Bacterial strains capable of assimilating methanol as the sole source of carbon and energy have been isolated. These bacteria produced phenazine methosulfate-dependent methanol dehydrogenase (primary alcohol dehydrogenase, EC 1.1.99.8). The enzyme activity was detected in electrophoresis on polyacrylamide gels by activity staining technique. The dehydro-genase can be grouped to at least three types based on their mobility in electrophoresis: (1) enzyme which migrates toward the anode slowly, (2) enzyme which remains at the top of the gel at pH 9.3 and (3) enzyme which migrates to the anode at faster mobility. The three bacteria were selected as producers of the respective three types of dehydrogenase. They were facultative methanol-utilizing bacteria and two of them were identified, one as Pseudomonas sp. No. 2941 and the other as Pseudomonas sp. S25. Most isolated bacteria produced the first type of enzyme and several strains produced the second type of enzyme. Strain. S50 was the only strain which produced the third type of enzyme. These groupings of methanol dehydrogenase may contribute to the classification of methylotrophic bacteria.

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α, β-Unsaturated aldehydes are formed by the aldol condensation reaction and lipic oxidation in various foods. The present investigation was undertaken to elucidate the initia reaction mechanism of crotonaldehyde (CA) or 2-methyl-2-pentenal (MP) with the amine group of glycine (Gly). The results are interpreted by assuming that the initial reaction of the CA-Gly system consists of competitive and/or consecutive pseudo first-order reactions as follows;
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The rate of the reaction of the MP-Gly system is very small compared with that of the CA-Gly system. The reactivity of α, β-unsaturated aldimine was discussed.

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A shaking apparatus that provides efficient aeration for test tube cultures is described. The shaking apparatus is characterized in striking the test tubes at a lower portion with reciprocally moving bars. The shaking efficiency was demonstrated to be higher than that of a commercial shaker in both oxygen absorption coefficient and sorbose production rate from sorbitol by Acetobacter suboxydans.

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The formation of rhodanese (EC 2.8.1.1) by mycelia of Trametes sanguinea was stimulated by glycylglycine (Gly•Gly). The Gly•Gly added to the medium containing sucrose was slowly metabolized, whereas the glycine added instead of Gly•Gly was metabolized more rapidly with less formation of rhodanese. The rhodanese formation was inhibited by 2, 4-dinitro-phenol, antimycin A, potassium cyanide, and cycloheximide, and was significantly depressed by several surface active agents. The fungal mycelia incubated with sucrose and Gly•Gly were; less active in isocitrate lyase and more active in NAD- and NADP-linked isocitrate dehydrogenase activities than those incubated with glucose and Gly•Gly. The results are discussed in regard to the mechanism of the rhodanese formation by the fungal mycelia.

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Anaerobic deamination of cyclohexylamine (CHA) by the intestinal microorganisms of rabbits was studied.
1. Urinary excretion of cyclohexanol (CHnol) in rabbits receiving CHA orally was suppressed by the oral administration of neomycin or metronidazole, although the effects were considerably different in individuals.
2. When CHA was incubated anaerobically with the cecal contents of rabbits receiving CHA in drinking water, the deaminated metabolites, cyclohexanone and CHnol were accumulated to various extents. Furthermore, cyclohexanone was converted to CHnol which was further metabolized to other compounds in the cecal contents.
3. Streptococcus faecalis, Escherichia coli, Clostridium sp. and Bacillus sp. were isolated from the cecal contents, and possessed the abilities to deaminate CHA or desulfate cyclamate anaerobically. These activities were maintained on subculturing and in the dried cells.

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To clarify the mechanism of electrochemical insolubilization of DNA, metal contents combined to DNA were examined, and the binding manner of the metals to DNA was pre-sumed from the sedimentation coefficient and ultraviolet absorption spectra of DNA insolu-bilized by electrolysis.
Copper content increased with an increase in current passed and decreased with an increase in temperature of DNA solution, but was not affected by pH.
The sedimentation coefficient revealed a slight cleavage of DNA.
The ultraviolet absorption maxima shifted to a shorter wavelength with an increase in copper content, and to the longer side with an increase in temperature of DNA solution.
On the other hand, nickel showed a similar behavior to copper except that their combined content was affected by pH. Iron content was only a paucity compared with the amounts of copper and nickel.
From these results, it was presumed that copper combined with the phosphate group of DNA when Cu-DNA was prepared from native DNA and with the bases of that from denatured DNA. In addition, it seemed that the binding manner of metals to DNA differed from one another with the kind of metal.

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A debranching enzyme was purified about 100-fold over the crude enzyme solution from non-glutinous rice seeds at the milky stage by using DEAE-, CM-cellulose, and then Sephadex G-100 column chromatography. This disc-electrophoretically homogeneous enzyme showed a specific activity of 16.5 pullulanase units/mg of protein (25°C) with its optimum pH at 5.6. The enzyme debranched the α-1, 6-linkages in pullulan or an α-amylolysis product from starch most favorably, several β-limit dextrins from starch or from amylopectin rapidly, and phytoglycogen β-limit dextrin and amylopectin moderately, while it was unable to debranch phytoglycogen. These substrate specificities were similar to those of the so-called “limit de0x-trinases” already reported with respect to several plant sources.

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Various fatty acids, some of which originally do not exist in the cell, were intactly incorpo-rated into cellular lipids, such as triglyceride and phosphatidylcholine, by a sake yeast, Kyokai No. 7 (K-7). Formation of flavor esters, such as isoamyl acetate, was linearly correlated to the melting points of the incorporated fatty acids, ranging from -20°C to 40°C, by semilogarithmic plotting.
The relation between isoamyl acetate formation and percentage of linoleic acid in cellular fatty acids was inverse proportion.
Clotrimazole stimulated the formation of isoamyl acetate by suppressed K-7 cells grown on linoleic acid.
It was ascertained that isoamyl acetate was biosynthesized inside the K-7 cell and excreted outside through cell membranes, and hence it was presumed that the formation of flavor esters by sake yeast was affected by some changes in permeability of cell membranes.

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Guanidinoacetate amidinohydrolase (glycocyaminase, EC 3.5.3.2) was purified 148-fold and crystallized from Flavobacterium sp. GE-1 grown in a medium supplemented with guanidinoacetate. The molecular weight of the enzyme was estimated to be about 281, 000 daltons by a gel electrophoretic procedure. SDS-polyacrylamide electrophoresis showed a single component of subunit with an estimated molecular weight of 70, 000 daltons, suggesting that the enzyme is composed of four subunits identical in molecular weight. These values are significantly distinct from those of an analogous enzyme from a Pseudomonas reported by us previously. The native enzyme was active without added divalent metals. Incubation of the enzyme with 1, 10-phenanthroline at an elevated temperature (50°C) resulted in almost complete inactivation. The activity of the inactivated enzyme was restored by incubation with Zn²⁺ or Co²⁺ at 50°C; the former cation was more effective. The enzyme was optimally active at pH 8.0_??_8.5. The apparent Km value for guanidinoacetate was 15 mat. 3-Guanidinopropionate and 4-guanidinobutyrate were hydrolyzed 19% and 9%, respectively, as fast as guanidinoacetate. PCMB was a potent inhibitor of the enzyme. The properties of the Flavobacterium enzyme were compared with those of the analogous Pseudomonas enzyme.

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The role of individual amino acid in diet was examined in regulation of some amino acid-catabolizing enzymes in the liver of protein-depleted rats. Feeding rats with a mixture containing 18 amino acids in equimolar proportion produced marked increase in activity of all enzymes examined; serine dehydratase (EC 4.2.1.13), glutamic-oxaloacetic transaminase (GOT, EC 2.6.1.1), branched-chain α-ketoacid dehydrogenase (BCADH, EC 2.6.1.42), gluta-mic-pyruvic transaminase (GPT, EC 2.6.1.2) and histidase (EC 4.3.1.3). The induction of serine dehydratase, GOT and histidase by complete amino acids mixture(CAA) could essentially be attributed to the action of tryptophan. The removal of non-essential amino acid mixture (NEAA) from the CAA in the diet caused enhancement of increase in the activity of serine dehydratase, GOT and GPT whilst it caused no further increase in that of both BCADH and histidase. These results suggest that NEAA in diet may inhibit the induction of some amino acid-catabolizing enzymes by essential amino acids (tryptophan and methionine). The results are discussed in relation to the nutritional role of non-essential amino acids.

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The diterpenes from the plants of the genus Isodon (Labiatae) exhibited a relatively specific growth inhibitory activity against lepidopterous larvae. The α-methylene cyclopentanone moiety (D-ring) was essential for this activity. In addition, these compounds strongly inhibited the oxidative phosphorylation in mitochondria isolated from silkworm midgut.

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5-Keto-n-gluconate reductase (EC 1.1.1.69) was purified and crystallized for the first time from cell-free extract of Gluconobacter suboxydans IFO 12528. Purification of the enzyme was successfully performed by column chromatography on DEAE-Sephadex A-50, bluedextran Sepharose 4B, followed by pH gradient chromatography on DEAE-Sephadex A-50. The enzyme was purified about 1200-fold with an overall yield of 40%. The enzyme was much stabilized against heating and storage by adding either D-gluconate or 5-keto-n-gluconate. 2-Keto-D-gluconate had no effect to stabilize the enzyme and the enzyme was confirmed to be a different entity from 2-keto-D-gluconate reductase stabilized by gluconate or 2-keto-n-gluconate but not 5-keto-D-gluconate. It was also confirmed with crystalline enzyme that 5-keto-D-gluconate reductase is considered to have a function to reduce intracellular 5-keto-D-gluconate to D-gluconate in combination with regeneration of NADP.

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A β-galactosidase producing strain was isolated. It belongs to the genus Bacillus from its morphological characteristics but it grows better in alkaline media. This strain produced a β-galactosidase inducibly and the induction of the enzyme was faster at pH 10.2 than at pH 7.2. At pH 8.5, intact cells exhibited higher enzyme activity than toluenized cells. At 60°C, pH 10.2 grown cells exhibited relatively higher enzyme activity than pH 7.6 grown cells. This difference in stabilization of the enzyme may be explained by the difference of outer surface under different culture conditions.

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Pulsed NMR was applied to measure the spin-lattice(T₁) and spin-spin(T₂) relaxation times of water protons in myosin suspension with or without sucrose during the course of its dehydration. T_l relaxation time curves showed two phase behaviour at the high moisture contents. Plots of T₁ as a function of moisture content showed minima in the multilayer region (in the range of 18.5_??_24% moisture contents), irrespective of the presence or absence of sucrose in the system. On the other hand, irrespective of the presence or absence of sucrose, T₂ relaxation time curves of myosin containing more than 89_??_91% of water exhibited a three phase behaviour, indicating the existence of three water fractions of different mobility (T_2-A, T_2-B and T_2-C). The amount of water in the bound fraction (T_2-B+T_{2_??_C}) showed a consistency until the free water fraction (T_{2_??_A}) is almost removed. T₂ values of three water fractions were found to decrease with a decrease in the moisture content, and these values as well as the populations of three classes of water protons did not show the significant difference between samples in the presence and absence of sucrose throughout the course of dehydration process.

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Proton NMR spectra of peracetylated methyl and phenyl glycosides of n-xylopyranose and L-arabinopyranose were measured in the presence of a europium shift reagent. The signals of H-2, H-3, and H-4 were more markedly shifted than others by the addition of the reagent and, in particular, the bound chemical shifts for H-4 in the α-D- and β-L-anomers and for H-3 in the β-D- and α-L-anomers were shown to be pronouncedly greater than those in the corresponding hexopyranosides. The lanthanide-induced shifts of most protons in equimolar mixtures of two sugars were also measured, and preferential coordination of the β-D-xylo-and β-L-arabinopyranosides with the europium was found to occur in comparison with the coexisting a-anomers or the corresponding hexopyranosides.

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The properties of curdlan gel were investigated by determining the theological behavior, the melting temperature, and the syneresis of gel. The modulus of elasticity of curdlan gel was found to be affected by the average molecular weight, pH, and the measuring temperature. The gel, which was set by heating the aqueous suspension of curdlan at 65°C and then cooling it below 40°C, melted when the gel was heated at 65°C. The melting energy of this gel was calculated to be about 25 Kcal/mol.

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Regulatory properties of the arginine biosynthetic enzymes in the L-arginine producers of Corynebacterium glutainicum were examined. D-Serine-sensitive L-arginine producer, DSS-8, had derepressed level of L-arginine biosynthetic enzymes, but had an N-acetylglutamokinase, the key enzyme of L-arginine biosynthetic pathway, sensitive to the feedback inhibition by L-arginine. D-Serine-sensitive and D-arginine-resistant L-arginine producer, KY10479, had an N-acetylglutamokinase released from the feedback inhibition by L-arginine.
Implications of these results are discussed in relation to L-arginine productivity, D-serine sensitivity, L-arginine analog resistance and regulation of L-arginine biosynthesis in these mutants.

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Methoxy-1, 4-benzoquinone and methyl gallate were isolated as cytokinin-induced betacyanin synthesis inhibitors from the fruits of Diospyrous kaki L. The inhibitory activities of these active components and their related compounds against Amaranthus caudatus L. were examined.

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Syntheses of navenones A, B, and C, trai-breaking alarm Pheromones of a sea slug, Navanas inermis, are described.

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The addition of trialkyltin chloride (TPT) to washed cell suspensions of E. coli and B. subtilis resulted in a decrease of 660 nm absorbance. The decrease was more marked with the higher TPT concentration. In the case of B. subtilis, cell lysis was found with microscopic observation. The decrease was dependent on the temperature of the reaction mixture; it was observed only at 30°C, however, it was not observed at 0, 60 and 100°C. TPT addition to the cell suspensions also resulted in a rapid release of such constituents as magnesium, protein, sugar and 260 nm absorbing materials. For both bacteria, the leakages of magnesium and protein were more rapid than that of other compounds. Very little leakage of phospholipids and inorganic phosphates was observed. The leakage of cell constituents was completely prevented by divalent cations.

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The fate of [4-¹⁴C]-cholesterol administered intraperitoneally or orally was compared in exercised (treadmill running for 14 days) and sedentary rats.
Plasma triglyceride, phospholipid and cholesterol decreased in exercised rats and this reduction lasted at least for 10 days after exercise was terminated.
When rats received [4-¹⁴C]-cholesterol intraperitoneally or orally, the turnover rate of serum cholesterol was considerably higher in exercised rats at the time shortly after the administration of the label. The radioactivity remaining in the liver was consistently lower in exercised rats, whereas that in extrahepatic tissues was the same between two groups.
Excretion into feces of the label as total steroids was moderately enhanced by exercise. This effect was almost entirely ascribed to the increase in output of the label shortly after the administration.
These results suggest that the mechanism responsible for cholesterol lowering effect of exercise is mainly attributable to the increase in turnover of cholesterol in the hepato-plasmic system. The moderate increase in fecal output of endogenous steroids may be the reflection of the increased turnover.

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A new synthesis of (-)-rotenone (1) has been accomplished. Decarboxylation and Vilsmeier formylation of (-)-tubaic acid (3) afforded (-)-tubaaldehyde (2) which was the key intermediate of (-)-rotenone synthesis. Condensation of 2 and Grignard reagent of phenyl 2-propynyl ether (5) and subsequent oxidation afforded (-)-1-(2, 3-dihydro-4-hydroxy-2-isopropenylbenzo[b]furan-5-yl)-4-(3, 4-dimethoxyphenoxy)-2-butyn-1-one (7). Thermal rearrangement of 7 gave (-)-6a, 12a-dehydrorotenol (8). 8 was smoothly converted to (-)-rotenone (1) in good yield by the treatment with weak bases such as pyridine or triethylamine in ethanol.

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The biological activities of gibberellins^** A₁, A₃, A₄ and A₇ were compared with 1β-iodo-gibberellinAl, 1β-iodogibberellin A₁ methyl ester and 1β-iodogibberellin A₄ in barley aleurone, lettuce hypocotyl and cucumber hypocotyl bioassays. Iodo GA₁ methyl ester was inactive in all three assays. However, both iodo GA₁ and iodo GA₄ showed considerable activity in the lettuce hypocotyl and cucumber hypocotyl assays and moderate activity in the barley aleurone assay. These results are discussed in terms of structure-activity relationships of the GAs. Some possible uses of iodo GAs are indicated.

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Two xyloglucan oligosaccharides were obtained by mild acid hydrolysis and cellulase-degradation from the arabinoxyloglucan, isolated from the midrib of the leaves of Nicotiana tabacum.
The structures of the oligosaccharides were determined by methylation, analyses before and after sodium borohydride reduction and by ¹H- and ¹³C-NMR spectroscopy as follows;
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Intermolecular transglycosylation of cyclodextrin glycosyltransferase from B. megaterium strain No. 5 was examined in the reaction system containing D-galactose and soluble starch. The yield of transfer products to D-galactose were 2 to 3 per cent of those to D-glucose. Four kinds of transfer products to D-galactose as the acceptor were obtained. These compounds were identified to be 1-O-α-D-glucosyl-D-galactose, 3-O-α-D-glucosyl-D-galactose, 2-O-α-D-glucosyl-D-galactose and 4-O-α-D-glucosyl-D-galactose. The transglycosylation to D-galactose was shown to occur at various hydroxyl groups of the acceptor; at Cl-, C3- and C2-(C4-) hydroxyl group in the proportion of 26:10:1.

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The two β-lactam antibiotics produced by Cephalosporium acremonium ATCC 14553, cephalosporin C and penicillin N, have been proposed to be biosynthesized through the peptide, δ-(L-α-aminoadipyl)-L-cysteinyl-D-valine.
Many β-lactam negative mutants were derived from the strain No. 52, which was a more potent producer of the f4-lactam antibiotics than the parent strain, C. acremonium ATCC 14553. Some of them were found to accumulate two sulfur-containing peptides. These compounds were isolated from the culture filtrate of one of the mutants, N-2 and determined to be the dimer of δ-(L-α-aninoadipyl)-L-cysteinyl-D-valine and the S-methylthio derivative of the tripeptide.

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A rapid and convenient procedure for the detection of inhibitors of DNA synthesis using starfish oocytes and sea urchin embryos at the 1-cell stage is described. The procedure is based on the findings that inhibitors of DNA synthesis block mitotic cell division of sea urchin embryos which requires DNA synthesis but do not affect meiotic maturational divisions of starfish oocytes which are independent of DNA synthesis. The efficacy of the method is verified by the isolation of aphidicolin, an inhibitor of DNA synthesis, from the culture filtrate of the fungus Harziella entomophila according to the bioassay procedure proposed in this study.

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