CS212149B1 - Microorganism strain of the penicillium chrysogenum ccm f-679 - Google Patents

Description

The vegetative inoculum was prepared submersibly in 500 ml beakers with 100 ml inoculum broth containing basic nutrients, ie carbon source (glucose and lactose), nitrogen (corn extract and ammonium sulfate) and salts. Vegetative flasks were inoculated with sloping agar spores and incubated on a rotary shaker for 48 hours.

The vegetative inoculum thus prepared was seeded with 500 ml fermentation flasks, 40 ml fermentation broth volume. The source of carbon was lactose and soybean oil, the source of nitrogen was Pharmamedia (globular protein of bleach seed) and ammonium sulfate, the soil further containing the appropriate precursor, calcium carbonate to buffer the soil, and the source of sulfur.

Vegetative inoculum preparation and fermentation were carried out on rotary shakers at 230 rpm, eccentricity 25 mm at 25 ° C. Isolates were evaluated twice before lyophilization, i.e. before preservation, two to three times after lyophilization, the best isolates multiple times. Lyophilized cultures serve for long-term preservation.

This evaluation system resulted in the strain to which the claim is claimed.

Fermentations are carried out for 10 days, samples for determination of efficacy are taken on days 8, 9 and 10 of fermentation. Maximum production is generally achieved on day 9 of fermentation.

The potency is determined by the hydroxamate method.

Fermentation procedure for penicillin V biosynthesis

Control strain 65/41 and strain 65 / 41-19 were fermented in 500 ml beakers with 40 ml of fermentation broth labeled on rotating shakers at 230 rpm, eccentricity 25 mm with oxygen transfer 0.81 ml Og / ml / h. The inoculation was performed with a 5% v / v vegetative inoculum. Composition of fermentation broth B ^ (in percent by weight):

lactose 15.0 Pharmamedia 2.5 ammonium sulfate 1.05 calcium sulfate kr. 0.5 sodium sulphate 0.4 potassium phosphate kys. 0.05 soybean oil phenoxy acetic acid dissolved 1.0 in sodium hydroxide solution 0.8

at 100 hours of fermentation, a 10% phenoxyacetic acid solution of 0.2% phenoxyacetic acid sol. in sodium hydroxide pH is not adjusted, after sterilization is 6.9 to 7.1 sterilization is performed for 20 min at 120 ° C

Fermentation procedure for penicillin G biosynthesis

The fermentation procedure for penicillin G biosynthesis is identical to that for penicillin V biosynthesis, except that the fermentation broth does not contain phenoxyacetic acid but sodium phenylacetic acid, which is dosed during the fermentation as a dragee every 24 hours starting at 24 hours and ends at 120 hours at 0.19% so that the total precursor concentration is 0.95%.

Production comparison:

strain fermentation product production j 65/41 IN 27 000 G 23 000 65 / 41-19 IN 30 000 G 25 000

The increase in production is about 10%.

Tribe description:

Strain 65 / 41-19 (CCM F-679) forms a colony of about 10 mm in size over 10 days of cultivation at 25 ° C on monosporic sowing on glycerin-molasses sporulation medium. The culture works well on different types of sporulating soils, especially on natural substrates such as millet, barley, corn, rice, etc. The color of the sporulated culture is deep green, gray after prolonged cultivation or storage at +5 ° C.

Strain 65 / 41-19 on natural substrates is stable for several months.

Strain Penicillium chrysogenum, labeled 65 / 41-19 was deposited in MS. collection of microorganisms (Czechoslovak Collection of Microorganism), University of J. E. Purkyns, Brno, Tr. Defenders of Peace 10, under no. CCM K-679

Claims (1)

SUBJECT OF THE INVENTION Penicillium chrysogenum strain CCM F-679 producing penicillin V and G.