RU2427644C1 - Bacillus subtilis BACTERIA STRAIN - FIBROBLAST GROWTH FACTOR PRODUCER - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: what is presented is a Bacillus subtilis 804 strain produced from a patient's unremarkable wound. The Bacillus subtilis 804 strain is deposited in the State collection of pathogenic microorganisms III-IV of L.A. Tarasevich State Research Institution of Standardisation and Control of Medical Biological Preparations, No. 288 and is a fibroblast growth factor producer in an environment.

EFFECT: preparations of the produced bacterial factor accelerate skin and mucosa wound healing.

Description

The invention relates to biotechnology, to bacteria producing fibroblast growth factor and can be used in medicine, veterinary medicine.

Currently, to obtain fibroblast growth factors, the gene responsible for its synthesis is isolated from an animal or plant cell, transplanted into a strain of Escherichia coli. The resulting gene-modified strain of Escherichia coli is grown on a nutrient medium (D.M. Ornitz, 2001). Fibroblast growth factor is isolated from it. As a result of such complex technology, the fibroblast growth factor produces a very small amount, and is very expensive. The cost of 1 mg reaches 40-70 thousand US dollars. This complicates its practical use both in biotechnology and in the creation of new drugs.

The strain Bacillus subtilis 804 obtained from the wounds of a patient who healed without complications. It is a natural strain of bacteria that produces fibroblast growth factor. Previously, natural strains producing fibroblast growth factor from bacteria of the genus Bacillus were not known. The strain Bacillus subtilis 804 was deposited April 13, 2010 in the State collection of pathogenic microorganisms of groups III-IV of the State Research Institute for Standardization and Control of Medical Immunobiological Preparations under number 288.

The strain of Bacillus subtilis 804 is characterized by the following properties:

Sticks, cell size (3-4) × (0.7-0.9) microns. Bacteria are mobile. Gram stains positively. Colonies on nutrient agar are white, rough, with scalloped edges, with a faint pink tint, with a diameter of 2-18 mm. The strain on meat and peptone broth at 37 ° C after 18-24 hours forms a whitish film on the surface and a small bottom sediment.

The strain has antagonistic activity against test strains of Staphylococcus aureus "Nikiforov", Staphylococcus aureus "Filippov", E. coli 157, Proteus mirabilis.

Causes hydrolysis of starch, reduction of nitrates. It breaks down glucose, sucrose, maltose, dulcite, galactose, lactose without gas evolution. Mannitol, rhamnose, lysine, arginine does not ferment. Hydrogen sulfide, indole does not form.

It is sensitive to streptomycin, lincomycin, erythromycin; not sensitive to benzylpenicillin, polymyxin.

The strain is non-toxic, non-toxic, non-virulent and harmless.

The product synthesized by the strain: fibroblast growth factor.

To obtain fibroblast growth factor, the strain Bacillus subtilis 804 is grown on a synthetic nutrient medium for 1-7 days. They carry out ultrafiltration, freeing themselves from living bacteria. Chemically pure growth factor from the metabolite is isolated by electrophoresis. From a liter of medium, 0.1 to 100 μg of growth factor is obtained.

The growth factor, which effectively stimulates the migratory and contractile activity of skin cells, as well as the proliferation of human fibroblasts, enhances the action of insulin as a growth factor, increases DNA synthesis. This is a complex of 4 proteins with a molecular weight of 11 to 14 kD. A dilution of 1:10 ⁹ contributes to almost the same increase in the number of fibroblasts in culture as the standard - 5% fetal calf serum. The growth factor is sensitive to proteolysis by trypsin, remains active after incubation at 100 ° C for 1 minute, and has an isoelectric point in the region of 9.2.-9.3. Preparations based on the obtained bacterial factor accelerate the healing of skin wounds and mucous membranes in experimental animals.

The cost of 1 mg of our growth factor is about 120 US dollars.

BIBLIOGRAPHIC LIST

1. Ornitz, D.M. Fibroblast growth factors. / D.M. Omitz, N. Itoh // Genome Biology - 2001 - No. 2 (3) - p. 3005.1-3005.12.

Claims (1)

The strain Bacillus subtilis, deposited in the State collection of pathogenic microorganisms III-IV of the State research institute for standardization and control of medical biological preparations named after L.A. Tarasevicha at number 288, which is a producer of fibroblast growth factor.