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M Kakuta

Publications and source records attributed to M Kakuta.

24 records · Page 2Linked to original sources

Chain conformation of deacetylated beijeran calcium salt.

A well-defined X-ray fiber diffraction pattern was obtained from a stretched film of the calcium salt of a new uronic acid containing polysaccharide designated as beijeran in the deacetylated form, poly[-->3)-alpha-D-GalUA-(1-->3)-beta-L-Rham-(1-->3)-alpha-D -Glc-(1-->]. The oriented film showed no diffraction spots, indicating it to be amorphous. However, when annealed at high temperature, the film exhibited high crystallinity. All the visible reflections could be indexed in terms of a monoclinic unit cell with the following dimensions: a = 1.297; b = 1.676; c (fiber axis) = 2.509 nm; and gamma = 106.50 degrees. The length of the fiber axis and the absence of meridional reflections at any odd layer line indicate that an extended two-fold helical conformation was made up of two trisaccharide residues.

Acylation↗

The alpha-mannosyl-binding lectin from leaves of the orchid twayblade (Listera ovata). Application to separation of alpha-D-mannans from alpha-D-glucans.

The carbohydrate-binding specificity of an alpha-D-mannose-specific lectin isolated from leaves of the orchid twayblade (Listera ovata) was elucidated by quantitative precipitation of mannose-containing polysaccharides and glycoproteins, hapten inhibition, and affinity chromatography on the immobilized lectin. L. ovata agglutinin (LOA) interacted with various alpha-mannans and galactomannans of yeasts, fungi and bacteria, but not with alpha-glucans, e.g., dextran and glycogen, as do mannose/glucose-binding lectins. This lectin, LOA, appears to be highly specific for alpha 1-3 mannosidic linkages. It reacted with a linear alpha 1-3-mannan (D. P. 15) and, surprisingly, even with a linear alpha 1-3-mannoheptasaccharide. The LOA/C. tropicalis mannan precipitation reaction was inhibited by alpha-linked mannooligosaccharides, in the order, alpha 1-3 > alpha 1-6 > alpha 1-2 linkages; alpha 1-3 [Man]4 and [Man]5 were the best inhibitors among various mannooligosaccharides tested, having 7-times greater potency than alpha 1-3 [Man]2, and 18-times that of methyl, alpha-mannoside. LOA/mannan interaction was also inhibited by periodate-oxidized and reduced alpha 1-3 [Man]5 which had an inhibitory potency similar to that of alpha 1-3 [Man]3, confirming that LOA also recognizes the internal alpha 1-3-mannosidic linkages of carbohydrate chains. Complete resolution of mannan and glycogen from yeast cells, by affinity chromatography on an immobilized LOA column, and retention of several high-mannose-glycoproteins suggest this lectin to be a useful tool for purification and structural investigation of alpha-mannosyl-containing polysaccharides and glycoconjugates.

Binding Sites↗

Structure of pestalotan, a highly branched (1----3)-beta-D-glucan elaborated by Pestalotia sp. 815, and the enhancement of its antitumor activity by polyol modification of the side chains.

Pestalotia sp. 815, a newly isolated fungus, produces extracellularly a highly (1----6)-branched (1----3)-beta-D-glucan in high yield when grown in a D-glucose-containing medium. This extracellular glucan, designated "Pestalotan", has [alpha]25D-0.1 degree (c 0.5, M NaOH) and a molecular weight greater than 2 X 10(6). Chemical and enzymic studies indicated that pestalotan has a very highly branched structure containing a back-bone chain of beta-D-(1----3)-linked D-glucosyl residues, and three out of five D-glucosyl residues are substituted at O-6, mostly with single D-glucosyl groups, and a very few with short beta-(1----6)-linked oligosaccharide units. This D-glucan becomes water-insoluble after isolation from the culture filtrate followed by dehydration, and shows moderate growth-inhibitory activities against mouse-implanted tumors. However, when the D-glucosyl groups of the side chains were modified by periodate oxidation and borohydride reduction, the resulting, still water-insoluble, D-glucan polyol exhibited potent antitumor activities, confirming that the attachment of many polyhydroxy groups to the beta-D-(1----3)-linked D-glucan back-bone gives a remarkable enhancement effect on the antitumor activity of the branched D-glucan. Prolonged treatment of the D-glucan polyol by ultrasonic irradiation afforded a low-molecular-weight D-glucan polyol (SD-pestalotan polyol), without alteration of its chemical structure. The water-soluble, SD-pestalotan polyol, having a molecular weight of 4.7 X 10(5), exhibited remarkable antitumor activities against both allogeneic and syngeneic, mouse-implanted tumors, at small dosages (1-5 mg/kg for 10 days) by intraperitoneal administration. A comparison of values of the molecular weight of SD-pestalotan polyol, estimated by 3-MPa l.c. for the aqueous solution and the dimethyl sulfoxide solution, strongly suggested that the D-glucan polyol must form a triple-chain conformation in aqueous solution.

Animals↗

Studies on interrelation of structure and antitumor effects of polysaccharides: antitumor action of periodate-modified, branched (1 goes to 3)-beta-D-glucan of Auricularia auricula-judae, and other polysaccharides containing (1 goes to 3)-glycosidic linkages.

Antitumor activities of two (1 goes to 3)-beta-D-glucans, isolated from the fruiting body of Auricularia auricula-judae ("kikurage", an edible mushroom), and other branched polysaccharides containing a backbone chain of (1 goes to 2)-alpha-D-glucosidic or (1 goes to 3)-alpha-D-mannosidic linkage [and their corresponding (1 goes to 3)-D-glycans, derived by mild, Smith degradation] were compared. Among these polysaccharides, a water-soluble, branched (1 goes to 3)-beta-D-glucan (glucan I) of A. auricula-judae exhibited potent, inhibitory activity against implanted Sarcoma 180 solid tumor in mice. The alkali-insoluble, branched (1 goes to 3)-beta-D-glucan (glucan II), a major constituent of the fruiting body, showed essentially no inhibitory activity. When the latter glucan, having numerous branches attached, was modified by controlled, periodate oxidation, borohydride reduction, and mild, acid hydrolysis, the resulting, water-soluble, regraded glucan, having covalently linked polyhydroxy groups attached at O-6 of the (1 goes to 3)-linked D-glucosyl residues, exhibited potent antitumor activity. Further investigations using the glucan-polyalcohol indicated that the attachment of the polyhydroxy groups to the (1 goes to 3)-beta-D-glucan backbone may enhance the antitumor potency of the glucan. On the other hand, partial introduction of carboxymethyl groups into glucan II (d.s., 0.47--0.86), which altered the insolubility property, failed to enhance the antitumor activity. The interrelation between the antitumor activity and the structure of the branched (1 goes to 3)-beta-D-glucan is discussed, on the basis of methylation and 13C-n.m.r. studies of the periodate-modified glucans.

Animals↗