CS199980B1 - Strain of microorganism penicillium chrysogenum - Google Patents

Description

The present invention relates to a new strain of Penicillium chrysogenum Thom 65/41 (CCM P-569), which produces penicillin G or V or 6-penicillanic acid in larger quantities than the presently used parent strain.

The production strains currently used to produce penicillin produce about 18,000 IU into the fermentation medium. / ml penicillin at a culture time of 120 hours. This yield is achieved by the parent strain of the designation 43/28, derived by multiple selection from the flood strain G-165/16. Penicillin permented with strain 43/28 takes place on a production site using sucrose as a carbon source, araid flour or soy flour as a nitrogen substrate, and other conventional fermentation substrate components, which are inorganic salts, precursors, antifoams, or antifoams.

In addition to optimum fermentation conditions, the length of the fermentation process with economically advantageous penicillin production is limited by strain swelling of the strain 43/28. The properties of strain 43/28 also make it difficult to prolong the fermentation time beyond 120 culture hours while maintaining economically advantageous penicillin production fineness under conditions of prieraysl production. Therefore, it is economically advantageous to orient the selection of strains obtained by active selection to strains achieving higher production per unit of time, or strains which maintain a high production rate of penicillin, even at prolonged fermentation times.

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The present invention, which is a new strain of the microorganism Řenicillium chrysogenum, deposited in the Seekoslovenskej mlkroorganizmov collection of University J, E. Furkyne in Brno, Class of Defenders of Peace no. 10, under the designation Thom 65/41 (CCM F -569).

The main advantage of the new strain is the increased production of penicillin, which is up to twice that of the parent, parent strain.

The predisposition of the new strain is its ability to grow 1 of production 1 at a high concentration of ammonia ions, represented in nutrients by nitrogenous substances, e.g. ammonium sulfate, urea and the like.

Among other advantages of the new strain is its ability to achieve high penicillin production even on the depot carbon source in the nutrient basin, e.g. on lactose. Thus, the control of the fermentation process greatly simplifies the dosing of eacharose as a carbon source, as opposed to the technology based on the continuous operation.

For breeding Penloilllum chrysogenum production strains, mutagenic agents N-mustard, ultraviolet irons, nitrosomethylurea, akriflavin and other cells alone or in combination with one another are used. These actively mutagenic interventions are often combined with β-passive selecel, possibly by passage on the pod by the addition of suitable members, e.g., precursors.

The novel strain of the invention was also isolated using these procedures. The basic starting material for its selection was strain 43/28 to which they were treated with ultraviolet radiation, selected isolates were coated with passage through ammonium phenyloctate containing medium, and after careful use of the ultraviolet light effects, they were used N-mustard. Selection of suitable mutants was accomplished by conventional routine methods. In the final part of the work we examined various other characteristics of the strain, in particular optimal technological conditions and possibilities of industrial utilization. The novel strain of the invention, Thom 65/41, has shown to be the most suitable.

The new strain ea is morphologically similar to the flood strain. On the peptone eporulation platform, it forms colonies of 8-12 mm in diameter with monoeporic. After 8 to 10 days, the colonies are eluted, eu radially wrinkled, have a white border, a circular crater in the center, the color of the colonies is green, after a long period of cultivation and changes to dark green. On Czapek - Dox by glucose, the growth lasts 14 to 16 days, the color of the colonies is evil yellow, the colonies do not resorble. On Czapek - Dox pod with a modified source of carbon (arabnose, lnozitol or mannitol) it takes up to 20 to 24 days, the colonies do not reporulate, they have a diameter of 5 to 9 mm. The percentage of colonies grown on Czapek - Dox by e erablnose versus e glucose control is 62.2%, inositol 31.1% and manltol 2.2%.

The microscopic image of the new strain is similar to that of the parent strain, but under the same culture conditions it has a smaller fiber depth. In contrast to the parent strain, the intercellular walls of the fibers were more clearly visible on the stained preparations, which is highlighted in older socks. Scores noticeable autolysis do not affect the penicillin product as much as new fibers grow in parallel.

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Example 1

Glucose inoculation was inoculated in the flasks with a spore suspension and cultured at 25 ° C on a rotary shaker for 40 hours. The grown vegetative inoculum was inoculated and cultured on a rotary shaker to produce nutrient flasks containing 15% lactose, 2.5% pharmarmedia protein extract, 0.8% phenoxyacetic acid, inorganic salts and soybean oil as an antifoam. In 215 hours of cultivation, ea produced 26,800 IU / ml penicillin V,

Example 2

Preparation of inoculum as in Example 1 · Growing vegetative inoculum of both the new strain and the control strain were inoculated and cultivated on a rotary shaker to produce flasks with nutrient-containing flasks containing 15% lactose, 2.5% soybean, 0.8% phenoxiacetic acid, inorganic salts and soybean oil. After 215 hours of cultivation, the new strain produced a production of 21.670 m / ml penicillin V,

Example 3

The fermentation of penicillin V was carried out in 20-liter laboratory fermentation tanks. The inoculum was also prepared on the following composition: 2.0% sucrose, 3.3% corn leach with 55% dry matter, inorganic salts and soybean oil. Growth of the inoculum was terminated at 33 hours of culture, with a 10% inoculum inoculating a production tank with a nutrient platform containing 3.2% pharma-medium, 1.6% corn steep liquor, 0.55% phenoxyacetic acid, inorganic salts and soybean oil. Aacharose was added in the amount of 8.5% during cultivation.

IU / ml penicillin V.

On the same plate with peanut instead of pharma-media, the production of penicillin was 17.285 IU / ml.

Example 4

The preparation of the inoculum for the penicillin V fermentation was the same as in Example 3. The production tank contained a bile pod consisting of 10.0% lactose, 3.2% pharma-media, 0.8% phenoxyacetic acid, inorganic salts and soybean oil. 6.0% lactose was added during the culture. After 175 hours of cultivation, 24,100 IU / ml penicillin V was produced.

Example 5

Penicillin V fermentation as in Example 4, however, the pharmaceutical media was replaced by cotton flour. After 175 hours of culture, 21,800 IU / ml penicillin V was produced.

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Example 6

Fermentation of penicillin V as in Example 4, the pharmermade of the sap was stolen with soybean. At 164 hours, cultivol e and the production of penicillin V, 23.945 a.i / ml resulted in 137.0% of the control strain production.

Example 7

The preparation of the inoculum for the fermentation of penicillin G was the same as in Example 3. The production tank contained a nutrient plate containing 10% lactose, 3.2% soybean meal, inorganic salts and soybean oil. Phenylootic acid was added in an amount of 0.8% during the cultivation. After 165 hours of culture, 27,140 IU / ml penicillin G was achieved,

Claims (1)

Penioilllum chrysogenum strain CCM F-569, producing penicillin V or G, or 6-aminopenillanic acid.