SU631529A1 - Method of obtaining 2-o-methyl-d-rhamnose - Google Patents

Description

The invention relates to the microbiological industry, namely to a method for producing monosaccharide-1 2-0-methyl-D-rhamnose. Known methods for producing monosaccharides are based on the use of cultures, for example / StVeptoni ces Slreptom ces aEbus.BacciEus ToruEopsis, Acelobaclep mboxidau & ClJ and C2 However, the known methods for obtaining d-ribose and b-sorbose are widely used in nature monosaccharides. Known is the production of 2-0-methyl-D-ramnose by cultivating its producer on a nutrient medium under aeration conditions containing sources of carbon, nitrogen and necessary mineral salts. As a producer of 2-O-methyl-D-rhamnose use a culture of microorganisms M Kobacteriuw 402 3. However, this culture is stored in UK collections and is difficult to access for use in our country. The purpose of the invention is to simplify the process. The goal is achieved by using the BoicctSue faecaDis aEoaPigerjes culture 6 as a producer. The specified strain was isolated from bile in 1960 and is stored in the Museum of Living Cultures of the Moscow State Control Institute of Medical Biological Preparations named after LA Tarasevich. Morphology and cultural and biochemical properties. The stick is short, sometimes mobilely gram-shaped, curved, O-size, 5-1-2M. On the broth grows in the form of a uniform turbidity with the formation of a film, wall ring and sediment. Aerobe. Optimal growth t -30-37 °. It grows well on synthetic media when used as a nitrogen source of amino acids, and is capable of growing on ammonium salts as a nitrogen source, if carbon is a source of acetate or other three carbon compounds. Strain well decomposes protein derivatives such as peptone with the formation of ammonia. Proteins do not break down Does not decompose most sugars, but consumes them in an oxidative way. The essence of the method is as follows:

The day of the preparation of 2-0-ethyl-D-rhamnose culture of Boccieus iaecaBia aEca & igenes strain 6 is grown on honey-peptone agar or broth for 1-2 days at 37. From the saline-washed bacterial mass, dried with acetone and ether known the specific polysaccharide is extracted by the method. The resulting polysaccharide is hydrolyzed with 1M sulfuric acid (0.5% solution of polysaccharide in acid) at 100 °, 1.5-2 hours. The hydrolyzate is neutralized with ion-exchange resin or with dry barium carbonate. From the resulting sugar mixture, 2-0-methyl-D-rhamnose is separated by separating the latter on a column of cellulose powder in ethanol-water solvent systems (95: 5) or butanol-water (95: 5). In addition, this sugar can be obtained from the mixture by preparative paper chromatography using the solvent p-butanol-pyridine-water (6: 4: 3).

The 2-0-methyl-D-rhamnose preparation obtained by the method does not contain any impurity. Other sugars are divided as follows; to the following solvent systems: rt-butanol-pyridine-water (6: 4: 3), P-butanol-water- (95 : 5), acetone-water (95: 5), ethanol-shod (95: 5), and its peak coincides when the chromatograms combine with the peak of this sugar in the hydrolysis of polysaccharide when the latter is separated by gas-liquid chromatography in the form of adsorbated aldononitriles and polyols studied sugars. The sugar yield is 5% of the dry weight of the Specific polysaccharide preparation.

Example. Culture Boss g iaeca2ts a ca igenes strain b is grown on mono-peptone agar at 37 for a day, washed and washed with saline and then with acetone and ether. The specific polysaccharide is obtained by hydrolysis of the bacterial mass with a hot Fuller form. The output of the drug is 5% of the dry weight of the bacterial mass. 100,500 mg of the polysaccharide is hydrolyzed with sulfuric acid 1Y (0.5% solution) for 1.5 hours at SO. The hydrolyzate is neutralized with Dowex 1x16 (HCOg), concentrated at 3540 ° and applied in a volume of 3-5 ml onto a 3.7x50 cm column packed with dry cellulose powder. The separation is carried out in a solvent ethanol-water (95: 5). The separation is controlled by staining individual fractions with anthrone reagent and chromatography on

paper.

Under these conditions, a reproducible separation of 2-0-methyl-D-rhamnose is achieved, and the resulting preparation was chromatographically pure when separated in various solvent systems: p-butanol-pyridine-water (6: 4: 3), p-butanol- acetic acid-water (4: 1: 5), p -butanol, suspended with water with 1% ammonia and others. In the development of chromatograms of various developing reagents: anilinphthalate, an alkaline solution of silver nitrate, iodic acid-benzidine, and others. also does not contain possible impurities

amino acids and was serologically active when braking a homologous precipitation reaction.

Claims (2)

1. US patent 3919046, cl.195-30 2. Author's certificate of the USSR 45№ 461116 Cl, C 12 D 13/00, 1973. Z.So1pas15ap JourdaE of Chemistrv, 1967, 45,1.987.