SU1735368A1 - Enzymatic nutrient medium for preparation of aminoglycoside antibiotic complex - Google Patents

Abstract

Usage: in the microbiological and medical industry to obtain aminoglycoside antibiotic complex, consisting of carbamoyltrabracin, carbamoylcanamycin and apramycin. The essence of the invention: the fermentation process producer Streptomuces cremeus subsp. nebramycini is carried out at 37 ± 1 ° C for 24 hours with stirring and aeration on a medium containing sources of carbon, nitrogen, phosphorus and mineral salts. The fermentation medium contains non-traditional raw materials: cotton flour and beet powder. The activity of the culture fluid is increased by 25%. 1 tab.

Description

The invention relates to the microbiological and medical industry, in particular, to a method for producing an aminoglycoside antibiotic complex consisting of carbamoyltrabracin (tobramycin, a medicine, is of interest in chemotherapy of infections caused by gram-negative bacteria, including Ps. aeruginosa), carbamoylcanamycin and apramycin (used in veterinary medicine in the treatment of gastrointestinal diseases of cattle and pigs).

Various fermentation media are known to produce an aminoglycoside ntibiotic complex containing tobramycin and apramycin.

Closest to the invention is a fermentation process using the producer Spteptomyces cremeus subsp. nebramycini, which is maintained on a fermentation nutrient medium of the following composition, wt%: soybean flour 3.5; glucose

1.5; animal fat 1.5; ammonium chloride 0.6; magnesium sulphate 1.1; chalk 0.7; tap water the rest.

The disadvantage of the prototype is the low antibiotic activity of the culture fluid at the end of the fermentation.

The aim of the invention is to increase the antibiotic activity of the culture fluid.

This goal is achieved by the fact that fermentation medium for deep cultivation of Streptomyces cremeus subsp. nebramycini additionally contains cotton flour and beet powder in the following ratio of components, may%:

Soya flour1.9 - 2.1

Cotton flour1,4-1,6

Beetroot

powder2,4-2,6

Magnesium sulphate 1.0-1.2

Ammonium Chloride 0.5 - 0.7

Mel0.6 - 0.8

fe

Vj

WITH

ate with

00

Animal fat1,4 - 1,6

WaterEverything

Cotton flour is a source of carbon, nitrogen, phosphorus, trace elements. Beet powder is predominantly a source of carbohydrates. The use of these two components within the specified limits allows to obtain the optimum carbon: nitrogen ratio in the fermentation medium, which leads to an increase in the antibiotic activity of the culture fluid.

The use of non-traditional types of raw materials allows to exclude food raw materials - glucose from the composition of the medium and reduce the content of soy flour in the medium.

Example 1 (by a known method). The culture of Streptomyces cremeus subsp. nebramycini (strains 2242, 9871). The vegetative inoculum is grown at 37 ± 1 ° C for 24 hours in flasks on a rocking chair with a speed of 240 rpm and in a seed drill with a capacity of 7 liters (filling factor 0.7) with a stirrer speed of 150 rpm per the medium of the following composition, wt.%: glycerin 2,0; corn extract 2.0; animal fat 0.75; ammonium chloride 0,2; sodium chloride 0.3; chalk 0.5; tap water the rest. The pH of the medium before sterilization is 7.4 ± 0.1.

The seed is used when fermentation is carried out in devices with a capacity of 30 liters (filling factor 0.5) and in flasks (VvsolOO ml of medium) on a swing with a speed of 240 rpm, adding it in an amount of 2% of the volume of the culture fluid. Fermentation is carried out at 37 ± 1 ° C on a regulated fermentation medium of the following composition, wt%: soybean flour 3.5; glucose 1,5; animal fat 1.5; ammonium chloride 0.6; magnesium sulphate 1.1; chalk 0.7; tap water the rest, pH of 6.2 ± 0.1. The amount of air supplied 1 volume to 1 volume of medium in 1 min; the operation of the stirrer is continuous, the speed of 150 rpm The duration of fermentation 72 hours

The antibiotic activity of the culture fluid was determined by diffusion into agar using a Va test culture. subtilis strain 6633 and expressed in units of 1 ml. The total antibiotic activity of the culture fluid is 2800 µg / ml for strain 9871 and 1400 µg / ml for strain 2242.

Example 2. The fermentation process of the producer of Streptomyces cremeus nebramycini is carried out according to example 1. As the fermentation medium, the medium of the following composition is used, wt%: soya flour 2.0; cotton flour 1,5; beet powder 2,5; magnesium sulphate 1.1; ammonium chloride 0.6; chalk 0.7; animal fat 1.5; tap water the rest.

The total antibiotic activity of the culture fluid is 3500 µg / ml for strain 9871 and 1750 µg / ml for strain 2242.

Example 3-14. The biosynthesis of the aminoglycoside antibiotic complex is carried out according to example 2.

The concentrations of the ingredients in the fermentation medium and the antibiotic activity values of the culture liquids are presented in the table.

Concentrations of chalk and fat in the range of 0.6–0.8 and 1.4–1.6 wt.%, Respectively, established on the basis of their optimal effect on stabilizing the pH and defoaming of the culture fluid, do not have a noticeable effect on the activity of the culture. fluid.

Claims (1)

From the table it follows that the following concentrations of ingredients are optimal, wt%: soybean flour 1.9-2.1; cotton flour 1.4-1.6; beet powder 2.4-2.6; magnesium sulfate 1,0-1,2; ammonium chloride 0.5-0.7. With these values of ingredients, the activity of the culture fluid increases by 25%, which leads to an increase in the yield of the antibiotic complex and, accordingly, to a decrease in its cost. In addition, the proposed environment allows the use of unconventional raw materials. Claims of the invention Enzymatic nutrient medium for the production of an aminoglycoside antibiotic complex using the producer Streptomyces cremeus subsp. nebramycini, containing soy flour, chalk, ammonium chloride, magnesium sulphate, animal fat, water, characterized in that, in order to increase the antibiotic activity of the culture fluid, it additionally contains cotton flour and beet powder in the following ratios of components, wt.% : Soyeva flour1.9-2.1 Cotton flour1,4-1,6 Beetroot powder2,4-2,6  Magnesium sulphate 1.0-1.2 Mel0.6-0.8 Ammonium chloride 0,5 - 0,7 Animal fat1,4-1,6 Water the rest.