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Kunio OHMIYA, Tsutomu KAJINO, Shoichi SHIMIZU, Kunihiko GEKKO
1989 Volume 53 Issue 1 Pages
1-7
Published: 1989
Released on J-STAGE: April 05, 2006
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The effects of pressure on the association states of enzyme-treated casein molecules were studied by monitoring the turbidity of solutions under high pressures up to 3, 000 kg/cm
2. β-Casein molecules partially degraded with immobilized trypsin were dissociated with increasing pressure up to a critical pressure (
e.g., 1.200 kg/cm
2 in Tris-HC1 buffer of pH 6.4 and ionic strength 1.0, at 40°C) and then reassociated under higher pressures up to 3, 000 kg/cm
2, following a parabola-like turbidity-pressure curve. The critical pressure for minimum turbidity increased with increasing degradation of the hydrophobic moiety of β-casein. In the case of chymosin-treated κ-casein, a similar pressure dependence of turbidity due to dissociation and reassociation was found in the same pressure region up to 3, 000kg/cm
2. However, chymosin-treated α
s1-casein was not reassociated even at 3, 500kg/cm
2. Based on these results, the pressure effects on the dissociation and reassociation of casein molecules are discussed in terms of the volume change of water around their hydrophobic moieties.
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Takuo SAKAI, Masahiko SAWADA, Tohoru KATSURAGI, Kenzo TONOMURA
1989 Volume 53 Issue 1 Pages
9-18
Published: 1989
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The extracellular production of a protopectin-solubilizing enzyme (protopectinase) of
Trichosporon penicillatum SNO-3 decreased when peptone was used as the nitrogen source. Peptone caused a morphological change in the microorganism. Cells cultured in the presence of peptone were elongated (E-cells), but those cultured in the absence of peptone were yeast shaped (N-cells). E-cells contained less cell wall mannan than N-cells. When cells were converted into protoplasts, protopectinase was liberated from N-cells together with polysaccharides, but not from E-cells. The sugar moiety of the protopectinase was identified. The enzyme contained 2 residues of glucosamine and 0.7 residues of mannose per molecule. The presence of protopectinase on the cell surface was monitored by means of immunofluorescence and enzyme immunoassays, which showed that the enzyme protein was present on the surfaces of both N- and E-cells. The results obtained suggested that cell wall mannan affects the secretion of this enzyme, and that E-cells cannot secrete the enzyme because they are deficient in cell wall mannan.
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Junji NAKAMURA, Kunio YAMANE, Koji YODA, Makari YAMASAKI
1989 Volume 53 Issue 1 Pages
19-24
Published: 1989
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An integration plasmid for
Bacillus subtilis, pTUE128 (8.2kb) which contained the
amyE'-'
bla fused gene, the
cat gene, and the DNA replication origin of the
E. coli plasmid μBR322, was constructed and was inserted into the
amyE region of the chromosome of a tunicamycin-resistant
B. subtilis mutant (
tmr A7 mutation). After cultivation of the transformants with 100μg/ml of chloramphenicol and/or 10μg/ml of tunicamycin, amplification of pTUE128 was analyzed by the increase of β-lactamase activity, the gene for which was included in pTUE128 and of α-amylase activity from the chromosomal gene, and by the method of DNA-DNA hybridization. This was done using the 24.5 kb amplification unit of
tmr A7 either by the addition of the high concentration of chloramphenicol or by the addition of tunicamycin.
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Hiroyuki UKEDA, Hideaki KAMIKADO, Kiyoshi MATSUMOTO, Yutaka OSAJIMA
1989 Volume 53 Issue 1 Pages
25-30
Published: 1989
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A new method is described for the co-immobilization of enzyme alcohol dehydrogenase (ADH) and coenzyme NAD onto Sepharose gel. In order to increase the degree of freedom of NAD, spacer molecules were introduced to the support by allowing it to repeatedly react with hexamethylenediamine and glutaraldehyde, and ADH and NAD were co-immobilized to the spacer-modified Sepharose. The results suggest that the re-usability of Sepharose-bound ADH and NAD (AN-Seph) depended both on the concentration and the reaction time of glutaraldehyde to a great extent. Furthermore, it is suggested that AN-Seph prepared under the optimum conditions might be applicable to the determination of ethanol.
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Norihiro AZUMA, Hiroaki MORI, Shuichi KAMINOGAWA, Kunio YAMAUCHI
1989 Volume 53 Issue 1 Pages
31-35
Published: 1989
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The DNA synthesis-stimulating activity of holo-lactoferrin from human milk was demonstrated using BALB/
c 3T3 cells. The stimulatory effect of lactoferrin was caused by the iron bound to the lactoferrin, because the lactoferrin molecule itself did not stimulate the DNA synthesis in the absence of iron. Previously, however, such stimulatory activity of lactoferrin had been thought to be restricted to human cell lines. The maximal degree of DNA synthesis attained in the presence of holo-lactoferrin was found to be about 4-fold greater than that attained in the presence of holo-transferrin. Transferrin had neither a synergistic nor inhibitory effect on the DNA stimulating activity of lactoferrin. From these results and the fact that the iron-binding affinity of lactoferrin is higher than that of transferrin, especially at low pH values, it is possible that the mechanism of iron-transportation is different between lactoferrin and transferrin. Lactoferrin from bovine milk was as effective as human lactoferrin.
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Masato YOSHIKAWA, Takashi SUGIMURA, Akira TAI
1989 Volume 53 Issue 1 Pages
37-40
Published: 1989
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Optically pure pahutoxin and its antipode were synthesized from (
S)- and (
R)-3-hydroxyhexadecanoic acids, which were obtained by the enantio-differentiating hydrogenation of methyl 3-oxohexadecanoate over asymmetrically modified Raney nickel catalyst. By an optical correlation with synthetic materials, dextrorotatory natural pahutoxin was determined to be of (
S)-configuration.
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Isamu SHIIO, Atsushi YOKOTA, Yasuhiko TORIDE, Shin-ichi SUGIMOTO
1989 Volume 53 Issue 1 Pages
41-48
Published: 1989
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Threonine-producing mutants of
Brevibacterium flavum that lacked dihydrodipicolinate synthase (DPS) activity but had wild-type homoserine dehydrogenase (HD) sensitive to feedback inhibition by threonine were isolated as strains resistant to α-amino-β-bydroxyvaleric acid (AHV). The growth of a mutant, DK131, derived from an aspartate-producing strain was slow and markedly enhanced by diaminopimelic acid (DAP), but not by L-lysine alone. In a medium containing 10% glucose and the optimum concentration of DAP, strain DK131 produced 13.7 g/l of threonine, which was comparable to that by the previously reported mutant, BB69, with a feedback-resistant HD (HD
R). The production was more strongly inhibited by lysine in strain DK131 than in strain BB69. DPS-defective mutants derived from a lysine producer with feedback-insensitive aspartokinase were selected as those which were resistant to AHV and which produced more threonine than lysine. A representative strain, DA105, produced 10.3 g/l of threonine and 3.5 g/l of lysine•HC1, while AHV-resistant mutant, BA68, with an HD
R, derived from the same parent, produced 9.7 g/l of threonine and 12.2 g/l of lysine•HC1. Strain DA105 grew well in the absence of DAP.
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Tamizi SUGIYAMA, Takeshi HASHIZUME
1989 Volume 53 Issue 1 Pages
49-52
Published: 1989
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The presence and levels of 14 cytokinin species were determined in the developing tuberous roots of sweet potato by mass spectrometry using deuterium-labeled standards. The following structures were assigned:
trans- and
cis-zeatin,
trans- and
cis-ribosylzeatin,
trans- and
cis-glucosylzeatin,
trans- and
cis-2-methylthioribosylzeatin, dihydrozeatin riboside,
N6-isopentenyladenine,
N6-isopentenyladenosine, 9-glucosyl-
N6-isopentenyladenine and
trans- and
cis-zeatin ribotide.
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Pichet ITKOR, Osamu SHIDA, Norihiro TSUKAGOSHI, Shigezo UDAKA
1989 Volume 53 Issue 1 Pages
53-60
Published: 1989
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Screening for raw starch digesting bacteria revealed that bacteria producing amylase, which hydrolyzes raw starch, are widely distributed in nature. From among 155 raw starch digesting bacterial isolates, 20 strains were selected and examined as to both the characteristics of their enzymes and some bacteriological properties. According to the action patterns of their enzymes on starch, these strains could be divided into two groups,
i.e., exolytic and endolytic amylase producers. The phenotypic properties of a representative bacterial isolate belonging to each group were examined. One of them (isolate TB1012) was assigned to
Bacillus polymyxa (Prazmowski) Mace, which produced an exolytic β-amylase. The other (isolate B1018) was identified as
Bacillus sp., which produced an endolytic amylase giving a single activity band corresponding to an approximate molecular weight of 75, 000 on SDS-polyacrylamide gel electrophoresis.
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Yohji EZURE, Shigeaki MARUO, Nobutoshi OJIMA, Kiyotaka KONNO, Hiroshi ...
1989 Volume 53 Issue 1 Pages
61-68
Published: 1989
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A transglycosylation reaction with
N-substituted moranolines was carried out with soluble starch as the glucose donor and cyclodextrin glycosyltransferase [EC 2.4.1.19] from
Bacillus stearothermophilus, the degree of
N-substituted moranolines that were converted to transglycosylation products being within the range of 60-81%. The resultant transglycosylation products were hydrolyzed by glucoamylase [EC 3.2.1.3] from
Rhizopm niveus, and the degree of
N-substituted moranolines that were converted to
N-substituted 4-
O-α-D-glucopyranosylmoranolines was within the range of 53-72%.
The inhibitory activities of
N-substituted 4-
O-α-D-glucopyranosyImoranolines against cyclodextrin glycosyltransferase were greater than those of the corresponding
N-substituted moranolines, and
N-substituted 4-
O-α-D-glucopyranosylmoranolines lost the inhibitory activities against glucoamylase. Among these
N-substituted 4-
O-α-D-glucopyranosylmoranolines, the N-methyl derivative was a very potent inhibitor.
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Kazuo TAKIGUCHI, Kunihiko YAMADA, Mamoru SUZUKI, Ken-ichi NUNAMI, Kimi ...
1989 Volume 53 Issue 1 Pages
69-76
Published: 1989
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A series of α-isocyanoacetic acid compounds was examined for their inhibitory activities against several plant-fungi. Among them, various α-isocyanoacetamides showed protective effects against
Sphaerotheca fuliginea,
Cladosporium fulvum and
Phytophthora infestans at the concentration of 1000 ppm. Particularly, α-isocyanoacetanilides (9 g and 9 h) exhibited more potent activity than maneb against
Cladosporium fulvum in tomato seedlings.
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Kazuo TAKIGUCHI, Kunihiko YAMADA, Mamoru SUZUKI, Ken-ichi NUNAMI, Kimi ...
1989 Volume 53 Issue 1 Pages
77-82
Published: 1989
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A variety of α-isocyano-β-phenylpropionamides were synthesized and their inhibitory activities examined against
Sphaerotheca fuliginea and
Cladosporium fulvum. Among them,
N-cyclohexyl-α-isocyano-β-phenylpropionamide (3c) was chosen as the first candidate for an anti-
Sphaerotheca fuliginea agent and exhibited about 2 times more potent activity than dinocap in a field test using cucumber plants.
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Zen-ichi YOKOYAMA, Hiroko HIRAI
1989 Volume 53 Issue 1 Pages
83-88
Published: 1989
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Two kinds of nucleotide-specific phosphatases were separated chromatographically on a DEAE-Toyopearl 650 M column, and designated as NSP-I and -II. NSP-II was further purified to apparent homogeneity using both affinity and gel-filtration chromatography, but NSP-I purified by the same procedures was not homogeneous on polyacrylamide gel electrophoresis. NSP-I and -II had many similar features in molecular size, optimum pH, heat stability, and substrate specificity. The molecular weight of each enzyme protein was estimated to be approximately 45, 000 by molecular sieve chromatography. The optimum pH of each enzyme was 6.5 and showed activity in the wide pH range of 5-9. Neither enzyme hydrolyzed bis(
p-nitrophenyl)phosphate, adenosine 2':3'-cyclic-monophosphate, or sugar phosphate, but both showed a marked preference for 5'-mononucleotides as substrate over
p-nitrophenylphosphate.
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Kunihiko GEKKO, Yoshiko IDOTA
1989 Volume 53 Issue 1 Pages
89-95
Published: 1989
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The solubilities of several amino acids and diglycine were measured in aqueous maltitol solutions at 25°C to evaluate the free energy of transfer of amino acid side-chains and peptide groups from water to aqueous maltitol solutions. The free energy of transfer was positive for most nonpolar side-chains, oppositely negative for most polar side-chains, and zero for the peptide group. The corresponding enthalpies and entropies of transfer, which were measured calorimetrically, were positive for nonpolar side-chains and negative for the peptide group. These results suggest that as well as other polyols and sugars, maltitol could stabilize protein structures through strengthening of the hydrophobic interaction. The thermal stability of a model protein, ribonuclease A, increased with increasing concentrations of maltitol, its stabilizing ability being comparable with maltose but less than glucose and sorbitol.
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Satoru ASAHI, Muneharu DOI, Yutaka TSUNEMI, Shun-ichi AKIYAMA
1989 Volume 53 Issue 1 Pages
97-102
Published: 1989
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The role of uridine and cytidine compounds in regulating pyrimidine nucleotide biosynthesis
de novo was studied with cytidine deaminase-negative mutants of
Bacillus subtilis. In the wild type strain, the formation of six enzymes for uridine S'-monophosphate (UMP) biosynthesis was severely repressed by exogenous cytidine or uracil, whereas the formation of the enzymes in a cytidine deaminase-negative mutant was repressed only by uracil. On the other hand, the formation of cytidine 5'-triphosphate (CTP) synthetase was not affected by uracil. This enzynme was repressed only when a cytidine deaminase-negative mutant was grown in the presence of excess cytidine. Studies on feedback inhibition also showed that the activity of CTP synthetase was inhibited by cytidine nucleotides, but not by uridine nucleotides.
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Tatsuo HORIUCHI, Toshiko KUROKAWA, Narimasa SAITO
1989 Volume 53 Issue 1 Pages
103-110
Published: 1989
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A new enzyme, fructosyl-amino acid oxidase (fructosyl-α-L-amino acid: oxygen oxidoreductase (defructosylating)) was found, which decomposes Amadori rearrangement compounds of α-L-amino acids to the corresponding α-ketoaldehydes and α-L-amino acids. The enzyme was purified from a strain of
Corynebacterium sp. about 38.9-fold to a single protein band with an overall yield of 35 % from the crude extract, and crystallized in rhombic plates. The molecular weight of the enzyme was about 88, 000 on gel filtration and 44, 000 on SDS-poIyacrylamide gel electrophoresis. Non-covalently bound FAD was the prosthetic group. Its isoelectric point was pH 4.6. The optimum pH of the enzyme reaction in potassium phosphate buffer was about 8.3. Fructosyl-α-L-amino acid was the substrate having the highest susceptibility to the enzyme, but
N-fructosyl derivatives of other materials, such as β-amino acids, L-imino acids, D-amino acids, alkyl amines, and ammonia, showed almost no susceptibility. The apparent
Km values for fructosyl-glycine and fructosyl-phen via la nine were 0.74 mM and 0.71 mM, respectively. The enzyme was inhibited by Hg
2+ and Pb
2+.
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Setsu KADOWAKI, Takeshi UEDA, Kenji YAMAMOTO, Hidehiko KUMAGAI, Tatsur ...
1989 Volume 53 Issue 1 Pages
111-120
Published: 1989
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A fungus isolated from soil was found to produce α-
N-acetylgalactosaminidase in culture, this enzyme being the dominant glycosidase. The fungus was identified as an
Acremonium sp., based on various taxonomical characteristics. The enzyme produced by the fungus was purified to electrophoretic homogeneity from the culture broth by procedures including ammonium sulfate fractionation, and chromatography on DEAE-Sephadex A-50, hydroxylapatite, Sephadex G-150 and Concanavalin A-Sepharose 4B. The molecular weight of the enzyme was estimated to be 55, 000 and 57, 000 by gel filtration and SDS-polyacrylamide gel electrophoresis, respectively, and the enzyme appeared to have a monomer structure. The enzyme was most active at pH 4.0-4.5, and was stable at pH 6.0-7.5 and below 40°C. The Michaelis constant for
p-nitrophenyl α-
N-acetylgalactosaminide was 1.3 mM. The enzyme liberated the
N-acetylgalactosamine from the bovine submaxillary glycoprotein, which had been desialyzed with neuraminidase. The enzyme also liberated the
N-acetylgalactosamine from various blood type A active substances, including porcine gastric mucin, horse gastric mucin and human salivary mucin, which was ascertained by a hemagglutination inhibition test. The enzyme could change blood type A erythrocytes into blood type O erythrocytes, which was confirmed by a hemagglutination test.
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Chiaki YOSHIDA, Gen-ichi DANNO
1989 Volume 53 Issue 1 Pages
121-128
Published: 1989
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The effects of the water soluble fraction and the dough mixing procedure on the formation of a viscoelasic gluten mass were examined by means of a reconstitution study. Gluten and the water soluble fraction were prepared by two methods, water extraction and dough mixing, and then lyophilized. With the dough mixing method, water soluble components, especially gliadin, were incorporated into the gluten mass. On the other hand, gluten prepared by water extraction was poor in gliadin and so, being elastic and less extensible, differing from gluten obtained from dough.
Gluten was prepared from reconstituted dough, in which lyophilized gluten, lyophilized water soluble fraction and starch were mixed in the original ratio. The reconstituted gluten with gluten obtained by dough mixing had the same rheological properties as those of gluten from flour. The same results were obtained when gliadin was used instead of the water soluble fraction. On the other hand, despite the same ratio of gliadin and glutenin, the reconstituted gluten with gluten obtained by water extraction was elastic and less extensible. These lines of evidence suggested that for the formation of a viscoelastic mass, gliadin must be well mixed with glutenin to form a homogeneous gluten network.
In this paper, we also found that gliadin was one of the factors causing breakdown and this phenomenon was related to the formation of a viscoelastic mass during dough mixing.
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Yoshi-hisa SUGITA
1989 Volume 53 Issue 1 Pages
129-133
Published: 1989
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L-Pyrrolidonecarboxylic acid (PCA) is a cyclic compound derived on intramolecular dehydration of L-glutamic acid. The conventional method used for the conversion of PCA to glutamic acid has been hydrolysis with strong acids or alkalis at high temperatures. On the reaction of metallic ions such as zinc with PCA, a chelate compound of glutamic acid, and not a metallic salt of PCA, was found to precipitate. This chelate compound dissolved in a dilute mineral acid and glutamic acid crystallized out at the isoelectric point. The yield achieved with this novel direct ring opening of PCA at neutral pH reached 93.1%.
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Shinsaku HAYASHIDA, Koichi NAKAHARA, Kazutaka KURODA, Toshiyuki MIYATA ...
1989 Volume 53 Issue 1 Pages
135-141
Published: 1989
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The raw-starch-affinity site of
Aspergillus awamori var.
kawachi glucoamylase I (GAI), that was proved to be essential for its adsorbability and digestibility on raw starch granules, was found to be located separately from the active site in the region corresponding to glycopeptide I (Gp-I) liberated from the glucoamylase I through the action of subtilisin. Gp-I consists of 45 amino acid residues, hydroxy amino acids being characteristically abundant, and 56 mannose residues. The sequence was determined with an automatic amino acid sequencer to be ATGGTTTTATTTGSGGVTST SKTTTTASKTSTTTSSTSCTTPTAV. A structure of parallelly arranged short mannoside chains linked
o-glycosidically to the successive sequence of hydroxy amino acid residues on Gp-I was revealed. On comparison of the amino acid sequence of Gp-I with those of three glucoamylases, from
Aspergillus awamori,
Aspergillus niger and
Rhizopus oryzae, significantly homologous regions (91%, 91% and 77% homology, respectively) were detected as the affinity site that could be functionally constrained and essential for raw-starch-digesting glucoamylase.
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Shinsaku HAYASHIDA, Koichi NAKAHARA, Werasit KANLAYAKRIT, Takahiro KAR ...
1989 Volume 53 Issue 1 Pages
143-149
Published: 1989
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The enzymatically inactive but raw-starch-adsorbable glycopeptide-I (Gp-I), which was obtained from raw-starch-adsorbable, raw-starch-digesting glucoamylase I (GA I) of
Aspergillus awamori var.
kawachi as a result of proteolysis with subtilisin and proved to contain the raw-starch-affinity site essential for raw-starch-digestion, was found to promote the digestion rate of raw corn starch with GA I, maximally to the extent of 2.5 times. The raw-starch-adsorbability of Gp-I was decreased by the partial removal of carbohydrate moiety from Gp-I. The carbohydrate-split GA I could hydrolyze gelatinized substrates almost at the same rate as the native one, but significantly decreased in the adsorbability and digestibility onto raw starch in comparison with the original one. From the function and the characteristic structure of the parallel short mannoside chains linked to the successive sequence of hydroxy amino acid residues of Gp-I, the "water-cluster-dissociating model" for the hypothetical mechanism of raw-starch-digestion by the adsorption of GA I at the affinity site onto starch was proposed.
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Kuni SUEYOSHI, Nagao OGURA, Hiroki NAKAGAWA
1989 Volume 53 Issue 1 Pages
151-156
Published: 1989
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Degradation intermediates of nitrate reductase (NR) were extracted from fresh spinach leaves in the presence of protease inhibitors, and concentrated by chromatography on an anti-NR IgG-conjugated Sepharose-4B column. Seven polypeptides, 120 (that of the intact subunit), 110, 100, 76, 64, 62, and 44 kDa, were obtained. Their cross-reactivities to mono-specific polyclonal antibody and monoclonal antibody prepared against NR subunit were assessed by an immunoblotting method. All polypeptides were recognized with mono-specific polyclonal antibody. On the other hand, four of them, at 120, 110, 100, and 44 kDa, were recognized with monoclonal antibody which specifically binds to the NADH-ferricyanide reductase domain of NR. The possibility that these polypeptides were artifacts in preparation was ruled out by an internal tracer experiment. Thus, it is concluded that spinach leaf NR
in vivo degraded
via putative intermediates catabolic products of 110, 100, 76, 64, 62, and 44 kDa.
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Shao-Yong WU, Akinori HIRASHIMA, Morifusa ETO, Kazunori YANAGI, Eriko ...
1989 Volume 53 Issue 1 Pages
157-163
Published: 1989
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Optically active salithion (>98%
e.e.) was obtained
via an L-proline methyl ester derivative (PS), whose diastereomers were easily separated, with subsequent sulfuric acid-catalyzed methanolysis. The configuration of each diastereomeric PS was determined by NMR and single-crystal X-ray analysis. *The optical purity of each PS and of the salithion enantiomers was measured by
1H-NMR and HPLC, respectively. The sulfuric acid-catalyzed methanolysis of PS to salithion was confirmed to have proceeded with an inversion of the configuration at the phosphorus atom by an experiment using model compounds.
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Shao-Yong WU, Akinori HIRASHIMA, Ryuko TAKEYA, Morifusa ETO
1989 Volume 53 Issue 1 Pages
165-174
Published: 1989
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Four optically active isomers (optical purity >94.6%
d.e.) of 2-methoxy-5-phenyl-1, 3, 2-oxazaphospholidine 2-sulfide (5-PMOS) were synthesized by a two-step phosphorylating method, and their insecticidal activities were determined against susceptible (SRS) and resistant (3-Y
F) strains of house flies by topical application. The relationship between the configuration and insecticidal activity was similar in the susceptible and resistant strains of house flies. The (R)
C(R)
P-isomer showed the highest insecticidal activity, followed by (R)
C(S)
P>(S)
C(R)
P>(S)
C(S)
P.
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Akinori HIRASHIMA, Isaac ISHAAYA, Ryohei UENO, Yoshifumi ICHIYAMA, Sha ...
1989 Volume 53 Issue 1 Pages
175-178
Published: 1989
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Salioxon enantiomers were obtained from salithion enantiomers by MCPBA oxidation. The larvicidal and adulticidal activities of salithion enantiomers, and the anti-cholinesterase activity of salioxon enantiomers were determined on house flies. On larvae, the potency of (
S)(-)-salithion (LC
50=1.1 ppm) was about two-fold higher than the (
R)(+)-enantiomer (LC
50 = 2.1 ppm). A similar ratio was obtained when these compounds were applied topically to female adults. The relative potency of these enantiomers correlated well with the anti-cholinesterase activity of their respective oxidative enantiomers, (
R)(+)-and (
S)(-)-salioxon, which are assumed to have been formed after their application to the insects.
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Tatsuyuki KAMIRYO, Yuji SAKASEGAWA, Hironobu TAN
1989 Volume 53 Issue 1 Pages
179-186
Published: 1989
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The genes
POX2 and
POX4, which encode the subunits (PXP-2 and PXP-4) of peroxisomal fatty acyl-coenzyme A oxidase of
Candida tropicalis, were introduced into the related yeast
Candida maltosa. The cells transformed with
POX2 or
POX4 gave much PXP-2 or PXP-4 in the purified peroxisomes. The polypeptides associated with the heterologous organelle were resistant to added protease, implying that they were transported into the peroxisomes. Genes for curtailed versions of PXP-4 were constructed
in vitro and introduced into the host cells. Peptide-C, the COOH-terminal two-thirds of PXP-4, was efficiently transported into the host peroxisomes, and the polypeptide containing the NH
2-terminal one-third was also, in much lesser amount. These and other results suggested that there were at least two regions of peroxisomal targeting information in PXP-4 and the primary information was internal. The deletions in Peptide-C inhibited the transport of many, but not all, of the host-cell peroxisomal polypeptides. This suggested heterogeneous transport systems on the peroxisomal membrane.
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Ning ZONG, Tsuneo YASUI
1989 Volume 53 Issue 1 Pages
187-195
Published: 1989
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An α-D-xylosidase from
Bacillus sp. No. 693-1 was purified by precipitation with ammonium sulfate and successive chromatography on DEAE-Toyopearl 650 and hydroxyapatite. The purified enzyme, with its isoelectric point at pH 4.25, had an apparent molecular weight of 82, 000 in SDS-polyacrylamide gel electrophoresis, and 400, 000 in gel filtration chromatography on Toyopearl HW60. The optimum pH for enzyme action was 7.5 and the optimum temperature was 45°C. The enzyme was stable from pH 6.0-8.5 and up to 45°C. The activity was inhibited by monoiodoacetic acid,
p-CNB, and metal ions such as Zn
2+, Cu
2+, and Hg
2+. The α-D-xylosidase was highly specific for α-xylosidic linkages, and hydrolyzed isoprimeverose [α-D-xylopyranosyl-(1→6)-D-glucopyranose] to produce xylose and glucose, and hydrolyzed tamarind seed polysaccharide (soluble fraction) to release xylose as well.
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Nobuaki HORI, Mutsumi WATANABE, Yoshinari YAMAZAKI, Yoichi MIKAMI
1989 Volume 53 Issue 1 Pages
197-202
Published: 1989
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Many thermophiles, which can grow at 65°C, were examined as to their ability to produce 5-methyluridine from inosine and thymine (5-methyluracil) in the presence of phosphate and cells as enzyme sources.
Bacillus stearothermophilus JTS 859 was selected as a strain that synthesized 5-methyluridine efficiently. The reaction is supposed to be carried out by a combination of a thermostable purine nucleoside phosphorylase and a thermostable pyrimidine nucleoside phosphorylase. Their halflives were 7200 hr and 400 hr at 63°C, and 148 hr and 14 hr at 70°C, respectively.
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Jun KAWABATA, Junya MIZUTANI
1989 Volume 53 Issue 1 Pages
203-207
Published: 1989
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Three novel sesquiterpene lactones having a unique lindenane skeleton were isolated from
Chlomnthus spp. (Chloranthaceae). The first one, shizukanolide D (1), was found in ether extracts of
C. japonicus Sieb., Hitorishizuka, and characterized to be an epoxy derivative of previously known chloranthalactone C (3). The other two compounds, shizukanolides E (4) and F (6), were isolated from ether extracts of
C. sermtus (Thumb.) Roem. et Schult., Futarishizuka, and their structures were determined to be hydroxy derivatives of shizukanolide C (2).
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Hitoshi OHMORI, Shigehiro TAKAHAMA
1989 Volume 53 Issue 1 Pages
209-213
Published: 1989
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We report the essential role of lipid in the development of a serum-free culture medium that supports the primary antibody response by murine lymphocytes
in vitro. In our preceding paper, we have reported that β-cyclodextrin (β-CD) is effective as a serum-substitute in murine lymphocyte cultures [H. Ohmori and I. Yamamoto,
Eur. J. Immunol., 17, 79 (1987)]. At first, we employed RPMI-1640 that was supplemented with fatty acid-free albumin, insulin, transferrin, and β-CD as the basal serum-free medium (BSF medium). BSF medium supported the antibody response less than 30 % as efficiently as 10% fetal calf serum (FCS)-containing medium. When ether-extracts of FCS (FCS ex.) was added to BSF medium, the antibody response was enhanced by approximately 2-fold. FCS ex. was purified by silica gel thin-layer chromatography. The analyses of the purified active component by high performance liquid chromatography and mass spectrometry revealed it to be cholesterol. Authentic cholesterol or low density lipoprotein enhanced the antibody response as efficiently as FCS ex. in BSF medium. The same level of the antibody response as that in 10 % FCS-containing medium was attained when low density lipoprotein was added to BSF medium in combination with i -a la nine, putrescine, and pyruvate.
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Hiromichi OHTA, Yoshitaka MIYAMAE, Gen-ichi TSUCHIHASHI
1989 Volume 53 Issue 1 Pages
215-222
Published: 1989
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Microorganisms that hydrolyze the one enantiomer of
dl-phenoxyacetaldehyde cyanohydrin acetate were screened, and
Bacillus coagulans isolated from soil was found to be the best. This bacterium was applied to the asymmetric hydrolysis of other aryloxyacetaldehyde derivatives to give satisfactory results. The effect of adding dimethyl sulfoxide to the medium is also described.
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Gentaro OKADA, Takehiro UNNO
1989 Volume 53 Issue 1 Pages
223-228
Published: 1989
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A glucodextranase was highly purified from a cell-free culture broth of
Arthrobacter globiformis I42. The glucoamylase to glucodextranase activity ratio was almost the same (
ca. 0.88%) at each purification step. The activity profiles of the stabilities of the glucoamylase to pH and temperature coincided well with those of the glucodextranase. Furthermore, the inactivation profiles of the glucoamylase with various metal ions and inhibitors were almost the same as those of the glucodextranase. From these results, it was strongly suggested that a single enzyme molecule was responsible for both the glucodextranase and glucoamylase activities.
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Shigeo YOSHIDA, Tadao ASAMI, Yasuko TSUCHIHASHI, Masami UJI-IE, Koichi ...
1989 Volume 53 Issue 1 Pages
229-233
Published: 1989
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A series of nitrophlorophenone derivatives of grandinol, a potent inhibitor of germination and photosynthetic electron transport (PET) in
Eucalyptus grandis, was synthesized and their activities on cress seed germination and on the PET system were determined. In general, the nitrophlorophenone derivatives were found to inhibit both cases. Studies on the structure-activity relationships of those compounds indicated that nitrophlorophenone derivatives with an alkyl group on the nuclus may interact with receptor sites in a similar manner to that of grandinol, whereas those without the alkyl substituent may behave in a different manner.
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Hiroyuki UKEDA, Kazuhiko ONO, Masatomo IMABAYASHI, Kiyoshi MATSUMOTO, ...
1989 Volume 53 Issue 1 Pages
235-237
Published: 1989
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Enamul HOQUE
1989 Volume 53 Issue 1 Pages
239-240
Published: 1989
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Kunio SUETSUNA, Yutaka OSAJIMA
1989 Volume 53 Issue 1 Pages
241-242
Published: 1989
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Masayoshi SAWAMURA, Tetsuya TSUJI, Shigeru KUWAHARA
1989 Volume 53 Issue 1 Pages
243-246
Published: 1989
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Masaaki YASUDA, Misao KUWAE, Hisako MATSUSHITA
1989 Volume 53 Issue 1 Pages
247-249
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Koichi MORITA, Kinya IDE, Yoshio HAYASE
1989 Volume 53 Issue 1 Pages
251-252
Published: 1989
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Katsuhiro AIKAWA, Koichi CHIKUNI
1989 Volume 53 Issue 1 Pages
253-254
Published: 1989
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Katsuichiro OKAZAKI, Susumu KIMURA
1989 Volume 53 Issue 1 Pages
255-257
Published: 1989
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Suguru TAKATSUTO, Kumiko OMOTE
1989 Volume 53 Issue 1 Pages
259-261
Published: 1989
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Suguru TAKATSUTO, Fumio FUTATSUYA, Kiyomi KOBAYASHI, Hitoshi SATOH
1989 Volume 53 Issue 1 Pages
263-265
Published: 1989
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Hidehiko YOKOGOSHI
1989 Volume 53 Issue 1 Pages
267-269
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Michiko KAWAKAMI, Hiromi UCHIDA, Akio KOBAYASHI, Tei YAMANISHI
1989 Volume 53 Issue 1 Pages
271-275
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Hiromasa MIYAJI, Tatsunari NISHI, Seiga ITOH
1989 Volume 53 Issue 1 Pages
277-279
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Hiromichi OHTA, Yoshitaka MIYAMAE, Gen-ichi TSUCHIHASHI
1989 Volume 53 Issue 1 Pages
281-283
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Akira YAMAMOTO, Toshinobu ISHIHARA, Takehiko FUKUMOTO
1989 Volume 53 Issue 1 Pages
285-287
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Youichi TAMAI, Takahiro ISHIKAWA, Shunnosuke ABE, Yasuo WATANABE
1989 Volume 53 Issue 1 Pages
289-291
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Yukio FURUICHI, Takao TAKAHASHI
1989 Volume 53 Issue 1 Pages
293-294
Published: 1989
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