SU1631083A1 - Strain of actinomycet streptomyces lavendulae subsparuptini for producing aruptin - Google Patents

Abstract

The invention relates to the microbiological industry and relates to the production of a highly active actinomycete strain, the producer of arubutin, intended for use in agriculture, namely in vegetable production for combating. Bacterioses of agricultural plants and poultry to improve the efficiency of feed use. The aim of the invention is to obtain a new strain with higher antibiotic activity. The strain of the actinomycete Streptomyces lavendulae subsp.aruptini K-4111 was deposited in the All-Union Collection of Microorganisms under the number Ac-1267D. This strain allows increasing the synthesis of aruptin from 300-348 µg / ml (known strain under K & 39) to 990-1120 µg / ml. 1 tab.

Description

The invention relates to the microbiological industry and relates to the production of a highly active actinomycete strain producing the antibiotic aruptin for use in agriculture, namely in vegetable production for controlling bacterioses of agricultural plants and in poultry farming for increasing feed utilization efficiency.

The purpose of the invention is to obtain a new strain with a higher antibiotic activity.

Strain acticomycete Streptomyces lavendulae suhsp.aruptini, obtained in. the result of its selection from the homologous series of spontaneous variants.

From the strain of actinomycete Streptomyces lavendilae subsp.aruptini assigned to the number K-4111, the strain deposited in the All-Union Collection of microorganisms under the number AC-1267D,

Cultural characteristics of the strain are presented in the table.

Morphological properties: sporonate - straight and hook-shaped or in the form of primitive spirals, spores - oval, the surface of the spores is smooth.

Physiological and biochemical properties: aerobic, optimal growth temperature of 26-28 ° C, starch hydrolyzes weakly, milk coagulates and liquefies gelatine on the 10th day of growth.

As the sole carbon source, it consumes glucose, xylose, arabinose, and inositol. Slightly uses fructose and sucrose, does not consume mannitol, rhamnose, raffinose.

It has a broad antimicrobial effect, suppressing growth of gram-positive (Staphiloccocus aureus, Bacillus mycoides. Bacillus subtilis

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and others) and gram-negative (Esch. coli and others) bacteria, yeast (Candida albicang, Monilia sp. and others), fungi (Fusariun, etc.) and actonomycetes (Streptomyces allae, Streptoiny- 5 ces levorts and others).

The strain is maintained on potato agar medium. For long-term storage, methods are used to preserve cultures of actinomycetes under liquid paraffin in the form of soil cultures or in a lyophilized state.

The process of biosynthesis of the antibiotic auptin is carried out by growing actinomycete for 3-4 days in flasks on a circular rocking chair or in a fermenter on a liquid nutrient medium of the following composition,%: corn flour 1.5-3; starch 1.0-2.0 :, ammonium 0 sulfate 0.5-1; potassium phosphate monosubstituted 0.02-0.05; sodium chloride 0.2-0.5; technical chalk 0.3-0.6; tap water, pH 6.5 before sterilization.

The drug aruptin is a heated at 70-75 ° C for 10 min and freeze-dried 3-4-day-old culture fluid Str.lavendulac subsp. aruptini 30 To determine the content of biotic in the preparation of auptin, the latter is extracted with 0.01 n. HCl for 2 hours. The antibiotic activity of the extract is determined by diffusion into agar. Citrate buffer was used to dilute the extract, and Bacillus subtilis 6633 spores were used as a test culture. For comparison, an antibiotic of grysin with an activity of 1000 µg / mg was used as a standard.

Example 1. Streptomyces la vendulae subsp. aruptini K-4111 is grown for 14 days at 28 ° C in test tubes on skew potato 45 agar, which is prepared as follows: 200 g of peeled and finely cut potatoes are boiled in 1 liter of water, the broth is cooled, filtered, 2% is added agar and sterilized 50 at 1 atm. 30 min.

In order to obtain a vegetative seed, an agar block of a 14-day culture is introduced into 750 ml flasks containing 100 ml of 55 liquid nutrient medium of the following composition,%: corn extract 1.0; technical chalk 0.5; glucose 0,5; starch 1.5: ammonium sulphate

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0.35; sodium chloride - 0.5. water - plumbing, p natural environment. Sterilization at 1 atm for 30 minutes

Inoculated flasks are placed on a shaker (200 rpm). The culture is grown at 28 ° C for 48 hours. The vegetative seed obtained in this way is used to inoculate the fermentation medium, which has the following composition,%: corn flour 2; starch 1.5: ammonium sulfate 0.7; potassium phosphate monobasic 0.03; sodium chloride 0.2; technical chalk 0.5; water is tap water, the pH of the medium before sterilization is 6.8-7.0. Sterilization mode 1 atm, 30 min.

Seed material in an amount of 5 ml was added to 750 ml flasks containing 100 ml of fermentation medium each. The flasks are placed on a circular rocking chair and incubated at 28 ° C for 3 days. After this period, the culture fluid is heated prr 70-75 ° C for 10 minutes and freeze dried.

Antibiotic activity of culture liquid of strain K-4111 Str.lavendulae subsp.aruptini 1120 µg / ml. The antibiotic activity of the culture fluid in the control (strain 39 Str.lavendilae subsp. Aruptim) was 300 µg / ml.

Example 2. Streptomyces lavendulae subsp.aruptini, grown on stubbled potato agar and in a liquid nutrient medium to obtain a vegetative inoculum as indicated in Example 1. 5 ml seed material was added to 750 ml flasks containing 100 ml of fermentation environment (composition according to example 1). The flasks are placed on a circular rocking chair and incubated at 28 ° C for 4 days. Next, the charged culture liquid is heated at 70-75 ° C for 10 minutes and lyophilized.

Antibiotic activity of the culture fluid of the proposed strain K-4111 Str.lavendilae subsp.arup-- tini 1040 µg / ml. The antibiotic activity of the culture fluid in the control (strain No. 39 Str.lavendulae subsp.aruptini) was 348 µg / ml. , Example 3. Streptomyces lavendulae subsp.aruptini is grown on beveled potato agar and in

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liquid nutrient medium to obtain a vegetative inoculum, as indicated in Example 1

Seeds in the amount of: 5 ml were added to 750 ml flasks containing 100 ml of fermentation time (for composition, see Example 1). The flasks are placed on a circular rocking chair and incubated at 28 ° C for 5 days. After this period, the culture fluid is heated at 70 ° C for 10 minutes and lyophilized.

Antibiotic activity of the culture liquid of the proposed strain K-4111 Str.lavenhulae subsp. aruptini 990 mcg / ml

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The antibiotic activity of the culture liquid in the control (strain No. 39 Str.lavendulae subsp.aruptini) was 300 µg / ml. ,

Thus, the strain of the actipomicit Streptorayces lavendulae subsp. aruptini K-4111 allows to intensify the process of antibiotic biosynthesis 3-3.5 times.,

Claims (1)

Invention Formula The Strepomyces lavendulae subsp.aruptini BKM Ac-1267D strain of actinomycete to obtain aruptin. Cultural characteristics of the strain Glucosomineral (CP-0 Oats on Yeast Extract Potato Maize number 2 Gause number 1 Soeva M Sapepton Agar Wort Agar the color of the mycelium is given according to the Bondartsev color scale.