SU881117A1 - Actinomyces grisesus vniigenetika-107f strain as grisin antibiotic producent - Google Patents

Description

I

This invention relates to the microbiological industry and relates to the preparation of industrial strains that produce a fodder antibiotic grisin.

Biologically active substance antibiotic grisin is widely used in agriculture as an animal growth stimulator. On the basis of a grizin, the feed preparation formogrizin is prepared using a filler. The grizin producer is the culture of Actinomyces griseus,

Of the known strains, the strain Ac.tinomyces griseus VNIIgenetica-115 has the highest activity, its activity after 48 hours of fermentation is 9600 units / ml 1. The VNIIgenetika-115 strain, as the most active, is considered as a prototype and as a basic sample.

However, these strains are characterized by sensitivity to production phages, as well as an insufficiently high growth rate and antibiotic formation.

The purpose of the invention is to obtain a culture strain of Actinomyces griseus, attributing to new actinophages isolated from the phagolysates of industrial crops during the production of the feedmogram, as well as having an increased growth rate and

10 antibiotic formation, and an increased ability to synthesize antibiotic grisin.

A new strain of VNIIgenetica-307F was obtained as a prototrophic revertant of the genes-dependent mutant of the Act nomyces griseus strain VNIIgenetica-115 against the background of 400 μg / ml of antibiotic grisin; .

Strain Actinomyces griseus VNIIGE20 notik-307F, producer of the antibiotic grisin is stored in the Central Museum of npoNJbmineHHbix microorganisms of the All-Union Scientific Research

Institute of Genetics and Breeding of Industrial Microorganisms jH has the registration number CMPM§ -5 2,

Morphological and cultural characteristics. Cells of filamentous mycelium, sporiferous spiral, oval spores of size 1, 8-0,9.MK Spore shell.

Potato agar. After 7 days, cultivation when forms: colonies with a diameter of 6-7 mm, rounded, slightly convex smooth, airy target gray, clearly expressed white border, opposite to the back of the colonies light gray, with a bright border.

Soyeva Wednesday. After 7 days, cultivated at 28 C forms colonies with a diameter of mm, round, smooth, aerial mycelium light gray. Colonies with white borders are on the opposite side of sulfur, with a bright border.

Maso-hepton agar. . After 7 days of cultivation., At 28 ° C, it forms colonies with a diameter of 3-5 mm, round, smooth, slightly convex, with poorly developed aerial mycelium.

Wednesday Gause 1, After 7 days of cultivation at 28 ° C forms colonies with a diameter of 7-8 mm, round, smooth Slightly convex, aerial mycelium is gray or light gray, along the edges of the colonies is white border, the reverse side of the colonies is dark sulfur, with light edging

On a 43-hour liquid procedural production environment, the culture forms large mycelial plexuses of freely growing hyphae.

Physiological and biochemical properties. Carbon sources are well absorbed by fructose, glucose, maltise, xylose, mannitol, mannose, rnose, starch, weakly arabinose, sucrose, do not absorb lactose, sorbitol, and do, lcite. Good growth in fiber is observed.

Sources of nitrogen S grows well on synthetic nutrient media containing mineral sources of nitrogen (ammonium nitrate, sodium nitrate). and on complex nutrient media with an organic source of nitrogen (peptone). Gelatin is diluted in the first days of growth. Peptonized milk on the seventh day of growth without coagulation,

Antagonistic properties. It has a broad antimicrobial spectrum.

inhibits gram-positive and gram-negative bacteria. The proposed Actlncxnyces qriseus strain VNIIgenetika-307F is resistant to Rd / 70, Rd antinophages, which lyse the production culture, has accelerated growth and antibiotic formation, in 43 hours

fermentation synthesizes on average

II000-12000 units / ml of antibiotic grisin. The maximum activity of the strain reaches 15000 units / ml.

When testing the VNIIGenetika-307F strain under experimental conditions at the Ungheni biochemical plant, antibiotic activity was obtained in the culture capacity from 4500 to 9000 units / ml. The average for 25 production operations m 6244 u / ml

Average activity of a known

Actinomyces griseus strain All-Union Scientific Research Institute of Genetics-115 under production conditions - 4000-4500 U / ml.

Strain Usage Examples

Actinomyces griseus VNII-GeneticsZrUF for the preparation of the antibiotic grisin a.

Example about Colonies of Actinofflvces gfriseus strain VNIIgenetica 307F grow an attraction for 7 days with potato agar medium and then pesesevat on spiked with the same medium. Culture on

Kokah grown in a thermostat for 7 days at 28 C. A seed culture of the strain is seeded with 750 ml Erlenmeyer flasks containing 50 ml of fermentation medium of the following composition,%: corn flour starch 1.5; ammonium nitrate 0.5; chalk 0.3; sodium chloride 0,2; monosubstituted potassium phosphate 0.02; magnesium sulfate 0.05 (pH 6.8-7.3) Fermentation is carried out on a circular rocking chair at -240 rpm and 28 ° C. After 43.h fermentation, the producer of the formogrizin Actinomyces gtlseiis, the NIIgenetics-307F strain, synthesizes in laboratory conditions an average of 11000Г2000 u / ml of antibiotic grisin. The content of antibiotic in the culture fluid is determined by the method of diffusion into agar using the test culture of Vas. subtTUs 6633.

Claims (1)

To test the VNIIGenetica-307F strain for resistance to antinophages isolated from production Fatalizates, pure cultures of 5 actinophages, designated Pg 70, Pg 1 15, are obtained. Propagated by the two-layer actinophagi, introduced into the upper layer of the medium with 0.8% agar (write the medium below) approximately 1-10 spores of a sensitive culture and an actinophage in an amount sufficient to form fusions from negative colonies. After 18–20 incubations, the top layer is removed with a spatula and centrifuged for 15–20 min. At 5,000 rpm. The test of grown culture .sht-amma VNIIgenetika-307F for resistance to actinophages was conducted on solid nutrient medium PKA-5 of the following composition,%: glucose 1.0; corn extract 0.5; hl. sodium sodium 0.5; calcium chloride 0.05; 1.5 agar. PRI me R 2. The medium of Example 1 is used to grow the antibiotic producer of grisin. Flasks with a volume of 750 ml and 100 ml of the medium are seeded with Actinomyces qrise us VNIIgenetika-307F. The culture in flasks is grown on rocking chairs at 240 rpm and 28 ° C for 24 hours. The seed grown in this way is transferred to the seed drill. The cultivation of seed in sowing machines is 18 hours at. The prepared seed in the amount of 10% was introduced into a 16 m fermenter with the fermentation medium of Example 1. The fermentation is done at 76 ° C. After 45 h of fermentation, the activity of the antibiotic was obtained in the culture fluid of 8100 units / ml. The use of the highly active phage-resistant strain Actincxnvces grtseus VNIIgenetika-307F, which produces the antibiotic grizin, makes it possible to carry out cost-effective large-scale production of the feed preparation formogrizin {as much as possible, to exclude the possibility of phagolysis fermentation. In addition, the use of the proposed strain under industrial conditions makes it possible to reduce the prime cost of the Onion product — the preparation of formogrizin; increasing its productivity — removing the product from 1 m per unit. Claims of the invention: Actinomyces griseus strain VNIIgenetika-307F, which produces the antibiotic grisin. The strain is stored in the Central Museum of Industrial Microorganisms of the All-Union Scientific Research Institute of Genetics and Breeding of Industrial Microorganisms and has a registration number CMPM5-512. Sources of information taken into account during the examination 1. USSR Author's Certificate No. 510512, cl. From 12 to 1/00. 1976 ..