RU2484129C1 - Method to produce biomass of aerobic microorganisms - Google Patents

Abstract

FIELD: biotechnologies.

SUBSTANCE: method includes inspection of an inoculator with process equipment for tightness, sterilisation of the inoculator with steam via an aeration device at the pressure of 0.20 - 0.25 MPa for 30…40 min., its filling with nutrient medium heated by steam to the temperature of 100°C. Then the temperature of the nutrient medium is increased to 121 - 123°C at steam pressure of 0.10 - 0.15 MPa, and the nutrient medium is maintained at these parameters for 15-60 min., afterwards the pressure in the inoculator is reduced down to 0.03…0.05 MPa. The nutrient medium is cooled down to cultivation temperature of 31…32°C with cold water with temperature of 7 - 10°C. After cooling of the nutrient medium, it is seeded with a seeding material with simultaneous mixing and aeration with sterile air. Cultivation of the produced liquid seeding culture is carried out at pH 4.2 - 4.5 and temperature of 31 - 32°C to achieve the phase of exponential growth for 12 - 14 hours. Then it is sent by means of displacement with sterile air from the inoculator into the prepared fermenter in the amount of 3…10% of the nutrient medium amount with its filling by 7/10 of its volume, and the microorganism culture is grown at fermentation temperature of 28 - 40°C for 96 - 120 hours with continuous aeration with sterile air, mechanical mixing and supply of warm water with temperature of 27 - 47°C into a heating jacket of the fermenter. After fermentation the cultural fluid with accumulated biomass is supplied into previously sterilised collectors of finished culture.

EFFECT: increased yield of cultural liquid with accumulated biomass of aerobic microorganisms, reduced specific power inputs and provision of environmental safety at all stages of production.

1 dwg, 2 ex

Description

The invention relates to the microbiological industry and can be used in the process of aerobic deep fermentation in the cultivation of cultures of microorganisms and producers of enzymes.

Known methods for the production of biomass of aerobic microorganisms [Fermentation apparatus for the processes of microbiological synthesis / A.Yu. Vinarov, L.S. Gordeev, A.A. Kuharenko, V.I. Panfilov // M .: DeLi print, 2005. - 191 p. .], including the preparation of a liquid nutrient medium, seed and the cultivation of microorganisms.

The disadvantage is that the known methods do not provide for the preparation of energy and their rational use in the cultivation of cultures of microorganisms and cannot be effectively implemented in conditions of decentralized heat supply systems, when thermal energy is generated directly at the production facility, which is typical for mini-production and low-power enterprises . At the same time, the possibility of using the heat of a low-temperature potential, in particular the waste heat of gas turbine plants and boiler units, is excluded, which does not allow us to fully solve the problems of energy conservation.

The closest in technical essence and the achieved effect to the proposed is a method for the production of biomass of aerobic microorganisms [Pat. RF №2322488 C2, C12N 1/00, C12M 1/00, Publ. 04/20/2008. Bull. No. 11, part III], which provides for the saturation of the culture fluid with an aerating agent, which is used as air, with exhaust air and the culture fluid with accumulated biomass.

The disadvantage is that in this method there is no system for the preparation of energy carriers, in particular “warm” and “cold” water in a closed thermodynamic cycle using a steam ejector chiller operating in the heat pump mode to stabilize the temperature conditions when preparing a liquid seed crop in an inoculator , when directly growing a culture of microorganisms in the fermenter and cooling the finished culture in receiving collections, which does not allow us to consider the known method as an ene energetically efficient, environmentally safe and provides a high yield of the culture broth with biomass accumulation.

An object of the invention is to increase the yield of culture fluid with the accumulated biomass of aerobic microorganisms, reduce specific energy consumption and ensure environmental safety at all stages of production.

To solve the technical problem of the invention, a method for producing biomass of aerobic microorganisms is proposed, characterized in that they first check for leak-proofness of the inoculator with a cooling jacket, with aeration and mixing devices, with exhaust lines for exhausted sterile air and for transferring liquid seed to the fermenter for aerobic deep fermentation, and then the inoculator is sterilized with steam through an aeration device under a pressure of 0.20 ... 0.25 MPa for 30 ... 40 minutes and is filled with nutrient Doy, which is heated with steam to a temperature of 100 ° C, bring the temperature of the nutrient medium to 121 ... 123 ° C at a steam pressure of 0.10 ... 0.15 MPa and maintain it at these parameters for 15 ... 60 minutes, then reduce the pressure in inoculator up to 0.03 ... 0.05 MPa, and in its cooling jacket serves “cold” water with a temperature of 7 ... 10 ° C and cool the nutrient medium to a cultivation temperature of 31 ... 32 ° C, after cooling the nutrient medium, the supply of sterile air to inoculator, close the exhaust line of exhaust sterile air, produce Acebo growth medium inoculum and ensure its uniformity by agitating the device with aeration with sterile air; the obtained liquid seed culture is cultivated at a pH of 4.2 ... 4.5 and a temperature of 31 ... 32 ° C until the exponential growth phase is reached within 12 ... 14 hours and then it is sent by squeezing sterile air from the inoculator into a pre-tested and sterilized fermenter with a heating jacket, with aeration and mixing devices in an amount of 3 ... 10% of the amount of the nutrient medium, and fill it with 7/10 of its volume and carry out the cultivation of a microorganism culture at a fermentation temperature of 28 ... 40 ° C within 96 ... 120 hours with continuous aeration with sterile air, mechanical stirring and the supply of "warm" water with a temperature of 27 ... 47 ° C in the heating jacket of the fermenter; after fermentation, the culture fluid with the accumulated biomass is fed into pre-sterilized collections of the finished culture, in which its temperature is maintained at 8 ... 10 ° C by means of a cooling system; for the preparation of “cold” and “warm” water, a steam ejector chiller is used, operating in the heat pump mode and consisting of an ejector, an evaporator, a cold receiver, a condenser, a thermostatic valve, a waste water collector, a steam generator with electric heating elements and a safety valve, and a water supply pump a steam generator, a refrigerant recirculation pump through a cold receiver, operating in a closed thermodynamic cycle; at the same time, working steam is obtained in the steam generator and, at a pressure of 0.05 ... 0.06 MPa, is supplied to the ejector nozzle, involving the ejected refrigerant vapor, which is used as water, from the evaporator and create a reduced pressure of 0.0009 ... 0.001 MPa in it with temperature boiling refrigerant 4 ... 7 ° C; due to the recirculation of the refrigerant through the cold receiver, “cold” water with a temperature of 7 ... 10 ° C is obtained by recuperative heat exchange between the refrigerant and the water and serves it in the cooling jacket of the inoculator and the cooling system of the collections of the finished culture; the heat of condensation formed after the ejector of a mixture of refrigerant vapor and working steam in the condenser is used to produce “warm” water, which is heated to a temperature of 27 ... 47 ° C by means of regenerative heat transfer and fed to the heating jacket of the fermenter; part of the water condensate formed after the condenser is sent through a thermostatic valve to the evaporator to replenish water losses in it, and the excess part is taken out of the closed cycle of the steam ejector refrigeration machine and, together with the waste water after the inoculator, fermenter and ready-made culture collectors, is fed into the waste water collector, from which one part of the water is sent to replenish the loss of water in the steam generator, and the other part is fed through two streams to the refrigeration receiver and the steam ejector refrigeration condenser The machines to form a closed loop.

The technical result of the invention is to increase the yield of culture fluid with the accumulated biomass of aerobic microorganisms, increase energy efficiency and environmental safety of production.

Figure 1 presents a diagram that implements the proposed method.

The diagram shows a fermenter 1 with a heating jacket 2, mixing devices 3 and aeration 4; inoculator 5 with a cooling jacket 6, mixing devices 7 and aeration 8; collections of finished culture 9 with a cooling system 10; a collection of waste water and condensate 11; a steam generator 12 with electric heating elements 13 and a safety valve 14; ejector 15; capacitor 16; thermostatic valve 17; evaporator 18; cold receiver 19; pumps 20, 21, 22, 23, 24; material flow lines: 0.1 - sterile air; 0.2 - exhaust exhaust sterile air; 1.0 - cold water; 1.1 - waste water; 1.2 - refrigerant recirculation through a cold receiver; 1.3 - water in the cold receiver; 1.4 - water to the condenser; 7.5 - warm water in the fermenter; 2.0 - working steam; 2.7 - a pair of inoculator and fermenter; 2.2 - ejected refrigerant vapor; 2.3 - a mixture of working and ejected vapor; 2.4 - condensate; 2.5 - pressure relief; 3.1 - nutrient medium; 3.2 - seed; 3.3 - squeezing liquid sowing culture; 3.4 - culture fluid with accumulated biomass of aerobic microorganisms.

The method is as follows.

The technological cycle for the production of biomass of aerobic microorganisms begins with the preparation of a liquid seed culture in the inoculator 5 with a cooling jacket 6, mixing devices 7 (two- or three-tier mixer) and aeration 8 (nozzles or bubblers).

After checking for leaks with sterile air supplied through line 0.1 at a pressure of 0.07 MPa and soap foam applied to the joints (cover, flanges, seals, valves, etc.), the inoculator is checked for leaks with steam supplied through the line 2.1 through an aeration device 8 with a closed exhaust line exhaust sterile air 0.2. If air or steam leaks are detected, as well as a decrease in pressure in the inoculator, the detected defects are removed and the leak test is performed again.

After checking for leaks, the inoculator 5 is sterilized with steam supplied through line 2.1 from the steam generator 12 through an aeration device 8 under a pressure of 0.20 ... 0.25 MPa for 30 ... 40 minutes, and then filled with nutrient medium along line 3.1, which is heated with steam to temperature 100 ° С with an open exhaust line 0.2. After that, close the exhaust line of exhaust sterile air 0.2 and bring the temperature of the nutrient medium to 121 ... 123 ° C at a steam pressure of 0.10 ... 0.15 MPa and maintain it for 15 ... 60 minutes, then reduce the pressure in the inoculator to 0, 03 ... 0.05 MPa, and in his cooling jacket 6 through the line 1.0 serves "cold" water and cool the nutrient medium to a cultivation temperature of 28 ... 40 ° C.

After cooling the nutrient medium, it is inoculated with seed through a seed hatch (not shown in the diagram) along line 3.2. Before inoculation, the sterile air supply to the inoculator is stopped along line 0.1 and at the same time the exhaust sterile air exhaust line 0.2 is closed. After seeding, the supply of sterile air is resumed, the exhaust line of exhaust sterile air 0.2 is opened, and a culture of microorganisms is grown.

The resulting liquid seed culture in the inoculator 5 is cultivated at a pH of 4.2 ... 4.5 and a temperature of 31 ... 32 ° C until the exponential growth phase is reached within 12 ... 14 hours, ensuring its uniformity by means of a mixing device 7.

After the cultivation time has passed, the liquid seed culture is crushed with sterile air through the crushing line 3.3 from the inoculator 5 to a preliminary tested for tightness and sterilized fermenter 1 in the amount of 3 ... 10% of the amount of nutrient medium, which is filled with 7/10 of its volume.

The check for leaks and sterilization of the fermenter is carried out in the same way as the inoculator.

In the fermenter 1, a microorganism culture is grown with a fermentation temperature of 28 ... 40 ° C for 96 ... 120 hours with continuous aeration with sterile air through an aeration device 3, mechanical stirring with a stirring device 4 and the supply of “warm” water through line 1.3 to the heating jacket 2.

The culture fluid with the accumulated biomass of aerobic microorganisms from the fermenter using the pump 22 is fed through line 3.4 to pre-sterilized collections of the finished culture 9 with a cooling system 10, where the culture temperature is maintained at 8 ... 10 ° C.

For the preparation of "cold" and "warm" water using a steam ejector refrigeration machine operating in the heat pump mode, consisting of an ejector 15; evaporator 18; cold receiver 19; capacitor 16; thermostatic valve 17; a waste water collector 11, a steam generator 12 with heat elements 13 and a safety valve 14; a pump for supplying water to the steam generator 23; refrigerant recirculation pump through a cold receiver 20 operating in a closed thermodynamic cycle.

At the same time, in the steam generator 12, by means of electric heating elements 13, working steam is obtained and, under pressure 0.05 ... 0.06 MPa, is fed through line 2.0 to the ejector nozzle 15, involving ejected refrigerant vapors along line 2.2, using water from the evaporator 18, and create in it a reduced pressure of 0.0009 ... 0.001 MPa with a refrigerant boiling point of 4 ... 7 ° C. Due to the recirculation of the refrigerant along line 1.2, cold water with a temperature of 7 ... 10 ° C is obtained through the cold receiver 19 through recuperative heat transfer between the refrigerant and the water supplied through line 1.3 to the cold receiver 19 from the collector 11 using pump 24.

The resulting "cold" water from the cold receiver 19 through line 1.0 is fed into the cooling jacket 6 of the inoculator 5 and the cooling system 10 of the collections of the finished culture 9.

The mixture of refrigerant vapor and working steam formed after the ejector 15 is sent to the condenser 16 through line 2.3. The condensation process is accompanied by the release of heat, while the condensation heat in the condenser 16 is used to produce “warm” water through the regenerative heat exchange between the water supplied from the collector 11 by the pump 24 through line 1.4 to the condenser 16, and the condensing vapors of the mixture in the condenser 16. The water heated to a temperature of 27 ... 47 ° C is fed through line 7.5 to the heating jacket 2 of the fermenter 1.

A part of the water condensate formed after the condenser 16 is sent through a thermostatic valve 17 via line 2.4 to the evaporator 18 to replenish the water loss in it, and the excess part of the condensate is removed from the closed cycle of the steam ejector refrigeration machine and together with the waste water after inoculator 5, fermenter 1 and ready-made collectors cultures 9 are fed through lines 1.1 to the waste water collector 11, from which one part of the water is sent via line 7.2 to replenish the loss of water in the steam generator 12, and the other part along two streams 1.3 and 1.4 with the pump 24 is fed into the cold receiver 19 and the condenser 16 of the steam ejector refrigeration machine with the formation of a closed cycle. With increasing steam pressure in the steam generator, a safety valve is activated, which relieves pressure.

Examples of the method.

A method for the production of biomass of aerobic microorganisms is carried out in the following sequence. First, prepare a liquid seed culture in the inoculator in accordance with the production cycle (in h):

washing and inspection 1,0 leak check 0.5 pressure check and sterilization 2,5 nutrient loading 0.5 medium sterilization 1,5 cooling and inoculation of the nutrient medium 2.0 growing liquid crops 12 ... 14 fermenter inoculum transfer 0.5

The process of growing microorganism cultures in an aerobic deep fermentation fermenter is carried out with aeration with sterile air using a bubbler and stirring using a mechanical stirrer, which consisted of a metered supply of flows of nutrient medium, inoculum (seed), sterile air, “warm” water into the heating jacket for ensuring high intensity mass and energy exchange of microbial cells of the inoculum with the nutrient medium due to stabilization of process parameters at the level required for optimality of the producer and the target product. The exhaust air, waste water and culture fluid with accumulated biomass are removed from the fermenter.

The production cycle of the fermenter was (in h):

washing and inspection 1,0 leak check 0.5 pressure check and sterilization 1,5 medium filling 4.0 fermenter cultivation 96 ... 120 culture fluid transfer in consumables 0.5

The fermentation process is carried out in a vertical fermenter company "Sartorius Stedim Biotech" series BIOSTATc with a working volume of 100 l, intended for growing microorganisms or cell cultures. Control over the process parameters is provided by the DCU (Digital Control Unit) microprocessor control system. To stabilize the temperature regimes when preparing a liquid seed culture in an inoculator, directly growing a culture of microorganisms in a fermenter and cooling the finished culture in receiving tanks, the preparation of “warm” and “cold” water is carried out using a steam ejector chiller operating in the heat pump mode with a technical characteristic :

cooling capacity, kW twenty boiling temperature: in the evaporator, ° С four in a steam generator, ° С 154 condensation temperature, ° С 127 water temperature at the inlet to the condenser, ° С fifteen ejection coefficient four heat exchange surface area cold receiver, m ² 8 heat transfer coefficient of the cold receiver, W / m ² · ° С 92 condenser heat exchange surface area, m ² 6 heat transfer coefficient of the condenser, W / m ² · ° C 49 refrigerant water

The design of the steam ejector chiller does not contain moving wear elements, which ensures trouble-free operation of the machine in long cycles without direct maintenance, while minimizing the volume of current repairs, the cost and need for spare parts and auxiliary materials.

Example No. 1

The enzyme preparation of inulinase obtained by the in-depth method using the micromycete producer Aspergillus awamori 2250 was used as an object of production.

An inoculator with a cooling jacket, with aeration and mixing devices, with exhaust lines for exhausted sterile air and squeezing liquid seed into an aerobic deep cultivation fermenter is checked for leaks, then the inoculator is steam sterilized under a pressure of 0.25 MPa for 40 minutes. Fill with a nutrient medium, which is heated with steam to a temperature of 100 ° C, bring the temperature of the nutrient medium to 123 ° C at a vapor pressure of 0.15 MPa and maintain it at these parameters for 40 minutes, then reduce the pressure in the inoculator to 0.05 MPa and cool the nutrient medium to a cultivation temperature of 31 ± 0.5 ° C and sow the nutrient medium with seed. A liquid seed culture is cultivated at pH 4.2 and a temperature of 31 ± 0.5 ° C until the exponential growth phase is reached within 12 hours. Then it is sent from the inoculator to a pre-tested for tightness and sterilized fermenter in the amount of 4% of the amount of nutrient medium, and it is filled at 7/10 of its volume and the microorganism culture is grown at a fermentation temperature of 31 ± 0.5 ° C for 96 hours with continuous aeration with sterile air, mechanical stirring. After fermentation, the culture fluid with accumulated biomass is fed into pre-sterilized collections of the finished culture, in which its temperature is maintained at 8 ° C.

The maximum yield of the target product in activity was achieved in the following cultivation mode:

the composition of the nutrient medium,%:

molasses concentration 5,0 concentration (NH ₄ ) ₂ HPO ₄ 1,0 the concentration of MGSO ₄ · 7H ₂ O 0.05 KH ₂ PO ₄ concentration 0.1 sterile air pressure in the fermenter, MPa 0,03 agitator rotation frequency, s ^-1 3,5 pH of the liquid phase 4.2 cultivation temperature, ° C 31 ± 0.5 solids content in the culture fluid,%. 7.0 ± 0.5 inulinase activity, units / cm ³ 25 ± 3 β-fructofuranosidase activity, units / cm ³ 100 ± 5 specific activity, u / g protein 1200 fermentation time, h 96

Example No. 2

The method was carried out analogously to example 1, but the aerobic deep cultivation was carried out by the micromycete Trichoderma harzianum F114 producer of the enzyme β-mannanase, which hydrolyzes mannans of vegetable carbohydrate-containing raw materials to mannose.

An inoculator with a cooling jacket, with aeration and mixing devices, with exhaust lines for exhausted sterile air and squeezing the liquid seed into an aerobic deep cultivation fermenter is checked for leaks, then the inoculator is steam sterilized under a pressure of 0.20 MPa for 30 minutes. Fill with a nutrient medium, which is heated with steam to a temperature of 100 ° C, bring the temperature of the nutrient medium to 121 ° C at a steam pressure of 0.10 MPa and maintain it at these parameters for 40 minutes, then reduce the pressure in the inoculator to 0.03 MPa and cool the nutrient medium to a cultivation temperature of 35 ± 0.5 ° C and sow the nutrient medium with seed. A liquid seed culture is cultivated at pH 4.5 and a temperature of 35 ± 0.5 ° C until the exponential growth phase is reached for 14 hours. Then it is sent from the inoculator to a preliminary tested for tightness and sterilized fermenter in the amount of 2% of the amount of nutrient medium, and it is filled at 7/10 of its volume and the microorganism culture is grown at a fermentation temperature of 35 ± 0.5 ° C for 72 hours with continuous aeration with sterile air, mechanical stirring. After fermentation, the culture fluid with accumulated biomass is fed into pre-sterilized collections of the finished culture, in which its temperature is maintained at 10 ° C.

The maximum yield of the target product in activity was achieved in the following cultivation mode:

the composition of the nutrient medium,%:

glucose 40,0 MgSO ₄ 0.5 NaNO ₃ 4.0 KH ₂ PO ₄ 1,0 Kcl 0.05 FeSO ₄ 0.1 sterile air pressure when feeding to the fermenter, MPa 0.04 agitator rotation frequency, s ^-1 3.6 pH of the liquid phase 4,5 cultivation temperature, ° C 35 ± 0.5 solids content in culture fluid,% 7.0 ± 0.5 β-mannanase activity, units / cm ³ 2400 ± 5 specific activity, u / g protein 11800 fermentation time, h 72

The rational use of heat and electric energy in a cold and heat supply system using a steam ejector chiller operating in the heat pump mode was considered in terms of increasing the yield of culture fluid with accumulated biomass of aerobic microorganisms and reducing the cost of the resulting target product.

Stabilization of the temperature regimes of the processes of preparing liquid seed crops and deep aerobic fermentation by reducing the temperature dispersion of energy carriers, which were used as “warm” and “cold” water in a closed thermodynamic cycle using a steam ejector refrigeration machine, allows to increase the yield of culture fluid with accumulated biomass of aerobic microorganisms .

The main solution to reduce energy consumption in the proposed method for the production of biomass of aerobic microorganisms is the optimal choice of temperature differences in the evaporator and condenser of the steam ejector refrigeration machine when receiving “cold” and “warm” water. Deviation from these values will inevitably lead to an increase in energy consumption: a decrease in the boiling point of the refrigerant in the evaporator by 1 ° C will lead to the need to increase the consumption of working steam in the ejector, and consequently, to an energy expenditure of 5 ... 7%, and an increase in the condensation temperature by 1 ° C will lead to an increase in energy consumption by 2.0 ... 2.5% [Thermal and structural calculations of refrigeration machines / E.M. Bambushek, N.N. Bukharin, E.D. Gerasimov and others; Under the total. ed. I.A. Sakuna. - L .: Mechanical engineering. Leningrad branch, 1987. - 423 p.].

The proposed method for the production of biomass of aerobic microorganisms using a steam ejector chiller extends the boundaries of energy-efficient pairing of objects of different temperature potentials based on the utilization and recovery of secondary energy resources. At the same time, a universal approach has been fully implemented in creating a competitive technology that provides heat and cold production for co-occurring processes for preparing a liquid seed culture in an inoculator, directly growing a microorganism culture in a fermenter, and cooling the finished culture in receiving collections.

Thus, the proposed method for the production of biomass of aerobic microorganisms allows:

- increase the yield of culture fluid with accumulated biomass of aerobic microorganisms;

- ensure environmental safety at all stages of the production of culture fluid with the accumulated biomass of aerobic microorganisms;

- allows to reduce specific energy consumption by 5 ... 7% by rationally incorporating the inoculator, fermenter and collections of the finished culture into the heat production scheme using a steam ejector refrigeration machine operating in the heat pump mode;

- to provide increased energy efficiency of fermentation processes through the use of heat of condensation of the refrigerant in the condenser of the chiller when water is heated, followed by its supply to the heating jacket of the fermenter and the potential of the refrigerant in the cold receiver during cooling of the water, followed by the supply of the inoculator to the cooling jacket and the cooling system of the collection tanks of the finished culture ;

- create real conditions for the disposal of low pressure steam;

- water vapor with a pressure of 0.05 ... 0.06 MPa is used as an energy carrier, due to which energy savings are achieved, which is spent only on the operation of control elements, refrigerant and water pumps, heat-generating elements of the steam generator.

Claims (1)

A method for the production of biomass of aerobic microorganisms, characterized in that they first carry out a leak test of the inoculator with a cooling jacket, with aeration and mixing devices, with exhaust lines for exhausting sterile air and transferring liquid seed to the fermenter for aerobic in-depth fermentation, and then the inoculator is steam sterilized through aeration device under pressure of 0.20 - 0.25 MPa for 30-40 minutes and filled with a nutrient medium, which is heated with steam to a temperature of 100 ° C, dov the temperature of the nutrient medium is up to 121-123 ° C at a vapor pressure of 0.10-0.15 MPa and is maintained at these parameters for 15-60 minutes, after which the pressure in the inoculator is reduced to 0.03-0.05 MPa, and cold water with a temperature of 7-10 ° С is supplied to its cooling jacket and the nutrient medium is cooled to a cultivation temperature of 31-32 ° С, after cooling the nutrient medium, the sterile air supply to the inoculator is stopped, the exhaust sterile air exhaust line is closed, the nutrient medium is inoculated seed and provide it about heterogeneity by means of a device for mixing with aeration with sterile air, the obtained liquid seed culture is cultivated at a pH of 4.2-4.5 and a temperature of 31-32 ° C until the exponential growth phase is reached within 12-14 hours and then it is sent by squeezing with sterile air from inoculator in a pre-tested for tightness and sterilized fermenter with a heating jacket, with aeration and mixing devices in the amount of 3-10% of the amount of nutrient medium, and fill it with 7/10 of its volume and carried out The culture of a microorganism is grown at a fermentation temperature of 28–40 ° C for 96–120 h with continuous aeration with sterile air, mechanical stirring and the supply of warm water with a temperature of 27–47 ° C to the heating jacket of the fermenter; after fermentation, the culture fluid with accumulated biomass is fed in pre-sterilized collections of the finished culture, in which its temperature is maintained at a level of 8-10 ° C by means of a cooling system, and a steam ejection chiller is used to prepare cold and warm water a machine operating in the heat pump mode and consisting of an ejector, an evaporator, a cold receiver, a condenser, a thermostatic valve, a waste water collector, a steam generator with electric heating elements and a safety valve, a water supply pump to the steam generator, a refrigerant recirculation pump through a cold receiver operating in a closed thermodynamic the cycle, while the working steam obtained in the steam generator under pressure of 0.05 ... 0.06 MPa is fed into the ejector nozzle with the involvement of the ejected refrigerant vapor, which is used as water from the evaporator, and a reduced pressure of 0.0009-0.001 MPa is created in it with a refrigerant boiling point of 4-7 ° C, due to the recirculation of the refrigerant through a cold receiver, cold water with a temperature of 7-10 ° C is obtained by regenerative heat exchange between refrigerant and water and feed it into the cooling jacket of the inoculator and the cooling system of the collections of the finished culture, and the heat of condensation formed after the ejector of the mixture of refrigerant vapor and working steam in the condenser is used to produce warm water, which is heated to a temperature of 27-47 ° C by means of recuperative heat transfer and fed to the heating jacket of the fermenter, part of the water condensate formed after the condenser is sent through a thermostatic valve to the evaporator to replenish the water loss in it, and the excess part is removed from the closed cycle of the steam ejector chiller and together with wastewater after the inoculator, fermenter and ready-made culture collectors are fed to the wastewater collector, from which one part of the water is sent for replenishment and water in the steam generator, and its other part in two streams is fed into the refrigeration receiver and condenser of the steam ejector refrigeration machine with the formation of a closed cycle.