The crystalline α-glucosidase from Mucor javanicus has a sedimentation constant (s⁰_{20, w}) of 6.1 S, a diffusion constant (D_{20. w}) of 4.8×10^-7 cm²•sec^-1, and an average molecular weight, as determined by two different methods, of 124, 600. The α-glucosidase is a glycoprotein containing the following constituents; tryptophan₂₃, lysine₈₁, histidine₃₉, arginine₃₄, aspartic acid₁₀₂, threonine₆₉, serine₄₆, glutamic acid₇₈, proline₅₅, glycine₇₈, alanine₅₅, half cystine₃, valine₅₃, methionine₁₇, isoleucine₅₈, leucine₆₁, tyrosine₅₁, phenylalanine₄₁, glucosamine₁₂, and mannose₃₈.
The low content of half cystine, the high contents of aspartic acid, lysine, and histidine, and the presence of mannose as the sole constituent of neutral sugar are the characteristics of this enzyme.

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The degraded products of β-cyclodextrin (β-CD) by γ-irradiation in aqueous solution were identified as six oligosaccharides, such as glucose, maltose, maltotriose, maltotetraose, maltopentaose and maltohexaose, by the gel-filtration method on Biogel P-2 at 60°C. Analysis of the hyperfine structure of the ESR spectrum observed in the oxidation of β-CD with Ti³⁺-hydrogen peroxide system indicated that a radical was formed mainly by hydrogen abstraction at C-5 of a glucose residue. A mechanism of the ring cleavage of β-CD by γ-irradiation in oxygen-free aqueous solution was proposed.

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Neutral sugar composition of cell walls of suspension-cultured tobacco cells was examined with the advance of culture age by an anion-exchange chromatography. Isolated cell walls gave on hydrolysis the following sugars: 2% of L-rhamnose, 6% of D-mannose, 26% of L-arabinose, 13% of D-galactose, 8% of D-xylose and 47% of D-glucose as neutral sugars. Little changes in composition of cell wall polysaccharides were recognized with the advance of culture age. Sugar composition of the extra-cellular polysaccharides was similar to that of hemicellulose fraction from cell walls. Pectinic acid gave on hydrolysis 2-O- (α-D-galactopyranosyluronic acid) -L-rhamnose, D-galacturonic acid and its oligosaccharides.

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An α-glucosidase which was homogeneous in disc electrophoretic and ultracentrifugal analysis was obtained from buckwheat seeds by fractionation with ammonium sulfate, chromatography on CM-cellulose and gel filtration on Bio-Gel P-150. The sedimentation coefficient (S_{20, w}) was estimated to be 6.2 S, and the molecular weight, 80, 000_??_88, 000. The ratio of initial velocity of hydrolysis for maltose (Km=2.13mg/ml) and soluble starch (Km=7.46mg/ml) was calculated to be 100:86. Both activities were competitively inhibited by Tris and erythritol, and stimulated approximately 1.5-fold in the presence of 25mM-K⁺ or Na⁺. The Ki values for Tris on hydrolysis of maltose and soluble starch were 0.78mM and 1.07mM, and those for erythritol, 77.7mM and 64mM, respectively.

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A strain of Bacillus sp. (Bacillus R-4) produces a protease and a chitosanase which have the ability of lysing Rhizopus cell wall. Some enzymatic properties of the protease purified to a homogeneous state were examined.
The molecular weight of the enzyme was estimated as 19, 000 and the isoelectric point as pH 8.65. The protease appeared to have a relative wide range of substrate specificity, hydrolyzing various proteins, such as gelatin, hemoglobin and protamine, and synthetic peptides, such as Z-Gly-Try-NH₂, Z-Gly-Ala-NH₂, Z-Ala-Leu-NH₂ and Z-Gly-Leu-NH₂. The activity lost by EDTA and by Hg²⁺ was restored by Zn²⁺ and reduced glutathione, respectively.

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Eucheuma muricatum mucilage which was extracted and purified after irradiation of the seaweed with γ-ray of ⁶⁰Co formed a complex with I^-₃, and exhibited a new absorption band at 555 nm. The absorbancy observed at that time depended on the concentration of urea and on the temperature. The curves representing relations between absorbancy at 555 nm and the above factors have two inflection points. The fact that their inflection points shift toward the lower temperature side with the increase in urea concentration suggests that the coloring phenomenon may relate closely to the sol^→_←gel transition of the mucilage. It was also found that the absorbancy at 555 nm depended on the content of pyruvic acid residue in the same mucilages, the absorbancy decreased with the increase pyruvic acid residues, and that the steric hindrance caused by a sugar residue of large demension affected the stable from containing viscous polysaccharide.

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Dextran was subjected to oxidative scission by periodate, followed by ring closure with nitromethane to form nitrodextran. The nitro group attached to the ring was reduced by LiAlH₄ to yield amino-polysaccharide of which the molecular weight was about 10, 000. It became clear that nitrodextran consisted of 3-deoxy-3-nitro-mannopyranoside, -glucopyrano-side, -galactopyranoside and -talopyranoside and their molar ratio was 6:5:1:2 as determined by column chromatographic separation and gas chromatographic analysis of the methanolyzate of nitro-dextran.

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Cholesterol ester hydrolase (EC 3. 1. 1. 13) was purified about 10 times from the culture supernatant of Pseudomonas fluorescens ATCC 21156 by a procedure involving fractionations with acetone and ammonium sulfate, chromatography on hydroxylapatite, gel filtration on Sephadex G-100, and separation into two isoenzymes, I and II, by isoelectric focusing.
The purified enzyme from the Sephadex G-100 fraction sedimented as a single peak on ultracentrifugation. The isoenzymes I and II showed isoelectric points of pH 3.8 and 4.9, respectively. They both had an apparent molecular weight of about 129, 000, hydrolyzed preferentially long-chain fatty acid esters of cholesterol, and exhibited a pH optimum at 7.3 with cholesterol linoleate as substrate. The Km values of I and II for cholesterol linoleate were 3.86×10^-5 M and 1.43×10^-4 M, respectively. The cholesterol esterase activity was remarkably stimulated when Triton X-100 was used to prepare the miscellar aqueous substrate, and further increase in activity was demonstrated by the addition of cholic acid to the miscellar system.

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A sex stimulant to the Periplaneta males was isolated from various plant leaves and identified as germacrene D. The compound was active with a threshold amount of 10μg to the males of Periplaneta americana L. and P. japonica Karny. The response elicited with germacrene D was identical with that caused by the female sex pheromone of the each species.

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The sex pheromone produced by adult females of the potato tuberworm moth was isolated from unmated female moths reared in the laboratory. The gas chromatographic and mass spectrometric data suggested the pheromone to be a tridecatrienyl acetate. The isolated pheromone was subjected to partial hydrogenation with hydrazine and hydrogen peroxide and subsequent ozonolysis to produce a mixture of ω-acetoxy-alkanals. They were identified by mass chromatographic technique as 4-acetoxy-butanal, 7-acetoxy-heptanal, and 10-acetoxy-decanal respectively. Consequently, the pheromone was identified as 4, 7, 10-tridecatrienyl acetate except the geometric configuration.

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Intracellular localization of several enzymes related to tricarboxylic acid cycle was investigated during the aerobic growth of Candida tropicalis on acetate, n-alkane and glucose. NADP-linked isocitrate dehydrogenase in acetate-grown cells was mostly found in S₂ fraction (20, 000×g supernatant fraction of protoplast lysate), whereas more than half of this activity in n-alkane-grown cells was recovered in P₂ fraction (20, 000×g pellet fraction). Large parts of NAD-linked isocitrate dehydrogenase and malate dehydrogenase were present in P₂ fraction, while NADP- and NAD-linked glutamate dehydrogenases were found preferentially in S₂ fraction, irrespective of the growth substrates used. Isocitrate lyase was detected in both fractions. Citrate synthase and aconitase in acetate-grown cells were almost particulate. Catalase activity recovered in P₂ fraction was far higher in alkane-grown cells than in acetateor glucose-grown cells.

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Partially purified preparations of lipoxygenase from the germinating barley embryos converted linoleic acid to 9- and 13-hydroperoxy linoleic acids in the ratio of approximately 3:1, while the similar preparations from the ungerminated embryos converted linoleic acid mainly to 9-hydroperoxy linoleic acid.
Isoelectric focusing of the partially purified preparations of the germinating embryos revealed the presence of the two lipoxygenase active peaks, having isoelectric point at pH 4.9 and 6.6, respectively. The former peak (barley lipoxygenase-1) was identical to lipoxygenase of the ungerminated embryos, but the latter peak (barley lipoxygenase-2) was found only in the germinating embryos. The newly found isoenzyme, barley lipoxygenase-2, converted linoleic acid mainly to 13-hydroperoxy linoleic acid, and could oxidize esterified derivatives of linoleic acid (methyl linoleate and trilinolein) much strongly than barley lipoxygenase-1.

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The structures of ezomycins A₁ and A₂, antifungal antibiotics produced by a strain of Streptomyces, were determined as 1 and 2, respectively, by degradative and spectrometric studies.

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Fusion reactions of the biologically important synthetic cytokinin N⁶-benzyladenine and fully acetylated D-ribofuranose (2), D-ribopyranose (7) and D-glucopyranose (6) have been carried out in the presence of bis- (p-nitrophenyl) hydrogen phosphate. N⁶-Benzyl-9-β-D-ribofuranosyladenine (5), N⁶-benzyl-9-β-D-ribopyranosyladenine (11), N⁶-benzyl-9-β-D-glucopyranosyladenine (9) and their acetates (4, 10, 8) were obtained in fairly good yields respectively. This provides an alternative synthetic route of N₆-benzyladenine-9-glycosides.

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Acids of lamina and midrib cigarette smoke were converted into trimethylsilyl derivatives and they were analyzed with glass capillary column gas chromatography. Then compositional differences of acids between lamina and midrib cigarette smoke were discussed. The concentrations of organic acids were higher for lamina cigarette smoke than for midrib cigarette smoke. Large concentration difference were found in formic, acetic, propionic, lactic, glycolic, furoic, benzoic, phenylacetic, fumalic and m-hydroxybenzoic acid. Succinic and methylsuccinic acid were similar in lamina smoke and in midrib smoke.
A large amount of 2, 3-dihydro-3, 5-dihydroxy-6-methyl-4 H-pyran-4-one (amino-carbonyl reaction product) was identified for the first time in lamina smoke.

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Fractionation of smoke condensate and a detailed analysis of phenols in the cellulose cigarette smoke were performed during the course of systematic compositional studies of the smoke. Nearly equal yields of phenolic, acidic and neutral fractions were obtained in fractionation of cellulose cigarette smoke condensate. Thirtyseven phenols were newly identified in the smoke of cellulose cigarette in addition to six phenols ever found in pyrolysis products of cellulose. Semi-quantitative determination of some of these phenols revealed that dihydric phenols, such as catechol, hydroquinone, resorcinol and their alkyl derivatives were the main phenols and monohydric phenols were the minor phenols.

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In the course of systematic studies of cellulose cigarette smoke, organic acids in the smoke were examined by glass capillary gas chromatography and 43 carboxylic acids were either tentatively or positively identified. Semi-quantitative estimation of some of these acids revealed that formic, acetic and glycolic acid are the major acids present in cellulose cigarette smoke and together these three acids accounted for more than 80% of total acids estimated. Mono- and di-basic short-chain acids were found to be present in much more amounts than acids possessing benzene or furan rings. The acids in cellulose cigarette smoke are compared with those in tobacco cigarette smoke and the differences between them are discussed.

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Wax bean hemagglutinin was dissociated by 8 M urea at pH 8.3 and fractionated on DEAF-cellulose in the presence of 8 M urea. Two out of the four fractions obtained were further fractionated on CM- or DEAE-cellulose in the absence of urea. By these fractionation procedures, fractions I, II-1, III-1, III-2 and IV were obtained and one of them, fraction III-2, possessed an enzyme activity, ribonucleolytic activity. These fractions were characterized by SDS-polyacrylamide gel electrophoresis. From the results, fraction II-1 was identified as a highly purified 20, 000 molecular weight subunit and fraction III-2 contained a 25, 000 molecular weight subunit predominantly.

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It was observed that the DL-dipeptide derivative was formed prodominantly over the LL-compound, only when L-Pro-OR was allowed to react as amino-component to the pseudooxazolone- (5), in contrast to the other L-amino acid esters. From the observation of the influence of the solvent, the added base and H-GIy-OMe, some of the mechanism in this reaction was discussed. The preparative isolation of the DL-compound from the reaction product, the synthesis of Tfp-L-Ileu-OH and the corresponding pseudooxazolone- (5) compound were also described.

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The accumulation of vitamin B₁₂ by Bacillus badius grown on hydrocarbon was investigated. The bacterium could assimilate n-alkanes of C₁₁-C₁₈, ethanol, fumarate, α-ketoglutarate and malate. n-Alkanes of C₁₆-C₁₈ were the best for vitamin B₁₂ production. The bacterium utilized well all of the nitrogen sources tested. Above all, ammonium dihydrogen phosphate was the best for the bacterial growth and vitamin B₁₂ production. Addition of organic nutrients such as malt extract and meat extract, and addition of metal ions such as ferrous and cobalt promoted the growth and vitamin B₁₂ production. Interestingly, vitamin B₁₂ was produced mostly in the supernatant. The cyanoform of the corrinoid predominantly formed in the supernatant would confirm the identity with cobalamin.

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A key intermediate, 2-isocyano-3-hydroxybutyrate (III) was isolated from a reaction of isocyanoacetate (I) with acetaldehyde (II) in the presence of Et₃N. It was found that III was readily converted into 2-isocyanocrotonate (V) and 2-isocyano-2-(1'-hydroxyethyl)-3-hydroxy-butyrate (VI) which are undesirable compounds for the synthesis of threonine. However, by use of a metal catalyst (e. g. NiCl₂ or PdCl₂), the isocyano-hydroxy compound (III) was selectively converted into 5-methyl-4-alkoxycarbonyl-2-oxazoline (IV) which is an important precursor of threonine. Furthermore, chemical properties of IV were examined; the results suggested that cis-oxazoline was relatively sensitive to acid, base and heat.
On the basis of these results, the reaction of I with II was carried out using Et₃N-PdCl₂ as a catalyst to obtain threo-threonine (85% purity) in a good yield (85%).

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The pyrolyzate of the nondialyzable melanoidin prepared from glucose-ammonia reaction system (kept in pH 5.3_??_6.0 during the reaction) was fractionated to volatile fraction and nonvolatile fraction. Among the volatile components, two pyridines and four alkylpyrazines were identified. On the other hand, one imidazole compound and two β-hydroxypyridines isolated from the nonvolatile fraction were identified as 4 (5) -methylimidazole, 3-hydroxypyridine and 2-methyl-5-hydroxypyridine, respectively. It is inferred that these compounds are not produced by the fission of the main skeleton in the melanoidin molecule, but formed by pyrolysis of the heterocyclic compounds present as a small moiety in the melanoidin.

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This investigation was carried out to demonstrate the occurrence of phosphonolipids in bovine gall bladder bile.
It has been demonstrated that the bovine bile contains three phosphonolipids (glyceryl-ciliatine, glyceryl-trimethyl-ciliatine and ceramide-ciliatine) with a combination of silicic acid column and silica gel thin-layer chromatographic behavior and chemical properties.

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Various physiological roles of mammalian aldehyde dehydrogenase had been anticipated because of its broad substrate specificity. In order to clarify roles of the enzyme and the regulation of aldehyde metabolisms in liver, the intracellular distribution and isozyme of beef liver aldehyde dehydrogenase were studied.
The presence of the mitochondrial, the microsomal and the cytoplasmic isozymes were proved by the isoelectric focusing. These isozymes were different from each other in pH-activity curve in the responces for steroid hormones and disulfiram.
It was suggested by comparing the reactivities of these isozymes for various aldehydes that particular aldehyde might be oxidized by a favorite isozyme at particular locality in the liver cells and that a share of physiological role among these isozymes is probable.

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Intracellular arylsulfatases from Klebsiella aerogenes W 70 cells grown in methionine medium (M enzyme) and inorganic sulfate medium containing tyramine (T enzyme) were purified respectively by fractionation with (NH₄)₂SO₄, followed by successive chromatographies on DEAE cellulose, hydroxylapatite, Sephadex G-100 and DEAE Sephadex A-25. On polyacrylamide gel electrophoresis, the two enzymes gave single bands with the same mobilities. Molecular weights of both, determined by SDS gel electrophoresis and by Sephadex G-100 chromatography, were 47, 000 and 45, 000, respectively. Their activities were maximal at pH 7.5. The affinities of the enzymes (M and T enzymes) for their substrate (Km) and the maximum velocity of hydrolysis (V_max) were enhanced by addition of electron withdrawing substituents. The enzymes were inhibited by inorganic phosphate, cyanide, hydroxylamine and tyramine. The inhibition by tyramine was competitive (Ki=1.0×10^-4 M). These results show that the two enzymes were identical. This was confirmed by the fact that mutant strains, which were unable to synthesize arylsulfatase when grown with methionine, could also not synthesize the enzyme when grown with tyramine.

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A choline dehydrogenase, which was present in the particulate fraction of the cell-free extract of Pseudomonas aeruginosa A-16, oxidized choline to betaine aldehyde without any dissociable coenzymes, while the enzyme, which was treated with Triton X-100, oxidized choline only with a supplement of phenazine methosulfate. The difference spectrum showed the presence of cytochrome-like components in the particulate. Km values for choline and phenazine methosulfate were 1.7×10^-3 M and 1.4×10^-4 M, respectively. The dehydrogenase was inhibited by SH-reagents such as p-chloromercuribenzoate and iodoacetic acid. Of a variety of substrates tested, only choline caused the enzymatic reduction of phenazine methosulfate. The estimation of choline was tried using the enzyme.

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Two nematicidal polyacetylenes, 3-cis, 11-trans- and 3-trans, 11-trans-trideca-1, 3, 11-triene-5, 7, 9-triyne, were isolated from flowers of Carthamus tinctorius L., by column chromatography and high speed liquid chromatography under the dark condition.
The nematicidal activities of 3-cis, 11-trans- and 3-trans, 11-trans-isomer to Aphelenchoides besseyi were 80% at 10 ppm and 95% at 1 ppm, respectively.

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The synthesis of 2, 3-(³H) -gibberellin A₉ (GA₉) with a specific activity of 47 Ci mmole^-1 is described. Δ^{2, 3}GA₉ methyl ester epoxide was converted to (³H)-GA₉ methyl ester epoxide using carrier-free tritium gas. This product was de-epoxidized then hydrolysed to yield (³H) -GA₉.

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