CS234589B1 - Microorganism strain Penicillium chrysogenum CCM F-753 - Google Patents

Abstract

A strain of the microorganism Penicillium chrysogenum CCM F-753 (Cu P-2) producing increased levels of penicillin V, penicillin G and 6-aminopenicillanic acid (6-APK). The advantage of this strain is its considerable resistance to mechanical stress during mixing in the fermentation vessel.

Description

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The invention relates to a strain of Penicillium chrysogene CCM F- (Cu P-2) producing penicillinV, penicillin G'a and 6-aminopanicillanic acid (6-APK). The prime of this strain is its considerable resistance to mechanical stress in admixture in the fexmentation vessel. The production of penicillin has been in production fermentation tanks for more than 30 years, and its volume is constantly increasing. The increase in production is related to the demand for the production volume of semi-synthetic penicillins derived from 6-aminopenicillanic acid. The increase in production can be realized by expanding the production fermentation plants or by increasing the yield of the fermentation broth. A second alternative, which does not require significant investment costs, is possible either by improving the fermentation conditions by improving the fermentation process or by increasing the production capacity of the strains used for biosynthesis. Breeding liens of penicillin biosynthesis strains have long been guided by a number of established procedures.

Penicillium chrysogenum strain CGM F- (Cu P-2) was a zinc from Penicillium chrysogenum strain number 2/170 by a series of repeated mutagenic interventions (nitrosomethylurea, nitrosomethylguanidine, UV-light, p-fluorophenylalanine) and subsequent rules passive selections. The isolates were evaluated according to the following scheme: - 2 - isolated colonies on the Petri dish and slanted agar vegetative inoculum 1 of the actual fermentation. 234 88 ·

Isolates were evaluated in two experiments before lyophilization (preservation) and in three experiments after lyophilization. · Lyophilized preservatives serve for long-term storage of spore material and allow preservation of appropriate growth and production properties.

Determination of penicillin efficacy was performed by an automated system using a hydroxamate polarimetric method · A strain was obtained with this evaluation system ·

The Penicillium chrysogenum strain OCM P-jfX (Cu P-2) forms a spontaneous spread on the sporulation medium of the glycerine-melasacolonia size of 7.5-10 mm during 10-12 days of culture at 25 ° C. Radial folds form at the edge, the center of the colony is depressed, the edge of the colony is lined with a white non-sporulating zone. Spores have a spherical to elipoid shape of the usual dimensions, they are colorless during microscopic observation. maize, oat groats tad ·), especially when enriching natural substrates with a source of nitrogen, preferably in the form of a 1% Pharmamedia solution · The color of a cultured culture is Teal ·

The Penicillium chrysogenum OCM P- (O 2 P-2) strain is sufficiently stable, both in vegetative passage (does not lose production capability of passages up to 4 generation) and in long-term storage in lyophilized form.

Biosynthesis of Penicillin V.

The vegetative inoculum was prepared submersely in 500 ml boiling flasks with 60 ml of inoculum, containing the basic nutrients, "ie carbon source (preferably sucrose), nitrogen (maize extract in combination with a yganic nitrogen source), a5sole · 234 589

The vegetative inoculum was inoculated with a spore agar loop and cultivated on a rotary shaker for 48 hours.

500 ml fermentation flasks with 40 ml of fermentation broth were seeded with the vegetative inoculum thus prepared. Sources of carbon Lactose and soybean oil in combination and addition of carbohydrate, the carbon source was soybean flour or Pharma-media, the soil also contained phenoxyacetic acid, as a side chain pre-cursor for penicillin V biosynthesis, chalk and sulfur source. the actual fermentations were carried out on rotary shakers at 230 rpm, 25 mm high at 25 ° C.

The production evaluation of the isolates was repeated 2 to 3 times, the best isolates were lyophilized and, after vaccination, repeatedly minimally 3 times production evaluated. These fermentation processes yielded the production of the use of the strain Penicillium chrysogenum CCM P- (Cu P-2) at a level of 20 to 26,000. penicillin V / ml according to the fermentation medium used and the fermentation process

Penicillin G Biosynthesis

Permentation of the Penicillium chrysogenum strain CCM P- (Cu P-2) for penicillin G bidsynthesis was performed analogously to penicillin V biosynthesis. The difference was only that the phenoxyacetic acid was not a side chain precursor but sodium phenylacetic acid which, due to higher toxicity, is dosed during fermentation in the form of a dragee or solution. Laboratory-scale production ranged from 17 to 19,000 µl of penicillin G / ml. Example 3Biosynthesis 6-APK.

The 6-APK biosynthesis fermentation process is analogous to that of Example 1. The only difference is that the soil did not contain any side chain precursor, ie, it does not contain 4 - 234 589 l and phenoxyacetic or phenylacetic acid. The obtained production is at the level of 13 to 15 0 0 0 6 6-APK / ml.

Claims (1)

5- 234 589 pS b ION I FALL. Penicillium chrysogenum strain of penicillin V, G and β-aminopenicillanic acid. «