CS242495B1 - Chlorotetracycline-producing Streptomyces aureofaciens CCM 3,767 strain - Google Patents

Abstract

The long-term high production of chlortetracycline in the production equipment, when all the conditions of the technological process are met, depends primarily on the stability of the production strain and at the same time on its resistance to possible adverse effects, including actinophage infection. The Streptomyces aureofaciens CCM strain is resistant to phage infection and extremely stable in subsequent spore generations.

Description

The invention relates to a strain of Streptomyces aureofaciens CCM 3,767 (internally designated UV 68 / 83-6) producing chlorotetracycline, characterized by resistance to phage infection and stability of properties in subsequent spore gererations.

Increasing industrial production of the chlorotetracycline antibiotic is most often based on the higher production capability of strains used for bipsynthesis. These strains must exhibit specific fermentation characteristics in the production apparatus. In particular, it is to maintain a high production capacity and resistance to possible adverse effects during fermentation, of which phage infection represents the greatest danger.

The strain Streptomyces aureofaciens CCM 3,767 was obtained by a series of the following mutagenic interventions and passive selections from the strain Streptomyces aureofaciens CCM 3,493. One strain in the genealogical line is also Streptomyces aureofaciens CCM 3,766 (inert designation 5/211). The last mutagenic intervention was made by UV light by energy fluctuation of 214 J / m ² and subsequent selection in liquid and solid medium containing the own metabolite - chlorotetracycline at a concentration of 100 µg and 400 μξ chlorotetracycline. per milliliter of media. The obtained sporulated cane colonies were inoculated onto sloping agar with a standard sporulation platform. The cultures thus obtained were the starting material for the production evaluation system which is carried out according to the following system:

vegetative generation - fermentation

In this way, a strain of Streptomyces aureofaciens CCM 3,767 was obtained.

In particular, the characteristics of the strain obtained are: increased production, stability of this production in subsequent spore generations, and resistance to phage infection. The shape of the colonies grown on the sporulating soil after monospore separation, their pigmentation and the color of the spores are the same as that of the initial strain Streptomyces aureofaciens CCM 3,493. cultures from fermentation of strain CCM 3,767 are characterized by the formation of thin, short and thick branched fibers. Sporulation of CCM 3,767 culture takes longer and requires stricter adherence to culture conditions compared to CCM 3,493.

A comparison of the strain properties with the properties of the starting strains CCM 3,493 and CCM 3,766 (5/211) is shown in the following table:

strain

Bank production wg / ml%

Resistance to phage

(NM XXIV / 52) CCM 3,493 5 740 (5/211) CCM 3,766 4 900 (UV 68 / 83—6) CCM 3,767 5 200

EXAMPLE

The vegetatively inoculum is prepared in 500 ml brewing flasks with 60 ml of inoculum containing the basic nutrients, i. j. source of carbon - sucrose, nitrogen - corn extract and soybean and salt. The vegetative inoculum medium is inoculated with spores from one third of the slant agar and inoculated for 22 hours. This vegetative inoculum was used to inoculate a 500 ml beaker with 60 ml fermentation broth. The carbon source is sucrose, the nitrogen source is corn extract, soy flour and the ammonium salt. Among other salts, the soil contains calcium carbonate. The preparation of the vegetative inoculum and the fermentation are carried out on a rotary shaker at 230 rpm, an eccentricity of 25 mm and a temperature of 29 ° C. Fermentation is carried out for 4 days, samples are taken + + - + + after 96 hours. The production of 5000-5,400 pg / ml of chlorotetracycline is achieved as described above.

Example 2

The procedure of Example 1 is followed except that inoculum preparation and fermentation are carried out in large-volume fermenters. Inoculum preparation is done in a 5 m ³ inoculation tank filled with 2 m ³ inoculation soil. Fermentation takes place in a 50 m ³ fermentation tank filled with 29 m ³ fermentation medium. The culture of the strain in the inoculation tank takes 26 hours and the fermentation 110 hours with standard mixing and aeration of the culture. The above procedure produces 88% of the CCM 3,493 production.

The advantage of using the strain Streptomyces aureofaciens CCM 3,767 is its ability to produce chlorotetracycline even under phage infection conditions and the stability of this property.

Claims (1)

OBJECT OF THE INVENTION A strain of Streptomyces aureofaciens CCM 3,767 producing chlorotetracycline and resistant to phage infection.