A microorganism which could utilize methanol as the sole source of carbon and excreted a new polysaccharide was isolated from soil. This isolate was a stalked bacterium which reproduced by a budding process, and could grow on only methanol, formaldehyde or methylamines as the carbon source. The most suitable nitrogen source for growth was the ammonium ion. The optimum pH and temperature for growth were about 7.0 and 30°C, respectively. The cell growth was inhibited by blue light irradiation. Amino acid composition and fatty acid composition of the cells and electrophoretic behavior of methanol dehydrogenase were also studied. On the basis of these properties as well as taxonomical studies, the isolate (strain JTS-811) was identified as belonging to the genus Hyphomicrobium. This strain had different characteristics as compared to those described for other Hyphomicrobium isolates. At present, it is difficult to give a specific name to this strain, because classification of hyphomicrobia is not clear.

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An extracellular acidic polysaccharide, hyphomicran, produced by Hyphomicrobium sp. JTS-811 from methanol contained a new monomethyl sugar in nature, which was identified as 1-O-methyl-D-mannose by spectrometry and synthesis. Hyphomicran was found to consist of D-glucose, D-mannose, 2-O-methyl-D-mannose and pyruvic acid residues in the relative proportions of 2:1:1:1. By methylation analysis, pyruvic acid was proved to be linked as a ketal to O-4 and O-6 of a terminal 2-O-methyl-D-mannopyranosyl residue.

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The release of intracellular pullulanase (IP) of Aerobacter aerogenes was investigated to study the production mechanism of extracellular pullulanase (EP). A factor, named inductive effector (IE), which released IP from the cells was found in the culture broth of EP production. For IE formation, an initial medium pH of 9.0 was suitable, and the desirable culture period was 4 days. The IE fraction (M_w < 500, free of pullulanase activity) was purified from the supernatant of the culture broth by ultrafiltration and ion exchange resins. IE acted on living cells of A. aerogenes and released intracellular protease from the cells in shaking cultivation. Cell-bound pullulanase may be released by the proteases. Commercial endoproteases and lysozyme helped to make IP release, however, the mode of their action differed from that of IE.

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The addition of 1, 8-pyrenequinone into the assay system containing rat liver homogenates (S9) caused an approximately 10-fold increase in the mutagenicity of 2-acetylaminofluorene (AAF) in the current Salmonella reversion assay system. Since no chemical reaction between 1, 8-pyrenequinone and AAF was observed, the in vitro effects of 1, 8-pyrenequinone on the metabolisms of AAF with S-9 mix were studied. The enhancement of mutagenicity by 1, 8-pyrenequinone was not dependent on the dose of NADPH under the present assay condition. The mutagenicity of AAF was increased approximately 4-fold by the addition of 1, 8-pyrenequinone into microsomes, whereas it remained at the spontaneous level in the presence of cytosol. However, by reconstituting microsomes with cytosol, the mutagenicity enhancing activity was recovered to the original level. Since 1, 8-pyrenequinone inhibited the AAF hydroxylase activity, chemical analysis of the incubation mixture of AAF was tried. This indicated that a higher amount of unmetabolized AAF remained and higher amounts of 2-aminofluorene and N-hydroxy-2-acetylaminofluorene were accumulated in the presence of 1, 8-pyrenequinone compared with those in the absence of 1, 8-pyrenequinone. From these results, it seems probable that 1, 8-pyrenequinone inhibits C-hydroxylation (the detoxifying pathway) and promotes N-hydroxylation (the activating pathway) as well as deacetylation in the AAF metabolism.

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The heating of 7S globulin caused changes in the intensities, but hardly affected the positions of the peaks and troughs of the second derivative absorption spectra at wavelengths below 270 nm. On the other hand, above 271 nm, changes were reflected both in the intensities and in the positions of peaks and troughs. The difference-second derivative absorption spectra indicated that 60 and 70 percent, respectively, of phenylalanine and tyrosine residues buried in the native 7S globulin remained as the buried form even after heating.
A Spectrofluorimetry and fluorescence-quenching study suggested that one residue of tryptophan in the 7S globulin tended to be transferred to the more hydrophobic interior on heating.

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The structure of ribocitrin was elucidated to be 2-(S)-[O-α-D-ribofuranosyl-(1→2)-O-α-D-ribofuranosyl-(1→3)-α-D-ribofuranosyloxy]-1, 2, 4-butanetricarboxylic acid. The inhibitory activities of its components on dextransucrase of S. mutans E49 were examined.

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Chromatin fractions were isolated from intact and wounded sweet potato root tissues. The synthesis of RNA by the chromatin fractions was dependent on four ribonucleoside triphosphates and a divalent cation such as Mg²⁺ and Mn²⁺, Mn²⁺ being most effective. Whereas phosphate did not interfere with the polymerase reaction, it was totally blocked by pyrophosphate. The reaction was inhibited by DNase and actinomycin D as well as RNase and trypsin. The RNA polymerases of sweet potato root needed SH-groups for catalysis. Activity of chromatin-bound RNA polymerases (EC 2.7.7.6) promptly increased in the 6hr after wounding and then decreased gradually up to 24hr. Under the present experimental conditions it was mostly due to the activity of RNA polymerase I. RNA polymerase II contributed only about 5 to 15% to the total activity. The increase in the activity after wounding was completely inhibited by cycloheximide. Plant hormones such as 2, 4-dichlorophenoxyacetic acid, gibberellic acid and dibutyryl cyclic adenosine 3'5'-monophosphate stimulated the increase in RNA polymerases three to four times after wounding. Ethylene partially suppressed the wound-induced increase of RNA polymerases.

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In low-phosphate medium, a butirosin A producer B. vitellinus produced two alkaline phosphatases. These enzymes were fractionated by DEAE-cellulose column chromatography. One phosphatase (Pho I) was eluted with the lower concentration of NaCl compared with the other phosphatase (Pho II). In the wild type strain, Pho I was completely repressed in the high-phosphate medium, but 30% of the fully-derepressed level of Pho II was still produced.
The phosphatase-negative mutant, P-15, that was shown to accumulate butirosin A-6'-N-diphosphate in our previous study, produced only one phosphatase (Pho I) under the low-phosphate condition. Therefore, P-15 was characteristic of the deficiency in Pho II synthesis.
The partially purified preparations of Pho I and II were characterized. Although both enzymes had a similar molecular weight, they could be differentiated in control of synthesis, heat stability, substrate specificity and other properties. Kinetic properties showed that Pho-II was more specific than Pho I to aminoglycoside-phosphates; butirosin A-3'-phosphate, butirosin A-6'-N-diphosphate and 6'-deamino-6'-hydroxybutirosin A-6'-O-diphosphate. The roles of the two phosphatases in butirosin A biosynthesis were discussed.

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The microbial transformation of sclareol, a tobacco constituent, by a soil bacterium ITS-162 was studied. Manool, manoyl oxide and a new compound; 14, 15-dinorlabd-8(17)-en-13-one were isolated from the culture broth as transformation products. In the presence of metabolic inhibitors such as α, α'-dipyridyl, a compound; labda-8(17), 14-dien-13β, 18-diol and two new compounds; 13β-hydroxylabda-8(17), 14-dien-18-al and 13β-hydroxylabda-8(17), 14-dien-18-oic acid were accumulated in the culture broth as transformation products. Based on the experiments with degradation sequence, the biotransformation pathway of sclareol by the bacterium was proposed.

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The specificity of highly purified alkaline proteinase B (EC 3.4.21.14) from thermophilic Streptomyces rectus var. proteolyticus was investigated with an oxidized insulin B chain. Hydrolysis of the oxidized insulin B chain in a 4-hr incubation was observed mainly at three peptide bonds (Phe²⁴-Phe²⁵, Leu¹⁵-Tyr¹⁶ and Leu¹¹-Val¹²) and additionally at six others (Leu⁶-CySO₃H⁷, Gln⁴-His⁵, Leu¹⁷-Val¹⁸, His⁵-Leu⁶, Glu¹³-Ala¹⁴, Asn³-Gln⁴).
Hydrolysis of angiotensin (formerly designated angiotensin II) was observed at the Tyr⁴-Ile⁵ bond. Hydrolysis of proangiotension (formerly designated angiotensin I) was observed at the Tyr⁴-Ile⁵ and Phe⁸-His⁹ bonds.

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When rats were fed a low protein diet containing 3% or more of phenylalanine, their growth rate and food intake were depressed, and eye and paw lesions which were similar to those in tyrosine toxicity developed in all rats. Their liver phenylalanine hydroxylase activity was depressed in proportion to the dietary phenylalanine content, and dihydropteridine reductase activity was in a great excess over hydroxylation activity, so phenylalanine hydroxylase activity seemed to be limited firstly in the degradation of phenylalanine. Excessive phenylalanine was accumulated, and the tyrosine concentration was higher than that of phenylalanine in the plasma and tissues of rats fed a diet containing 2% or more of phenylalanine. When p-Cl-phenylalanine (p-Cl-Phe) was injected to the rats fed excess phenylalanine, the phenylalanine hydroxylase was depressed, the concentration of tyrosine in the body was lowered, and the development of eye and paw lesions was prevented completely. The development of eye and paw lesions seemed to be associated with the extremely elevated tyrosine concentration in the body.

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Bilirubin oxidase was purified from the culture filtrate of Myrothecium verrucaria MT-1 by a procedure involving ammonium sulfate precipitation, charcoal treatment, and QAE-Sephadex A50 and Sephadex G-100 column chromatographies. The purified enzyme was homogeneous on disc gel electrophoresis.
Copper and carbohydrate were contained in the enzyme. The enzyme was inhibited by Fe²⁺ and compounds that complex with copper. Bilirubin, biliverdin, hemin and chlorophyllin which consist of tetrapyrrole, and substrates of laccase were oxidized by the enzyme. Bilirubin was oxidized more rapidly than other substances. Bilirubin oxidase differed from laccase in reactivity with substances consisting of tetrapyrrole. Substances consisting of tetrapyrrole were oxidized only a little by laccase but rapidly oxidized by bilirubin oxidase. The apparent Km value for bilirubin was calculated to be 190μM.

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The thermal behavior of the starch structure was determined successfully under low moisture conditions by differential thermal analysis (DTA) using an anti-leak cell (resistant to an internal pressure of about 40 kg/cm²). Three kinds of starch, potato, kuzu and corn, with a low moisture content less than 12% indicated two strong endothermic peaks, one at the lower temperature (A) and the other at the higher temperature (B) in a temperature range approximately from 20 to 250°C. From microscopic observations and determinations of intrinsic viscosity, blue value and the solubility of heated starch, it is considered that peak A reflects an endothermic process accompanied by dextrinization involving the collapse of the ordered structure of starch with a limited breakdown in the polysaccharide chain. On the other hand, peak B corresponds to a thermal decomposition of the glucosyl residues, that is caramelization. The temperature at which such dextrinization takes place is markedly influenced by both the starch species and the moisture content of the starch.

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Mass spectra of N-trifluoroacetyl n-butyl ester (BTFA) of ornithinoalanine (OAL) and lanthionine (LAN) were compared with those of the BTFA derivatives of lysinoalanine (LAL) and the related amino acids. A difference of m/z 14, corresponding to one methylene group, was found in each pair of characteristic fragments between BTFA-OAL and BTFA-LAL. A temperature-programmed gas-liquid chromatography and gas chromatography-mass spectrometry were developed to analyze BTFA-LAN, BTFA-OAL and BTFA-LAL. More LAL and LAN were formed in α-lactalbumin than lysozyme by high-temperature treatment in water. OAL was detected in lysozyme treated at 100° and 120°C in alkali solution, but not in α-lactalbumin.

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A ³H-labeled bicyclic phosphate (BP), 4-[2, 3-³H]propyl-2, 6, 7-trioxa-1-phosphabicyclo[2.2.2]octane 1-oxide, bound specifically to rat brain synaptic membrane preparations. The apparent dissociation constant and the quantity of the BP bound at saturation were estimated to be about 30 μM and about 2 pmol per mg of protein, respectively. BP analogs except 4-amino BP, which is extremely low in toxicity, inhibited the binding but no obvious correlation was observed between the toxicity and the inhibitory potency of the BP analogs. The binding was also inhibited by high concentrations of GABA and picrotoxinin, whereas it was enhanced by bicuculline methiodide.

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Some toxic bicyclic phosphates. (BPs) inhibited acetylcholinesterases (AChEs), but the activity was very weak. Even the most potent inhibitor, 4-nitro BP, inhibited bovine erythrocyte and housefly head AChEs by only 37 and 38 per cent, respectively, at 1.5 HIM. Kinetic analysis indicated that the poor inhibitory activity of 4-nitro BP is ascribed not only to the low affinity for AChEs but also to its poor phosphorylating ability. Similar findings were obtained in the case of the reaction of BPs with the serine enzyme α-chymotrypsin. Despite the relatively high reactivity in an alkaline solution, BPs are much less active than other bioactive organophosphorus esters in phosphorylating a general-base-catalyzed hydroxyl group. This fact suggests that the toxic action of BPs does not result from the phosphorylation of a critical site in biological systems.

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Absorption of methionine and its sulfoxide was investigated in vitro with everted sacs and in insitu with circulated loops of rat small intestine. Transmural transport and tissue accumulation of methionine sulfoxide in the everted sacs were in fair agreement with those of methionine. Apparent kinetic parameters for the difference of transmural transport in the absence and presence of 10^-5M carbonylcyanide m-chlorophenylhydrazone, i.e. for the energy-dependent active transport, were similar for both methionine and its sulfoxide. Methionine was found at a low level in the serosal fluid of the everted sac on incubation with methionine sulfoxide. It was attributed to the methionine leaked out from the tissue but not to that formed by reduction of methionine sulfoxide during the course of intestinal transport. Similar transport was also observed in situ in circulated intestinal loops for methionine and its sulfoxide. The absorption efficiency of methionine sulfoxide in the small intestine is not the reason for the decreased nutritional availability of the most likely oxidation product of methionine.

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A bacterial strain, which was isolated from soil, and identified as Citrobacter sp., showed an inducible gallic acid decarboxylase activity producing pyrogallol from gallic acid. The strain also decarboxylated protocatechuic acid, pyrocatechuic acid, 3, 5-dihydroxybenzoic acid and m-hydroxybenzoic acid as well. The pyrogallol and pyrocatechol produced were isolated from the cultured broths to which gallic acid and protocatechuic acid were added, respectively.

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The effect of protein depletion on the metabolism of body collagen and muscle protein has been investigated in young male rats fed with a protein-free diet for 14 and 28 days.
During the protein depletion, . the protein content of the liver, intestine and skin decreased significantly, but the decrease of proteins was very little in the carcass, tail and bone (ossa cruris). An increase of tissue collagen in protein depletion was found in the carcass, bone, tail, skin and liver, while muscle protein in the carcass was evidently lost at a later stage of protein depletion. The increase of calcium in the bone was parallel to the increase of collagen, indicating continuous growth of the bones in spite of protein depletion. These results may indicate that the young animals continuously synthesize collagens of their special tissues from other tissue proteins even with severe protein deficiency. The metabolic responses of body collagens to dietary protein depletion in young rats have been discussed and compared with those in adult rats reported previously.

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Three flavanols, which inhibit root growth in the rice seedling test, have been purified. One was (+)-afzelechin (1) and the other two, named Pla and PIb, were new biflavanols shown by the structures 2 and 3 on the basis of chemical and spectroscopic evidence. Pla inhibited the root growth of the peach seedlings. In addition, Pla was found in the soil in the area where the peach trees were growing. The flavanols, therefore, are suggested to be one of the chemical factors causing soil sickness often encountered in peach cultivation.

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Twenty bacterial strains, grown on aniline as a sole source of carbon and nitrogen, and isolated from soil, were identified by morphological and biochemical tests, and cell wall analyses. Eight isolates were Rhodococcus erythropolis (Gray and Thornton) Goodfellow and Alderson, five were Pseudomonas maltophilia Hugh and Ryschenkow and seven were DAB (diaminobutyric acid)-type coryneform bacteria. The Rhodococcus spp. maintained the ability to assimilate aniline, but the pseudomonads and coryneform bacteria readily lost the ability, when grown in the absence of aniline.

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O, O-Dimethyl phosphorodithioate (DMDTP) is an initial breakdown product of organophosphorus pesticides in fields. DMDTP is also released to natural environments by pesticide manufacturers. DMDTP-degrading microorganisms were not known. We isolated two bacteria from activated sludge. One of them, strain TK-1 identified as Thiobacillus thioparus, utilized DMDTP as a sole energy source and produced dimethyl phosphate (DMP) and sulfate. The other, strain AK-2 identified as Pseudomonas sp., utilized DMP as a sole energy and carbon source and degraded DMP to inorganic orthophosphate (P_i). DMDTP was degraded to P_i by the coaction of the two bacteria.

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Asymmetric hydrolysis of the acetates of racemic secondary alcohols related to synthetic pyrethroids by Bacillus subtilis var. niger (IFO 3108) yielded optically active acetates and alcohols of varying optical purities.

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Conversion of 1, 8-cineole (1) by a strain of Aspergillus niger was investigated in connection with the biomass utilization of Eucalyptus plants. Two novel alcohols, 3-exo-(5) and 3-endo-hydroxycineole (6) were isolated from the culture broth and identified by interpretations of their spectral data. Furthermore, hydrogenolysis of 6 afforded p-methane-3, 8-e"-diol (8) which has been isolated as a plant growth inhibitor from Eucalyptus citriodora.

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