CS248901B1 - Method of fungicide production - Google Patents

Abstract

BACKGROUND OF THE INVENTION The present invention relates to a method for producing fungicidin by culturing Streptomyces noursei in liquid nutrient medium containing sources of assimilable carbon and nitrogen; and: mineral nutrient salts, under aerobic conditions. Dried as nitrogen sources yeast, or extract thereof, and / or soybean flour and / or corn starch and / or peanut flour and / or wheat meal and / or malt scrap in an amount of 0.2 to 2.0 wt. volume.

Description

The invention relates to a process for the production of fungicidin by culturing a microorganism Streptomyces noursei in a liquid culture medium containing sources of assimilable carbon and nitrogen and a mineral nutrient salt under aerobic conditions.

The present method of producing said antibiotic is used as a nutrient constituent at various stages in the preparation of cultures of Streptomyces noursei, such as millet, amino acids such as tyrosine, nicotinic acid, asparagine, glutamic acid, L-cysteine (corn) extract, calcium carbonate or micronized limestone, glucose, ammonium nitrate and sodium chloride (for the preparation of vegetative inoculum).

For the preparation of the actual production soil, maize extract is usually used as a source of organic nitrogen and growth factors such as nutrient salt, calcium carbonate or micronized limestone, as a source of inorganic nitrogen ammonium salts and as a source of carbon glucose, fructose syrup, or. starch. On nutrient media of this composition, a certain amount of yield is obtained, which, depending on the conditions, is in the range of 25,000 to 30,000 IU / ml of fungicidin.

It has been found that fungicidin yields in fermentation on soils of conventional composition can be increased by 30 to 60% by the measure of the present invention, which consists in using wholly or partially dried yeast as a source of assimilable nitrogen of organic origin and / or yeast and / or soy flour and / or corn meal and / or peanut flour and / or wheat meal and / or malt meal extract, in an amount of 0.2 to 2.0% by weight.

Dried yeast can be advantageously replaced by a mixture of amino acids used in the preparation of spore focula so far, in the soil for the preparation of vegetative inoculum it can be replaced by corn extract and in production non-suspending soil can be replaced by corn extract in whole or in part, optionally with the addition of other substances.

The process according to the invention entails considerable economic benefits in the fermentative production of fungicide, in particular by substantially increasing yields. It is easy to implement without any modification of the equipment and without any special demands on the operator.

Further details of the process according to the invention can be seen from the following examples.

Examples

He did

60 ml fermentation broth containing 0.5% corn extract, 0.5% dried yeast, 2.0% glucose, 8.0% fructose syrup, 0.25% soya flour (or equal weight of peanut flour) and 1 0% micronized limestone was inoculated with 3 ml of a vegetative inoculum of a 28 hour old Streptomyces noursei strain.

The vegetative inoculum was prepared on inoculation liquid broth of the following composition: corn extract 0.25%, soy flour 1%, glucose 4.4%, ammonium nitrate 0.5%, micronized limestone 0.5% and sodium chloride 0.2%. Simple canned vaccines were used, where the amino acids were replaced by 1.0% of dried yeast.

The fermentation was carried out for 120 hours on a reciprocating shaker at a temperature of 28 ° C. At the end of the fermentation, the soil contained 38,000 IU / ml fungicidin. Control fermentation, carried out with a seed inoculum containing 0.01% tyrosine, asparagine, glutamic acid, nicotinic acid and L-cysteine, yielded 26,000 IU / ml of fungicidin.

Example 2

60 ml fermentation broth containing 0.5% corn extract, 0.001% acid potassium phosphate, 0.5% dried yeast, 0.25% soy flour, 2.0% glucose, 8.0% fructose syrup, 0.5 % ammonium nitrate and 1.0% micronized limestone were inoculated with 3 ml of a 28 hour old vegetative inoculum of Streptomyces noursei strain prepared on soil in which the corn extract was completely replaced by dried yeast.

Inoculation soil composition: 1.0% dried yeast, 0.5% soy flour, 4.4% glucose,

0.5% ammonium nitrate, 0.5% micronised limestone, 0.2% sodium chloride. Fermented on a reciprocating shaker at 28 ° C. At the end of the fermentation, the soil contained 44,000 IU / ml of fungicidin. ·

Control fermentation with soil containing 1.18% corn extract, 2.0% glucose, 8.0% fructose syrup, 0.5% ammonium nitrate and 1.0% micronised limestone, inoculated with a vegetative inoculum of Streptomyces noursei, 28 hours old prepared on inoculum broth containing 0.25% corn extract, 1.0% soy flour, 4.4% glucose, 0.5% micronised limestone and 0.2% sodium chloride yielded 25,000 IU / ml fungicidin .

Example 3

60 ml fermentation broth containing 1.0% dried yeast, 0.001% acidic potassium phosphate, 0.5% soya flour, 2.0% glucose, 8.0% fructose syrup, 1.0% micronised limestone and 0.5 % ammonium nitrate was inoculated with 3 ml of a 28 hour old Streptomyces noursei vegetative inoculum prepared for inoculated soils with dried yeast. Fermented for 120 hours on a reciprocating shaker at 28 ° C. At the end of the fermentation, the soil contained 50,000 IU / ml fungicidin.

Control fermentation carried out on a maize extract broth, inoculated with a 28 hour inoculum, yielded 28,000 IU / ml fungicidin.

Example 4

60 ml fermentation broth containing 1,0% dried yeast, 0,001% acidic potassium phosphate, 0,5% maize meal, 2,0% glucose, 8,0% fructose syrup, 1,0% micronised limestone and 0% 5% ammonium nitrate was inoculated with 3 ml of a 28 hour old vegetative inoculum of Streptomyces noursei strain. Fermented for 120 hours on a reciprocating shaker at 28 ° C. After fermentation, the soil contained 56,000 IU / ml of fungicidin.

The congressional fermentation carried out on the maize extract broth, inoculated with a 28 hour inoculum, yielded a yield of 31,000 IU / ml of fungicidin.

Example 5 ml of fermentation broth containing 1.0% dried yeast, 0.5% wheat meal,

2.0% glucose, 8.0% fructose syrup, 1.0% micronised limestone, and 0.5% ammonium nitrate were fermented under the same conditions as in Example 4 to produce 50 ml of a vegetative inoculum of S. noursei strain. 000 IU / ml fungicidin. Control fermentation yielded 28,000 IU / ml fungicidin.

Example 6 ml of fermentation broth containing 1.0% dried yeast, 0.001% acidic potassium phosphate, 0.25% malt meal, 6.0% soluble starch, 1.0% micronized limestone and 0.5% ammonium nitrate were inoculation of 3 ml of a vegetative inoculum of the S.nourse strain was fermented under the same conditions as in Example 4, producing 38,000 IU / ml of fungicidin. Control fermentation yielded 26,000 IU / ml of fungicidin.

Example 7 ml of fermentation broth containing 0.5% yeast extract, 0.5% soy flour,

2.0% glucose, 0.001% potassium potassium phosphate, 6.0% fructose syrup, 0.5% calcium nitrate, and 1.0% micronized limestone were fermented after inoculation with a 3 ml vegetative inoculum of the S.noursei strain under the same conditions as in the of Example 4 with a production of 44,000 IU / ml of fungicidin. Control fermentation yielded 30,000 IU / ml fungicidin.

Example ml of fermentation broth containing 1.0% dried yeast, 0.5% soy flour, 0.5% ammonium nitrate, 7.0% amylase liquefied starch, 1.0% micronized limestone and 0.001% potassium phosphate acid were inoculated 3 ml of vegetative inoculum of Streptomyces noursei strain fermented under the same conditions as in Example 4, producing 48,000 IU / ml fungicidin. Control fermentation produced 30,000 IU / ml fungicidin.

Claims (1)

A method for producing fungicidin by culturing Streptomyces noursei in a liquid culture medium containing assimilable carbon and nitrogen sources and mineral nutrient salts, under aerobic conditions, characterized in that fully or partially dried yeast and / or extract of organic origin is used as the assimilable nitrogen source of organic origin. % yeast and / or soya flour and / or corn meal and / or peanut flour and / or wheat meal and / or malt meal, in an amount of 0.2 to 2.0 wt. vol.