SU1144378A1 - Leuconostoc dextranicum lu-122 strain - producer of dextran possessing immunity-stimulating effect - Google Patents

Abstract

The strain Leuconostoc dextranicum Lu-122 VKM V1526D (All-Union Collection of Microorganisms at the Institute of Biochemistry and Physiology of Microorganisms of the Academy of Sciences of the USSR) is a producer of dextran, which has an immunostimulating action.

Description

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00 1 The invention relates to microbiological susceptibility and relates to the preparation of dextran, which has an immunostimulating effect. Known shtamm Leuconostoc mesenteroides SF-4 - producer of dextran, which has an immunostimulating action. However, the known strain produces, dextran, has a weak immunostimulating effect. The purpose of the invention is a strain producing dextran with an increased immunostimulating action. This goal is achieved using Leuconostoc dextranocum LU-12 strain. Leuconostoc dextranicum strain Lu-122 was isolated from grape fruits Deposited by the All-Union Collection of Microorganisms of the Academy of Sciences of the USSR under the number VKM V1526D and is characterized by the following features. Cultural and morphological features. Small cocci 0.5-0.6 1.2-1.6 microns are assembled in pairs or chains of 3-8 cells. Gram-positive. On mediums containing sucrose, form a mucous capsule, immobile. On medium containing 0.5 g of yeast extract, 0.25 g of tryptone, 0.25 g of K, jHP04, 10 g of glucose, 2 g of agar and 100 ml of distilled water form smooth grayish-white rounded colonies with a diameter of 0.7- 1.0 mm On a medium of the same composition with 2 g of sucrose (without glucose), forms mucous, turbid-white hemispherical colonies with a diameter of 1.5-2.0 mm. Physiological and biochemical signs of Aerobe. Optimum temperature 23. The optimum pH of the medium is 7.0-7.2. It grows on media containing peptone, yeast autolysate and vitamins. On-potatoes do not grow, Starch does not hydrate. Hydrogen peroxide does not decompose. Gelatin and agar does not dilute. Glucose, sucrose, fructose, mannose ferments to acid. Does not ferment mannitol and sorbitol. Assimilates without the formation of acid arabinose, xylose, maltose, lactose, galactose, melezitose, raffinose Example. For the preparation of dextran, 20-30 ml of a live culture was transferred from 782 tons of medium of the following composition, g: KCf-0.1; 0.1 j KH2P04 1.75; 12HjO 4; P 0.5; Fe (NH4), 504-0.01; para-aminobenzoic acid 0.05; peptone 1.0; yeast autolysate 0.1 ml, sucrose 100; distilled water up to 1 l; The pH of the medium was adjusted to 7.2 with 1H. NaOH and grown at 25 ° C for 48-72 hours. During the growing process, the pH of the medium drops to 4.2, which is a sign of the end of biosynthesis. To isolate the dextran, the culture liquid was diluted with water twice, ethanol was added to a concentration of 25% by volume and the mixture was left in the refrigerator for 3 hours. The mixture was centrifuged (1800 g 30 min), the precipitate was discarded, and ethanol was added to the solution to 40 % by volume and left overnight in the refrigerator. Degrantran precipitates are separated by centrifugation, washed with 40% ethanol, repotted twice, and lyophilized. The yield is 15-20 g / 30-40% / of the theoretical. The resulting dextran is highly soluble in J8 water. At a concentration of 1-5% gives bluovay opalescent solution. Specific rotation / d / (C 1.0 in formamide). When fully hydrolyzed, only glucose is formed. According to methylation and C-NMR, 75% of the glucose spectroscopy contains ot -1,6-glyco-residues, linked by sidine bonds and 25% of glucose residues, connected by -1,2,2-glycosidic bonds located at the branch points of the molecule. The immune stimulating action of dextran is tested according to two criteria, 1. An increase in the phagocytic, activity of macrophages. Dextran at a dose of 0.2 mg is administered intramuscularly in the belmus weighing 1820 g. After 1,3.5 and 10 days, macrophages of the abdominal cavity are removed and incubated with a daily culture of Staphyfococcus. For each term take 5 experimental and 5 control animals. After 3 days, after dextran injection, an increase in the phagocytic activity of macrophages by 3.9 times compared with the control is observed. The phagocytic index increases 4.6 times in the same period. 2. Increased nonspecific resistance to infection. Dextran is administered intraperitoneally to white bottles weighing 18-22 g, 0.4 mg in 0.2 ml of saline. A control group of mice injected with saline in the same volume. In 24 hours, both groups of mice were infected with a pathogenic strain of E. coli E. coli 094 per dose, resulting in the death of 95% of the animals in the control group. The death of animals is observed for 5 days. In the experimental group of 110 mice survived 85, 25 died. The percentage of surviving bridges in this group was 77 ± 4%. 784 In the control group of 90 children, 4 survived, 86 died. The survival rate in this group was 4 + 2%. Testing the effect of the resulting and well-known dextrans on non-specific resistance to infection in mice shows that the survival of the mice under the effect of the resulting dextran is increased by 19% (25 + 2% against 6i1% of the prototype). The use of the invention allows to increase the immunological activity of dextran, as well as its output.

Claims (2)

The strain of Leisopois DehSgapsit Lu-122 VKM V1526D (All-Union Collection of Microorganisms at the Institute of Biochemistry and Physiology of Microorganisms of the Academy of Sciences of the USSR) is a producer of dextran with an immunostimulating action. 1 1144378 2