An endo β-1, 3-glucanase which is able to disrupt the cells of living yeast has been purified in homogeneous state from the culture filtrate of Flavobacterium dormitator var. glucanolyticae. The molecular weight of the enzyme was estimated to be 17, 000_??_22, 000. The mode of enzyme action has been suggested to be a “random” type of β-1, 3-glucanase. The enzyme preferes larger chains saccharides as substrate for its action, however, smaller oligosaccharides such as laminaritriose and laminaribiose are also decomposed by the enzyme. The Km values of the enzyme for laminarin, laminarihexaose, and laminaritetraose were determined to be 0.26, 1.18, and 2.00g/liter, respectively. The ability of this enzyme to disrupt the cells of living yeast is its remarkable point, since endo β-1, 3-glucanase of a smaller oligosaccharide-producing type from most sources has been recognized to be inactive (or very weakly active) on living yeast cells.

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Microbiological conversion of α-terpineol to borneol and terpinolene was studied with a newly isolated microorganism. Borneol is assumed to be an end-product of the degradation of α-terpineol by the organism, but with terpinolene this is not the case because of its poor reproducibility. The strain through the results of physiological tests was tentatively identified as Ps. aeruginosa.

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A phenanthrene-assimilating bacterium which belongs to the genus Aeromonas was isolated from soil. The cells which adapted to phenanthrene required a growth lag time on a naphthalene medium. The cells oxidized 1-hydroxy-2-naphthoate (1H2NA), 2-carboxybenzaldehyde (2CBAL), o-pbthalate (OPA) and protocatechuate (PCA) but did not oxidize salicylalde-hyde (SAL), salicylate (SA) and catechol (CAT) which are intermediates in naphthalene catabolism. Using the cell-free extract, the same results were obtained in oxidative capacity. The intact cells metabolized 1H2NA and 2CBAL without the lag time, giving 2CBAL and PCA, respectively. The ammonium sulfate-treated extract prepared from the cells grown in phenanthrene medium, converted 1H2NA to 2CBAL and 2CBAL to OPA. It was suggested that the Aeromonas sp. degraded phenanthrene through OPA.

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Differential thermal analysis (DTA) was used for study of milk protein denaturation. Protein solutions produced an endothermic peak of characteristic shape and temperature of peak minimum. The peak minimum is considered the coagulation temperature of the protein.
The influence of pH and additives such as sugars and NaCl was clearly observed on the thermograms of β-lactoglobulin solution. Addition of κ-casein to β-lactoglobulin solution showed an inhibitory effect on the heat coagulation.
Solid proteins produced two-stage exothermic peaks between 200°C and 400°C.
DTA was a useful method in the study of heat denaturation and degradation of protein.

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An extracellular alkaline proteinase produced by Candida lipolytica was purified through iso-propanol and ammonium sulfate precipitation, decolorization with DEAE-cellulose, gel filtration with Sephadex G-100 and ion-exchange chromatography on DEAE-Sephadex A-50. The optimum pH of its caseinolytic activity was 9.0, and this activity was completely inactivated with DFP but not with chelating reagents, PCMB, STI, TLCK, TPCK, or SSI. This enzyme also hydrolyzed salmin and synthetic esters, such as Bz. Arg. OEt, Bz. His. OMe, Tos. Lys. OMe or Ac. Tyr. OEt, and the optimum pH of its esterase activity was 8.0. The molecular weight of the enzyme was estimated to be about 30, 000 by the gel filtration method. These facts indicated that this enzyme was distinguishable from other microbial alkaline proteinases so far studied.

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Clostocin O is an inducible, phage tail-like bacteriocin produced by Clostridium saccharoperbutylacetonicum N1-4 (ATCC 13564). Added clostocin O was adsorbed within 5 min to the sensitive bacteria, C. saccharoperbutylacetonicum No. 8. The adsorption number of clostocin O per cell was about 150 particles. The adsorption sites of clostocin O were at the cell poles, the subpolar position and the division plain of the sensitive organism. The protoplasts lacking cell wall could not adsorb clostocin O. It is seemed that the receptors of clostocin O exist in the part of cell wall, especially in newly synthesized cell wall.

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Clostocin O had a remarkable lytic action toward the exponentially growing organisms of Clostridium saccharoperbutylacetonicum No. 8. The cellular lysis was inhibited by addition of heavy metal cations such as Cu²⁺, Ni²⁺, and Cd²⁺, or fradiomycin and RNase, which had been reported to be the inhibitors for lytic enzymes such as some of clostridial phage-endolysin and clostocin O-endolysin. The formalin-treated organisms and antibiotic-treated organisms, of which autolysin activity was inhibited, were also lysed by clostocin O. The induced cellular lysis by clostocin O was thought to be due to the lytic enzyme attached to clostocin O.

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Clostocin O is a phage tail-like bacteriocin produced by Clostridium saccharoperbutyl-acetonicum N1-4. One particle of clostocin O had an activity to kill one sensitive organism. Clostocin O had also the lytic activity, but this lytic activity was not an essential action of clostocin O, because clostocin O was able to show a sufficient killing activity even under the condition to inhibit its lytic activity. The biosynthesis of macromolecules (protein, RNA and DNA) in sensitive organisms was inhibited by clostocin O infection. The amounts of macromolecules of the infected organisms were held at the initial level.

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The structure of a new neolignan, piperenone, isolated as an insect antifeeding substance from Piper futokadzura Sieb. et Zucc. has been determined on the basis of chemical and spectral evidence. It is shown to be 5-allyl-3a, 7a-dimethoxy-2-(3, 4-dimethoxyphenyl)-3-methyl-2, 3, 3a, 6, 7, 7a-hexahydro-6-oxobenzofuran (I). Both the absolute configurations at C-2 and C-3, two of four asymmetric carbons in I are deduced to be S-arrangements.

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The metabolic fate of the carbon skeleton of L-serine-U-¹⁴C has been investigated, in vivo and in vitro, in growing rats and chicks fed the diets with various protein calories percents (PC%) at 410 kcal of metabolizable energy.
The incorporation of ¹⁴C into body protein at 12 hr after the injection of serine-¹⁴C was about 49% of the injected dose in rats fed the 10 or 15 PC% diet, though the value was reduced in rats fed lower and higher protein diets. The ¹⁴CO₂ production was smaller in rats fed the 10 and 15 PC% diet, and it showed an inverse pattern to that of the ¹⁴C incorporation into body protein. Urinary excretion of ¹⁴C was higher in rats fed 10 and higher PC% diets, whose growth rate and net body protein retention were maximum.
In contrast to the case of rats, the incorporation of ¹⁴C into body protein of chicks at 6 hr after the injection was rather reduced in the 15 PC% group. The proportion of ¹⁴C excreted as uric acid was remarkably increased above the 10 PC% group, and about 19% of the injected dose was recovered in the 50 PC% group.
The catabolic rate of serine in the liver slices of rats and chicks was increased by high protein diets.
These results support the concept that the nutritional significance of metabolism of the carbon skeleton of serine in growing rats and chicks is different from each other, especially at high protein diets.

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2-Arylamino-4-fluoroarylthiazoles have been synthesized by the condensation of fluoroketones with arylthioureas in presence of iodine. The resulting compounds were mercurated with mercuric acetate. 2-Arylideneamino-4-fluoroarylthiazoles and 2-arylsulfonamido-4-fluoroaryl-5-H/alkyl thiazoles have been synthesised by the condensation of 2-amino-4-fluoro-aryl-5-H/alkylthiazoles with aldehydes and halogenated benzenesulfonyl chlorides respectively. 2-Acetamido-4-fluoroaryl-5-aryloxythiazoles have been synthesized from 2-acetamido-4-fluoroaryl-5-bromothiazoles and sodium salts of phenols. In all sixty two new fluorinated heterocyclic compounds have been prepared. Out of these thirty six were screened against Aspergillus niger and Aspergillus flavus for their antifungal activity. Most of them showed promising results.

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4, 6-Dichloro-s-triazinyl Duolite A7 (CC-A7) as a carrier of immobilized enzymes was studied.
The reactivity of the second and the third organic chlorines of s-triazines attached to the carrier toward amino acids or proteins was investigated. The results show that CC-A7 can bind a large quantity of amino acids or proteins and these compounds seem to be bound covalently via the second organic chlorine of s-triazine of the carrier under the usual condition.
It is also shown that this carrier can bind a sufficient amount of protein and enzyme activity of aminoacylase and retains a high activity in column operation.
These results indicate that 4, 6-dichloro-s-triazinyl Duolite A7 is a convenient and useful carrier for the industrial application of immobilized enzyme.

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Monomer solution containing enzyme was frozen to a small bead shape by cool solvent and polymerized by Co-60 gamma ray with a dose of 125_??_850 Krad.
Thus, bead shaped immobilized enzymes with various diameters (0.02_??_15mm) were obtained.
The bead had spongy structure and large surface area. Consequently, even large sized bead shaped immobilized enzyme showed high enzymic activity.
Retained activity of bead shaped immobilized invertase was 16_??_73% depending on bead size.
Bead shaped lipase (1mm diameter) showed 13.5_??_42.5% of retained activity depending on monomer combinations.

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Invertase was immobilized by rediocopolymerization of some synthetic monomers which were mixed in various combinations. Irradiation was conducted aerobically while the mixture of monomers and enzyme was frozen. Retained activity was 51_??_76%. Immobilized invertase shifted its optimum pH by about 0.7 to the acidic site.
The optimum reaction temperature of enzyme beacame a little higher (Ca 5°C) by immobilization. Heat stability was improved by immobilization. Release of fixed enzyme was found to be considerably low (1.2_??_4.1%) and release of several immobilized proteins decreased as the molecular weight increased.

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Cathepsin B [EC 3. 4. 22. 1] from the liver of squid, Dorytheuthis bleekri, was purified by the following steps: homogenization, ammonium sulfate fractionation, CM-cellulose column chromatography and rechromatography, Sephadex G-75 column chromatography and isoelectric focusing. The purified enzyme appears to be homogeneous by polyacrylamide gel electrophoresis at both pH 4.3 and pH 9.5 and has an isoelectric point of 6.8. The molecular weight of the enzyme was determined to be 13, 600 by Sephadex G-75 column chromatography. The optimum pH for hydrolysis of α-N-benzoyl-L-argininamide (BAA), α-N-benzoyl-DL-arginine-2-naphthylamide (BANA) was 4.5, and it was 4.2_??_4.7 for N, N'-dimethyl hemoglobin. The value of Km for the hydrolysis of BANA was estimated to be 1.25mM.

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The amylase inhibitor produced by Streptomyces sp. consisted of, at least, three kinds of inhibiting materials (inhibitor A, B and C). These inhibitors were separated by the chromatographic techniques on Amberlite CG-120 and Dowex 1×2, and by paper chromatography etc.
Inhibitor A, B and C seemed to contain both carbohydrates and amino acids, but the specific activities (I. U./mg glucose) of the inhibitors A, B and C were different, that is, 18, 800, 45, 700 and 59, 000, respectively. Also their ratios of amino acids to neutral sugars (expressed by moles of leucine per those of glucose) were approximately 0.09 (inhibitor A), 2.77 (inhibitor B) and 0.58 (inhibitor C).

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On incubation with rat liver microsomes plus NADPH, Denmert (S-n-butyl S'-p-tert-butylbenzyl N-3-pyridyldithiocarbonimidate) underwent at least two different types of oxidation; hydroxylation and sulfoxidation. Hydroxylation of Denmert at the tert-butyl group was one of the major metabolic attacks in mammalian metabolism. Sulfoxidation of Denmert gave two isomers of Denmert sulfoxides which were intermediates in the metabolism and readily transformed into 2-(3'-pyridylimino)-4-carboxylthiazolidine (HM) in the presence of L-cysteine without enzymatic mediation. This type of conjugation with cysteine appears to be a new class of metabolic reactions in mammals. Denmert S-oxide showed increased fungicidal activity when assayed in liquid cultures, but not on plant leaves.

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Three acidic growth inhibitors were isolated from leaves of peanut and were identified as ferulic acid, p-cumaric acid and salicylic acid, respectively.

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A selective extraction procedure of the diapause hormone-B (DH-B) from male silkworm adult heads is described. By this new method a highly active extract (1 DH unit in 30_??_60μg) can be obtained easily without chromatographic purification. It was further purified by successive gel permeation chromatographies to give finally pure DH-B having an activity of 1 DH unit in 2 μg.

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Antitumor effect of Lipopolysaccharide (LPS) from Proteus mirabilis RMS-203 was evaluated.
Minimal effective doses detected by the Pigment test of LPS from both wild type and heptoseless mutant against solid-type Ehrlich carcinoma was 0.005mg/kg given by either intravenous or intraperitoneal administration. Besides Ehrlich carcinoma, solid-type tumors, such as Sarcomas 180 and 37, Meningeal sarcoma MS-147, Walker carcinosarcoma 256 and Adenocarcinoma 755, were also sensitive to LPS, but several other solid-type tumors, such as Yoshida sarcoma and C3H spontaneous sarcoma, as well as all ascites form of tumors were insensitive.
Ehrlich carcinoma and Sarcomas 180 and 37 in mice were completely regressed, but other three types of tumors sensitive to LPS by the Pigment test were not cured because of regrowth of the tumors.
The blockage of blood stream into the tumor tissue after administration of LPS to the mice bearing solid-type tumors was clearly demonstrated by exclusion (‘Lock out’) or by defect in exudation (‘Lock in’) from the tumor tissues of the injected materials. Using this action of LPS, the mice bearing Meningeal sarcoma MS-147 or Walker carcinosarcoma 256 was successfully treated with combined use of LPS and a small amount of antitumor agent, such as 6-mercaptopurine or cyclophosphamide.

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Production of fairly stable radical intermediates was detected in an early stage of reaction of DHA with primary amines in ethanol. These intermediates showed characteristic hyperfine esr signals apparently different from those formed by reaction of DHA with amino acids. These were separated as blue spots on tlc plates, and their Rf values suggested presence of amine residues in their molecules. Esr examination of aqueous extracts showed their common
basic compositions as of 2 units of DHA and 1 unit of amines. In the case of reaction of secondary or tertiary amines with DHA, no characteristic esr spectra could be detected other than the signal probably due to the radical product by the reduction of DHA.

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The aggregation behavior was investigated by the method presented in the previous paper, with the glutens separated from ten kinds of wheat flours different in the mixing property. It was shown that the rate constant of aggregation, k, increased in the order of the weak, medium and strong flour. This is an additional evidence for the correlation between the aggregation behavior of the separated gluten and the mixing property of the flour, which was suggested in the previous paper with four kinds of flours. The aggregation behaviors of glutenin and gliadin were also investigated after fractionation of gluten. A good correlation was observed between the τ₁₀/C values of gluten and glutenin separated from it, showing that the difference of the aggregation behavior of gluten is mainly due to the nature of glutenin.

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The specificity of acid proteases from a strain of Scytalidium lignicolum was investigated using oxidized insulin B-chain.
These enzymes were considerably specific for the bonds involving aromatic or hydrophobic amino acid residues. These specificities were similar to those of usual acid proteases. However, acid proteases A-1 and A-2 specifically hydrolyzed Cys (7)-Gly (8) bond, and acid protease B Tyr (26)-Thr (27) bond; these peptide bonds were not attacked by any acid proteases ever studied.
As expected from the results of investigation using various synthetic peptides, acid proteases A-1, A-2, and B thus exhibited distinguishable specificity on the oxidized B-chain with the usual microbial acid proteases which were inactivated by specific inhibitors such as S-PI, DAN, and EPNP.

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A cell-free protein synthesizing system was obtained for the first time from the pollen of Pinus densiflora. This system consisted of pollen ribosomes and partially purified pollen polypeptide elongation factors. Procedures were described for the complete resolution and partial purification of the two elongation factors (EF-1 and EF-2). One of the factors, EF-1, catalyzed a GTP-dependent binding of aminoacyl-tRNA to ribosomes. The second factor EF-2 catalyzed the polyphenylalanine synthesis in the presence of EF-1. Complete exchangeability was observed between pollen EF-1 and the silk gland EF-1, or between pollen EF-2 and the silk gland EF-2. Pollen EF-1 was heterogeneous in size; i. e. multiple forms. A larger form of EF-1 had a molecular weight of about 160, 000 and the smaller one had molecular weight of about 60, 000. The former was converted into the low molecular weight form in the process of germination and purification.

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