Two bacterial strains capable of growth on methanol, methylamine, formaldehyde and formate, as sole sources for carbon and energy have been isolated and designated as Pseudo-monas 1 and Pseudomonas 135. These bacteria are gram-negative, motile rods. The mean generation time of Pseudomonas 1 is 4 hr on 0.3 % methanol at 32°C, and that of Pseudomonas 135 is 5 hr. Extracts of Pseudomonas 1 and Pseudomonas 135 possess activities of methanol, formaldehyde and formate dehydrogenases when grown on methanol; and methylamine de-hydrogenase when grown on methylamine. While no hexulose phosphate synthase activities were detected in extracts of these bacteria, high activities of hydroxypyruvate reductase and methylene tetrahydrofolate dehydrogenase were found indicating the assimilation of C₁-compounds via the serine pathway and not via the ribulose monophosphate pathway.

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The number of extracellular and intracellular C_x enzyme components and their interrelations were studied using enzymes from strains, N-1 (H 373) and C-3 (N 87), of P. oryzae.
When P. oryzae was grown on carboxymethyl cellulose (CMC) as the carbon source, high activities were detected in C_x enzyme and β-glucosidase, with essentially no C, enzyme activity. Purification of extracellular and intracellular C_x enzymes showed that the former consisted of three components (Ex-I, Ex-II and Ex-III) and the latter of two components (In-I and In-II).
When the culture filtrate was incubated for 11 days at 28°C under sterile condition, the amount of component Ex-III, which was undetectable within the cell, increased with the concomitant decrease of that of component Ex-II. Thus, component Ex-II was suggested as converting to component Ex-III.

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One hundred and four acid amido compounds have been prepared by condensing 2-mercapto benzoic acid and its 5-bromo, 3, 5-dibromo and 3, 5-dichloro derivatives with 2-amino-4-substituted thiazoles and their 5-halogenated derivatives via the acid chloride. All these compounds have been tested against Helminthosporium. Four of the compounds were tested for their antibacterial, antihelminthic activities and also for their hypoglycemic activity.

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To investigate the aroma components characteristic of spring green tea, analysis of the aroma concentrates of green tea and fresh tea-leaves was accomplished. The research on the changes in aroma constituents of spring green tea and the aroma concentrate from fresh tealeaves during storage, showed that cis-3-hexenylhexanoate and cis-3-hexenyl-traps-2-hexenoate contributed to the typical fresh aroma of spring green tea. Twelve isomers of hexenol esters were synthesized for comparison of the fresh green note.

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The linear correlation was found by the stepwise multiple regression analysis between the sensory test of soy sauce flavor and the gas chromatographic (GLC) data transformed with arcsine and logarithm. GLC data will possibly be used for objective evaluation of soy sauce flavor. A multiple correlation coefficient or of a determination coefficient of more than 0.9 was respectively obtained at the 5 th or 10 th of 47 steps. The fact that the minimum standard error of an estimate was found at the 24 th step suggests the importance of selecting proper peaks from the whole gas chromatogram. High estimated accuracy was acquired by application of GLC data to the calculated multiple regression model.

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Two distinct forms of β-glucosidase, A and B, were found to occur in the cells of Pseudomonasfluorescens var. cellulosa: A was membrane-bound, while B cytosolic. They differed also from each other in some properties, such as molecular size, kinetic parameters, and susceptibility to various compounds. β-Glucosidase B was partially purified and studied especially of its substrate specificity. The results indicated that it may be an atypical β-glucosidase which possesses a certain character of exo-cellulase.

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3Z-Hexenal, 2E-hexenal (leaf aldehyde) and 1-undecene were first found in Farfugium japonicum Kitamura (japanese silver) leaves. On blending the leaves in the presence of oxygen, 2E-hexenal was enzymatically generated via a similar biosynthetic pathway to that in Thea sinensis leaves: via 3Z-hexenal from linolenic acid. However, unlike Thea sinensis leaves neither 3Z-hexenol (leaf alcohol) nor 2E-hexenol was found in F. japonicum leaves.

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A mutant (RAN 1) which produced extracellular soluble alkaline phosphatase was isolated from B. subtilis 6160-BC6 that constitutively produced phosphate-repressible alkaline phosphatase. The mutant produced the enzyme after the cell growth reached maximum and most of the enzyme produced was secreted into the culture medium at every culture period tested. The distribution of the enzyme in culture fluids and in cells of the mutant was quite similar to that of α-amylase while alkaline phosphatase in B. subtilis 6160-BC6 was particulate and located mainly in cell membrane.

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L-Leucine-α-ketoglutarate (α-KGA) transaminase from Acetobacter suboxydans was purified to the state of homogeneity by the criteria of ultracentrifugation and electrophoresis on a cellulose acetate membrane. The molecular weight was about 80, 000 and one mole of pyridoxal 5'-phosphate was bound per mole of enzyme as a coenzyme. The enzyme exhibited absorption maxima at 280, 337 and 414 nm.
The branched-chain amino acids and αa-KGA were specific as amino donors and an acceptor. L-Leucine-α-KGA transaminase is suggested to correspond to the enzyme so-called Transaminase B.

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By treatment with EDTA, all transport reactions tested with succinate-grown cells of Agrobacterium tumefaciens were simultaneously reduced; the tested transports were mannose, fructose, glucose, ribose, rhamnose, leucine and proline entry reactions. To the EDTA-treated cells, a supplement of D-glucoside 3-dehydrogenase (G3DH) induced the simultaneous restoration in all of the entry transports, which were previously reduced, with different degrees in different transport reactions. The functions of G3DH for transport reaction was discussed. The conclusion was that G3DH is a common factor having two functions: (1) a member of the energy-supplying system which is selectively coupled with transport reactions, and (2) an effector on maintenance of membrane structure being proper status for transport reactions.

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As one of the steps to elucidate the induction mechanism of encephalomalacia by feeding dilauryl succinate (DLS), possibility on inhibition of intestinal absorption of vitamin E was investigated from four points of view by checking the digestive and absorptive ability of the chicks fed DLS. 1) It was observed that preventive effect of injected vitamin E against encephalomalacia was about four times higher than that orally administered, while that of injected ethoxyquin, one of synthetic anti-oxidants, was much lower than that orally administered. 2) Increase in weights of the duodenum, jejunum, and liver was caused by feeding DLS or lauryl alcohol, i.e., one of hydrolysates of DLS, and that by the latter was significantly larger than that by the former. 3) Apparent digestibility of fat and protein of the chicks fed DLS was normal. 4) Sodium taurocholate had little effect to protect chicks from encephalomalacia. It was suggested that DLS feeding caused the change in weight of the tissues related to digestion, but did not affect the digestive function of chicks. Therefore, vitamin E deficiency induced by DLS might not be attributable to inhibition of intestinal absorption of vitamin E. Relationship among DLS, unsaturated fatty acids, and selenium is discussed in the text from the point of effectiveness of anti-oxidants.

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We have previously reported the antitumor activity of some streptovaricin C derivatives selected for their ability to inhibit preferentially the growth of virus transformed cells and human derived cancer cells in vitro at relatively low concentrations. n-Butyl ether, 2-thenoyl-methyl ether and the oxime of acetonyl ether derived from damavaricin C showed antitumor activity against ascites and solid Sarcoma 180 in vivo.

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In a continuing study of the enzymes involved in the 2, 3-diphosphoglycerate metabolism of mammalian erythrocytes, we report that 2, 3-diphosphoglycerate in pig erythrocytes is probably metabolized by one multifunctional enzyme.
Diphosphoglyceromutase, 2, 3-diphosphoglycerate phosphatase, and phosphoglyceromutase were simultaneously purified from pig erythrocytes. Three fractions (peaks I, II, and III) which had all three activities in different ratios were obtained by column chromatography. Peak I was extremely rich in phosphoglyceromutase, containing more than 90% of the total activity of this enzyme. In contrast, peak III was active in metabolizing 2, 3-diphosphogly-cerate, containing about 95 % of both the diphosphoglyceromutase and the 2, 3-diphosphogly-cerate phosphatase activity. It seems likely that peak I functions as phosphoglyceromutase and that peak III functions in the 2, 3-diphosphoglycerate metabolism.
The homogeneity of peak III was established by disc gel electrophoresis in the presence and the absence of sodium dodecyl sulfate, as well as by ultracentrifugation. The three activities of peak III were lost at the same rate on thermal inactivation. These results indicate that the two enzyme activities for metabolizing 2, 3-diphosphoglycerate, which were believed to be due to different proteins, are attributable to one protein, along with some phosphogly-ceromutase activity. The diphosphoglyceromutase activity of this protein was inhibited by the product, 2, 3-diphosphoglycerate, as well as by hydroxypyruvate phosphate, 2-phosphogly-colate, inorganic phosphate, and bisulfite. The 2, 3-diphosphoglycerate phosphatase activity was enhanced by 2-phosphoglycolate and hydoxypyruvate phosphate, and by the coexistence of chlorine ion and bisulfite, while it was inhibited by monophosphoglycerates. The native peak III protein had a molecular weight of 59, 000 as determined by equilibrium centrifugation. Disc gel electrophoresis in the presence of sodium dodecylsulfate yielded a single protein band with a molecular weight of 28, 000, indicating that this protein was composed of two subunits with a similar molecular weight.
Occurrence of multifunctional proteins in pig erythrocytes is compared with that in human erythrocytes from the standpoint of the universal existence of such proteins.

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The solution conformations of protodestruxin, desmethyldestruxin B and destruxin B, insecticidal cyclodepsipeptides, in DMSO-d₆ have been studied through the use of 100-MHz proton NMR spectroscopy. Proton-deuteron exchange and temperature dependence of the peptide proton chemical shift were used to delineate peptide protons in terms of exposure to solvent. The results defined the β-alanine and isoleucine peptide protons of desmethylde-struxin B and destruxin B as solvent-shielded. A rigid conformation of desmethyldestruxin B and destruxin B, in which there are two hydrogen-bonded rings of ten atoms, is proposed. Protodestruxin seems to have a mobile conformation in DMSO-d₆. The results indicate that the biological N-methylation of the alanine peptide proton in protodestruxin decreases the number of possible conformations which are stabilized by intramolecular hydrogen bonding and, as the result, fixes the molecule in a specific conformation.

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Strawberry seeds are shown to contain at least two alcohol dehydrogenases; one is NAD specific and reacts with ethanol and allyl alcohol, and the other is NADP specific and reacts with benzyl alcohol and geraniol. These two alcohol dehydrogenases were distinguished on disc electrophoresis. Their properties were different each other in ammonium sulfate fractionation, optimum reaction pH and thermostability.

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Thermonsenstivie division mutants were derived from Bacillus subtilis Marburg 168 thy trp₂ by means of membrane filtration after nitrosoguanidine mutagenesis. Among them, ts 42 requiring uracil for normal growth at 48°C was investigated.
In the absence of uracil, the mutant cells grew normally at 37°C and stopped dividing after temperature shift to 48°C resulting in filaments of two to four times length of normal rods. The total cell number after temperature shift from 37 to 48°C, increased two to three fold in 90min and remained constant thereafter. The viable count after the temperature shift to 48°C, increased 1.5 to 2 fold in initial 60min and then decreased exponentially. A rapid restoration of colony forming ability was shown when the mutant cells were shifted back to the permissive temperature after 120 to 180min of incubation at 48°C or when uracil was introduced to the culture at 48°C. This recovery of viability was partly observed even in the presence of chloramphenicol. The synthesis of RNA of this mutant was shown to decline 20min after the temperature shift to 48°C whereas the syntheses of DNA and protein proceeded for more than 80min at that temperature.
No newly isolated uracil requiring mutants formed filaments in the medium lacking uracil or showed growth pattern like ts 42.

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A thermosensitive uracil requiring mutant of Bacillus subtilis Marburg 168 thy trp₂ ts 42 was examined as to the colony forming ability at the permissive and nonpermissive temperatures. The viability of the mutant cells decreased rapidly at the restrictive temperature in the modified Woese's (MW) medium. However, the cells retained viability when sodium succinate or potassium chloride was added to the medium at that temperature although uracil deficiency was unchanged. A little but significant incorporation of adenine-8-¹⁴C into RNA still con-tinued even after the incorporation of N-acetyl-³H-D-glucosamine into acid insoluble fraction of the cells terminated in the MW medium at 48°C. Both incorporations as well as increase of absorbance were slowed down in the presence of sodium succinate at 48°C. This mutant, ts42, was more sensitive to deoxycholate (DOC) than the parent strain. The restoration of colony forming ability after the temperature shift back from 48 to 37°C was suppressed by the addition of DOC to the medium. However, the cell became resistant to DOC when uracil was added to the medium prior to the temperature shift.

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(1) Both glutaminases A and B of Pseudomonas aeruginosa are inactivated by urea and guanidine hydrochloride, and the activities are partially restored by removal of the denaturants, while sodium lauryl sulfate denatured irreversibly the isozymes. (2) Glutaminase A consists of 4 identical subunits (mol. wt., 35, 000) and B is composed of one polypeptide chain (mol. wt., 67, 000). (3) Glutaminase A, which catalyzes the hydrolysis and also the hydroxylamino-lysis of L and D isomers of glutamine and asparagine, does not act on γ-N-substituted glutamine e. g., γ-glutamylhydrazide. Some L- and D-γ-glutamyl derivatives, e. g., L- and D-γr-glutamyl-hydrazide, L- and D-γ-glutamylmethylester, and L-γ-glutamyl-L-alanine are substrates for glutaminase B, which does not catalyze the hydrolysis and hydroxylaminolysis of asparagine. α-Amino adipamic acid and α-amino substituted amino acids are inert for both the isozymes. (4) The acylation step is rate-limiting in the catalytic reactions by both the isozymes.

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A new polyacetylene separated from Solidago altissima L., methyl 10-(2-methyl-2-butenoyl-oxy)-(2Z, 8Z) -2, 8-decadiene-4, 6-diynoate, was first synthesized. With respect to the con-figuration of 2-methyl-2-butenoyloxy moiety, (E) and (Z) esters were prepared from tigloyl chloride and mixed anhydride of angelic acid respectively and the (Z)-configuration of the naturally derived product was confirmed by the synthetic evidence.

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Phosphatidylcholine was associated with soybean protein in two types of interaction. One is due to hydrophobic interaction between a phosphatidylcholine molecule and hydrophobic regions of the protein. The other is due to the binding of phosphatidylcholine lamellae to the protein surface.

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(S)-(-)-Frontalin (I), the natural form of the pheromone of Dendroctonus Bark Beetles, was synthesized from D-glucose.

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A one-step synthesis of dimethyl 3-substituted pyrrole-2, 4-dicarboxylates by the reaction of methyl isocyanoacetate with various aldehydes in the presence of DBU was carried out. Furthermore, the reaction mechanism was investigated; it was found that the methyl α-isocyanoacrylate compound is a key intermediate of the pyrrole compound.

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The enzyme uridine diphosphate N-acetylglucosamine pyrophosphorylase was purified about 330-fold from an extract of baker's yeast by the treatment with protamine sulfate and column chromatographies on DEAF-cellulose, hydroxylapatite and Sephadex G-150. The purified enzyme was proved to be homogeneous by disc gel electrophoresis. The molecular weight was determined to be approximately 37, 000 by gel filtration. The enzyme had an optimum reactivity in the pH range of 7.5_??_8.5 and was stable at 4°C in potassium phosphate buffer, pH 7.5, containing 0.1mM dithiothreitol, but was unstable when stored at -20°C. The addition of dithiothreitol also increased the thermal stability of enzyme. The enzyme was specific for UDP-N-acetylglucosamine as substrate, and none of the other sugar nucleotides could serve as nucleotide substrate. The estimated values of Km were 6.1×10^-3M for UDP-N-acetylglucosamine and 5.0×10^-3M for inorganic pyrophosphate. The enzyme required some divalent cations for activity. Magnesium ion was the most effective among the cations tested. The enzyme activity was highly stimulated by the addition of dithiothreitol or dithioerythritol.

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Chavicol was isolated as a drosophila larva-growth inhibitor from the leaves of Viburnum japonicum Spreng. The inhibitory activities of chavicol and its related compounds against drosophila larvae and adults were examined.

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Six strains of floc-forming bacteria belonging to Flavobacterium were isolated from activated sludge which were deflocculated by Pronase treatment. The flocculated cells of the strain B, one of the isolates, was deflocculated not only by Pronase, but also by ethylenediaminetetraacetate. Growth was stimulated when Pronase was added in the medium. An adequate amount of calcium ion in the medium was required for flocculation. No flocculation was observed, however, when calcium was added to the cells grown with a low level of calcium. Deflocculation was observed at the late stationary phase and the onset of deflocculation depended on the concentrations of calcium in the medium. The higher concentrations delayed the deflocculation. The flocs formed in the presence of calcium over 0.5mM in the medium became resistant to the Pronase treatment.

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