CS277576B6 - Microorganism strain Acremonium chrysogenum CCF-2035 - Google Patents

Abstract

The invention relates to a strain of the microorganism Acremonium chrysogenum CCF-2035 producing cephalosporin C. This strain was obtained from the strain Acremonium chrysogenum CCM F-750 by breeding aimed at increasing the production of cephalosporin C.

Description

The invention relates to a cephalosporin C-producing strain of Acremonium chrysogenum CCF-2035 (hereinafter CFS-C). This strain was obtained from Acremonium chrysogenum strain CCM F-750, by breeding aimed at increasing CFS-C production.

The starting strain achieved a production activity in the flasks of 10,200 ug / ml and in the tank of 22,400 ug / ml.

The described strain of the microorganism Acremonium chrysogenum CCF-2035 was obtained by subjecting the parent strain of Acremonium chrysogenum CCM-740 to mutagenic UV light.

Breeding was performed by sporulating the culture of the parent strain on SP-S slant agar composed of: peptone 1%, maltose 3.8%, liquid raffinate / heavy juice / containing 60%, sucrose 4%, agar 2.4% with washed with 10 ml of sterile water, filtered through a cotton plug placed on a pipette, in order to. separation of hyphae and arthrospores from conidia. Petri dishes with SP-S pod were inoculated from such a conidia suspension, suitably diluted.

Mutagenesis was performed using UV light with a Philips TUV 30 W germicidal lamp with a spectral energy distribution - wavelength of 253.7 nm to 93.5%, and wavelengths of 6.5%. The distance of the dish from the radiation source was 30 cm. After irradiation, the plates were immediately transferred to an incubator at 28 ° C, where they sporulated for 11 days. The recommended colonies were isolated under aseptic conditions and transferred to SP-S slant agar. IP-V soil was used as the propagation submerged medium with the following composition: sucrose 1.5%, C ₂ H ₇ O ₂ N 0.5%, DL-methionine 0.1%, KH ₂ PO ₄ 0.027%, corn leachate / 100 % / 0.8%, methyl oleate 0.08%. After culturing for 72 hours on a 4 Hz rotary shaker at 27 ° C, a vegetative inoculum (1st vegetative stage) was obtained.

ml of this inoculum were inoculated into 500 ml boiling flasks containing 60 ml of IP-V pod and cultured at 27 ° C on a rotary shaker for 48 hours / II. vegetative degree.

From the grown inoculum, 4 ml were inoculated into a 500 ml boiling flask filled with 40 ml of PP-II / as production medium consisting of: corn starch 3.2%, dextrin 6%, sucrose 1.5%, glucose 0.5%, CaCO ₃ 1 , 1%, / NH ₄ / 2SO ₄ 1.5%, KH ₂ PO ₄ 0.5%, MgSO ₄ 0.3%, corn extract 100% 8.7%, soybean oil 1.9%. We do not add DL-methionine to the soil, because the demands of the production microorganism on this amino acid are sufficiently ensured by other components of the soil, eg corn steep liquor.

Example 1

Conidia of the parent strain Acremonium chrysogenum CCM F-750 aseptically washed from the End tube with 10 ml of sterile water and diluted 1000-fold were inoculated with 0.5 ml onto SP-S-coated petri dishes. The plates were exposed to UV light for 1 minute in an aseptic box. After irradiation, the plates were immediately transferred to an incubator, and after 11 days, 55 separate colonies were inoculated onto SP-S slant agar slant. After growing for 11 days, the isolates thus obtained were first tested in a one-pot production test. The 5 best isolates from this set were subjected to the second stage of selection in a 3-pot test. An isolate was obtained from them, which had about 29% higher production! activity as the initial parent strain.

This strain was sent to the collection of microorganisms to the Department of Botany of Lower Plants, Faculty of Science, Charles University, Prague 2, Benátská 2, where it was assigned the number CCF-2035.

Example 2

The production strain of Acremonium chrysogenum CCF-2035 was gradually propagated from a lyophilized can onto SP-S slant agar in a Roux bottle. The stem forms cream-colored colonies with a wrinkled surface, with a crater of various sizes in the middle.

The conidia from the Roux bottle were propagated into a vegetative form in a two-step inoculation procedure. The Chemap 0030 laboratory tank was inoculated with 1500 ml of grown inoculum. After 136 hours of cultivation, a production activity of 35,800 ug / ml of cephalosporin C was reached, the volume discharged being 26,000 ml.

Claims (1)

A cephalosporin C-producing strain of Acremonium chrysogenum CCF-2035.