SK147694A3 - Strain of penicillium vermiculatum ccm 8194 microorganism and preparation method of macrolide antibiotic of vermiculine - Google Patents

Abstract

This patent describes the microorganism family called Penicillium vermiculatum CCM 8194 together with the procedure closed to macrolide antibiotics related to vermiculine and this microorganism is prepared based on the cultivation of Penicillium vermiculatum CCM 8194 family with the use of the soil containing carbon, nitrogen, compounds of phosphorus, magnesium, copper and the substance affecting transfer of electrons in biological systems.

Description

Strain of Penicillium vermiculatum CCM 8194 microorganism and process for preparation of macrolide antibiotic vermiculin

Technical field

The invention relates to a strain of the microorganism Penicillium vermiculatum CCM 8194 and to a process for the preparation of the macrolide antibiotic vermiculin.

BACKGROUND OF THE INVENTION

Penicillium vermiculaturn was isolated from the wood reinforcement of the Jeroným Mine Uranium Mine in Jáchymov. The isolate obtained, originally designated Penicillium sp. 51 Ci, identified as Penicillium vermiculatum Dangeard s / (Fasaatiová O. and Hartmanová V .: Cesk. Mykológie 26, 114, 1972) was deposited in the Czechoslovak Collection of Microorganisms under the designation CCM F-276 and in the American Type Culture Collection as Talaromyces virus var. (Klocker) Stolk et Samson ATCC 26105. Later the strain was purified and the isolate obtained was stored in Cs1. collection of microorganisms under the code CCM F-748 (Fuska J., Khandlova A., Fuskova A .: Cs.pat. 236 637).

The macrolide antibiotic vermiculin was first isolated by Fuška et al. (J. Antibiotics 25, 208, 1972) from Penicillium vermiculatum Dang. CCM F-276 Vermiculin is a white crystalline substance, molecular formula C20H240g, molecular weight 392.41, t. t. 175-177 ° C, [α] 20 °, -12.5 °, well soluble in chloroform, hot acetic acid, ethyl acetate, butyl acetate and dimethylsulfoxide. This antibiotic acts both antitumorally in vitro and in vivo (Fuška J. et al. J. Antibiotics 32, 667, 1979), inhibiting the proliferation of the protozoa Trypanozoma cruzi and Tricnomus sioetus (Fusková A. et al. Foil Microbiol. 23, 389 (1978), has an immunosuppressive effect in vivo (Horakova L. et al. Foil Biol. 26, 312, 1980) and has a curative effect on adjuvant arthritis in mice (Rovensky J. et al. Int.J. Immunopathol. Pharmacol. 3 35, 1990).

P. vermiculatum strains hitherto used for industrial production of vermiculin were not suitable due to the low production of the desired macrolide metabolite vermiculin. In view of the expected introduction of vermicillin into medical practice, it was necessary to prepare a production based on a new, high-production strain of P. vermiculatum and to develop the biosynthesis conditions of this substance suitable for economically advantageous production.

SUMMARY OF THE INVENTION

SUMMARY OF THE INVENTION The present invention is based on the use of a novel, active breeding strain Perticillium vermiculatum CCM 8194, which forms solid sporulating yellow colony with low pigment production on solid culture media, capable of producing increased amounts of vermiculin which is cultivated on sucrose-containing soil. , nitrate salts and corn steep liquor as sources of nitrogen, magnesium and phosphorus salts and electron transfer substances in biological systems, preferably Krebs acid, lactic or ascorbic acid, with stirring at 26-28 ° C. Using a new strain under the above conditions yields 3-8 times more vermiculin than by culturing the strain

Penici 11ium vermiciatum CCM F-748 The increased concentration of vermiculin in the soil after cultivation provides the opportunity to utilize more efficient methods of isolation.

Methods for preparing vermiculin are shown in the following examples.

Examples

Example 1

Soil containing (g / 100 ml) is prepared in 500 ml flasks: sucrose 9, NaNC> 2 0.2, corn steep liquor (50% dry matter) 1.0, KH ₂ PO ₄ 0.1, KCl 0.05, MgSO ₄ . 7Hz 0 0.05, CuSO ₄ . 5H ₂ 0 0.0025, make up to 100 ml with tap water, adjust pH to 6.3. After sterilization of the soil (20 min at 120 ° C) and cooling, the soil is inoculated with 1036 vegetative inoculum of Penicillium vermiclatum CCM 8194. Cultivated on a rotary shaker (220 rpm) with stirring and aeration at 25-30 ° C for 144 min. - 196 h. Vermiculin production is 550 mg / L, ie more than 3 times the production of the original strain.

Example 2

To flasks containing 500 ml soil were prepared containing (g / 100 ml) sucrose 9 0.2 NaNO, corn steep liquor (50% solids) 1.0 kHz 0.1 PO 4, KC1 0.05, MgS0 ₄ .7H ₂ O 0.05, CuS0 _fourth 5Ha0 0.0025, lactic acid 0.5, make up to 100 ml with tap water, adjust pH to 6.3. The further procedure is the same as that given in Example 1. Vericulin production is 1250 mg / L, ie 8 times higher than the original strain and process.

Example 3

Proceed as in Example 1, but fumaric acid, 0.25 g / 100 ml, was added to the soil. After 168 hours of cultivation, the vermiculin content is 950 mg / l, i. approximately 6 times more than with the original strain.

Example 4

Procedure as in Example 1, but 0.5 g / 100 ml of fumaric acid is added to the soil. The production of vermiculin is 1180 mg / l, i. about 8 times higher than the original strain and process.

Industrial usability

The invention is useful in the pharmaceutical industry in the manufacture of a new medicament.

Claims (2)

PATENT CLAIMS A strain of the microorganism Penici22ium vermiculatum CCM 8194 producing the macrolide antibiotic vermiculin. A process for the preparation of a macrolide antibiotic vermiculin, characterized in that the Penicillus vermiculatuin CCM 8194 strain is cultivated on a soil containing a source of carbon, nitrogen, sulfur, magnesium, copper and one of the electron transfer substances in biological systems, preferably acid. Amber, fumaric, dairy or ascorbic in an amount of 0.1 - 1 g / 100 ml of soil.