CS244573B1 - Acremonium chrysogenum ccm f-752 microorganism strain - Google Patents

Abstract

Microorganism strain Acremonium chrysogenum Cephalosporin-producing CCM F-752 C. The strain of the invention was buffered (by miutagens and qiplacivive passive selekciamii. The advantage of the strain according to of the invention is the increased production of cephalosporin c (2x), ability to maintain high production velocity in extended cultivation and the ability to produce cephalosporin C on non-containing production soils methionine.

Description

(54) Strain of Acremonium chrysogenum CCM F-752

Acremonium chrysogenum strain CCM F-752 producing cephalosporin C. The strain according to the invention has been withdrawn (by miutagens and qiplactive passive selection. and the ability to produce cephalosporin C on melamine-free production soils.

The invention relates to the top of the microcirganism of Acremonium chrysogenum CCM F-752 (internally designated BB-IV / 82-D), producing cephalosporin C.

This strain was obtained from Acremonium chrysogenum ATCC 48272 by mutagenesis and repeated passive selections.

The production strains currently used to produce cephalosporin C produce about 12,000 µg of cephalosporin C in a submerged fermentation medium in a fermentation medium at a cultivation time of 120 to 180 hours. The decisive way in increasing production and improving its profitability is to increase production using new, higher-production production strains. In the case of new strains obtained by active and passive selection, it is advantageous to focus on strains which achieve higher production per unit of time or on strains which maintain the production rate of cephalosporin C even at prolonged cultivation times.

These advantages also have a new strain of microorganism Acremonium chrysogenum according to the invention, deposited in the Czechoslovak Collection of Microorganisms of the University of J. E. Purkyně in Brno, under the reference CCM F-752 (working designation BB-IV / 82-D).

The advantage of the new strain is the increased productivity of cephalosporin C, which is up to twice the production of the previous strains.

An important property of the strain is its ability to maintain high production rates with extended cultivation times.

Another economically significant property of the strain is its ability to produce cephalosporin C on methionine-free production stages. In the previously known strains, methionine was a critical and determinant of nutrients for the production of cephalosporin C.

In the breeding of production strains, currently known methods are used which combine actively mutagenic interventions with passive selection. Among the most known mutagenic agents, ultraviolet radiation, nitrosomethylurea, N-mustard, acriflavin and others are used either alone or in combinations. Furthermore, these interventions are also combined by passing the culture on stage with the addition of suitable, specific reagents.

The novel strain of the invention was also obtained and isolated using these procedures.

The basic starting material for the noble process was the ATCC 48272 strain, to which we treated alternately with UV light and nitrosomethylurea. The selected isolates were further subjected to passage through media with reduced to zero methionine content. After reuse of UV light and further passage isolates were obtained which were subjected to verification of their properties for suitability for industrial use.

At the same time, this strain has been selected for the strain to which we claim patent.

In addition, cephalosporin C poodiication was used as a standard biological method using Bacillus subtilis and Alcaliges faecalis test strains, spectrophotometric method and high pressure liquid chromatography method.

The new strain has the same characteristics with the original strain. Upon monosporic expansion of the peptone nucleus below, it forms round colonies with a diameter of 6 to 8 mm. After 13 to 15 days, colonies are sporulated. They have blele color with a touch of rye, creamy. By aging, the color gradually turns into creamy. Colonies are radially, irregularly wrinkled, in the middle they rise to a slight point. On other types of sporulation platforms, the rise time is prolonged and colonies do not have all the characteristics. In submerged cultivation, the fibers at the propagation stage are thin, long, sometimes forming clumps. In the production stage, they gradually shorten to spherical forms.

Conventional forms of long-term preservation of the strain are not very effective. The freeze-dried culture retains spores germination for a maximum of three months, after which time this form of preservation does not guarantee a further culture life. Unambiguous and reproducible results in strain conservation are given by the method of permanent selection by monosporic selection from a cultured 3 to 4 week old culture.

Similar to the parent strain, the vegetative form of submerged culture is demanding for a sufficient concentration of dissolved oxygen in the culture medium. If there is insufficient oxygen transfer to the ipod and its concentration drops to near zero, growth and culture production will stop.

A very important characteristic of the strain is that its production of cephalosporin C is not undermined, nosťoiu metionirau.

Example 1 '·

The inoculum pod in the flasks was inoculated with a spore suspension and cultured at 28 ° C on a rotary shaker for 47 hours. The vegetative inoculum thus prepared was seeded with nutrient-containing flasks containing corn steep liquor, peanut flour, glucose, starch, calcium carbonate, salts and soybean oil. Cultivation on a rotary shaker was carried out at 25 ° C for 144 hours and a cephalosporin C production of 15.560 (Ug / ml) was achieved.

Example 2

The fermentation of cephalosporin C was carried out in laboratory fermentation tainto contents of 20 liters. Vegetative inoculum was also prepared on this underfloor apparatus containing mainly corn steep liquor, sucrose, ammonium acetate, calcium carbonate and soybean oil. Growth of the inoculum was stopped at 69 hours of culture. The inoculum at 10 ° / o was inoculated with a production tank containing a nutrient platform containing mainly corn steep liquor, soybean oil, calcium carbonate and a mixture of salts. Soybean oil, ammonium sulfate and ammonia water were added during the cultivation. After 151 hours of culture, production of 26,920 µg / ml cephalospor C was achieved.

Claims (1)

SUBJECT INVENTION Acremonium chrysogenum strain CCM F-752 producing cephalosporin C.