic substrate saturation curve for UDP-glucose, obeying the ordinary Michaelis-Menten type reaction. Manganese chloride had no effect on the Km for UDP-glucose, the S_0.5 for fructose-6-P and the n value of the enzyme from potato tuber tissues.

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An endo-polygalacturonase from culture extracts of Aspergillus japonicus was purified about 34-fold by ammonium sulfate fractionation, SE-Sephadex column chromatography and gel filtration. The purified enzyme was homogeneous on ultracentrifugation and disc electrophoresis. Using gel filtration a molecular weight of 35, 500 was estimated for the enzyme. The enzyme rapidly reduced the viscosity of pectic acid and released reducing groups in a random manner, yielding a mixture of mono-, di- and trigalacturonic acids as end products. The pH optimum of the enzyme for viscosity-reducing activity was 4.5 with pectin and pectic acid as substrates, and that for releasing reducing groups was also 4.5 with various pectic substances. The purified enzyme was able to macerate various kinds of plant tissues by itself.

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The lipase from Candida paralipolytica required activating factors for the hydrolysis of synthetic triglycerides, methyl esters of fatty acid and so on. Of the saturated monoacid triglycerides tested, tricaprylin was hydrolyzed most quickly. On the other hand, the lipase was hardly able to hydrolyze methyl butyrate, methyl caproate, monoolein, Tween 20 and Span 20.
Human blood plasma did not act as an activator, but act rather as an inhibitor of the lipase. Therefore, it seems that the lipase does not belong to the group of lipoprotein lipase.

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A biologically active substance which inhibits spawning of the starfish, Asterina pectinifera, has been isolated from gonads of the same organism and identified as L-glutamic acid.

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An acid DNase^* was purified about 700-fold from the pupae of Bombyx mori by ammonium sulfate fractionation, gel filtration, and chromatography with CM-cellulose. The optimal pH and temperature of the enzyme were found at about 5 and 50°C, respectively. Requirement of magnesium ion for the enzyme activity was not absolute. The enzyme liberates acid-soluble nucleotides from heat-denatured DNA slightly faster than from native DNA. The action of the DNase was endonucleolytic. Mono- and oligonucleotides of various sizes bearing 3'-phosphate were detected among the digestion products. These properties substantially resemble those of mammalian DNases of type II.

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In view point of the basic research for the enzyme properties and the development of utilization of lipase, a thermophilic fungus which could produce a remarkable amount of thermostable, alkalistable and extracellular lipase has been isolated from the compost soil. The taxonomical characteristics of this thermophilic fungus were examined and it was identified as Humicola lanuginosa S-38.

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The cultural conditions for the production of thermostable lipase by a thermophilic fungus Humicola lanuginosa S-38 were investigated. The optimal cultural conditions to obtain the maximum yield of thermostable lipase with a 600-liter stainless steel fermentor were as follows: optimal medium-2.0% soluble starch, 5.0% corn steep liquor, 0.2% K₂HPO₄, 0.1% MgSO₄•7H₂O, 0.5% CaCO₃, 0.5% soybean oil, 0.005% deforming agent (Adecanol LG-109); optimal fermentation conditions- temperature 45°C; rate of agitation 300rpm; initial pH 7.0; rate of aeration 1/1 volume per volume of medium per minute. The optimal pH of the crude lipase preparation for the hydrolysis of the polyvinyl alcohol-emulsified olive oil was 8.0 and the optimal temperature was 60°C. It retained 100% of activity with the heat treatment at 60°C for 2 hr, but at 70°C for 20min only 35% activity retained.

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Pestalotin, a gibberellin synergist, was isolated from culture broth of a fungal strain, Pestalotia cryptomeriaecola. The structure of the compound has been established as I on the basis of chemical and spectral data.

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A novel synthesis of dl-C₁₇-Cecropia juvenile hormone (I) was accomplished.

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The herbicide, pentachiorophenol decomposes within a few weeks after its application in rice fields. A reductive dechloronation was revealed as one of its decomposition pathways, in which some microorganisms play a dominant role and other soil chemical factors in a reduced condition are relatively of little importance. The stable decomposition products found in the paddy soil were 2, 3, 4, 5-, 2, 3, 5, 6- and 2, 3, 4, 6-tetrachlorophenol, 2, 4, 5- and 2, 3, 5-trichlorophenol, 3, 4- and 3, 5-dichlorophenol, and 3-chlorophenol.

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Two crystalline and one highly purified galactanases were obtained from the culture broth of Bacillus subtilis var. amylosacchariticus (1043) and their chemical and enzymatic properties, especially, their specificities were comparatively studied. Their molecular weights were almost the same, but the isoelectric points were considerably different from each other. The galactanases were sensitive to metal chelators and stabilized by Ca²⁺. The pH optimum of the enzymes were between 6.0 and 7.0. All the galactanases investigated here attacked soybean arabinogalactan without liberation of arabinose, though they were inactive against coffee bean arabinogalactan. In digestion of soybean arabinogalactan, all the galactanases purified here formed galactose, galactobiose and galactotriose whereas the galactanase previously isolated from Bacillus subtilis K-50 produced galactobiose as the main final product.

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Kinetics of photochemical reaction between uridine and sulfhydryl compounds were investigated in phosphate buffer (pH 7.0) and in unbuffered aqueous solution under aerobic condition. The results obtained clearly demonstrate that the photo-induced hydration or reduction of the uridine molecule was significantly influenced by the amount of sulfhydryl group present in the reaction medium. Reaction on uridine (1mM) was observed to lead to photohydration with pseudo first order rates, independently of the presence of cysteine (1mM or 2mM), while in the presence of dithiothreitol (DTT, 1mM to 10mM) both photoreduction and photohydration of uridine were observed. The rate of photoreduction came to predominate as the amount of DTT increased. The reaction was discussed from the view point of food chemistry as well as reaction pathways.

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In order to have increased extracellular production of cellulase by Trichoderma viride ITCC 1433, the organism was grown on various growth factors. Cellulose Powder -123 was found to be the best C-source while amongst raw materials, alkali-treated rice straw gave the best yield. A combination of peptone, urea and ammonium sulphate gave better production of cellulase than when a single nitrogen source was used. Sugars when added into the cellulose medium, generally suppressed the yield. When the organism was grown on sugars as the sole source of carbon, only lactose and maltose induced any cellulase production. Acetate and ascorbate were conspicuous in increasing cellulase production and when given together they had a cummulative effect and the yeild was doubled.

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An investigation of succinic acid production from n-paraffin under various culture conditions was carried out with Candida brumptii IFO 0731. Ammonium nitrogen was required for both cell growth and succinic acid production. Favorable culture conditions for succinic acid production were ascertained. The productivity was markedly increased by the additions of CaCO₃ and organic nutrients. Under the best condition, the largest quantity of succinic acid production, 23.6mg/ml, was obtained in a 67% yield from super heavy n-paraffin after 8 days cultivation.

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A mixture of two thionocarbamates was subjected to the acid-catalyzed rearrangement. A sample of the reaction mixture was analyzed by glpc and resolved into four components. From the cross-over result, it has been concluded that the acid-catalyzed rearrangement of alkyl thionocarbamates into the isomeric thiolcarbamates proceeds by an intermolecular alkylating mechanism. This conclusion was supported by the detection of a transalkylated intermediate.

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The reactivity of sphingosine bases as substrates in the enzymatic synthesis of ceramide was in decreasing order of erythro-4-sphingenine, erythro-sphinganine, threo-4-sphingenine and threo-sphinganine in the microsomal fraction obtained from chicken liver. Unsaturated base-containing ceramide was enzymatically formed from 4-sphingenine and saturated basecontaining ceramide from sphinganine. The formation of unsaturated base-containing ceramide proceeded similar to that of saturated base-containing ceramide. The data explain in part, the fact that 4-sphingenine is generally superior to sphinganine as the constituent sphingosine base in sphingolipids, the biochemical derivatives of ceramide.

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Addition of lysine to the culture medium of rumen ciliates increased the amount of pipecolate in the medium to the level over the control.
L-Lysine- U-¹⁴C was partly incorporated into ciliate protein unchanged, and partly converted to radioactive pipecolate. The autoradiogram of amino acids in supernatant fluid of the medium revealed the traces of two unidentified spots other than lysine and pipecolate as a main metabolite.
Since the radioactive pipecolate was not changed further more by ciliates, it seemed to be the end product in lysine degradation.
After addition of L-lysine-α, _??_-¹⁵N to the medium, ¹⁵N was detected mainly in the fractions of pipecolate and ammonia and a little in that of glutamate, alanine and aspartate of the medium.

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Synthesis of several prostaglandin-F₁, related compounds utilizing bicyclo(2, 2, 1) heptene derivatives as key intermediates were investigated.

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Biologically active substances which inhibit spawning of mature starfish, Asterias amurensis, have been isolated from ovaries of the same organisms and identified as asterosaponins A and B.

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