Journal of Applied Glycoscience
Online ISSN : 1880-7291
Print ISSN : 1344-7882
ISSN-L : 1344-7882
Volume 47, Issue 2
Displaying 1-18 of 18 articles from this issue
  • Xiaoming Qin, Ryo Yamauchi, Koichi Aizawa, Takahiro Inakuma, Koji Kato
    2000 Volume 47 Issue 2 Pages 155-161
    Published: June 30, 2000
    Released on J-STAGE: June 28, 2010
    JOURNAL FREE ACCESS
    Two kinds of arabinogalactan-protein (Cp-1-C and -D) were obtained from the fruit of Lycium chinense Mill. In Cp-1-C, the carbohydrate was composed of arabinose and galactose at a ratio of 3:1, and O -glycosidically linked to both the serine and threonine residues of protein. Its molecular weight was 42, 000. In Cp-1-D, the ratio of arabinose to galactose was 1:1, and the O -glycosidical junction between the carbohydrate and protein was composed of serine residue. The molecular weight was 23, 000. From the results of structural characterization, both Cp-1-C and -D were assigned to type II arabinogalactan-protein (AGP), where a (1→6) linkage of galactose occurred mainly at the branching point in the case of Cp-1-C and mainly at the linear part in Cp-1-D.
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  • Takayuki Uchibori, Koki Yokotsuka, Tsutomu Takayanagi
    2000 Volume 47 Issue 2 Pages 163-168
    Published: June 30, 2000
    Released on J-STAGE: February 23, 2011
    JOURNAL FREE ACCESS
    An induced chitinase was purified from Koshu grapes elicitated with glycolchitin. The protein fraction obtained by salting out with ammonium sulfate (80% saturation) from the extract solution of the grapes was separated by successive chromatographies on Chitopearl BL-3 and Q-Sepharose HP (HiTrap Q) columns. The induced chitinase was homogeneous on polyacrylamide gel electrophoresis. The optimum pH of the induced chitinase was 4.0, and the enzyme was stable under acidic conditions between pH 5.0-6.5. The optimum temperature was 40°C, and the enzyme was stable below 40°C. The N-terminal amino acid sequence of the induced chitinase was NH2-GTITVYXGQNGN, which is highly homologous with that of the class III chitinase of Vitis vinifera. The induced chitinase inhibited the growth of Botrytis cinerea, which causes grey mold disease, strongly suggesting that it plays an important role in the defense mechanism against phytopa-thogens in mature grape.
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  • Kinetic Study of the Sulfite Dehydrogenase-catalyzed Reaction in the Microbial State of Thiobacillus thiooxidans
    Takashi Matsumoto, Terutaka Yazaki, Masatake Ohnishi
    2000 Volume 47 Issue 2 Pages 169-175
    Published: June 30, 2000
    Released on J-STAGE: July 01, 2011
    JOURNAL FREE ACCESS
    A highly sensitive and automatic microbial sensor apparatus has been constructed with Thiobacillus thiooxidans and employed for sulfite analysis of food materials in practical fields including the starch industry, thus we tried to obtain enzymatic bases on the T. thiooxidans-catalyzed reaction. Based on the spectrophotometric observation of ferricyanide and the proportional dependency of the initial velocity on the concentration of T. thiooxidans, the sulfite oxidation was confirmed to be a microbial dehydrogenase-catalyzed reaction, of which the optimal pH is 7.5. The kinetic parameters, the Michaelis constant Km and the maximum velocity V were evaluated to be 1.0±0.1 mM and 0.065±0.032 mM/min/mg protein, respectively. The concentration of ferricyanide was proportional to the initial velocity in the range of 0 to 2 nix. These results prove that the microbial sensor apparatus developed is satisfactory to determine the enzymatic bases for sulfite oxidation.
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  • Supannee Kansarn, Naoyoshi Matsushita, Toshiaki Kono, Gentaro Okada
    2000 Volume 47 Issue 2 Pages 177-185
    Published: June 30, 2000
    Released on J-STAGE: July 01, 2011
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    An endo-cellulase from a commercial cellulase preparation of a filamentous fungus Acremonium cellulolyticus was separated and extensively purified to essential homogeneity by using consecutive column chromatographies of QAE-Toyopearl 550C, DEAE-Toyopearl 650S, Butyl-Toyopearl 650M and Bio-Gel P-100, and was designated as cellulase IV The purified enzyme was homoge-neous on both Native- and SDS-PAGE, and was completely free from β-glucosidase. The enzyme showed high specific activity for CMC and an extremely low specific activity for Avicel (a mi-crocrystalline cellulose powder), 106 and 0.08 U/mg of protein, respectively. The molecular mass of the enzyme was estimated to be about 38 kDa by SDS-PAGE. The enzyme was an acidic protein with a pI value of≤3.4. The N-terminal amino acid sequence from the 2nd up to the 27th residue of the purified cellulase was Gln-Ala-Ser-(Cys)-Phe-Glu-Trp-Phe-Gly-Ser-Asn-Glu-Ser-Gly-Ala-Glu-Phe-Gly-Ser-Gly-Asn-Ile-Pro-Gly-Val-Glu-. The optimum pH and temperature were 4.0 and 60-C, respectively. The enzyme was completely stable over the range of pH 5.0-12.0 at 4-C for 24 h and at temperatures below 50-C. Cellulase IV was characterized as a medium endo -type cellulase on the basis of its action on CMC and cellooligosaccharides. The enzyme was completely inhibited by 5 mM Hg2+, Fe3+ and KMnO4. Cellulase IV split various cellulosic substrates to pro-duce predominant cellobiose and a small amount of glucose as the final hydrolysis products.
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  • Masakuni Tako
    2000 Volume 47 Issue 2 Pages 187-192
    Published: June 30, 2000
    Released on J-STAGE: June 28, 2010
    JOURNAL FREE ACCESS
    The non-Newtonian behavior and dynamic viscoelasticity in an aqueous solution of rice starch (Yukihikari) were measured using a rheogoniometer. The gelatinization of Yukihikari starch oc-curred above 3.0% after heating at 105°C for 20 min. The flow curves, at 25°C, of Yukihikari starch showed shear-thinning behavior at 2.0, but plastic behavior above 3.0%. The viscosity of Yukihikari starch decreased gradually with an increase in temperature. The dynamic modulus of Yukihikari starch showed a large value during the increase in the temperature. The tan 6 was 0.23 in the low-temperature range and stayed at a low value with increasing temperature at a concentration of 4.0%. A slight decrease in dynamic modulus of Yukihikari starch was observed during the increase in temperature upon the addition of urea (4.0 M). The dynamic modulus, however, in-creased with increasing temperature up to 50°C, then it decreased rapidly with further increases in the temperature after storage at 20 and 4°C for 24 h in urea solution. The dynamic modulus of Yukihikari starch stayed in the low-values in 0.05 M NaOH solution even at low temperature range. The results supported a gelatinization mechanism of rice starch previously proposed.
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  • Tetsuya Ishii, Morimasa Tanimoto, Yasuharu Itagaki, Yoshihiko Honda, T ...
    2000 Volume 47 Issue 2 Pages 193-196
    Published: June 30, 2000
    Released on J-STAGE: February 23, 2011
    JOURNAL FREE ACCESS
    To synthesize nonreducing oligosaccharide derived from trehalose, enzymatic transgalactosylation was investigated using lactose and trehalose as substrates and a commercially available β-ga-lactosidase, which has a tendency to transfer the galactosyl residue to the C4 position of acceptor sugars, from Bacillus circulans. The galactosylation successfully resulted in the efficient production of O-β-D-galactopyranosyl-(1-4)-α, α-trehalose. This trisaccharide showed good stability against acidic treatment.
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  • Masayasu Takada, Koichi Ogawa, Takeomi Murata, Taichi Usuil
    2000 Volume 47 Issue 2 Pages 197-200
    Published: June 30, 2000
    Released on J-STAGE: July 01, 2011
    JOURNAL FREE ACCESS
    Enzymatic transformation of 42 -O-β-D-galactosylmaltose (1) into 42 -O-β-D-galactosylmaltobi-onolactone (2) was carried out by the use of oligosaccharide oxidase (OSOD). The anomer hy-droxyl group of glucose residue at the reducing end of 1 was readily oxidized to the corresponding lactone on a large scale, and the product was conveniently isolated by the sequential column chromatographies of Amberlite IR-120B and Toyopearl HW-405. The desired compound, 2, was obtained in a high yield (80%) based on the addition of 1. The jar-fermentor was included in this reaction system to facilitate the maintenance of pH and supply of air to the reaction mixture. As a result, 2 obtained enzymatically was shown to be identical to that obtained chemically.
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  • Kazunori Takamine, Jun-ichi Abe, Amane Iwaya, Kaori Shimono, Syunsaku ...
    2000 Volume 47 Issue 2 Pages 201-206
    Published: June 30, 2000
    Released on J-STAGE: June 28, 2010
    JOURNAL FREE ACCESS
    The conditions for selective and effective extraction of pectin from sweetpotato residue were es-tablished. Sodium or potassium phosphate solutions of pH 7 or above were suitable for the extraction. The gelatinization of starch in the residue, which interfered with the extraction, was minimized by extraction at 63°C or below. The extract was fractionated into 4 fractions, starch, arabinogalactan and pectin of two types, by DEAE-Toyopearl 650M ion-exchange chromatography. One pectin fraction had two peaks and their molecular weightes were 795, 000 and 167, 000, and that of the other pectin fraction was 278, 000. The uronic acid contents of the two pectin fractions were 82.4 and 92.1%, respectively.
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  • [in Japanese]
    2000 Volume 47 Issue 2 Pages 207
    Published: June 30, 2000
    Released on J-STAGE: June 28, 2010
    JOURNAL FREE ACCESS
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  • Yoshio Tsujisaka
    2000 Volume 47 Issue 2 Pages 209-216
    Published: June 30, 2000
    Released on J-STAGE: June 28, 2010
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  • Takehiko Yamamoto
    2000 Volume 47 Issue 2 Pages 217-219
    Published: June 30, 2000
    Released on J-STAGE: June 28, 2010
    JOURNAL FREE ACCESS
    This is to write the essentials for organization of The Amylase Research Society of Japan by Dr. Juichiro Fukumoto in 1965, in relation to his works and organization of the society on the memo-rial symposium.
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  • Shigetaka Okada
    2000 Volume 47 Issue 2 Pages 221-226
    Published: June 30, 2000
    Released on J-STAGE: July 01, 2011
    JOURNAL FREE ACCESS
    Recently, syrups of various homo- and hetero-oligosaccharides are on the market for the food in-dustry. Production and application of some oligosaccharides are reviewed in this report. (1) Coupling sugar: The syrup which contains glucosyl-sucrose, maltosyl-sucrose, etc. as the main compo-nent was produced by using the action of a transfer enzyme. The syrup inhibited the synthsis of in-soluble glucan from sucrose using Streptococcus mutans. This finding proved the syrup is a sweetener which hardly induces dental caries. (2) Fructooligosaccharides: This syrup contains fructofuranosylsucrose oligomers. The administration of these oligosaccharides has been reported to be effective in stimulating the growth of bifidobacteria in the human intestinal canal. (3) A novel enzymatic production system of trehalose was developed. Industrial production of trehalose has become possible using this system. (4) Branched cyclodextrins containing heterogeneous sugars and larger cyclodextrins (cycloamylose), which have more than 17 glucose units in the ring, have been prepared by some transfer enzymes. (5) For the increase of bioavailability of dietary calcium, phosphoryl oligosaccharides from potato starch have been studied.
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  • Noboru Taji
    2000 Volume 47 Issue 2 Pages 227-234
    Published: June 30, 2000
    Released on J-STAGE: February 23, 2011
    JOURNAL FREE ACCESS
    Being engaged in the enzyme industry, we should know the enzymes which are of interest to the sugar industry. The following three matters, which have been of interest for the past two decades, are described in this paper. 1) Starch liquefaction: Two-step liquefaction was applied in the past, and then a thermostable a-amylase was developed. This enzyme made it possible to perform more efficient liquefaction using a jet-cooker. Further improvement, such as a new liquefying a-amylase used under acidic coditions, is desired. 2) Starch saccharification: At first, a glucoamylase from Rhizopus was used for starch saccharification, and then an economical glucoamylase from Asper-gillus niger was developed and gained popularity in use. Finally, improvements such as the re-moval of transglucosidase and addition of pullulanase were developed and have contributed to high yields of glucose production. Further improvement is required. 3) Conversion of glucose into fruc-tose: At first, batch conversion was performed with intact cells. Development of an immobilized enzyme made it possible to perform continuous conversion. Further improvements, such as a new enzyme which converts to fructose over 50%, and works at acidic condition, are required.
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  • Atsuo Kimura, Toshiyuki Nishio, Wataru Hakamada, Tadatake Oku, San San ...
    2000 Volume 47 Issue 2 Pages 235-241
    Published: June 30, 2000
    Released on J-STAGE: February 23, 2011
    JOURNAL FREE ACCESS
    The suicide substrate of isomalto-dextranase was designed, and the ω-epoxyalkyl α-glucosides, of which the alkyl carbons were three to six, were synthesized. The 2', 3'-epoxypropyl α-glucoside did not decrease the activity. ω-Epoxyalkyl α-glucosides with four to six alkyl carbons resulted in inactivation which followed the pseudo-first order kinetics. A detailed kinetic analysis on inactivation was done, and the results obtained supported that the compounds were suicide substrates for isomalto-dextranase. All ω-epoxyalkyl α-glucosides synthesized inactivated β-amylase. The C5-al-kyl compound was the most effective for the inactivation of isomalto-dextranase, but β-amylase was strongly inactivated by the C4-alkyl compound. The discrimination of inhibitors by the two en-zymes was considered to be based on the difference in their substrate recognition, in which β-amy-lase recognizes the maltosyl residue and isomalto-dextranase recognizes the isomaltosyl residue having one methylene group longer than the maltosyl residue. No inactivation for α-amylase was observed with the ω-epoxyalkyl α-glucosides. The 3', 4' epoxybutyl α-glucoside did not show any effect on the substrate binding. Conduritol B epoxide, a suicide substrate for α-glucosidase, also caused no inactivation or no inhibition of α-amylase activity. These findings were applied to the α-amylase assay in the enzyme mixture coexistent with β-amylase and α-glucosidase. Since 3', 4' ep-oxybutyl α-glucoside was hydrolyzed by α-glucosidase, a mixture of three enzymes was incubated with conduritol B epoxide at first, and then treated with 3', 4'-epoxybutyl α-glucoside, resulting in the complete inactivation of α-glucosidase and β-amylase. This procedure was shown to be useful in the determination of α-amylase in preparations containing α-glucosidase and β-amylase.
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  • Susumu Ito, Tohru Kobayashi, Katsuya Ozaki
    2000 Volume 47 Issue 2 Pages 243-251
    Published: June 30, 2000
    Released on J-STAGE: February 23, 2011
    JOURNAL FREE ACCESS
    A very large amount of enzymes is used in the detergent industry, as well as in the starch, food and fabric industries. Today's world use of detergent enzymes has reached over 500 billion dollars, and has a growth of 10-15% per year. This high growth is mainly due to the new functionalities provided by cellulase and lipase, to the development of compact-type heavy-duty detergent, and the addition of the automatic dishwasher market. We have isolated many alkaliphilic Bacillus strains that produce alkaline cellulases, alkaline proteases, and amylolytic enzymes to incorporate them into compact-type heavy-duty detergents and automatic dishwasher detergents. We have succeeded in large-scale industrial production of some of these enzymes by hyperproducing mutants or by recombinant Bacillus subtilis cells. In this report, we describe the alkaline enzymes for use in detergents, with emphasis on their catalytic properties, genetics and improvements by protein engineering.
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  • Takeomi Murata, Taichi Usui
    2000 Volume 47 Issue 2 Pages 253-260
    Published: June 30, 2000
    Released on J-STAGE: February 23, 2011
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    With the growing biological importance of oligosaccharides involved in glycoconjugates, practical synthetic methods for obtaining oligosaccharides have been required. Enzyme-catalyzed oligosaccharide synthesis has been shown to be one of the most effective methods for large-scale oligo-saccharide synthesis. This review describes enzyme-catalyzed synthesis of di, tri, and tetrasaccharide units, which are related to biological roles: (1) galactose-containing disaccharides, (2) fucosecontaining trisaccharides, (3) core oligosaccharides in mucin-type carbohydrate, and (4) human milk oligosaccharides. Such well-defined oligosaccharide units are useful as substrates for glycosidases and glycosyltransferases, and as starting materials for artificial glycopolymers, which are valuable as probes of carbohydrate-protein interaction.
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  • Hirofumi Nakano, Taro Kiso, Sumio Kitahata
    2000 Volume 47 Issue 2 Pages 261-267
    Published: June 30, 2000
    Released on J-STAGE: February 23, 2011
    JOURNAL FREE ACCESS
    Specificities of glycosidases were studied from the viewpoint of the synthesis of oligosaccharides and glycosides that are not naturally occurring. Trehalase from Lobosphaera sp. catalyzed conden-sation not only of glucose (Glc) but also of 2-deoxy glucose (dGlc), yielding trehalose and dideoxy-trehalose, respectively. Deoxy-trehalose was also synthesized in a mixture of Glc and dGlc. Activi-ties for trehalose derivatives decreased considerably because of their deoxy site. Glucoamylases from Aspergillus niger and Rhizopus niveus, and α-glucosidase from A. niger afforded deoxy-oli-gosaccharides by the condensation of dGlc, while α-glucosidases from Torula and Saccharomyces gave them in low yields. Deoxy-glucobioses with α-1, 3-, 1, 4- and 1, 6-linkages were identified as the products of glucoamylase and α-glucosidase. The hydrolysis rates of these glucobioses and the transfer action of α-glucosidase seemed to affect the product composition, especially in the early stage of the reactions, and afterwards, the 1, 6-linked saccharide accumulated predominately. Strains of Clavibacter michiganense and Flavobacterium johnsonae produced novel β-glucosidases that hydrolyzed the β-glucosyl ester linkage of steviol glycosides. Transglucosylation occurred when this linkage was degraded, ρ-Hydroxybenzoyl glucose was chemically synthesized as a substrate containing the β-glucosyl ester linkage. Several β-glucosidases, such as that from Caldocellum saccharolyticum, hydrolyzed the substrate. ρ-Hydroxybenzoyl galactose with the β-galactosyl ester linkage was also synthesized, and was examined as a substrate for β-galactosidases. The activities showed great diversity depending on the enzyme origin. The enzyme from Penicillium multicolor showed the highest activity among those tested. β-Galactosidases catalyzed galactosylation not only of a hydroxyl group but also of a thiol group in the condensation of galactose and 2-mercap-toethanol. The enzymes from A. oryzae and P. multicolor were effective for the synthesis of thio-galactoside. The usual galactoside was synthesized more rapidly than the thin-galactoside, although the latter accumulated predominately as the reaction proceeded.
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  • Jun-ichi Abe, Kazuhiro Yoshinaga, Yasuhito Takeda, Susumu Hizukuri, Ma ...
    2000 Volume 47 Issue 2 Pages 269-273
    Published: June 30, 2000
    Released on J-STAGE: February 23, 2011
    JOURNAL FREE ACCESS
    α-Glucan lyase was purified from a seaweed, Graciralia chorda, by means of starch adsorption, ion-exchange chromatography and gel-filtration. The enzyme was stabilized and activated in the presence of Ca2+ and CI- ions. Chemical modifications showed that a carboxyl residue was essen-tial to the activity and a tryptophanyl residue(s) was probably involved in substrate binding. The enzyme degraded waxy rice amylopectin from its non-reducing ends and left the outermost chains as glucosyl residues. 1, 5-Anhydro-D-arabino-hex-2-ulose (1, 5-anhydro-D-fructose) was prepared by reacting the enzyme on waxy corn starch with 50% yields. The sugar served as an antioxidant on the oxidation of linoleic acid and its ability was -2-fold as high as that of ascorbic acid.
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