SU899646A1 - Process for preparing alkaline protease - Google Patents

Description

() METHOD OF OBTAINING THE NON PROTEASIS

one

The invention relates to the enzyme industry of the microbiological industry, in particular, to a method for producing an alkaline protease.

A method of producing an alkaline protease is known by growing it with {{centered} bacteria from the group Bacillus subfills or Bacillus mesentericus Ha in a nutrient medium containing sources of carbon, nitrogen, mineral salts and stimulators of the biosynthesis of enzymes at a constant temperature, pH, aeration, followed by isolation enzyme from the culture fluid. During the cultivation of producers during the whole cultivation process in the free space of the fermenter, an overpressure is maintained at a level of 0.3-0.5 kgf / cm in order to eliminate the ingestion of extraneous micropores from the external environment l,

The closest to the proposed technical essence and achievable

A positive effect is a method of obtaining a complex of enzyme preparations by growing their producers, bacteria from the Bacilus subfilis group, on a nutrient medium containing a constant carbon source, nitrogen, mineral salts and enzyme biosynthesis stimulants under controlled conditions of temperature, pH, aeration, followed by the release of enzymes from the culture fluid, in which the cultivation process starts at an elevated temperature and then reduced to 37 ° C, aeration is carried out step 15, but The amount of air supplied from 0.35 to 1.0 min., the pH of the medium, in the case of an alkaline protease, is maintained in the range of 6, t6, 3, and the concentration of a kind of carbon source is at a level of 20 degrees. Growing Bacillus subfnis-12 under the specified conditions allows to obtain a proteolytic activity of 12000 units / ml. according to method 3 of the FOLL (pilot industrial regulations for the production of granular alkaline protease, which is in force at the Ladyzhinsky enzyme preparation plant) 21. However, the cultivation of alkaline protease producers by a known method does not provide optimal conditions for mass exchange, oxygen sorption and desorption of carbon dioxide at various stages of growth. The purpose of the invention is to increase the activity of the enzyme. The goal is achieved by the fact that according to the method of obtaining an alkaline protease by growing its producers - bacteria from the Bacillus subfilis group, on a nutrient medium containing a constant carbon source, nitrogen mineral salts and stimulators of enzyme biosynthesis under controlled conditions of temperature, pH, aeration, followed by separation of fermentation from the culture fluid, in the process of cultivation, additionally regulate the pressure in the fermenter by maintaining it during the period of intensive growth of the culture until the beginning of the stationary phase in the range of 2.0-0.8 kgf / cm, in the period of active synthesis of farms, reducing it to 0.50, 2 kgf / cm.

Example 1. In a 63 m fermenter, 31.5 m of sterile nutrient medium is loaded, containing starch, casein, BVK, corn flour, corn extract and disubstituted ammonium phosphate. Bring the medium temperature to 1800 air to aeration, sowing of the culture of Bacillus subfilis-12, and raising the pressure in the fermenter to 1.0 kg / cm. During the first 2 hours, the pressure in the fermenter is maintained at a level of 1.0 kgf / cm and the pressure is reduced to 0.3 kgf / cm after 2 hours, leaving it that way until the end of cultivation. Activity at the end of the growth unit / ml (according to the FOLL method). Without stepwise formula of the invention

A method of producing an alkaline protease by growing its bacteria producers from the Bacillus subfilis group on a nutrient medium containing a constant carbon source, nitrogen, mineral salts and enzyme biosynthesis stimulants under controlled temperature, pH, aeration conditions, followed by release of the enzyme from the culture fluid , characterized in that, in order to increase the activity of the enzyme, the culture process additionally regulates the pressure in the fermenter by maintaining it during the period of intensive growth culture before the start of the stationary phase within the pressure range, the activity of the culture liquid at the end of fermentation is 12000 units / ml. Example 2. A 63 m fermenter was charged with 31.5 m of sterile nutrient medium containing starch, casein, BVK, cornmeal, corn extract and disubstituted ammonium phosphate. The temperature is adjusted to 1800, air is fed to aeration, the culture of Bacillus subfiIis-72 is sown and the pressure is raised in an enzyme of 2.0 kg / cm. During the period of active biomass growth, before the beginning of the stationary phase during the first 2k hours, the pressure in the fermenter is maintained at 2.0 kgf / cm, after 2 hours the pressure is reduced to 0.2 kgf / cm, leaving it that way until the end of the cultivation. Activity at the end of cultivation is 13,500 units / ml (according to the FOLL method). Without a stepwise change in pressure, the activity of the culture at the end of the fermentation liquid is 12000 units / ml. Thus, the implementation of the invention makes it possible to increase the activity of the alkaline protease by 1520. The expected economic effect from the implementation of the invention with the planned release of 750 commodity tons per year will be 1, .32 million rubles.

5889b4b

2.0-0.8 kgf / cm, and during the ak-1 period. Kalun nts KA, Golger L.I. active synthesis of enzymes of reduced production of enzyme preparations up to 0.5-0.2 kgf / cm from fungi and bacteria. M., 1970.

Sources of information, s No. 39511, cl. C 12 D 13/10, 1975 taken into account during the examination (prototype).

2. USSR author's certificate

Claims (2)

Claim A method of producing an alkaline protease by growing its bacteria producers of the Bacillus subfills group on a nutrient medium containing a constant concentration carbon source, nitrogen, mineral salts and enzyme biosynthesis stimulators under controlled conditions of temperature, pH, aeration, followed by isolation of the enzyme from the culture fluid, characterized with the fact that, in order to increase the activity of the enzyme, in the process of cultivation, the pressure in the fermenter is additionally regulated by maintaining it during a period of intensive growth to ultures before the onset of the stationary phase within 2.0-0.8 kgf / cm *, and during the period of active synthesis of enzymes, reducing to 0.5 “0.2 kgf / cm ^g .