RU1692144C - Method of preparing substance showing lytic action - Google Patents

Abstract

FIELD: microbiology, biotechnology. SUBSTANCE: strain Xanthomonas campestris ВКПМ B-4102 is cultured on the nutrient medium containing the sources of carbon, nitrogen and mineral salts, and the end product is isolated by common procedures. Prepared substances show lytic action against pathogenic microorganisms showing resistance to antibiotics. EFFECT: improved method of substance preparing.

Description

 The invention relates to the microbiological industry, and in particular to methods for producing a lytic drug used in medicine.

 An example of the method.

Culture of Xanthomonas campestris were grown in a medium containing 1.5% yeast extract "Difeo", pH 7.6, for 26-28 hr at ²⁸ ° C on a rocking medical flasks 750 ml.

 Cells are separated by centrifugation in the cold at 45,000 g for 40 minutes. The resulting culture fluid is subjected to ultrafiltration. Ammonium sulfate is added to the concentrated solution in the cold, the precipitate is removed by centrifugation (1 h at 45000 g).

 Ammonium sulfate was added to the supernatant to 80% saturation, and the precipitated protein was collected by centrifugation in the cold (1 h at 45,000 g). The pellet was resuspended in a small volume of 0.01 M Tris-HCl buffer, pH 8.5 and dialyzed against 0.001 M Tris-HCl buffer on a magnetic stirrer for 12-18 hours with frequent buffer changes.

 The resulting strain is characterized by the following properties.

 Morphological and cultural characters.

 Gram-negative bacilli, not forming spores, 0.5-0.7x2.0-5.0 microns with growth on agar media. When cultivated in liquid nutrient media with aeration, it forms filament-like cells from 10 to 20 microns. Flagella absent. The cells are motionless.

 It forms a pigment of two types: a pale greenish-yellow carotenoid nature that does not stand out in the medium, and brown, melanin-like, diffusing into the medium.

 The stroke on the IPA is greenish-yellow, shiny, slimy. Growth is good. Bacterial mass is poorly captured by the loop.

 Colonies on MPA after 18-24 hours 2-3 mm in diameter, convex round with a smooth edge, do not shine in transmitted light.

 With growth in BCH forms a film, precipitate, turbidity.

 Physiological and biochemical properties. As a source of nitrogen, it uses ammonium salts, peptone.

 As sources of carbon, carbohydrates, organic acids, grow well in rich organic environments.

Carbohydrate metabolism is oxidative. It forms acid in mineral media with arabinose, glucose, galactose, fructose, sucrose, lactose, trehalose, cellobiose. It does not use acetic and oxal fiber, agar does not destroy, starch does not hydrolyze, it dilutes gelatin, milk coagulates and pentonizes, it does not form hydrogen sulfide on media with peptone, it does not form nitrites from nitrates. Catalase activity is weak, 0.1% tetrazolium bromide inhibits growth. The temperature optimum of 28 ^° C does not grow at 37 ° ^C. The pH optimum of 7.0. Strict aerob grows in the pH range of 6.6-9.0. The percentage of G + C in DNA is 66.0. Stored on MPA. Due to vigorous autolysis, it requires frequent reseeding in 3-5 days.

 The lytic effect of the isolated preparation was carried out on 99 non-pathogenic cultures of gram-positive and gram-negative bacteria and 39 representatives of yeast from the collection of IBPM Academy of Sciences of the USSR. The drug has an active lytic effect on all gram-positive bacteria selected for testing and on some yeast.

Claims (1)

 METHOD FOR PRODUCING A LYTIC ACTION SUBSTANCE on pathogenic microbially resistant to antibiotics microorganisms, characterized in that the strain Xanthomonas campestris VKPM B-4102 is cultured on a nutrient medium containing carbon, nitrogen and mineral salts, and the target product is isolated by cultivation using known methods.