SU1081207A1 - Culture medium for culturing bacillus thuringiensis - Google Patents

Abstract

NUTRIENT FOR CULTIVATION Bacillus thuringiensis, containing yeast, source of carbohydrates, magnesium sulphate or manganese sulphate, calcium salt and tap water, is characterized in that, in order to accelerate the sporulation of Bacillus thuringiensis, the ammonium phosphorus additionally contains an phosphorus, and it also contains ammonia phosphorus. the calcium salt is calcium chloride or calcium carbonate, and the source of carbohydrates is hexose-containing wood or peat hydrolyzate in the following ratio of components, wt.%: Hexose-sodium Laughing hydrolyzate of wood or peat,% of Sakha (С py0.7-1.25 (L Yeast BVK0.8-1.0 Ammonium phosphate or ammonium sulfate 0.15-0.25 Calcium carbonate or calcium chloride 0.15-0 , 2 Magnesium sulphate or manganese sulphate0, 1-0.15 Water plumbing Else

Description

1 The invention relates to a microbiological industry and can be used in the production of entomopathogenic bacterial preparations based on Bacillus thuringiensis. The corn-glucose culture medium for the cultivation of Bacillus thuringiensis lj is known. However, the known medium is not very productive. A nutrient medium for the cultivation of Bacillus thurin giensis, containing feed yeast, a source of carbohydrates (cornmeal, starch, bran or others), sodium chloride, calcium chloride, magnesium sulfate or manganese sulfate and tap water Y, is also known. However, the medium is not available sporoobrazovanie in a short time. The aim of the invention is to accelerate the sporulation of Bacillus thuringiensis. This goal is achieved in that the nutrient medium for cultivation of Bacillus thuringiensis, containing yeast, a source of carbohydrates, magnesium sulfate or manganese sulfate, calcium salt and water into a supplementary one, additionally contains ammonium phosphate or ammonium sulfate, as calcium salt — calcium chloride. carbonic acid and, as a source of carbohydrates, a hexose-containing wood or peat hydrolyzate with the following ratio of components, wt. Hexose-containing wood or peat hydrolyzate,% by sugar 0.7-1.2 Yeast BVK0.8-1.0 Ammonium phosphate or ammonium sulfate0, 15-0.2 Calcium carbonate or calcium chloride 0.15-0.2 Magnesium sulfate or manganese sulfate0, 1-0.15 Water plumbing Remaining The medium is prepared as follows: 1% aqueous suspension of yeast BVK is heated to, held at this temperature for 30.60 minutes, then cooled to 50 ° C and 07 the rest of the components are introduced into it. Calcium salt is introduced last. The medium is stirred, the pH before sterilization is set within 7.4-7.6, sterilized at 120-130 C for 30-40 minutes. The pH after sterilization is 6.8-7.2. Example 1. Preparing the medium of the following composition, wt%: Hexose-containing wood or peat hydrolyzate,% sugar 0,7 Yeast BVK 0,8 Ammonium phosphate or sulfuric acid 0,15 Calcium carbonate (chalk) or chloride 0,15 Sulfuric magnesium or manganese 0,1 The water is tap Else The medium after sterilization and cooling is seeded with Bacillus thuringiensis, cultivated for 37 hours at. On a medium with a specified minimum concentration of components, the yield of the target product (concentration of spores and crystals) is 3.2 ppm. in 1 ml of culture fluid with virulence LK5-0 0,003%. Example 2. A medium of the following composition is prepared, wt%: Hexose-containing wood or peat hydrolyzate,% by sugar1, 0 Yeast BVK0.9 Ammonium phosphate or sulfate 0, 2 Potassium phosphate, 0.07 Calcium carbonate (chalk) or chloride 0.18 Magnesium sulphate or manganese 0.13 Water tap Else On a medium with a specified optimum concentration of components, the yield of the target product (concentration of spores and crystals) is 3.7 billion in 1 ml of culture fluid with nKf virulence (7 0.001%. Example 3. Ready t Wednesday of the following composition, yas.%: Hexose-containing wood or peat hydrolyzate,% by sugar1, 25 Yeast BVK1.0 Ammonium phosphate or sulfate0, 25 Potassium phosphate 0.1 Calcium carbonate (chalk) or chloride 0.2 Magnesium sulphate or manganese 0.15 Water tap Water Remaining medium with specified The maximum concentration of components, the yield of the target product (concentration of spore and crystals) is 3.3 billion in 1 ml of culture fluid with a virulence of 0.002%. Example 4, Hexose-containing wood hydrolyzate is prepared from wood chips by treating it with 0.5% sulfuric acid, first at 140-160 ° C, then at 180-190 ° C. After the end of the hydrolysis, the excess sulfuric acid is neutralized lime milk. After filtration, the hydrolysis of at is purified from volatile and non-volatile impurities (formaldehyde, furfural, humic substances, etc.) and then evaporated in a vacuum evaporator at 7Q-8cPc. After evaporation, the hydrolyzate should contain at least 20-25% of reducing substances, which are glucose (80-90%) and a small amount of other sucrose: galactose, mannose, xylose, arabinose. In addition to the sehare, the hydrolyzate contains mineral components: salts of potassium, sodium, magnesium, iron, copper, and others in microscopic amounts. Peat hydrolyzate obtained from a hydrolysis plant contains 16.3% of reducing substances. Data on the use of hydrolysates for growing Bacillus thuringiensis is presented in Table 1,

With woody guid39 roller

With peat

49 hydrolyzate

The data of Table 1 shows that on a medium with peat hydrolyzate, the culture of strain 49 grows and develops normally with the formation of spores and crystalline ticks. The culture fluid has a high insecticidal capacity. Some decrease in productive0, 001 Spores, crystals (1: 1), opted cl. 5% vegetative cells. one%

0,003

2.6

Also

culture on a medium with peat hydrolyzate is associated with less thorough cleaning of the hydrolyzate.

Comparative data on the time of fermentation and sporulation are given in Fable. 2

Optimum47

Minimum37

Optimum37

Maximum37 Table data. 2 shows that the proposed medium allows for a shorter time to reach 100% sporulation of the Bacillus thuringiensia culture.

T a b; i and c a 2

90

100 100 100 The use of a nutrient medium makes it possible to accelerate the sporulation of Bacillus thuringiensiS, to intensify production, improve the quality of biomass, and reduce the cost of production.

Claims (1)

Nutrient for cultivation of Bacillus thuringiensis, containing yeast, a source of carbohydrates, magnesium sulfate or manganese sulfate, calcium salt and tap water, characterized in that, in order to accelerate spore formation of Bacillus thuringiensis, it additionally contains ammonium phosphate, calcium sulfate or - calcium chloride or calcium carbonate, and as a source of carbohydrates - hexose-containing hydrolyzed wood or peat in the following ratio, wt.%: Hexose-containing hydrolyzate of wood or peat,% sugar BVK Yeast Ammonium phosphate or ammonium sulfate Calcium carbonate or calcium chloride Magnesium sulfate or manganese sulfate Tap water 0.7-1.25 0.8-1.0 0.15-0.25 0.15-0.2 (/) 0.1-0.15 Rest 1081207 ’