Polysaccharide S-60 and bacterial fermentation process for its preparation

• US 4,326,053 A
• Filed: 08/14/1980
• Issued: 04/20/1982
• Est. Priority Date: 12/04/1978
• Status: Expired due to Term

• Current Assignee: Cp Kelco (JM Huber Corporation)
• Original Assignee: Merck & Co., Inc.
• Inventors: Veeder, George T., Kang, Kenneth S.
• Primary Examiner(s): Brown, Johnnie R.
• Application Number:
  US06/178,054
• Time in Patent Office: 614 Days
• Field of Search: 536/1, 536/114, 536/119, 260/112 R
• US Class Current: 536/123
• CPC Class Codes: A23G 2200/06 containing beet sugar or cane sugar if specifically mentioned or containing other carbohydrates, e.g. starches, gums, alcohol sugar, polysaccharides, dextrin or containing high or low amount of carbohydrate
• CPC Class Codes: A23G 3/346 Finished or semi-finished products in the form of powders, paste or liquids A23G3/343 takes precedence
• CPC Class Codes: A61L 9/048 air treating gels
• CPC Class Codes: C08B 37/006 Heteroglycans, i.e. polysaccharides having more than one sugar residue in the main chain in either alternating or less regular sequence; Gellans; Succinoglycans; Arabinogalactans; Tragacanth or gum tragacanth or traganth from Astragalus; Gum Karaya from Sterculia urens; Gum Ghatti from Anogeissus latifolia; Derivatives thereof
• CPC Class Codes: C12N 1/00 Microorganisms, e.g. protozoa; Compositions thereof medicinal preparations containing material from protozoa, bacteria or viruses A61K35/66, from algae A61K36/02, from fungi A61K36/06; preparing medicinal bacterial antigen or antibody compositions, e.g. bacterial vaccines, A61K39/00; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
• CPC Class Codes: C12N 1/205 Bacterial isolates
• CPC Class Codes: C12P 21/005 Glycopeptides, glycoproteins
• CPC Class Codes: C12R 2001/38 Pseudomonas
• CPC Class Codes: Y10S 516/02 Organic and inorganic agents containing, except water
• CPC Class Codes: Y10S 516/06 Protein or carboxylic compound containing
• CPC Class Codes: Y10S 530/825 Bacteria

Abstract

A novel polysaccharide S-60 is disclosed composed of principally carbohydrate, 10-15% protein, and 3-4.5% acetyl groups as the O-glycosidically linked ester. The carbohydrate portion contains about 11% glucuronic acid (based on wt. gum) and the neutral sugars rhamnose and glucose, the latter having an approximate molar ratio of 3 to 2. This polysaccharide is produced by a new Pseudomonas species, P. elodea, in a suitable fermentation medium.

Claims

1. Heteropolysaccharide S-60, the carbohydrate portion of which contains about 6-9% (wt./wt.) O-acetyl groups as the O-glycosidically linked ester, about 22% (wt./wt.) glucuronic acid, and the neutral sugars rhamnose and glucose in the approximate molar ratio 3:

2, said rhamnose and glucose sugars being primarily 1,4 β

-linked, said heteropolysaccharide being further characterized in that it is anionic, forms elastic, thermoreversible gels which melt and set in the range of about 65°

-70°

C., and forms aqueous solutions whose 0.1% viscosity is 40-80 cP, measured on a Brookfield LVF viscometer, spindle 2, 60 rpm, 25°

C.

2. S-60 of claim 1 comprising about 50 % (wt./wt.) carbohydrate and about 50% (wt./wt.) of insoluble material of which 20-30% (wt./wt.) is protein.

3. Heteropolysaccharide S-60 produced by a process comprising incubating the organism Pseudomonas elodea in a fermentation medium containing a carbon source, source of potassium ions, a source of nitrogen, trace inorganic elements, and water at a temperature of 28°

-32°

C. and pH of 6-8 for 40-60 hours, and recovering the heteropolysaccharide by precipitation with a suitable lower alcohol.

4. S-60 of claim 3 wherein the organism is ATCC 31461.

5. S-60 of claim 3 wherein the nitrogen source is corn steep liquor and the alcohol is isopropanol.