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Method for producing biologically active polysaccharide RON substance

  • US 5,332,667 A
  • Filed: 09/02/1993
  • Issued: 07/26/1994
  • Est. Priority Date: 02/08/1989
  • Status: Expired due to Fees
First Claim
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1. A process for the preparation of a biologically active polysaccharide RON substance which comprises(a) culturing a microorganism belonging to the genus Leuconostoc, which has an ability of producing a physiologically active polysaccharide RON substance, said microorganism being at least one strain of Leuconostoc mesenteroides subsp. dextranicum, said strain selected from the group consisting of strain BL-75 (FERM BP-2242), strain NCFB 517 (FERM BP-2711), strain NCFB 531 (FERM BP-2712), strain NCFB 861 (FERM BP-2713), strain NCBF 864 (FERM BP-2714), strain 46-1 (FERM BP-2670), to form a polysaccharide RON substance having the following properties:

  • (1) form;

    the substance is a white amorphous powder having no taste and no smell;

    (2) solubility;

    the substance is soluble in water, but the solution becomes milky white and viscous when the concentration is elevated;

    the substance is soluble in formamide and dimethylsulfoxide, but the substance is insoluble in alcohols, acetone, benzene, ethyl acetate, hexane, chloroform and carbon tetrachloride;

    (3) pH of aqueous solution;

    the aqueous solution is neutral to weakly acidic;

    (4) constituent saccharide;

    the substance comprises solely glucose;

    (5) elementary analysis values;

    the substance comprises 44.0 to 45.0% of C and 6.1 to 6.3% of H;

    (6) structure;

    the substance is an α

    -1,6-glucan and comprises a small amount of 3,6-branched side chains;

    (7) proteins;

    the substance comprises almost no proteins;

    (8) molecular weight;

    the substance does not permeate through a dialysis membrane and the molecular weight is presumed to be higher than 10,000 daltons;

    (9) specific rotation;



    )D25 =+190°

    ˜

    +220°

    (C=0.5, formamide);

    (10) color reaction;

    the substance is positive to an anthrone sulfuric acid reaction and a phenol sulfuric acid reaction, but negative to a biurette reaction, the Lowry-Folin reaction, the Elson-Morgan reaction and an iodine reaction;

    (11) melting point;

    the substance has no definite melting point;

    (12) ultraviolet absorption spectrum;

    the substance has no characteristic absorption;

    (13) infrared absorption spectrum;

    the substance shows a characteristic absorption of α

    -glucan;

    (14) 13 C-NMR spectrum;

    the substance shows characteristic chemical shift values of α

    -1,6-glucan in the main signals; and

    (15) biological activity;

    the substance has an antitumor activity against mice having a transplantable tumor, and(b) collecting said substance from a culture broth.

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