CS244963B1 - Streptomyces aureofaciens strain CCM 3,766 producing chlortetracycline and resistant to phage infection - Google Patents

Abstract

Infection of a culture of a chlortetracycline-producing Streptomyces aureofaciens strain with an actinophage results in the lysis of the susceptible population of the strain. The lysis ranges from partial disintegration of the mycelium biomass to its complete disappearance. The degree of damage to the producing mycelium corresponds to disturbances in the fermentation process and lower antibiotic production compared to an uninfected culture; with complete lysis, the culture is devoid of antibiotic production. The Streptomyces aureofaciens CCM 3 766 strain is resistant to the actinophage and infection of a culture of this strain does not result in mycelium lysis and subsequent undesirable changes in the fermentation process.

Description

The present invention relates to a strain of Streptomyces aureofaciens CCM 3 766 (Intra-label 5/211} producing an antibiotic chlorothetracycline and characterized by resistance to phage infection. During the industrial production of chlorotetracycline, its production is constantly increasing. Higher production is achieved either by expanding the fermentation plants or The second method, unlike the first one, does not require significant investment costs: it is based on better production ability of the strain used for biosynthesis, or on better fermentation conditions resulting from improved fermentation control. which results in the cessation of the production of vines for a long time. phag particles which cannot be removed from the manufacturing equipment by conventional sterilization methods.

Streptomyces aureofaciens strain CCM 3766 was obtained from the strain Streptomyces aureofaciens CM3493 by not conducting spontaneous resistance to the phage, i.e. by growing the strain. Streptomyces aureofaciens CCM 3,493 in the presence of phage and subsequent selection of a resistant microorganism from the fermentation broth containing predominantly mycelium. The spores of the strain obtained were sown on a Petri dish with phage-containing sporulation-containing soil, and on a Petri dish with phage-free sporulation soil. Identical growth and sporulation demonstrated strain resistance to phage infection. In this way, the strain to which the claim is claimed has been obtained.

The main feature of Streptomyces aureofaciens CCM strain is its resistance to phage infection. The shape of the colonies in monospore sowing, their pigmentation and the color of the spores are the same as in the initial strain of Streptomyces aureofaciens CCM 3 493. The growth of Streptomyces aureofaciens CCM 3 766 is slower on stiff and liquid soils than on the initial strain. Also, sporulation starts later and the Streptomyces aureofaciens CCM 3,766 strain is more sensitive to sporulation conditions. The production ability of Streptomyces aureofaciens strain CCM 3,766 is 10-15 ° C lower than that of Streptomyces aureofaciens CCM 3 493. Example 1

According to the prior art fermentation of Streptomyces aureofaciens strain CCM 4993, the inoculum is vegetatively prepared submerged in 500 ml boiling flasks with 60 ml of inoculum, which contains basic nutrients, namely carbon source sucrose, nitrogen sources corn extract, ammonium sulfate and soybean sulphate and salts. The soil for the preparation of the vegetative inoculum is inoculated with spores of about one third of the culture on an agar slope and incubated on a rotary shaker for 22 hours. A 500 ml fermentation flask with 60 ml of production broth is inoculated with the vegetative inoculum thus prepared. The source of carbon is sucrose, the source of nitrogen is corn extract, soybean and ammonium salt. The soil further comprises calcium carbonate and other salts. At the start of the fermentation, a sterile isothiocyanate solution is added to the soil. The preparation of the vegetative inoculum and the actual fermentation are carried out on a rotary shaker at 230 o / min -1, 25 mm eccentricity and 29 ° C. Fermentation lasts for 4 days when maximum antibiotic production is achieved. The above procedure yields 5,500 to 6,000 (Wg / ml with an average of 5,737 µg of chlortetracycline / ml of soil corresponding to an index of 100%).

According to the prior art fermentation of Streptomyces aureofaciens strain CCM 4993 in the flasks of Example 1, the vegetative inoculum and the actinophage-infected vaccine production soil are vegetatively inoculated. Phage lyses mycelial cells of a sensitive population of the production strain. Under these conditions, production of 0-4000 µg / ml is achieved with an average production of induction cultures of 3,626 µg chlorothetracycline / ml of soil corresponding to an index of 63 ° / o. Example 3

The subject matter of the invention in fermentation of Streptomyces aureofaciens strain CCM 3766 in flasks under the conditions of Example 2 does not lysate myeloma cells of the strain by actinophage. The production achieved is 4,500 to 5,500 µg / ml with an average of 4,900 µg of chlortetracycline / ml of soil corresponding to an index of 85%. The advantage of using the strain Streptomyces aureofaciens CCM 3,766 is its resistance to phage infection while maintaining the ability to produce chlorotetracycline.

Claims (1)

SUBSTITUTE The Streptomyces aureofaciens strain CCM 3766 producing chlortetracycline and resistant to phage infection.