Galactosylsucroses contained in soybeans are not digestible. Thus we wished to detect α-galactosidase (EC 3. 2. 1. 22) in intestinal bacteria. The strain of E. coli in the title was found to produce considerably this enzyme adaptively. We could prepare rather pure solution of the enzyme from the sonicate of the strain. It was purified about 142-fold. It showed optimum pH and temperature at 6.8 and 37°C, respectively, with the substrate p-nitrophenyl-α-D-galactoside (PNPG). Dilute enzyme solutions were very unstable even at 0_??_5°C. However, concentrated solutions were considerably stable. The Michaelis constant (M) was 1.07×10^-4, 2.33×10^-3, and 3.65×10^-1 for PNPG, melibiose, and raffinose, respectively, The maximum velocity (mole/min/mg protein) was 2.72×10^-5, 2.67×10^-5, and 2.04×10^-5, respectively for the same three substrates. This enzyme had a weak transferase action.

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A microorganism M-2 was isolated as a strain capable of converting (-)-menthone to other compounds. The strain was identified as Pseudomonas fluorescens by taxonomical investigation. The conversion products of (-)-menthone were determined to be (-)-t-4-isopropyl-3-oxo-γ-1-cyclohexanecarboxylic acid, ^* (+)-c-4-isopropyl-3-oxo-γ-1-cyclohexanecarboxylic acid^* and (+)-t-3-hydroxy-t-4-isopropyl-γ-1-cyclohexanecarboxylic acid.^* As the main pathway, it was proposed that (-)-menthone was oxidized to a keto acid which was successively reduced to a hydroxy acid.

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Lipases from Aspergillus niger and Rhizopus delemar hydrolyzed triolein and produced 1, 2 (2, 3)-diolein and 2-monoolein. These two lipases appears to have strong specificity towards the outer chains of the triglyceride. Comparing the proportions of fatty acids in position 1 (3) of cocoa butter with proportions of fatty acids liberated after limited hydrolysis of cocoa butter, it becomes clear that these two lipases do not hydrolyze the ester bond in position 2 of the triglyceride.
On the other hand, lipases from Geotrichum candidum Link and Penicillium cyclopium Westring attacked the fatty acid chains regardless of their positions. Geotrichum candidum lipase liberated oleic acid and palmitic acid in preference to stearic acid from cocoa butter.

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A lytic enzyme which was capable of lysing cells of Streptococcus mutaus was purified from the culture filtrate of Streptomyces griseus H-402 by Amberlite CG-50 treatment, CM-cellulose and hydroxylapatite column chromatographies, and Sephadex G-150 gelfiltration, The lytic enzyme was obtained in a crystalline form which was homogeneous in polyacrylamide gel electrophoresis. The molecular weight was estimated to be 2×10⁴ by the thin-layer gel-filtration method on Sephadex G-75, and 2.3×10⁴ by the method of polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. The enzyme was found to be a N-acetylmuramidase whose activity was lost by N-bromosuccinimide as an inhibitor.

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An α-glucosidase has been isolated from the mycelia of Penicillium purpurogenum in electrophoretically homogeneous form, and its properties have been investigated. The enzyme had a molecular weight of 120, 000 and an isoelectric point of pH 3.2. The enzyme had a pH optimum at 3.0 to 5.0 with maltose as substrate. The enzyme hydrolyzed not only maltose but also amylose, amylopectin, glycogen, and soluble starch, and glucose was the sole product from these substrates. The Km value for maltose was 6.94×lO^-4M. The enzyme hydrolyzed phenyl α-maltoside to glucose and phenyl α-glucoside. The enzyme had α-glucosyltransferase activity, the main transfer product from maltose being maltotriose. The enzyme also catalyzed the transfer of α-glucosyl residue from maltose to riboflavin.

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The changes during growth and sporulation in activities of cells of Bacillus subtilis to incorporate various amino acids were investigated with wild-type strain and its asporogenous mutant. In the case of wild type strain the uptake of valine, phenylalanine, and proline was largest during the logarithmic growth period. The uptake of these amino acids decreased rapidly during the early stationary phase. The uptake of valine and cysteine increased again to some extent just prior to the forespore stage. The uptake of glycine and serine, however, was largest at the forespore stage at which the formation of spore coat took place. From these observed phenomena it was assumed that the remarkable incorporation of glycine and serine into the wild type strain during sporulation was closely related to the formation of spore coat.

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Rapid absorption and degradation of chlomethoxynil were observed in seedlings of rice and barnyard millet. The amounts of absorption by rice and barnyard millet, at the 2-leaf and 4-leaf stages of plants, were determined in water and soil cultures.
The major metabolic products of chlomethoxynil were 2, 4-dichlorophenyl 3'-hydroxy-4'-nitrophenyl ether, 2, 4-dichlorophenyl 3'-methoxy-4'-aminophenyl ether and their conjugates, and 2, 4-dichlorophenyl 3'-methoxy-4'-acetylaminophenyl ether. Several minor unknown substances were also detected. A major metabolic pathway of chlomethoxynil in plants was postulated.

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Glycopeptides from the pronase digest of a 7 S protein have been separated into five fractions (peaks IIIa, IIIb, IIIc, IIId and IIIe) on Dowex 50-X 2 with 13.7mM pyridineacetic acid buffer (pH 3. 10). With the exception of the last peak (IIIe), the peaks were tripeptide-carbohydrates. All the fractions had asparagine as the N-terminal amino acid residue to which the carbohydrate moiety was attached. By amino acid sequence analysis, two tripeptides were found; Asp-Gly-Thr in peaks IIIa, IIIb and IIIc and Asp-Ala-Thr in peaks IIIb, IIIc and IIId. Peak IIIe was a mixture of three peptides. The tripeptides had two glucosamine and seven to nine mannose residues per aspartic acid, showing the same molar ratio as the L-β-aspartamido-carbohydrates. The same three types of carbohydrate groups probably resulted from paucidispersity.

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Synthesis of twentyone 4-oxa-farnesane derivatives with a variety of substituents at C₁ and C₁₁ is described. Some of these compounds are more active than farnesyl methyl ether against Dysdercus koenigii nymphs.

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Some proteolytic properties of neutral proteinases I and II (N-1 and N-II), zinccontaining metalloenzymes, from Aspergillus sojae were investigated.
Both of the enzymes acted on proteins as endo-type proteinase, and did not show any esterase-, amidase- or exopeptidase-activities. However, they were distinguishable in some properties as follows. (A) N-I digested various proteins effectively, especially milk casein and soybean protein, while N-II was specifically active on basic nuclear proteins such as protamine and histone. (B) The di-substituted di-peptides such as Z-Gly-Y-NH₂ (Y=Phe or Leu) were hydrolyzed by N-I at the peptide bond of Gly-Y, while N-II was completely inactive on these substrates. (C) The peptide bonds in oxidized insulin B-chain susceptible to the action of N-I and N-II were quite different from each other.

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A minor carotenoid in integuments of orange pupae of a swallowtail, Papilio xuthus, was found also in carapaces of a crab, Paralithodes brevipes (hanasakigani). This new carotenoid showed a visible absorption spectrum similar to that of papilioerythrin (=α-doradexanthin) and was named papilioerythrinone. Reduction of this compound with sodium borohydride occurred in 2 steps and gave products identical with the reduction products of papilioerythrin. Therefore, the structure 3-hydroxy-β, ε-carotene-4, 3'-dione (IV) is ascribed to papilioerythrinone.

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Fatty acid compositions of intracellular and extracellular lipids from Mycotorula japonica grown on glucose or individual n-alkane (C₁₀-C₁₆) as a sole source of carbon were comparatively studied.
In neutral lipid fraction of intracellular lipids, an increase of fatty acid having the same chain length as that of n-alkane substrate was observed when cells were grown on n-alkane, while significant amounts of total C₁₇ fatty acid (heptadecanoic and heptadecenoic acids) were produced when cells were grown on odd-carbon number n-alkane (C₁₁, C₁₃ and C₁₅).
In phospholipid fraction of intracellular lipids, an increase of short chain length fatty acids (C₇-C₉) was found when cells were grown on individual n-alkane (C₁₀-C₁₆); especially in case of C₁₄-C₁₆, being more than 70%, while the amount of short chain fatty acids was little when cells were grown on glucose.
In extracellular lipid found in glucose medium, a greater proportion of short chain fatty acids (C₆-C₉) was observed, although such short chain fatty acids were contained in small quantities in intracellular lipid. In n-alkane medium, such short chain fatty acids were also observed.

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Ribonucleic acid prepared according to modified Kirby's phenol method from 14 day old chick embryos was fractionated into three peaks by MAK column chromatography. To the peak eluted in the range from 0.65 to 0.7M NaCl, the name “Peak II” was given tentatively. Purified Peak II was homogeneous upon ultracentrifugation, polyacrylamide gel electrophoresis and elution patterns of chromatographies.
The peak gave a positive result with the orcinol reaction. Furthermore, the hydrolyzate by DNase and snake venom was active in promoting the growth of L. leichmannii 7830. The peak was not disrupted by treatment with RNase, but was with DNase. Peak II treated with heat gave the same fractionation profile as the native peak by gel filtration on Sephadex G-200.
By treatment with 0.3 N KOH at 37°C for 18 hr, Peak II was resolved into two peaks, Peak II-1 and Peak II-2. It was found that Peak II consisted of the susceptible and the non-susceptible components to the alkali. From the susceptible component (Peak II-2) the usual ribonucleotides were detected. Moreover, the four usual deoxyribosides were recognized from the component digested by DNase and snake venom.
The susceptible component was more active in promoting the growth of L. leichmannii even without enzyme digestion. On the other hand, the non-susceptible component (Peak II-1) was not active, The digest of the non-susceptible component gave the similar results as calf thymus DNA digested by DNase and snake venom, while the susceptible one gave different results in the activity of promoting the growth of L. leichmannii

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A new synthetic approach of 2-hydroxy-5, 5-dialkylcyclopent-2-en-1-ones was investigated. The reaction of cyclotene (2-hydroxy-3-methylcyclopent-2-en-1-one) (1) and 1, 3-propanedithiol in chloroform gave the corresponding 1, 3-dithiane derivative (2) in good yield. The methylation of 2 afforded 2, 2-dimethyl-5, 5-[propanediyl-(1, 3)-dithio] cyclopentan-1-one (3). Similarly, 2 was alkylated with ethyl iodide, n-propyl bromide, allyl bromide, and benzyl chloride to yield the corresponding 2, 2-dialkyl-5, 5-dithiocyclopentan-1-one derivatives (4_??_7). Hydrolysis of the thioketal function in 3 to give the corresponding enolizable 1, 2-diketone (8) was performed by the use of N-chlorosuccinimide in the presence of silver ion.

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Sodium dodecyl sulfate (SDS)-insoluble proteins from wheat flour were solubilized by the reduction of their disulfide linkages with 2-meracaptoethanol. The polypeptide compositions of the reduced SDS-insoluble proteins were compared with those of the reduced glutenin by SDS-polyacrylamide gel electrophoresis, isoelectric focusing and amino acid analysis. SDS-polyacrylamide gel electrophoretic patterns of the reduced SDS-insoluble proteins almost coincided with those of the reduced glutenin. Seven major bands (Band 1_??_7) were obtained from both samples of the reduced proteins. These protein bands were subjected to analysis of amino acid compositions and isoelectric focusing, and similarities between polypeptides of the SDS-insoluble proteins and the glutenin were observed in their amino acid compositions and isoelectric focusing patterns. The results obtained suggested that the preparation of the reduced SDS-insoluble proteins might be used as a simple and rapid method to obtain the glutenin subunits.

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In the course of the development of new herbicide (MK-616: N-(p-chlorophenyl)-3, 4, 5, 6-tetrahydrophthalimide), the structure-activity relationship of cyclic imide herbicides was investigated. Sixty N-substituted phenyl-3, 4, 5, 6-tetrahydrophthalimides and related compounds were prepared and their herbicidal activity were examined. The results indicate that, for high inhibitory activity, lipophilic tetramethylene moiety, double bond or nitrogen atom (s) in bridgehead position and N-(p-substituted) phenyl moiety are essential in these cyclic imides. New 4-(p-chlorophenyl)-1, 2-tetramethyleneurazole and 3 (p-chlorophenyl)-1, 5-tetramethylenehydantoin were found to be herbicidally active.

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A cyclodextrin glycosyltransferase-producing bacterium was isolated from soil in a high alkaline pH medium containing 1% of Na₂CO₃. The isolated alkalophilic microorganism was identified as a member of the genus Bacillus. Highest yield of the enzyme was obtained by using the following medium; 1% soluble starch, 5% corn steep liquor, 0.1% K₂HPO₄, 0.02% MgSO₄•7 H₂O and 1% Na₂CO₃. The crude enzyme showed a very broad pH-activity curve and had two optimum pH ranges at 4.5_??_5.0 and 7.5_??_8.5. The crude enzyme also catalyzed the formation of cyclodextrins at both of the optimum pH ranges.

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Several 5-aryl-2-triazolyl oxadiazoles, 5-aryl-2-oxadiazolylamino/thiadiazolylamino oxadiazoles, have been prepared by cyclisation of 1-aroyl-4-oxadiazolyl thiosemicarbazides, and 5-aryl-2-thiatriazolylamino oxadiazoles were prepared by diazotisation of 4-oxadiazolyl thiosemicarbazides. These compounds have been screened against two fungal species, for their fungicidal activity.

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The relationship between structure and activity in bacteriostasis was compared to the relationship between structure and reactivity in metal chelate formation for various kojic acid analogs. The metal chelating property with Cu²⁺ and Cd²⁺ was closely related to the acid dissociation constant. Since structure-activity analyses showed that the neutral molecule is the active form, and since the pK_{_??_} value as the electronic parameter is related to the activity only for the neutral molecule but not for the ionized form, it was postulated that metal chelate formation is not significant for manifestation of the bacteriostatic activity of this series of compounds. This postulation was confirmed in experiments under conditions in which the metal was depleted or excessive. The bacteriostatic activity of kojic acid analogs against such bacteria as S. aureus 209 P, E. coli K-12 and Ps. fluorescens was neither retarded nor enhanced.

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Mutants, which fail to grow on glucose medium but can grow on succinate medium, were isolated by treatment with N-methyl-N'-nitro-N-nitrosoganidine from the wild-type strain of Agrobacterium tumefaciens, and were found to lose growth on several hexoses and three-carbon intermediates. The revertant mutants, which recovered the ability to grow on glucose medium, simultaneously regained the ability to grow on hexoses and three-carbon intermediates. By comparison of biochemical properties of the wild-type, the mutants and the revertant mutants, two mutant strains were characterized to be pyruvate carboxylase-deficient. Then, we concluded that these mutants might be induced by a single mutation at a genetic locus of pyruvate carboxylase and that the deficiency in the enzyme gave a pleiotropic effect on the ability to grow on hexoses and three-carbon intermediates. Some properties of pyruvate carboxylase of this bacterium were also presented.

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A mutant of Agrobacterium tumefaciens was isolated by treatment with N-methyl-N'-nitro-N-nitrosoguanidine as selective markers of succinate-negative and glucose-positive. This succinate-negative mutant was found unable to grow on pyruvate or any intermediate of the tricarboxylic acid cycle as the sole carbon source but able to grow on hexoses. Comparison of biochemical properties of wild-type and mutant strains demonstrated a very low level of succinate dehydrogenase activity in the mutant strain. Reverse mutation from succinate-negative to succinate-positive spontaneously occurred at a frequency of about 10^-9 . All strains of the revertant isolated recovered their activity of succinate dehydrogenase approximately to the level of the wild-type strain. Thus we presumed that the deficiency of succinate dehydrogenase in this mutant gave a pleiotropic effect on the ability to grow on tricarboxylic acid cycle intermediates.

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Incorporation of the carboxyl group of uronic acids and polyuronic acids into a triazine ring was studied. Following s-triazine derivatives were synthesized from uronic acids: 5 (R)-(2-amino-4-dimethylamino-1, 3, 5-triazin-6-yl)-L-arabopyranose (I) and methyl 5 (R)-(2-amino-4-dimethylamino-1, 3, 5-triazin-6-yl)-β-L-arabopyranoside (II) from galacturonic acid, 5 (R)-(2-amino-4-dimethylamino-1, 3, 5-triazin-6-yl)-α-D-xylopyranose (III) from glucuronolactone, and methyl 5 (R)-(2-amino-4-dimethylamino-1, 3, 5-triazin-6-yl)-α-D-lyxopyranoside (IV) from mannuronolactone respectively.
s-Triazine derivatives of polyuronic acids such as pectin and alginic acid were prepared similarly. After hydrolysis, triazinylated monosaccharides were isolated and identified with authentic derivatives.

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N-Blocked amino acids were condensed with o-phenylene diamine in the presence of DCC at 0°C to give derivatives of monoacyl-o-phenylene diamine. The ring closure of monoacyl derivatives thus obtained was then carried out by refluxing them for 10_??_200min in toluene, xylene or acetic acid. The dehydrating cyclization by triphenylphosphine was suitable for glutamine, asparagine, and threonine. By these methods, the carboxyl group of naturally occurring amino acids was all cyclized to benzimidazole-ring.
Among these new compounds, there are some biological active compounds as regulators for growth of plants or fungi.

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