Sixteen fluorine containing 2-(N-arylamino)/2-methyl-4-aryl thiazoles have been synthesised and characterized by spectral (IR and NMR) studies. Some representative compounds have also been screened for their bactericidal activity.

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The appearance of reducing power during incubation of silk gland homogenates of the silkworm, Bombyx mori, is due to the same manner as observed with an enzyme-substrate system. The silk gland cells play the role of an enzyme, and the liquid silk serves as a substrate. The appearance of reducing power from silk gland homogenates is due not to the specific physiological properties of the liquid silk, but to those of the silk gland cells of the silk glands during metamorphosis of the silkworm. A material with reducing power appearing during incubation of the silk gland homogenates was identified to be glucose.

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Physicochemical and immunochemical comparisons were made on the two acid proteases Mucor pusillus rennin (MPR) and Mucor miehei rennin (MMR). The molecular weight and isoelectric points of the two enzymes were similar: MPR (M. W., 38, 500 and I. P., 3.9) vs. MMR (M. W., 42, 000 and I. P., 4.1). D-Galactose and D-glucosamine, which had been found in MMR, were also detected in the carbohydrate moiety of MPR with gas-chromatography and amino acid analyses. The structural similarity between MPR and MMR was investigated utilizing peptide mapping techniques, which showed a close correlation between the two enzymes. Furthermore, immunodiffusion test using antisera prepared against each enzyme suggested the presence of a common antigenic structure. The results confirm, that a high similarity exists between MPR and MMR, which has been supposed from the morphological identity of the two enzyme-producing fungal strains.

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The glycomoiety of invertase of Candida utilis was suggested to be localized at certain sites of the enzyme protein. The oxidation of sugar moiety with periodate to a certain extent had no effect on the enzyme activity, but the stability of the enzyme was decreased. This partially oxidized enzyme bound hide powder or nylon powder and the enzyme activity was retained.
The invertase was shown to be divided into glycoprotein subunits of a similar molecular size when heated in the presence of SDS. The dissociation of the enzyme did not occur by reducing agent such as mercaptoethanol, and the results are discussed.

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The extracellular protease of Euglena gracilis z was purified to a single protein. It was an endopeptidase as found by the Nunokawa's method, and showed optimum pH for the proteinase, esterase and amidase activities at 7.3, 7.0 and 6.3, respectively. It had a molecular weight of 41, 000 and isoelectric point of 8.3. The bleached mutant of E. gracilis produced higher activity of extracellular protease than the wild strain, and supplementation of peptone to the growth medium augmented the enzyme production in both green and bleached cells. The Euglena extracellular protease was markedly inhibited by diisopropylfluorophosphate and Streptomyces subtilicin inhibitor, and to lesser extents by EDTA and p-chloromercuribenzoate. The enzyme was potentiated by some sulfhydryl compounds, activated greatly by Fe²⁺ and stabilized by Ca²⁺ and K⁺.

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The following three new analogs of naringin (I) were synthesized. Naringein-7-[6'-Omethyl-O-α-L-mannopyranosyl-(1→2)-β-D-glucopyranoside] (II), naringenin-7-[6'-Omethyl-O-α-D-mannopyranosyl-(1→2)-β-D-glucopyranoside] (III) and naringenin-7-[O-α-L-rhamnopyranosyl-(1→6)-β-D-galactopyranoside] (XVI).
Compounds II, III, narirutin (naringenin-7-[O-α-L-rhamnopyranosyl-(1→6)-β-n-glucopyranoside] (XV) and XVI are 0.10, 0.13, 0.05 and 0.08 times as bitter as I, respectively. From the above organoleptic result it was concluded that the methyl group of L-rhamnose in I is one of the essential factors for intense bitterness. Attachment of α-L-rhamnose to the_??_-position of D-glucoside or D-galactoside of naringenin does not abolish the taste of the resulting glycosides but elicits a moderate bitter taste.

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Buckwheat α-glucosidase showed especially high activity toward maltooligosaccharides, phenyl-α-maltoside and nigerose. Its activities toward isomaltose and phenyl-α-glucoside were very weak. The ratios of the maximum velocities for maltose, nigerose, kojibiose, isomaltose, phenyl-α-glucoside and phenyl-α-maltoside were estimated to be 100:101:17:2:4:106 in this order; those for maltose, malto-triose, -tetraose, -pentaose, -hexaose, -heptaose, -octaose and maltodextrin (DP_n=13), 100:110:106:121:131:109:111:86.
The Michaelis constants Km, the molecular activities ko and the ratios kolKm for a series of maltooligosaccharides were determined. From the experimental data, the subsite affinity of each subsite in the enzyme was evaluated according to the subsite theory. The difference in the substrate specificities between glucoamylase and α-glucosidase was interpreted by the relative value of subsite affinities of the first and the third subsites counting from the terminal (the first subsite) to which the nonreducing end of substrate is bound.

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The three microbial alkaline proteinase inhibitors (alpha, beta and gamma-MAPI) have been obtained as needle crystalline forms by a combination of various adsorption and ionexchange column chromatographies.
The crystalline inhibitors showed the same inhibitory spectrum but different inhibitory potential. They strongly inhibited various microbial alkaline proteinases, such as fungal alkaline proteinases, and also inhibited α-chymotrypsin, papain, ficin, bromelain and cathepsin B.
The three MAPI gave positive Rydon-Smith, Sakaguchi and 2, 4-dinitrophenyihydrazine reactions but were negative to ninhydrin reaction. From the amino acid analysis, three MAPI contained arginine, phenylalanine and some aliphatic amino acids; aliphatic amino acid of alpha-MAPI was valine only, that of beta-MAPI was probably valine (small amount of leucine and isoleucine was detected) and that of gamma-MAPI was leucine plus isoleucine (trace amount of valine was also detected).

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6β-Bromo-4, 4-dimethyl-lα, 2α-epoxy-3α, 5α-oxidocholestane (1) was derived from 4, 4-dimethyl-lα, 2α-epoxy-5-cholesten-3α-ol (6). The A/B ring of the bromo-epoxy-oxide (1) was found to be transformed into a 5/7 membered ring (3a) on acidic treatment. From the product (3a), the 2α, 5β-dihydroxy-3α, 6α-oxide (16) and the 3α, 5β-dihydroxy-2α, 6α-oxide (20) were synthesized. The mechanisms of the transformation were discussed.

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Volatile components of peel oil and juice from Citrus unshiu were studied. The peel oil and aroma concentrate from the juice were separated into hydrocarbons and oxygenated compounds by column chromatography. These fractions were analyzed by gas chromatography and combined gas chromatography and mass spectrometry.
Ninety-eight percent of the peel oil consisted of hydrocarbons such as d-limonene, r-terpinene, myrcene and a-pinene. The aroma concentrate from the juice was composed of the same hydrocarbons (60%) and oxygenated compounds (40%) such as 3-methylbutanol, cis-3-hexenol, trans-2-hexanal and n-hexanal. Ninety percent of the oxygenated compounds was alcohols and aldehydes.

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The regulatory mechanism of homoserine dehydrogenase and dihydrodipicolinate synthetase was investigated in Brevibacterium lactofermentum. Brevibacterium lactofermentum AJ 3585, which is a mutant resistant to S-(2-aminoethyl) L-cysteine (AEC) and α-amino-β-hydroxyvaleric acid (AHV), simultaneously produced 13 mg/ml each of L-threonine and L-lysine•HCl. Homoserine dehydrogenase of AJ 3585 was genetically desensitized to feedback inhibition by L-threonine and L-isoleucine, and its affinity for asparto β-semialdehyde was increased about 8-fold compared with the parental strain. However, L-threonine production and homoserine dehydrogenase formation of AJ 3585 were inhibited by L-methionine. We tried to derive the mutant resistant to S-methylcysteine sulfoxide from AJ 3585 to release the repression of homoserine dehydrogenase. Derepressed mutant, M-15, produced 17.4 mg/ml of L-threonine and 8.6mg/ml of L-lysine•HCl, and the homoserine dehydrogenase level was increased about 2-fold as compared with AJ 3585. On the other hand, L-lysine production and dihydrodipicolinate synthetase formation were inhibited by L-leucine. On the contrary, L-threonine accumulation increased remarkably in the presence of excess L-leucine. The data provides significant evidence that the level of homoserine dehydrogenase and dihydrodipicolinate synthetase in Brevibacterium lactofermentum affects the production of L-threonine and L-lysine.

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D-Glucose dehydrogenase [β-D-glucose: NAD(P) oxidoreductase (EC 1.1.1.47)] was synthesized derepressively in a mutant of a Bacillus species which was isolated as an improved strain for D-ribose production. The enzyme was very unstable and inactivated during storage or column chromatography. The inactivation was prevented in the presence of NAD⁺, NADP⁺ or certain salts. The inactive enzyme was reactivated by the addition of NAD⁺, NADH, NADP⁺, NADPH, AMP, ADP, ATP or certain salts. The molecular weights of the inactive and active form of the enzyme were estimated to be about 45, 000 and 80, 000, respectively, by Sephadex G-150 gel filtration. Thus, it seems that the enzyme activity is regulated by monomer-dimer interconversion of the enzyme molecule.

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The intermediates in the browning reaction of triose reductone (I) with guanine, guanosine, 2'(3')- or 5'-guanylic acid were isolated. The reaction of I with guanine in 4 N HCI at 65°C for about 1 hr produced only a brown tricyclic compound, 1, N²-(2-hydroxypropenylidene)-guanine or 7-hydroxy-10-oxo-9, 10-dihydropyrimido[1, 2-a] purine (III) with the analogous structure to natural occurring Y bases, whereas that at room temperature yielded a labile light yellow intermediate, N²-(3-oxo-2-hydroxypropenyl)guanine (II) with the enaminol structure as a mixture with III. The ratio of II to III was about 1:1. The isolation of II in pure form was difficult because of its instability.
On the other hand, the reaction of I with guanosine, 2'(3')- or 5'-guanylic acid in 4 N HCl at room temperature gave the reductive intermediate with the same enaminol structure as II, N²-(3-oxo-2-hydroxypropenyl)guanosine (IV), N²-(3-oxo-2-hydroxypropenyl) 2' (3')-guanylic acid (V) or N²-(3-oxo-2-hydroxypropenyl) 5'-guanylic acid (VI), respectively.

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Alginate lyase from Turbo cornutus acted on sodium alginate and resulted in a rapid decrease in viscosity and a gradual increase in reducing power of the substrate solution. The result of gel filtration of the reaction products indicated that oligouronides were liberated as the final product. The fact that the enzyme attacked preferentially on the mannuronaterich moieties of alginate molecule was demonstrated by an analysis with paper electrophresis of the reaction products after acid treatment, which separated the oligouronides from the polyuronides.

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Two kinds of new tetrasaccharide were isolated from cottonseed and their structures were established as 1^F -β-D-fructofuranosylraffinose; O-α-D-galactopyranosyl-(1→6)-O-α-D-glucopyranosyl-(1→2) [O-β-D-fructofuranosyl-(2→1)]-β-D-fructofuranoside and 4^G-α-D-galactopyranosylraffinose; O-α-D-galactopyranosyl-(1→6) [O-α-D-galactopyranosyl] (1→4)]-O-α-D-glucopyranosyl-(1→2)-β-D-fructofuranoside, respectively.

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After irradiation with sub-lethal doses of γ-rays, cells of M. radiodurans stop temporarily the incorporation of DNA precursors into acid-insoluble fraction no matter how much the dosage. However, the length of lag period to restore its normal rate of DNA synthesis depends on the dose of irradiation.
In permeable cells prepared after irradiation and reincubation in a growth medium, nonconservative-type DNA synthesis was inhibited while the stimulated level of DNA synthesis remained in the permeable cells prepared from the cells that had been reincubated at 0°C in the presence of EDTA. These results suggest involvement of some regulatory system for DNA synthesis in M. radiodurans that might play an important role in its high radioresistance.

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With an aim of developing menthol derivatives having good solubility in water, the glycosidation reaction of menthols was extensively investigated, such as the condensation of menthols with acetobromoglucose by the method of Koenig-Knorr and that with fully acetylated sugars in the presence of acidic catalysts. The water-solubility of menthyl monosaccharides was generally poor, but that of menthyl disaccharides was good enough for practical usage as food additives and medicines. Furthermore, these glycosides derivatives were advantageously utilized for optical resolution of dl-menthol because of their high crystallinity.

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Raw solid swine manure was extracted with dichloromethane. The extracted solution was mixed with distilled water after concentration and steam-distilled. The distillate was extracted with dichloromethane and fractionated into carboxylic acid, phenol, basic, and neutral fractions and all fractions had a characteristic and strong odor. Quantification and characterization of odorous components were performed by GC, GC-MS, NMR, and IR. Ten compounds were detected from the carboxylic acid fraction and three compounds from the phenol fraction. The two main components in the neutral fraction were mesityl oxide and diacetone alcohol.

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Glycolaldehyde dehydrogenase was partially purified from Bacillus subtilis IFO 3007. The enzyme was separated into two fractions by DEAE-cellulose column chromatography. These fractions behaved differently in substrate specificities and other properties.
Both the change of vitamin B₆ content (ΔB₆/Δt) and the activity of glycolaldehyde dehydrogenase reached simultaneously to the maximum values at the early stationary phase. Although none of compounds of vitamin B₆ family affected the enzyme activity, the formation of the enzyme was repressed by the addition of pyridoxal 5'-phosphate. The repression increased with the increased concentration of pyridoxal 5'-phosphate and reached about 30 at the concentration of 20 μM. These results that glycolaldehyde dehydrogenase of B. subtilis is closely related to vitamin B₆ biosynthesis.

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The effect of heating on the functional properties of ovotransferrin greatly varied with both the pH and ionic strength of the solution. When small amounts of inorganic salts (μ<0.1) were present in the ovotransferrin solution of which the pH was between 6.0 and 8.5, both the solubility and foaminess greatly decreased upon heating at a temperature above 60°C for 5 min. However, when the amounts of inorganic salts were larger (μ≥0.3), no decrease in either property was observed. These phenomena were assumed to be caused by either cation binding (at a low concentration of salts) or anion binding (at a high concentration of salts) of ovotransferrin. Among the anions tested, both phosphate and citrate which are assumed to have a rather high affinity for ovotransferrin did not cause any heat-induced defect at any of the salt concentrations tested.

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Serum lipoproteins in rats with fatty liver, which was induced by feeding an amino acid imbalanced diet containing 8% casein supplemented with 0.3% of DL-methionine, were analyzed. Three classes of serum lipoproteins, VLDL, LDL and HDL, were isolated by flotation in the ultracentrifuge and their concentration and chemical composition were determined. In serum VLDL of the imbalanced group, the concentration of TG was significantly higher than those for the control group, and that of apoprotein was maintained at the normal level.
These results suggest that this type of fatty liver is not due to a block in the transport of hepatic TG to the plasma. On the contrary, the administration of DL-ethionine resulted in a reduced level of TG of VLDL.

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An aqueous extract of culture of Trichoderma viride Y244-2 on wheat bran showed antitumor activity against L5178Y mouse leukemic cells in vitro and L1210 mouse leukemia in vivo. The principle of the antitumor activity was proved to be an L-lysine-oxidizing enzyme. Oxygen consumption was accompanied by the formation of α-keto acid, ammonia and hydrogen peroxide in the enzyme reaction with L-lysine. The ratio of amount of oxygen consumed to those of α-keto acid and ammonia formed in the presence of catalase was approximately 1:2:2. This enzyme is an L-amino acid oxidase which is highly specific for L-lysine. Thus, we designated it as L-lysine oxidase. Identity of the antitumor substance with the enzyme was confirmed by DEAE-cellulose column chromatography: the elution pattern of growthinhibitory activity against L5178Y cells coincided with that of L-lysine oxidase activity.

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The molecular weight of quinolinate phosphoribosyltransferase from Aicatigenes eutrophus nov. subsp. quinolinicus (IAM 12305) was estimated by gel filtration and ultracentrifugation respectively to be about 210, 000 and 222, 000. The enzyme contains eight identical subunits with a molecular weight of 27, 500 each. Optimum pH (constant concentration) and isoelectric point were 8.5_??_9.0 and 5.3, respectively. Optimum temperature was 60°C. H₂PO₄-inhibited the activity. The enzyme activity was also inhibited by increase in ionic strength. At constant ionic strength, the maximum activity was obtained at pH 9.5. The kinetic data of the reaction are consistent with the formation of a ternary complex comprising the enzyme, quinolinic acid and 5-phosphoribosyl-l-pyrophosphate. Reaction product was identified as l-nicotinic acid mononucleotide.

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The molecular weight of crystalline quinolinate phosphoribosyltransferase from hog liver was re-evaluated at 202, 000 and 210, 000 by the methods of sedimentation equilibrium and Sephacryl S-200 gel filtration, respectively. Since the molecular weight of its subunit was 34, 200, this enzyme was considered to be composed of six identical subunits. The enzyme required Mg²⁺ for its activity but it was found that the crystalline enzyme did not contain any detectable amounts of Mg, Mn, Fe, Cu, Zn and Ca, by using an atomic absorption spectrophotometer. In the liver enzyme reaction, Mn²⁺ could fully replace Mg²⁺. The reaction mechanism of the enzyme was like ping-pong. The enzyme activity was inhibited by various carboxylic acids, but this inhibition was suppressed by increasing the Mg²⁺ ion concentration. The enzyme activity was inhibited by the addition of glycerol and the inhibition increased as the pH was raised.

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The lateral diffusion coefficient in phosphatidylcholine membranes was determined using pyrene as a fluorescence probe, and the effect of α-, γ- and σ-tocopheryl acetates on the liposomes and the interaction between tocopheryl acetates and cholesterol in the liposomes were studied. α-, γ- and σ-Tocopheryl acetates reduced the diffusion of pyrene in the liposomes of dipalmitoyl, egg yolk and soybean phosphatidylcholines. α-Tocopheryl acetate was the most effective compound among the three tocopheryl acetates to reduce the diffusion coefficient of pyrene, and the degree to reduce the diffusion of pyrene by each tocopheryl acetate correlated well with its vitamin E activity. Tocopheryl acetates and cholesterol independently reduced the diffusion of pyrene in the membranes. The results suggest that tocopherols may physically stabilize biological membranes.

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Rhodotorucine A, which induces mating tube formation in Rhodospridium toruloides, was found to be a novel peptide composed of eleven amino acid residues with a lipophilic group at the C-terminus. The amino acid sequence of the peptide was determined by Edman degradation and enzymatic hydrolyses. The C-terminus of rhodotorucine A was revealed to be S-farnesylcysteine by proton magnetic resonance, mass spectrometry and chemical synthesis. We proposed the following structure for rhodotorucine A.
H-Tyr-Pro-Glu-Ile-Ser-Trp-Thr-Arg-Asn-Gly-Cys(S-farnesyl)-OH.

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A mutant of E. coli (MP2) sensitive to an antitumor antibiotic of high molecular weight and to proteinase was isolated and its characteristics were studied. The sensitivity of mutant MP2 to neocarzinostatin was greatly influenced by the environment in which MP2 was exposed to NCS. MP2 was also sensitive to a certain extent to some hydrolytic enzymes added exogenously such as pronase, DNase, and RNase. The sensitivity of the parental spheroplast to NCS and pronase was still lower than that of intact MP2. Proteinase-induced inactivation of ornithine transcarbamylase of the intact cells occurred only with MP2, but it did not with the parental cells, while it occurred with the spheroplasts of both strains.
From the analysis of membrane proteins by polyacrylamide gel electrophoresis, MP2 seemed to have a considerable change in the outer and inner membranes. The mutations responsible for these characters located in two separate regions of chromosome; i.e., one near his locus and the other between the point of origin of Hfr AB312 and the marker lys.

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Pyrethrolone was synthesized in a practical manner from readily available allethrolone.

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