The film strain of Saccharomyces grows through non-film and film stages. The differences in cell surface hydrophobicity were examined at both stages. The degree of hydrophobic was quantitatively determined by comparing distribution ratios of cells between buffered aqueous and organic solvent phases. The cell surface in the film stage was more hydrophobic than that in the non-film stage, whereas the inherent non-film strain of Saccharomyces always showed low hydrophobicity. These results indicate that the change from non-film to film stage was due to a change in cells from hydrophilic to hydrophobic. The effects of growth conditions on hydrophobicity were further examined with the film strain Saccharomyces bayanus. Ethanol as sole carbon source more efficiently increased hydrophobicity than glucose. The increase in hydrophobicity seemed to depend upon respiration accompanying assimilation of ethanol. It was also found that the addition of a limited amount of biotin, as well as higher pH in medium lowered hydrophobicity. Variation in degree of pellicle formation was positively related to that of cell surface hydrophobicity.

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Cell surface hydrophobicity, which is necessary for pellicle formation, keeps film yeast cells floating on the medium surface. The substance responsible for hydrophobicity on Saccharomyces cell surfaces was examined. Cells treated with pentane, a chloroform: methanol, surface active agents and sodium hydroxide showed low hydrophobicity. Treatment with 0.5N sodium hydroxide was especially effective in lowering hydrophobicity to naught. This treat-ment removed only fatty acids, mainly palmitoleic and oleic acids from the cell surface. The same results were obtained with the cell wall. The amount of fatty acids extracted from the wall was nearly equal to that from the surface. This demonstrates the presence of fatty acids on the cell surface. As the fatty acid content of the Saccharomyces cell wall increased, stronger hydrophobicity was shown. Similar tendencies were seen on the cell surface. It was concluded from these findings that hydrophobicity extensively depended largely upon fatty acids on the cell surface. Similar results were found in several film strains other than Saccharomyces. Thus, variations in hydrophobicity with changes in carbon source were due to differences in fatty acid content at the cell surface.

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An interpretation was attempted of previous results that the method of reduced variables could not be applied to the relaxation curves of mochi cake. Temperature dependence of the apparent specific volume or the apparent density of mochi cake was determined. The correction factor by density Tρ/T₀ρ₀ was very small and ignored with usual materials. Ineffecti-veness of WLF Eq. to the temperature dependence of relaxation modulus of mochi is attributed to the decrease of viscosity, which might be derived from free space on the mobility of polymer segments. It is suggested that free space corresponds to fine air bubbles mixed in mochi cake during its preparation which causes a decrease in viscosity and relaxation modulus.

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4-Chloro-2-butynyl N-(3-chlorophenyl) carbamate (Barban) is an herbicide whose alkaline hydrolysis leads to the release of the chlorine atom of the ester group. The dechlorination kinetics were investigated in alkaline media at 25°C and followed spectrophotometrically or colorimetrically. The rate law of this reaction is complex and shows that the substrate as well as its anion are the reactive species. The pK_a value determined spectrophotometrically was 13.50.

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The complete separation of diastereomers of some Tfa-tetrapeptide-OMe by gas chromato-graphy of a packed column provided a simple method for assessing racemization during peptide synthesis using well known coupling reagents and various model reactions. The coupling method with diethyl phosphorocyonidate in dimethyl formamide was confirmed to be almost entirely free of racemization.

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Candida salmenticensis possesses an inductible inulinase of β fructosidase type, also active on raffinose and saccharose. The enzyme resides in the cell wall but is easily excreted into the culture medium during growth. Its principal characteristics for inulin are: optimum pH 3.5-4, optimum temperature 46°C, good stability at 50°C and Km=1.7×10^-2M.

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A variety of halophilic bacteria were isolated from various saline materials of different salt concentrations. Enriched cultures were used with 4M NaCl Sehgal and Gibbons complex medium conventionally applied for cultivation of extremely halophilic bacteria. The isolates were classified into nine types from halotolerant to extremely halophilic, based on the salt-response pattern of growth. Among 168 isolates, 75 isolates belonged to type III-A that was moderately halophilic and grew well in added NaCl but not with KCl. This type of bacteria was distributed widely in salted fish, dried fish, salt farm samples, crude salt for soy-sauce making, soy-sauce mashes, seasands and seaweeds. Very high bacterial rates were found in the first three materials. When two enrichment culture media containing 4M NaCl and 4M KCl were used in parallel, isolates of different salt-response types were obtained from the same material. No isolate requiring KCl specifically was found, even with 4M KCl medium.

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Changes in aggregation and properties of alkali-treated soybean 7S and 11S globulins depend upon protein concentration during alkali-treatment. Such variables were investigated by viscosity, electrophoresis, circular dichroism, pulsed NMR, emulsion capacity and CaCl₂ precipitation measurements. In lower protein concentrations, the intrinsic viscosity decreased and the penetrative fractions into electrophoresis gel increased. The reduced contacts of proteins during neutralization resulted in smaller aggregates. Also specific fractions which were more sensitive to protein concentration on aggregation were observed for 11S globulin. The quantity of bound water depended only on pH at 7% concentration treatment. When the gel was formed, the bound water of protein increased, e.g., 0.085g and 0.135g/g protein at pH 10.6 and 13.2 treatment, respectively, whereas at 1% treatment, bound water showed almost no pH dependence (about 0.13g/g protein). Furthermore, proteins prepared at higher protein concentrations were characterized by higher emulsion capacity and CaCl₂ pre-cipitation ability. However, no protein concentration dependence was seen in the secondary structure of the aggregates.

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Mixed culture of Methylomonas flagellata, a methane-utilizing bacterium, and Alcaligenes eutrophus, a hydrogen-utilizing bacterium, was attempted in order to use the carbon dioxide produced from methane by the former bacterium as the carbon source for the autotrophic growth of the latter. In mixed culture with methane and hydrogen as the carbon and energy sources, carbon dioxide produced by M. flagellata was utilized almost completely by A. eutro-phus, and recycling of gaseous substrates was possible in a closed cultivation system. However, growth of the mixed culture was poor compared to growth of M. flagellata and A. eutrophus in pure culture. During growth of the mixed culture, a predominance of A. eutrophus and decay of M. flagellata were observed. A. eutrophus seemed to inhibit the growth of M. flagellata by its preferential uptake of carbon dioxide and by production of a growth inhibitor.

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Strain 9-5, which was newly isolated from soil as a hydrogen bacterium, was found to be a new species of amylolytic pseudomonad and named Pseudomonas hydrogenovora. The (G+C) content of the DNA of P. hydrogenovora was 62.3-62.5 mole%.
Genus Pseudomonas is divided into four sections in “Bergey's Manual.” P. hydrogenovora showed characteristics intermediate between section II and section III types. Similarities were present in the properties of Strain 9-5 and Pseudomonas saccharophila, an amylolytic hydrogen-oxidizing pseudomonad, but detailed studies revealed many important differences between these two strains. The distribution of hydrogenase in this strain was also examined and found to differ from that in P. saccharophila. P. hydrogenovora showed membrane-bound hydrogenase and very weak, almost negligible soluble hydrogenase activity to reduce NAD.

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Bacillus subtilis strains M15, TM23 and T2N26 produced α-amylase at rates of 0, 50 and 350 units/hr/ml, respectively. RNA extracted from cells of these strains were used as messenger sources. These sources stimulated the incorporation of labeled amino acids in a heterologous cell-free protein synthesizing system prepared from Escherichia coli. The amounts of mRNA specific for α-amylase were estimated in vivo and in vitro by precipitation of radio-labeled pro-teins with anti-α-amylase serum. The results suggest that the capability of α-amylase synthesis in B. subtilis largely reflected the content of mRNA for α-amylase.

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α-Amylase-like proteins were synthesized in a heterologous cell-free protein synthesizing system prepared from Escherichia coli. The proteins were precipitable with anti-α-amylase serum and detected only when RNA extracted from α-amylase producing Bacillus subtilis cells was used as messenger. The in vitro α-amylase-like products seemed to consist of two components having molecular weights of 30, 000 and 13, 000.

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The major wax component secreted by Anomoneura mori Schwarz nymph was identified as lacceryl laccerate, n-dotriacontyl n-dotriacontanoate. Its content in the wax was 93.8%.

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Carbohydrate analogs of N-acetylmuramyl-L-alanyl-D-isoglutamine, such as 2-acetamido-2, 6-dideoxy-3-O-(D-2-propionyl-L-alanyl-D-isoglutamine)-D-glucopyranose (11), 2-acetamido-6-amino-2, 6-dideoxy-3-O-(D-2-propionyl-L-alanyl-D-isoglutamine)-D-glucopyranose (12), and 2-acetamido-2-deoxy-3-O-(D-2-propionyl-L-alanyl-D-isoglutamine)-D-xylopyranose (18) were synthesized, and their immunoadjuvant activities were examined to clarify their structural requirements for activity in the carbohydrate moiety.

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A variety of 6-amino- and 6-(acylamino)-6-deoxy carbohydrate analogs of N-acetylmuramyl-L-alanyl-D-isoglutamine, N-acetylmuramyl-L-valyl-D-isoglutamine, and N-acetylmuramyl-L-seryl-D-isoglutamine were synthesized starting from benzyl 2-acetamido-6-azido-3-O-(n-1-carboxyethyl)-2, 6-dideoxy-4-O-tetrahydropyranyl-α-D-glucopyranoside (1), and their biological activities were examined.

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A radiation-resistant red bacterium isolated mainly from rice in 1965 was identified as a new species in the genus Pseudomonas in 1971. This bacterium easily accumulated poly-β-hydroxy-butyrate granules as intracellular carbon reserve, and the GC content of DNA on the type strain No. O-1 was ca. 65%. All of the isolates were strictly aerobic and decomposed glucose oxidatively, never fermentatively. Its taxonomic characteristics were found to be different from all described species in the genus Pseudomonas to warrant its description as a new species named previously as P. radiora.
The catalase activity of this species varied considerably among strains, and it was not possible to directly correlate with radiation resistance. However, some reduction in oxygen effect by strains of each catalase activity suggested that H₂O₂ was involved in the expression of this oxygen effect.

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The apparent viscosities of glycinin (11S), β-conglycinin (7S) and acid-precipitated protein (APP) of soybean suspending systems and their heated gels were investigated using a modified coaxial cylinder viscometer. A hybrid program was established: (i) cyclic temperature test (20→90→20°C) under constant shear rate and (ii) cyclic shearing test (48.7→243.7→48.7 sec^-1) under isothermal conditions. The viscosities of the soy protein suspending system (12%, w/v) gradually decreased with increasing temperature to about 70°C. Thereafter, charac-teristic behavior depended upon the sort of soy protein. The apparent viscosity of 7S globulin increased considerably, especially at lower ionic strength (μ=0.01). However, the viscosity of 11S globulin remained almost unchanged under the different temperatures and ionic conditions used. A considerable increase in viscosity was found in the cooling process rather than in the heating process with 7S globulin and APP. A noticeable thioxotropic characteristic was found in all gels and also in unheated 11S globulin at low ionic strength.

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Radioactive substances were identified in urine of rats fed browned casein, which had been labeled with U-¹⁴C-L-lysine. When browned casein was ingested by growing rats, high radio-activity was found in urine taken for 24 hr after feeding. Urinary recovery of radioactivity and specific radioactivity were about 9-times as high as those of the control. The radioactive substances were separated by Sephadex gel filtration and ion-exchange chromatography. 75-83% of the total radioactivity was recovered in the first peak of Sephadex gel filtration. The material with radioactivity was separated into two fractions by ionexchange chromato-graphy. The ratio of radioactivity of these peaks on the chromatogram was about 30 to 70. The main peak was identified as ε-fructoselysine with an amino acid autoanalyzer. Urinary ε-fructoselysine content of 24 hr after a single dose feeding of 600mg browned labeled casein was 13-18mg per head. The relationship between ε-fructoselysine content as an absorption delayed-material in the small intestinal lumen and the amount excreted in urine was explained in a scheme together with results from previous studies.

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The structure and absolute configuration of a metabolite isolated from Aspergillus silvaticus was studied by an X-ray diffraction method. Crystals were orthorhombic, with space group P2₁2₁2₁, and a=9.894 (5), b=6.803 (4), c=22.519 (6)Å, and Z=4. The structure was solved by a direct method and refined by a block-diagonal least-square method to R=0.07 for 1143 non-zero reflections. The metabolite was identified as naphthalic anhydride obtained from atrovenetin. The absolute configuration determined by the Bijvoet method is consistent with that shown chemically. The molecule has two intramolecular hydrogen bonds.

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Glycyl-L-leucine is one of the best substrates for peptide hydrolases in the intestinal mucosa. Its absorption and hydrolysis were investigated in epithelial cells isolated from the rat intestine in the presence of bestatin, a specific inhibitor of certain peptide hydrolases. Bestatin competi-tively inhibited dipeptide hydrolase activities in isolated cells with a K_i value of 10^-8M, but noncompetitively inhibited, and less significantly, the dipeptide absorption by isolated cells. At 10^-4M bestatin inhibited half the dipeptide absorption, but only minimally inhibited the absorption of constituent amino acids. In the presence of bestatin at substantial concentrations the isolated cells took up a significant amount of the intact dipeptide, which otherwise appeared entirely in the form of free amino acids. These results are interpreted to substantiate a notion that a dual mechanism is operative for the absorption of readily-hydrolysable peptides: the peptide hydrolysis followed by uptake of thereby released amino acids, and the peptide transport followed by cytosolic hydrolysis.

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Another pectate lyase was purified to a nearly homogeneous state from the culture filtrate of Streptomyces nitrosporeus. The molecular weight was estimated to be about 41, 000. Iso-electric point was pH 4.6. The enzyme was most active at pH 10.0 and 50°C, and was relatively stable at a pH range of 4-11 (at 2°C for 48 hr) and below 40°C (at pH 7.0 for 10min). Ca²⁺ was required for maximum activity. The enzyme was an endo-pectate lyase which was more active on low methoxyl pectin than on polygalacturonic acid and had macerating activity on potato tissue and Ganpi bark.

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New ingenol-esters were isolated from leaves of Euphorbia cotinifolia L. (Euphorbiaceae) as piscicidal constituents. On the basis of spectral properties, the structures were established as 3-O-propionyl-20-O-(S)-(2'-methyl) butyryl-ingenol (1), 20-O-isobutyryl-ingenol (2), 3-O-propionyl-20-O-isobutyryl-ingenol (3), and 3, 20-O-di-isobutyryl-ingenol (4).

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Funicin, a new antimicrobial substance, C₁₇H₁₈O₅, was isolated from mycelial mats of Aspergillus funiculosus. The structure was elucidated to be ethyl-2-hydroxy-4-(3-hydroxy-5-methylphenoxy)-6-methylbenzoate from chemical and spectroscopic evidence. Funicin showed inhibitory activity against bacteria and fungi.

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An analytical method was developed for determining the four optical isomers of a new synthetic pyrethroidal insecticide, α-cyano-3-phenoxy-benzyl 2-(4-chlorophenyl)isovalerate (Fenvalerate, S-5602). The cyano group in S-5602 molecule was converted into l-menthyloxy-carbonyl group by heating S-5602 with l-menthol and hydrochloric acid. The obtained diastereoisomer derivative (l-menthyl ester deriv.) was chromatographed on a μPorasil column (4mm i.d.×0.3m) using a solvent system of hexane-ethyl acetate (500+7, v/v) as a mobile phase, and the four isomers of S-5602 were separated from one another within 20min. The compositions of S-5602 isomers were determined from their peak areas.

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A bioassay survey was conducted of constituents of leaves of Viburnum odoratissimum Ker., a “fish poison” plant. The bioassays led to the isolation of a new piscicidal compound vibsanine A (TL_m against Oryzias latipes, 0.18μg/ml), a new plant growth inhibitor vibsanine B (IC₅₀ against root growth of rice seedlings, 57μg/ml) and related compounds vibsanines C, D, E and F.

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Prorennin-specific messenger ribonucleic acid (mRNA) has been purified by a combination of sizing techniques, including Sepharose 2B chromatography and sucrose density gradient centrifugation, and affinity chromatography with poly (U)-Sepharose, from total nucleic acid extracted from dry ice-pulverized, fourth stomach of a calf. This mRNA bound to poly (U)-Sepharose, indicating that it contained a poly (A) sequence. The total translation product in the mRNA-dependent wheat germ system, upon addition of this mRNA, was identified as authentic prorennin by gel electrophoresis. The molecular weight of this mRNA was about 3.5×10⁵ as determined by gel electrophoresis. These results indicate that the synthesis of prorennin is directed by this mRNA 1, 020 nucleotides in length and requires the full coding capacity of the molecule.

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Dimethylglycine oxidase was purified to homogeneity from the cell extract of Cylindro-carpon didymum M-1, aerobically grown in medium containing betaine as the carbon source. The molecular weight of the enzyme was estimated to be 170, 000 by the gel filtration method and 180, 000 by the sedimentation velocity method. The enzyme exhibited an absorption spectrum characteristic of a flavoprotein with absorption maxima at 277, 345 and 450nm. The enzyme consisted of two identical subunits with a molecular weight of 82, 000, and contained two mol of FAD per mol of enzyme. The flavin was shown to be covalently bound to the protein. The enzyme was inactivated by Ag⁺, Hg²⁺, Zn²⁺ and iodoacetate. The enzyme oxidized dimethylglycine but was inert toward choline, betaine, sarcosine and alkylamines. Km and Vmax values for dimethylglycine were 9.1mM and 1.22μmol/min/mg, respectively. The enzyme catalyzed the following reaction: Dimethylglycine+O₂+H₂O→sarcosine+form-aldehyde+H₂O₂.

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Sarcosine oxidase was purified to homogeneity from the cell extract of Cylindrocarpon didymum M-1, aerobically grown in medium containing choline as the carbon source. The molecular weight of the enzyme was estimated to be 45, 000 by gel filtration method and 48, 000 by the sodium dodecylsulfate disc gel electrophoresis method. The enzyme exhibited an absorption spectrum with maxima at 277 and 450nm and shoulders at 370 and 470nm. The anaerobic addition of sarcosine to the enzyme resulted in the disappearance of the peak at 450nm. The enzyme contained one mol of covalently bound FAD per mol of enzyme. Enzyme activity was inhibited by Ag⁺, Cu²⁺, Hg²⁺, p-chloromercuribenzoate and iodoacetate. The enzyme oxidized sarcosine but was inert toward choline, betaine, dimethylglycine and N-methyl amino acids. Km and V_max values for sarcosine were 1.8mM and 26.2μmol/min/mg, respectively. The enzyme catalyzed the following reaction: Sarcosine+O₂+H₂O→glycine+formaldehyde+H₂O₂.

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Transformation of maridomycin III, a macrolide antibiotic, by maridomycin (MDM) III-insensitive streptomycetes was examined. Three main transformation products were obtained. In comparison with authentic samples, these transformation products were identified as 18-dihydro MDM III, 4''-depropionyl MDM III and 18-dihydro-4''-depropionyl MDM III. Reduction of the C-18 position of the macro lactone moiety of the antibiotic was thought to be a detoxication mechanism of the antibiotic by these insensitive strains.

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