In the course of our examination for the alkaloid productivities of Streptomyces strains, Streptomyces sp. NA-15 was found to produce a new alkaloid, pyrindicin, in the culture medium. The strain NA-15 was found to be a variant of Streptomyces griseoflavus and was designated as S. griseoflavus var. pyrindicus nov. var. After the culture conditions for pyrindicin production were studied, pyrindicin was obtained as its hydrochloride (mp 145°C, decomp.) from the cultured broth. The compound was shown to possess weak antimicrobial and several pharmacological activities. The LD₅₀ of the hydrochloride (ip, in mice) was 87mg/kg.

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Alkyl aryl esters, dialkyl esters and diaryl esters of N-methyl-N-phenylphosphoramidic acid were synthesized and tested for cross-resistance and jiont action with phosphorothiolate (PTL) fungicides using rice blast fungi, Pyricularia orzae. Negative correlation in crossresistance was found between PTL fungicides and most of the amidates. Synergism in the fungicidal action was also found between them fro wild types of the fungi, when tested by crossing filter paper strips impregnated with fungicides on agar plates inoculated uniformly with the test fungi.

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The possibility of lysine formation from α, ε-diaminopimelate (DAP), acetate, aspartate or α-aminoadipate (AAA) in rumen ciliates was examined. DAP-1, 7-¹⁴C added to the medium was decarboxylated and converted to radioactive lysine in great amounts and radioactive pipecolate in small amounts by rumen ciliates. Difference of the ability to form lysine from DAP between genus Entodinium and Diplodinium was not observed. With sodium acetate-U-¹⁴C, amino acids fraction of the supernatant fluid of the incubation medium and ciliates contained only 0.56 and 0.59% of the total radioactivity, respectively. In the case of L-aspartate-U-¹⁴C, 95.1% of the radioactivity of the supernatant fluid desalted and 62.2% of the radioactivity incorporated into ciliates (1.5% of the total radioactivity) remained as aspartate. Autoradiograms revealed the negligible spots of lysine in ciliates in both cases. AAA-6-¹⁴C remained almost unchanged, even after incubation with rumen ciliates.

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Cell walls containing α, ε-diaminopimelate-1, 7-¹⁴C (DAP) was prepared from Escherichia coli isolated from the rumen. After incubation of ciliates with the cell walls, 22.0% of DAP contained in cell walls of E. coli was converted to lysine and pipecolate. Heat-treated mixed rumen bacteria and heat-treated cell walls of mixed rumen bacteria added to the culture medium of rumen ciliates increased 0.572 and 0.934μmole/ml of sum of lysine and pipecolate, respectively.
From these results, it is clear that rumen ciliate protozoa can form lysine from DAP contained in the mucopeptide of bacterial cell walls. One of the nutritional significance of inhabitation of ciliates in the rumen was revealed.

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A cyclic dipeptide with a bitter taste, compound B-1, was isolated from sake and its properties were characterized.
Compound B-1 was chromatographically purified by the use of Amberlite XAD-2, silicic acid, Dowex 50-X4 (H type), Amberlite IR-45 (OH type) and silicic acid again, and crystallized from ethanol.
From the decomposition point (159_??_165°C) and the ultraviolet and infrared spectra, the compound was identified as L-prolyl-L-leucine anhydride.
The content of this compound in sake samples aged under various conditions was determined by gas chromatography and it was found that the amount of this compound increased with the time of storage of sake.
Furthermore, the contribution of the bitter compound to the flavor of sake was evaluated from its content and the threshold taste value.

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Aspergillus niger NRC-A-1-233 was cultivated by the shaking method. The optimal cultural conditions for ribonuclease (RNase) production were: composition of medium: sucrose, 15%; NH₄NO₃, 0.2%; KH₂PO₄, 0.1%; MgSO₄•7 aq., 0.025 %; initial pH, 2.2; shaking conditions: 50ml of medium /500ml flask; cultivation time, 120hr. The RNase was purified by acid clay treatment and chromatography on DEAE-cellulose and Sephadex G-75 columns. The purified RNase was homogeneous by ultracentrifuge and disc electrophoresis.
The molecular weight of the RNase was estimated to be 28, 500 on SDS-polyacrylamide gel and its isoelectric point was 2.8 by Ampholine electrofocusing method. Digestion rate of RNA by the RNase was 100%. The RNase did not have an exact base specificity and produced four kinds of 3'-nucleotides from yeast RNA.

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Starch granules from Chlorella, Chlamydomonas and Scenedesmus, grown heterotrophically in a medium containing organic carbon sources, were isolated by means of the toluol treatment of the sonicate of alga. The toluol treatment separated the starch granules in the water layer from the cells and cell debris coagulated in the upper toluol layer.
The starch granules of Chlorella vulgaris and Chlamydomonas sp. were composed of amylose (12 to 3%) and amylopectin. The amylose content of the starch granules of Scenedesmus basilensis was 22%. All the X-ray diffraction patterns of algal starch obtained in this investigation were of the A-type, identical to that of corn starch.

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A lipase with a high molecular weight was purified from Chromobacterium viscosum by chromatography using the Amberlite CG-50 and Sephadex G-75. The purified lipase (Lipase A) was found to be homogeneous by disc electrophoresis.
Lipase A had an optimum pH around 7 for lipolysis of olive oil and the enzyme was stable at the range of pH 4 to 9 and below 50°C. Zn²⁺, Cu²⁺, Fe³⁺ and high concentrations of L-cysteine, iodoacetic acid and NBS had remarkable inhibitory effects. Bile salts were activator. Lipase A was more active on water insoluble esters than water soluble esters. The isoelectric point of the enzyme was pH 4.7.

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An enzyme which catalyzes a decomposition of α-aminoisobutyrate (AIB) was purified and its kinetic properties were investigated. Michaelis constants for AIB decomposing reaction are able to be calculated by Ping Pong initial velocity equation. This enzyme catalyzes also L-alanine: α-ketobutyrate transamination as well as AIB decomposing reaction. Approximately equal values of Michaelis constants were obtained for α-ketobutyrate and pyridoxal 5'-phosphate (PLP), which are common substrates of both reactions.
In higher concentration of the enzyme, transamination between PLP and AIB or L-alanine was detected, whereas the reaction between pyridoxamine 5'-phosphate and pyruvate was not observed. These results are probably ascribed to a difference in affinity of two coenzymes for the enzyme.

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The lytic enzyme complex produced by Bacillus circulans WL 12 upon induction with the mycelia of Piricularia oryzae P₂ was analyzed by large scale polyacrylamide gel electrophoresis. Six β-1, 3 glucanases were separated. Activity profiles were obtained of these multiple β-1, 3 glucanases against P. oryzae P₂ cell walls, Saccharomyces cerevisiae was, laminarin, oat glucan, Phytophthora glucan and pachyman.

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Bacillus circulans WL 12 was grown on the cell walls of Piricularia oryza P₂ and Saccharomyces cerevisiae and on pachyman. Decomposition of the three substrates by the crude enzyme complexes induced by these substrates was compared. Patterns of the multiple β-1, 3 glucanases were analyzed by large scale polyacrylamide gel electrophoresis Characteristic patterns of the multiple β-1, 3 glucanases were obtained which were distinctly different from each other according to the inducer used.

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F-I_a which showed the highest lytic activity toward Piricularia oryzae P₂ cell walls among the six multiple β-1, 3 glucanases produced by Bacillus circulans WL 12 was purified and some of its properties were studied. The preparation was homogeneous by disc polyacrylamide gel electrophoresis at pH 9.4 and 4.0. The molecular weight was estimated to be around 48, 000. F-I_a hydrolyzed laminarin by a random mechanism. The enzyme liberated glucose, oligosaccharides and a high molecular weight heteroglycan from Piricularia oryzae P₂ cell walls. Heteroglycan showed a singles edimentation peak in ultracentrifugation and contained mannose, glucose and galactose.

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Several isobutenyl- and isobutyl-cyclopropanecarboxylates were synthesized. The insecticidal activities against the housefly of their 5-benzyl-3-furylmethyl or allethronyl esters were tested. Among the substituents on the cyclopropane ring, a methyl group cis to the ester linkage has been found to be essential for toxicity against the housefly and a trans-isobutenyl group greatly enhances the toxic activity.

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A callus was induced from the veins of a leaf of Symphytum officinale, comfrey, on a medium containing the inorganic elements reported by Murashige and Skoog with addition of 3% sucrose, 0.5mg/liter 2, 4-D and 0.3_??_3.0mg/liter kinetin.
Suspension cultures of this cell line obtained from the callus were shown to accumulate a large amount of L-glutamine intracellularly. The effect of growth hormones and nutrients on accumulation of the amino acid has been examined in suspension cultures. The most suitable concentrations of 2, 4-D and kinetin for glutamine accumulation were 0.3mg/liter each. The presence of potassium nitrate as a nitrogen source was beneficial for growth and ammonium nitrate stimulated the accumulation of glutamine. High levels of these nitrogen sources in the medium were required for obtaining a high level of glutamine. The concentration of glutamine accumulated reached to approximately 20% of dry cell weight when S. officinale was incubated in the medium containing 0.495% of ammonium nitrate and 0.57% of potassium nitrate which corresponded to three times higher levels than those in a Murashige and Skoog's medium.
Most of the amino acid was found intracellularly but a small amount was excreted into the medium in the later stages of the incubation. Addition of a cationic surfactant, cetyltrimethylammonium bromide, to the cultures caused to increase the amount of the amino acid in the culture filtrate.
The contents of free amino acids in leaves of S. officinale were compared with those in the callus. The level of glutamine in the callus was 260 times higher than that in the intact plant.

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Homoserine dehydrogenases and aspartokinases in L-threonine- or L-threonine and L-lysine-producing mutants derived from Corynebacterium glutamicum KY 9159 (Met-) were studied with respect to the sensitivity to the inhibition by end products, L-threonine and L-lysine. The activities of homoserine dehydrogenases in the mutants which produced L-threonine or L-threonine and L-lysine were slightly less susceptible to the inhibition by L-threonine than the activity in the parent strain, KY 9159. The aspartokinases in the threonine-producing mutants, KY 10484 and KY 10230, which were resistant to α-amino-β- hydroxylvaleric acid (AHV, a threonine analog) and more sensitive to thialysine (a lysine analog) than the parent, were sensitive to the concerted feedback inhibition by L-lysine and L-threonine by about the same degree as KY 9159. The aspartokinase in an AHV- and thialysine-resistant mutant, KY 10440, which was derived from KY 10484 and produced about 14mg/ml of L-threonine in a medium containing 10% glucose was less susceptible to the concerted feedback inhibition than KY 10484 or KY 9159, although the activity was still under the feedback control. In the parent strain, L-threonine activated aspartokinase activity in the absence of ammonium sulfate, an activator of the enzyme, but partially inhibited the activity in the presence of the salt. On the other hand, the enzyme of KY 10440 was activated by L-threonine either in the presence or in the absence of the salt. In another AHV- and thialysine-resistant mutant, KY 10251, which was derived from KY 10230 and produced both 9mg/ml of L-threonine and 5.5mg/ml of L-lysine, L-threonine and L lysine simultaneously added hardly inhibited the activity of aspartokinase.
Implications of these results are discussed in relation to L-threonine or L-lysine production, AHV or thialysine resistance and regulation of L-threonine biosynthesis in these mutants.

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Acidic polysaccharide, PLS F-II, was prepared from Serratia piscatorum polysaccharide, PLS N-I, by a sequence of ultrasonication and gel filtration and was examined for chemical composition and biological activity.
The purified PLS F-II preparation was shown to be homogeneous by ultracentrifugation, zone electrophoresis and column chromatography. The molecular weight was estimated to be about 2×10⁴ by the Archibald method. PLS F-II was composed of L-rhamnose, D-galactose and D-galacturonic acid in the molar ratio of 2:1:1 and was partially acylated on the galacturonic acid residues.
PLS F-II was found to enhance the antibody formation in mice, although it showed no anti-inflammatory activity.

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It was indicated from ultraviolet difference spectra and ultracentrifugal experiments that associations occurred between two casein components (α_s and, κ-caseins, β- and κ-caseins and α_s- and β-caseins) at lower CaCl₂ concentrations (2_??_3mM) and that aromatic amino acid residues participated in the associations. Chemical modification studies with 2-hydroxy-5-nitrobenzylbromide indicated that tryptophane residues of each casein component were not essential for these associations. It was also demonstrated by nitration of tyrosine residues with tetranitromethane that tyrosine residues of k-casein were essential for α_s•κ-association and for κ-association and that tyrosine residues of as casein were important to α_s•βassociation.
Interactions between casein components were also studied at higher CaCl₂ concentration (10mM) which is enough for micelle formation. It was found that tyrosine residues of _κ-casein played an important role for the stabilization of as and β-caseins. Properties of the nitrated-β-casein were almost the same as that of the native β-casein except the absorption spectrum. _??__s•β-Interaction in the presence of 10mM CaCl₂ was investigated by use of the nitrated-β-casein instead of the native β-casein. It was proved that α_s-casein was stabilized by the nitrated-β-casein and that precipitation of the nitrated-β-casein increased in the presence of α_s casein.
The mechanism of interactions between casein components at higher CaCl₂ concentration (10mM) are discussed in connection with the associations at lower CaCl₂ concentrations (2_??_3mM).

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The distribution of the enzyme activities relating to CoA biosynthesis from pantothenic acid in various microorganisms and the effect of CoA on these activities are described.
High activities of partial reactions involved in CoA biosynthesis were surveyed in various type culture strains involving bacteria, actinomycetes, lactic acid bacteria, molds, and yeasts. Generally, higher activities were found in bacteria. CoA inhibited the phosphorylation of pantothenic acid, and resulted in a decrease of CoA production in all the CoA producing strains, while only a little inhibition by CoA was observed in the other reactions, and CoA production from pantothenic acid 4'-phosphate by Brevibacterium ammoniagenes IFO 12071 was not repressed even in the presence of 4mM of CoA. Extracellular excretion of the enzymes of CoA biosynthesis was observed when cells were in contact with sodium lauryl sulfate. Degrading activity against CoA and that against AMP were relatively lower in CoA producing strains when compared with those in other strains. It was confirmed that Brown's route of CoA biosynthesis operates in Brevibacterium ammoniagenes IFO 12071.

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An extracellular amylase from a bacterium, Bacillus megaterium strain No. 32, was purified over 2600-fold by precipitation with ammonium sulfate, column chromatography with SE-Sephadex and gel-filtration with Sephadex G-100. The enzyme was most active at pH values around 6.5, and was stable in pH range between 5 and 7.5. The enzyme activity was inhibited by p-chloromercuribenzoate and was restored completely by the addition of cystein. The isoelectric point of the enzyme was pH 9.1. Results of experiments in which maltooligosaccharides terminated at the reducing end by radioactive glucose were used as substrates for the enzyme, showed that the enzyme removed two glucose unit (maltose) from the nonreducing end. From these results, the enzyme resembled the higher plant β-amylase in the action.

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Medicarpin-β-D-glucoside (II) was isolated from roots of alfalfa, Medicago sativa L. Its structure was established by chemical and physicochemical evidences.

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The life of m-tolyl N-methylcarbamate (Tsumacide^®) in rat body is very short. Depending on the sex, from 78 to 96.6%, of an administered dose of ¹⁴C-Tsumacide is eliminated in the urine by 24hr after treatment. In rats, Tsumacide is mainly metabolized by oxidative reactions at m-methyl group and 4-position of the aromatic ring. Hydrolysis of the ester linkage occurs only to a minor extent. The resulting oxidation products and phenols are partly conjugated as sulfate and glucuronides. The major metabolic pathways are identical in male and female rats.
From 78.7 to 85.8% of the radiocarbon is rapidly lost from the bean plants treated with ¹⁴C-Tsumacide and -Meobal by 24hr after treatment. Approximately 70% of the dose is also lost within 24hr after topical application of ¹⁴C-Tsumacide to houseflies. Oxidation of m-methyl group, N-methyl group and the aromatic ring at 4-position occurs with Tsumacide in houseflies and bean plants. Meobal undergoes hydroxylation at 4-methyl group and N-methyl group in bean plants. The oxidation products are conjugated as glucuronide and glucoside in houseflies and as glucosides in bean plants.
Tsumacide metabolites formed by oxidation are less active than the original compound in anticholinesterase activity.

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The survey of the presence of chemical resistant factors in plants against the larvae of Spodoptera litura F. was examined. In addition, the antifeeding diterpenes were surveied from thirteen species of plants that belong to Verbenaceae family. Finally isolated thirteen antifeedants and one derivative were examined the antifeeding activity for the larvae by the leaf disk method.

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A procedure was elaborated to prepare LMM fragment of myosin by means of treatment with neutral protease from Bacillus polymixa (BPNP). The LMM obtained by digestion with BPNP proved to be similar to the same fragments obtained by treatment with cyanogen bromide (LMM-C) or trypsin (LMM Fr 1), as regards amino acid composition and helical content.
The sedimentation coefficient, s°_20W, and the intrinsic viscosity of this LMM were determined as 2.95 S and 1.02dl/g, respectively, the molecular weight was estimated to be about 140, 000 by sodium dodecyl sulfate gel electrophoresis and gel filtration on Sepharose-6 B in 5M guanidine hydrochloride. The particle lengths measured on electron micrographs were 590, 635 and 652 Å for number average, weight average and peak length, respectively. Those molecular characteristics indicated that BPNP-treated LMM (LMM-BPNP) was the smallest LMM among three LMMs examined.
While LMM-BPNP was prepared by proteolysis like LMM Fr 1, it possessed thermostability very similar to that of LMM-C.

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Oxidation of D-limonene with selenium dioxide-hydrogen peroxide affords (+)-1-hydroxyneodihydrocarveol as the major product formed via cis- and trans-limonene epoxide. Hydrolysis of the former epoxide is much faster than that of the latter, which can therefore be obtained in almost quantitative yield on acid hydrolysis of a mixture of cis- and translimonene epoxide (1:1) under mild condition.
Minor significance of oxygenation in an allylic position to a trisubstituted double bond and the difference of accessibility of an allylic position to di- and trisubstituted double bond toward the oxidant were also observed.

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Chlorination of DDT gave unsymmetrical tetrachloro derivative which was converted into 4, 4'-dichlorobenzil in a single step of rearrangement and hydrolysis by heating with p-toluenesulfonic acid or monochloroacetic acid containing a little water. The optimum condition for the conversion of 4, 4'-dichlorobenzil to chlorobenzilic acid is also described.

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A strain of Erwinia aroideae produces a remarkable amount of pectolytic enzyme when the organism was induced by nalidixic acid for the bacteriocin production. This pectolytic enzyme was purified approximately 60-fold from the induced medium by carboxymethylcellulose and Sephadex G-75 gel column chromatographies after batchwise treatment with carboxymethyl- and diethylaminoethyl-celluloses. The purified enzyme was almost homogeneous on sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis, a molecular weight of about 28, 000 to 32, 000 was determined for this enzyme. The optimum pH of the enzyme activity was about 8.0 to 8.2. The purified enzyme produced reaction products from pectin and methoxylated pectic acid which had a strong absorption at 235nm indicating a trans-eliminase reaction. Pectin or pectic acid with higher methoxyl content was a good substrate for this enzyme, while no significant activity was observed when pectic acid was a substrate. The limit of degradation of pectin and pectic acid with higher methoxyl content (90% esterified) by the enzyme were 6.5% and 43%, respectively. It was concluded that the enzyme is a new endo-pectin trans-eliminase from bacterial origin

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