SU1232681A1 - Corynebacterium ch-1 strain strian - producer of l-valine - Google Patents

Abstract

The Corynebacterixjm glutamicum CH-l strain (collection of the industrial microorganism museum of the VNIIgenetika Institute, collection number CMPM-B-31139) is the producer of the L-roller. (L with to her yu O5 oo

Description

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The invention relates to the microbiological industry and is the Corynebacterium .glutamicum CH-1 Hsv strain, which has the property of producing L-valine, which is used in medicine for the preparation of amino acid mixtures for parenteral administration, in the chemical-pharmaceutical industry For synthesis of protein hormones of nature; ( iysulin, growth hormone, etc., as well as in microbiology and biochemistry for research purposes.

The purpose of the invention is to obtain a Corynebacterium glutamicum CH-i strain that produces L-valine with greater activity, expands the collection of strains - Valine Producers, which allows to simplify the process of obtaining valine on industrial scales due to a significant decrease in the number of coenzymes of the fermentation medium.

Scheme for the preparation of the strain of Bone-.terium glutamicum CH-1, C.glutamicura

ATCC 13032 (original strain). (biotin7 thiamine)

selection of auxotrophic mutants

after treatment with N-methyl-K-nit G-nitrosoguanidine (chemical mutagen), acetrophic mutants

i selection for productivity on L-valine With .glutamicum-78 (biotin, thiamine, arginine, 8-10 g / l L-valine)

G selection of mutants resistant to o-aminobutyric acid.

one

Mutants Resistant to Ottamino Acid Acid

I selection for productivity on L-vapina

C. glutamicum AN-10 (biotin, thiamine, arginine, resistant to oi-aminobutyric acid 14-15 g / lL-valine)

selection of mutants resistant to S-2-aminoethylcysteine. Mutants Resistant to S-2-Amino Tylcysteine

 on productivity by

1- .- valine

C. glutamicum CH-1 (biotin, thiamine, arginine, resistant to oi - aminobutyric acid

four

I

And S-2-aminoethylcysteine, 20-22 r / j L-valine).

The strain has the following morpho-cultural signs. Cells are oval, short sticks of sizes 0.6–1.2 microns, uncontrolled, gram-positive, immobile. On the second day of growth, m on septal agar forms colonies with a diameter of 2.0–3.0 mm, round with smooth edges, opaque, shiny, cream colored. The growth of the stroke for 2 days is moderate, the surface is smooth, the color is cream. There is no surface film on the m-peptone broth, it forms a precipitate.

Physiological properties. Aerobe, grows well at 29 - 30 ° C, pH 6.8 - 7.8. Uses nitrates, urea, ammonium salts. Assimilates glucose, sucrose, maltose, xylose, galactose, mannauo, sodium acetate, weakly uses ethyl alcohol, does not ferment lactose.

The auxotrophin strain for arginine, biotin, thiamine, at stochiv o-aminobutyric acid and S-2-aminoethyl-cysteine, on a nutrient medium containing molasses and hydrolyzed BVK, synthesizes 20-22 g / l L-valine.

Example 1. The culture of Corynebacterium glutamicum CH-1 is grown in test tubes with beveled agar (1.2% co-peptone agar) at 28 for 18-24 hours. 5% of seed inoculated with sterile water from agar sprays , are introduced into a 250 ml flask containing 10 ml of fermentation medium of the following composition, wt%: molasses 20; BVK 15; CaCO 2, pH 7.2-7.4. The flasks are placed on a circular rocking chair (rotation speed of 180 rpm) and fermented for 72 hours at 28-30 ° C. The content of L-valine in the culture fluid of 20 g / l.

Example 2. The process is carried out analogously to example 1, only the washing of the culture from beveled agars is transferred to 750 ml flasks containing 30 ml of the inoculum of the following composition, wt%: molasses 4, BBN 4.8, pH 7.2 - 7.4. The flasks are placed on a circular rocking chair with a rotation speed of 180 rpm and are grown for 16-18 hours, 2 a-30 ° C. 5% of the seed thus prepared is seeded in fermentation

312326814

flasks (250 ml) containing 10 ml of raral liquid is 22 g / l of the fermentation medium, which is 8-12% higher than and then the process is carried out as in the method, which is e1 "1M with active Re 1. The content of L-roller in the culture of a known strain.

Claims (2)

Strain Sogupe'as Segtit βΐυίβπάсит СН-1 (collection of the Institute of Industrial Microorganisms of the Institute * 'VNIIgenetika ”, collection number CMPM-B-3139) - producer of C-valine. zi „„ 1232681 > one 1232681 2