RU2142995C1 - Smaller fermentation plant (versions) - Google Patents

Abstract

FIELD: fermentation plants used in scientific research institutes, microbiological, medical, biochemical, pharmaceutical and food-processing field. SUBSTANCE: plant is made f form of compact including modules of automated monitoring systems and box. Arranged in box are detachable fermenter with flow-type loop and sampler provided with connector sterilized by electric heating. Sterilizing of fermenter and sampler is effected in autoclave. Fermenter is provided with inlet and outlet gas sterilizing filtration system secured inside it with surface and depth aeration selector, deaeration centrifugal pump driven by shaft of mechanical foam suppressor and detachable flow-type loop. Loop includes nephelometer dish, cell for reserve pH and EH electrodes and connectors for aseptic connection/disconnection of loop and sampler which are sterilized by electric heating. EFFECT: extended functional capabilities, reduced overall dimensions, ease in servicing possibility of maintenance of conditions aggressive for any microorganisms and aseptic conditions in box and in room. 4 cl, 9 dwg

Description

 The invention relates to fermentation machines, namely to small (laboratory production and research) fermentation units (MFPs), designed to study fermentation processes and obtain various products of microbiological synthesis with desired properties, and can be used in research institutes and in enterprises of microbiological, medical , biochemical, pharmaceutical and food profiles.

Known MFPs / 1-18 /, consisting of one or more stands (modules, blocks, places), which include a basic fermenter unit (mainly a fermenter with a capacity of up to 25 dm ³ with a mechanical mixing device), contain equipment and automated control and regulation systems (ASKR) hydrodynamic, heat and mass transfer conditions in the fermenter and the dynamics of the fermentation process using parameters such as stirrer speed (n _m ), working fluid level (H), working foam level (H _p ), pressure (p), temperature (t), heat dissipation rate (q ^T ), aeration air flow rate (Q _in ), hydrogen index (pH), redox potential (eH), partial pressure of oxygen and carbon dioxide in the culture fluid (QO) (pO ₂ and pCO ₂ ), the optical concentration of cells in the QOL (OK), the concentration of O ₂ and CO ₂ in the gases leaving the fermenter (Co ₂ and Cco ₂ ), as well as derivatives of these parameters and biological parameters determined by calculation and laboratory analysis of QL samples .

 Unlike pilot and industrial fermentation plants, multifunctional devices, which are mainly used for research purposes, contain or should contain the maximum possible amount of ASKR, and their small fermenters contain appropriate sensors and units for introducing additives, for sampling and draining of QOL. At the same time, the technical level of a particular MFP is determined, in addition to general quality indicators, including reliability, the number of ASKR, ease of maintenance and ensuring aseptic conditions under which contamination of the product (QL) by extraneous microflora and leakage (emissions) are simultaneously eliminated to a certain extent. product or producer from the fermenter to the environment (into the workroom and beyond). Moreover, the provision of aseptic conditions is a key requirement for fermentation plants, since the quality of research and products, the safety of work and a clean environment depend on these conditions in the aggregate.

To the greatest extent, contamination and emissions, as is known / 19-27 /, are characteristic of fermentation plants, including MFPs, with the following symptoms (drawbacks of known MFPs):
a) the presence of many detachable connections and locking nodes on the technological equipment;
b) the use of rubber (silicone) tubes as process pipelines;
c) the use of compounds sterilized above the flame of the burner;
d) the use of movable pressurized joints (gland, contact or mechanical seals of the shafts of the drives of the mixer, mechanical defoamer, etc.);
d) the use of compressed air for aeration of QOL and the maintenance of excess pressure in the fermenter;
e) lack of reliability of the sterilization system of the inlet and outlet gases during aeration or the lack thereof in the installation;
g) clutter of the superfluid space in the fermenter with sensors, pipes that prevent mixing, free rise and effective quenching of the foam and contribute to the hanging of the foam on them;
h) the location of commonly used detachable joints and valves below the QOL level in the fermenter and especially on its bottom;
i) the inconvenience and complexity of the quality preparation of the fermenter to work (washing, checking for leaks, sterilization);
j) inconvenience, complexity, and duration of aseptic connection operations — disconnection of process pipelines and equipment;
k) the use of steam and compressed air supply systems (sources of increased emergency danger) connected to the fermenter;
m) the lack of ensuring normal (comfortable) working conditions for maintenance personnel in terms of temperature, humidity, sound pressure, as well as simplicity and ease of maintenance of the installation.

 The listed disadvantages to varying degrees have all known analogues, and their constructive diversity is, in part, a consequence of the use of various methods and means to eliminate these shortcomings when creating an MFP. Moreover, the uncompromising exclusion of any one drawback causes the aggravation of others and the emergence of new ones. For example, in order to exclude many detachable connections, including rubber-tubular ones (see paragraphs a, b, c, k), installations are created with a stationary small fermenter, which is connected to support systems (filters, dispensers, etc.) mainly by welded pipelines, for which it is washed, checked for leaks and sterilized in place (at the installation stand). Compared with a multifunctional device with a removable fermenter, such an installation is more complicated and inconvenient in preparation for work, and the spent next cultivation cycle of the fermenter cannot be replaced by a backup one prepared in advance. In this regard, long downtime of such an installation between successively conducted cycles of cultivation is inevitable.

 Or, for example, the use of a small excess pressure in the fermenter, if it reduces the likelihood of contamination (although, as practice shows, the main causes of "dirt" is poorly sterilized compounds, additives and a nutrient medium), but contributes to the release (leakage) of the product into the environment, as well as the occurrence of failures such as locking the supply filter of the fermenter through a bubbler.

 Taking into account possible emergency situations, such as depressurization of the fermenter and connecting devices during the operation of the installation, all known analogues do not provide aseptic conditions. Therefore, to work with highly pure products or biologically active products using such multifunction devices, specially equipped isolated rooms are required, in which aseptic conditions must be maintained through periodic hygienic treatments of rooms and installations and the use of special hygienic or protective clothing by personnel. According to this method of isolation of a small fermenter is very irrational, since it does not provide either sufficient aseptic conditions or the reliability of the installation and is expensive. At the same time, working conditions for personnel are deteriorating sharply.

The only effective way to ensure aseptic conditions is the local isolation of the fermenter, samplers and their compounds, including external ones, from the environment in a special biologically protective box / 28-31 /. Known MFP / 32 /, which used this method of isolation. The fermenter of this installation, with a capacity of 15 dm ³ , is located inside the insulating box. The latter has an upper chamber for the fermenter and a lower one for two vessels (with a capacity of 20 dm ³ ) for the nutrient medium (PS) and QOL. The fermenter is connected to these vessels by means of rubber tubes. To prevent product leakage into the working room, the box has systems that maintain a vacuum in its cavity, and in case of a violation of the depressive regime, it includes ventilation, which creates an upward flow of air that flows from the lower chamber to the upper through a sterilizing filter, then passes through the box another similar filter enters the processing zone located outside the working room. All operations in the box are carried out through rubber gloves (sleeves) mounted in the front panel. The installation has an autonomous fermenter sterilization system in place, including a filter on the air supply line to the fermenter. The latter has a water shirt. The stirrer drive is located on top, the shaft is sealed with a double contact seal, "lubricated" with a water-formalin mixture. There is no mechanical antifoam (PGM).

 This installation has a highly specific purpose, contains an insufficient amount of ASKR. It is not possible to aseptically connect to other installations (apparatus, equipment), for example, to conduct the process in flow mode. Used detachable rubber-tubular connections, the design of the fermenter and the box do not provide the ability to maintain constant aseptic conditions in the cavity of the upper chamber of the box. The latter performs only one-sided function - protecting the environment from product leakage, but is not an inactivating (sterile) barrier between the fermenter and the environment, that is, it is not a secondary barrier providing aseptic conditions in emergency situations. The operations of preparing for the operation of the fermenter and servicing this unit due to boxing are significantly more complicated than non-boxed MFPs.

The need to ensure ease of maintenance by compactly placing equipment in the box, reducing the number of operations performed in it, as well as the need to ensure aseptic conditions in the box during operation, eliminate the possibility of using known designs of insulating boxes, fermenters, samplers, and sterilized connecting devices when creating a boxed installation. ,
Known insulating boxes / 33, 34 / are designed to work with open research objects (products, producers, substances, etc.), the leakage of which into the environment is unacceptable. The design of such a box is a sealed chamber, made primarily of stainless steel, having glove openings on the front panel and an overview windshield. On the side walls from the outside, transmission devices are mounted - prechambers with hermetic doors. One or two inlet filters and two exhaust filters are mounted outside the box, respectively, bottom and top. In addition to special communications, the boxes have ventilation systems and maintain a depressive regime, feed and drain the inactivating solution, nozzles for spraying it in the box chamber and in the chambers. Aseptic transfer devices are additionally used hydrolock ("slicking") chambers filled with inactivating solution, as well as autoclaves connected to the box. Under isolation conditions, such a box must be permanently closed and under vacuum.

 The disadvantages of these boxes, in relation to the MFP, are the location of the prechambers on the side walls, which creates significant inconvenience when opening the internal doors and the transfer of equipment from the box, as well as the lack of primary exhaust filters that are replaced during operation from the inside of the box, with their transmission through the pre-chambers. In addition, it is unacceptable to use pre-chambers or a special enlarged pre-chambers, as well as “diving” cameras and an autoclave for transferring the fermenter, since in this case the designs and maintenance of the box and the boxed installation as a whole are complicated and their dimensions are prohibitively increased due to what would be problematic is maintaining sterility in boxing. In well-known boxes there are no devices for making external aseptic compounds, for example, compounds of portable sterile vessels located in different, not adjacent boxes.

 Taken together, these shortcomings do not allow to realize the idea of universal use of boxing for multifunction devices, namely, as an insulating and inactivation chamber, which simultaneously performs the function of a box and the function of a prechamber for a fermenter, samplers, and their compounds, including external ones.

Fermenters of known MFPs / 1-18 /, having an average capacity of 20 dm ³ (from 3 to 25 dm ³ mainly), for the most part have: a stirrer and an MPG, respectively, with lower and upper drives and transmission of rotation through magnetic couplings; sensors of parameters t, pH, eH and (or) pO ₂ , operating levels of QL, foam and emergency foam level; bubbler; siphon; heat exchanger; nodes for connecting dispensers and sampler. At the same time, small (used in MFPs) fermenters, in contrast to pilot and industrial ones, are divided, as noted above, into stationary and removable (portable) ones.

 The main reason for creating an MFP with a stationary fermenter is the need to exclude sterile compounds with a sterilizing filter system for incoming and outgoing gases, including a supply filter connected to a bubbler and an exhaust filter connected to the air cavity of the fermenter through a moisture trap (condenser) and an air heater (dehumidifier) eliminating condensation of residual moisture in the exhaust filter.

 It is practically impossible to fix such a set of devices in a removable small fermenter without a significant increase in the complexity of its design and dimensions. Therefore, devices such as individual elements are connected to the removable fermenter sterile.

 At the same time, removable fermenters sterilized in an autoclave provide higher reliability of MFPs due to redundancy, that is, the use of a removable fermenter and other removable equipment, which allows the cultivation process to be carried out at the same time and a stand-by set of equipment ready for operation. In addition, a removable fermenter allows you to better wash it (a very important requirement for small fermenters), check for leaks, and autoclaving is a more reliable way of sterilization. For a removable fermenter, as well as for removable sterilized equipment connected to it, unlike stationary ones, increased strength and special thermal insulation, as well as backpressure devices for sensors, are not required. Installations with a removable fermenter are more convenient for maintenance. However, the need and complexity of performing many sterile compounds before work and aseptic disconnections after reduce the reliability of ensuring aseptic conditions and put removable fermenters on the set of shortcomings on a par with stationary ones.

 The disadvantages of removable fermenters can be eliminated by reducing the number of sterile (aseptic) compounds and equipment with their quick-acting, including fast-sterilizing connectors. It is possible to significantly reduce the number of sterile compounds, for example, by equipping a removable fermenter with a stationary supply and exhaust filters.

 Known MFPs containing removable fermenters / 14 /, equipped with a stationary supply filter, mounted on the side above the cover, on the bubbler tube exiting from it. The disadvantages of this are the clutter of the lid and the increase in size of the fermenter. The irrational location of the stationary supply filter and the lack of exhaust do not give a significant effect, especially since any individual filter can be connected to the bubbler fitting before sterilization in an autoclave.

 From the point of view of providing asepsis, a common significant drawback of most known removable fermenters is the use of multiple rubber tubes as process pipelines connected above the burner flame. Because of this, a number of maintenance difficulties arise. Firstly, this method of sterile tube connection requires great skills, and secondly, it is not suitable for disconnections if product emissions are excluded. Thirdly, a special binding of the tubes on the fittings, as well as the correct fastening and arrangement of the tubes, excluding their compression, bending and twisting, are required.

 The use of needle-membrane connectors / 35, 36 / in the multifunction device, consisting of a syringe needle or a fitting needle with a plug for the needle hole and a response membrane unit, including a rubber membrane or a compressed rubber ring, allows for aseptic connection-disconnection of the connector under the layer of inactivating solution filled into the membrane unit, as well as reduce the length of the rubber tubes and, in some cases, refuse to use the latter. But due to the small cross section of the needle hole, such connectors are used and can only be used to connect dispensers and a sampler to the fermenter, that is, they are not suitable for connecting main pipelines (supply of a nutrient medium, drainage of QOL, etc.).

 Known devices (connectors) for sterile connection of pipelines (equipment) with steam sterilization of the connecting "deaf" cavity / 37-46 / or with washing the cavity with a chemical inactivating solution / 47 /. A connector of this type consists of two hermetically mating locking nodes (for example, valves), one of which contains a connecting chamber. Pipes for supplying steam and condensate discharge or supplying and draining the washing solution are connected to the latter.

 The disadvantages of these connectors are the bulkiness of the structures, the duration of the sterilization of the connection, as well as the need to use additional systems to discharge and inactivate condensate or drains, provided that these connectors are used for aseptic disconnection of equipment. In this regard, such devices are not suitable for use on small removable fermenters, and even more so in boxed multifunction devices.

 The placement of many sensors and connecting nodes (sockets) on a small MFP fermenter without cluttering the fermenter lid and super-fluid space (that is, ensuring ease of maintenance, aseptic conditions and effective defoaming) is possible with a partial placement of these sensors and nodes on the fermenter body or in special flow-through ( circulation) circuits through which QOL is constantly pumped. At the same time, to ensure aseptic conditions, the attachment points of the sensors and the connectors of the circuit should be located above the QOL level. In compliance with this requirement and in the presence of short sensors, placing them on the case at an angle is not acceptable. In addition, this arrangement of sensors can significantly increase the size of the small fermenter and create additional inconvenience in maintenance.

 Known fermenter / 48 / containing a circulation (flow) circuit for sensors of the physico-chemical state of the culture. The inlet (discharge) pipeline of the circuit is fixed at the bottom of the fermenter and is made curved at a right angle to the opposite direction of rotation of the mixer, and the outlet (drain) pipe is fixed in the center of the bottom and brought under the mixer (into the suction zone).

 Such a fermenter, which does not have reflective walls, is very specific in purpose. The placement of the circuit below does not meet the requirements for ensuring aseptic conditions and ease of maintenance. A variant of such a circuit is possible by mounting a special centrifugal pump on the shaft of the mixer, in which the circuit can be connected to the pump through the cover. However, even the most significant drawback remains, namely, the limited lower limit of the agitator speed control as one of the main parameters for controlling mass transfer and the need to use an additional complex and cumbersome throttle control system or the impossibility of such control when automatically controlling (changing) the agitator speed and flow rate air during aeration. The use of pumps with stand-alone drives for a small fermenter is probably also unacceptable or irrational due to the inevitability of complicating the design of the fermenter and the introduction of additional detachable and movable sealed joints and (or) rubber-tubular joints.

 The invention is aimed at solving the problem of simultaneously improving the aseptic conditions, convenience and service conditions of the MFPs by improving the quality of the fermenter and means to prevent product contamination in the fermenter by extraneous microflora and product leakage into the environment.

 The indicated means, which are of equal importance in ensuring aseptic conditions, include an insulating box, a sterilizer system for incoming and outgoing gases of the fermenter, a sampler, and units for aseptic connection-disconnection of this equipment and their pipelines. At the same time, the main significant indicators of the quality of these tools and the installation as a whole are: complexity (total number of elements and connections), mass and compactness of structures, speed of connections, including sterilized ones, as well as reliability indicators, including the ability to reserve the above elements, assemblies, assemblies MFP systems and the ability to quickly replace them in case of failure.

 The need to simultaneously address issues of improving aseptic conditions, ease of maintenance and service conditions is due to the presence of causal relationships between these properties of MFPs. And since the MFP is an assembly complex, which includes the main and backup set of assembly units (units, systems, units) and parts (that is, devices that include other devices that contain other devices, etc.), so far as these properties for the MFPs are the totality of such properties of its assembly units. In this regard, this application belongs to a group of inventions that form a single inventive concept, which consists in creating an aseptic boxed multifunction device with a quick-detachable sterilizable autoclave fermenter, samplers, the insulating box of which is not a significant obstacle to maintenance, is a small biologically isolated, enclosed space, separating the sterilized equipment and its compounds from the environment, other equipment of the MFP and in this space during operation of the MFP aggressive (inactivating) conditions for any microorganisms are kept.

The set of essential distinguishing features of the claimed invention and the achieved technical results sufficient to solve this problem are as follows:
1. The proposed MFP, containing an insulating box with a fermenter in it with a mechanical mixing device and sensors located in the lid, providing control of temperature, heat dissipation rate, stirrer speed, oxidation-reduction potential, hydrogen index, optical cell concentration, pressure in the fermenter, levels of foam and culture fluid, as well as with equipment and automated systems located outside the isolation box that provide control ue culturing process in flow modes "q ^T -CTAT", "eH-STAT", "pH-stat", "OK-STAT", unlike analogue formed as a single prefabricated stand area of not more than 0.5 m ^2, a height of not more than 2 m, inside of which are removable, autoclavable fermenters with a flow path, a sampler; the stand additionally contains a control panel and a base table, on which the module blocks of automated systems and connectors for connecting sensors are fixed, an insulating box is located between the panel and the table, the volume of which is not more than 0.1 m ³ , but not less than three volumes of the fermenter, having a lower chamber equipped with a sealed door with a window, made in such a way that when the fermenter is placed in it, its lid and the bottom of the upper chamber are located at the same level with the formation of a countertop, for viewing, lighting and access to Each box has one frontal and two side windows, as well as two glove openings in the front panel and two prechambers designed to transmit the flow path and the sampler, and are located outside at the corners of the rear boxing machine, while the inner doors of the prechamber, sampler, fermenter cover with the flow path and their connecting nodes are located between the glove openings at a distance of not more than 0.3 m from them; removable fermenter with flowing circuit and a sampler are connected in the box with each other and with the stand by means of connectors, sealed valve self-locking connectors and sealed electrical contact connectors. The combination of these features provides small-sized and convenient maintenance of the box and installation, the possibility and convenience of aerosol treatment of the entire cavity of the box and its prechambers. In this case, autonomous lighting of the box cavity is not required.

 2. A small fermentation unit containing a removable, autoclavable fermenter equipped with a stirrer, a mechanical defoamer, a heat insulating jacket, a tubular heat exchanger, a bubbler and a siphon built in through the cover, as well as connection units for dispensers, a sampler and sensors, characterized in that the fermenter is additionally contains inside the system of sterilizing filtration of gases entering and leaving during aeration, including a supply filter with a bellows valve or solenoid a valve connected to a bubbler through a through hole above the seat in the valve or valve body, as well as an exhaust filter hermetically connected to the upper cavity of the fermenter through its jacket and a radial channel hole in the cover, each filter having a glass-like housing with a hermetically screwed cover containing a cylindrical filter cartridge, filter housings and valve or valve housing are vertically fixed in the lid of the fermenter and are located under the lid in its upper cavity, and the filter lids and the flywheel valve or a solenoid valve disposed outside the lid of the fermenter, and the filter cover and protruding from the lid of the fermenter heat exchanger tubes have similar fittings on the self-closing valve connectors, whereby the fermenter is connected with an aeration and temperature control installation systems.

 In aggregate, these signs make it possible to exclude the use of a supply filter, a condenser and a moisture collector of exhaust air separately sterilized and sterilely connected to it, as well as an exhaust filter with a heater, and use the fermenter jacket simultaneously as a heat-insulating device and as a condenser and moisture collector from exhaust air, and also to exclude special heaters using the existing negative temperature gradient of the medium and cavity with fermenter lid (heat of the fermentation process, dissipation of mixing energy and heat flux from bearings and electric motors of working drives of the mixer and mechanical defoamer). In addition, the valve (valve) connecting the supply filter through the housing with a bubbler provides the possibility of deep (through bubbler) or surface aeration, and when the valve is open (open valve using a permanent magnet or an electromagnetic coil), it is impossible to lock the supply filter during inspection the tightness of the fermenter with compressed air (if there is liquid in the fermenter), as well as urgently stop intensive foaming from deep aeration by manual or automatic switching m to the surface, without changing the specified flow rate of air supplied to aeration during cultivation, while ensuring (during the decline in the level of the foam) effective ventilation of the liquid space and surface aeration of the liquid with fresh air.

3. Small fermentation unit, characterized in that it contains a device for aseptic connection-disconnection of pipelines, including a sterilizable electrical connector, consisting of a tightly interconnected through a heat-resistant gasket with a union nut or using flanges of a bellows valve having a connecting chamber with an electric heating element, and a spring valve made of stainless steel, a welding transformer with connectors, while the valve has a lock outside the seat and stem with a spacer ring and with a pusher for sequential opening of the seat and the spring valve connected to the valve, a cylindrical shell of the connecting chamber located under the seat, made as a unit with the valve body and containing contact rings from two ends, is used as an electric heating element to which a secondary circuit is connected and short-circuited via flexible insulated cables with spring-loaded contact pliers or via busbars with clamps transformer failure, moreover, the spring valve, geometric, thermal and electrical parameters of the connector sterilized by electric heating, and the transformer with connectors are selected in such a way that when the power is turned on for a predetermined time, but not more than 30 s, the cavity of the connecting closed chamber of the connector heats up from the shell to a temperature of at least 300 ^o C with the release of excess pressure through the valve, and the valve body and valve are heated with no more than 90 ^o C.

 The combination of these features ensures the small size of the connector (diameter not more than 40 mm, length not more than 120 mm, with a nominal diameter of not less than 6 mm) and the ability to quickly aseptically connect and disconnect equipment or pipelines, and the connector does not require the use of rubber tubes and can be used in the composition of both stationary and removable equipment with direct mounting of the valve and valve on the pipes to be connected by welding. The valve, in which the compression force of the rod spring is adjusted to a predetermined pressure, provides excessive steam pressure from the connecting "deaf" chamber of the connector into the connected pipeline.

 In FIG. 1 shows a general view of the MFP, FIG. 2 shows a box of an installation, FIGS. 3 a, b, c shows a general view of the fermenter and sections; 5 is a general view of a connector sterilized by electric heating (RSEN).

The proposed MFP, FIG. 1, consists of two main units - removable, autoclavable fermenter 1 (working capacity 12 dm ³ , full - 20 dm ³ ) and stand 2 (height 1.9 m, width 0.7 m, length 0.6 m) . Stand 2 includes a base table 3, an insulating box 4 (volume 0.1 m ³ ) and a control panel 5. The latter contains quick-detachable modules 6-14 (connected to the circuit, inserted at the front of the stand and fixed in place using multi-pin connectors) providing respectively control and regulation of such parameters as: temperature (t) and heat dissipation rate (q ^Т ); agitator rotation frequency (n _m ); air flow during aeration (Q _in ); redox potential (eH); hydrogen indicator (pH); optical particle concentration (OK); working and emergency foam levels (Np ₁ and Np ₂ ); pressure in the fermenter (P _f ) and box (P _b ); the duration of heating (τ _n ) and the current strength (Itr ₁ ) in the primary winding of the transformer - the power supply of the connector of the connectors sterilized by electric heating (RSEN), as well as the volumetric flow rate of the inactivating solution (Q _r ) supplied to the nozzle of the box. Block 15 provides programming and the inclusion of flow-through cultivation modes, such as: "q ^T -STAT"; eH-STAT, pH-STAT, OK-STAT. Block 16 is a common power supply. Secondary modules of blocks 6-11 are similarly placed in the support table 3, which ensures compactness of stand 2. Executive mechanisms and support systems are mounted below, on the rear wall and on top of the box hatch 4. These include: electric stirrers 17 and mechanical defoamer 18; an electric control valve 19 and a rotameter 20, equipped with a photoelectric device for controlling the level of the float and the mechanism of movement of the flow regulator Q _V ; solenoid valves 21-25, welded on pipelines designed to supply inactivating solution and compressed air to the nozzle, for supplying cold and hot water to the fermenter heat exchanger from a centralized system; solenoid valves 26-29 - actuators of dosing systems for acid and alkali solutions, chemical antifoam.

 A resistance thermometer for the fermenter and two electromagnetic coils 30 from the solenoid valves that are part of the removable pipettor designed for connecting the fermenter to other vessels are hermetically introduced into the cavity of the box. On the back wall of the box above the level of the table and the lid of the fermenter, the connectors plugs for connecting pH and eH sensors, plugs of sealed electrical connectors for connecting a nephelometer, level sensor are hermetically fixed. Moreover, all the connectors and connectors used in the box are stable and designed to work in aggressive conditions.

 Box 4 (Fig. 1, 2) has: a lower chamber 31 with a sealed door for the fermenter; two side and frontal viewing windows 32 and 33; two 34 pre-chambers (aperture 180 mm high, 120 mm wide) with hermetic quick-release doors suspended on hinges; one inlet filter 35 with a spring valve for regulating the vacuum in the box; two primary filters 36 and two secondary 37, hermetically fixed inside and outside the box, respectively. Moreover, each primary filter 36 is fixed in a narrow-profile chamber, the bottom of which is the roof of the prechamber. The nozzle for processing the inactivating solution (aerosol) of the cavity of the box and the equipment located in it is connected via vacuum tubes to the fittings of the solution and compressed air supply pipelines included in the box.

 The tightness of the power transmission and isolation of the drive 17 from the cavity of the box is ensured by a magnetic coupling (rotation transmission through the bottom of the box and the bottom of the fermenter), and the insulation of the connection of the PGM drive: electric motor - an intermediate telescopic shaft-magnetic coupling, is provided by a telescopic tube casing 38 (see Fig. 1 ), in which the intermediate shaft is mounted with the leading magnetic coupling half, while the rotation of the magnetic coupling is transmitted through the bottom of the casing and the cover of the fermenter.

 The fermenter (figure 3, 4) is a sealed cylindrical vessel 1 with a lid 2 and contains a mixing device with two turbine 6-blade stirrers 3, a defoamer with a working body in the form of a turbine, closed by two disks, 4-blade stirrers 4, having as devices for power drives, magnetic couplings 5 and 6. In addition, the fermenter includes: a tubular annular bubbler 7 with a valve 8 and a filter 9; siphon 10 (pipe for filling-emptying with valve 11; tubular heat exchanger 12 with fittings 13; two valves 14 - for connecting an external (removable) flow path; condensate jacket 15 and filter 16 for drying and sterilization of the exhaust gases; side chamber 17 for attaching electrodes 18 (pH and eH); a combined sensor 19 of the working liquid, foam and emergency foam levels; a centrifugal pump 20 designed for deaeration and pumping of QOL through the flow path and mounted on the upper supporting crosspiece 21; windows 22 23 - for illumination and viewing of the fermenter cavity; four reflective partitions 24 (plates welded to the tubes of the heat exchanger, siphon and bubbler); sleeve 25 - for the temperature sensor. As support bearings used: for the MPG shaft - rolling bearings 26; for the mixer shaft - plain bearings 27, 28. Equipment 3-14, 16, 19-21, 24-28 is included in the assembly of the cover 2, the rest is in the assembly of the vessel 1.

 The filter housings 9, 16 are closed by sealed covers, on the bottom of which filter cartridges are attached, and on top there are fittings 13 (as on the heat exchanger).

The filter 9 is connected to the bubbler 7 through a through hole under the seat in the housing of the bellows valve 8, so that when the valve is open, the air supplied to the aeration does not enter the fermenter (filled with liquid) from the bubbler, but from the valve. Instead of a valve, a solenoid valve can be mounted in its body, if necessary manually opened using a permanent magnet. First, the aerating air from the fermenter enters the jacket 15 through an opening in the casing under the lid 1 (where moisture condensates under the influence of the negative temperature gradient of the outer wall of the jacket and the exhaust air), then leaves the jacket through the sealing boss located on the opposite side (see Fig. .3, section AA), passes through a radial hole in the lid and enters the filter 16 through the connecting pipe and enters the vacuum line (see Fig. 4). Moreover, since the temperature of the lid, the connecting pipe with the filter is higher than the temperature in the jacket, condensation of residual moisture in the filter is excluded 16. Since the optimum temperature of most processes is more than 28 ^o C, the normal temperature in the working room is 18-22 ^o C, and the temperature of the lid and a filter 16 above the cavity temperature with a constantly running PGM (heating from friction in bearings 26).

 The fitting 13 is the counterpart of the self-locking spring-loaded valve connector. Valve 11 and valve 14 are the counterparts of the connector sterilized by electric heating (RSEN), through which dispenser pipelines and a flow circuit are connected to the fermenter.

The RSEN connector (Fig. 5) consists of two main parts - a bellows valve 1 and a spring valve 2, hermetically connected to each other through a heat-resistant gasket 3 with a union nut 4 or flanges (possible). The valve and valve can be connected to the equipment by welding. The valve can be mounted on equipment according to options I or II. The valve 1 has a lockable seat on the outside, a stem with a spacer ring 5 and with a triangular pusher 6, for sequential opening of the seat and a spring valve connected to the valve, adjusted to a given pressure. Under the valve seat there is a cylindrical shell 7, forming a connecting "deaf" chamber. The shell 7, which performs the function of an electric heating element in the connector, is made integrally with the valve body and has slip rings 8 that can be locked through it, to which the secondary winding of the welding transformer is connected via flexible insulated cables with spring-loaded contact clamps or via busbars with clamps - a power source mounted on the rear wall of the MFP box. When the transformer installation is turned on from the control panel for the time specified by the relay (30 s) in the short-circuited through the shell, the heating element of the circuit generates an electric current of about 700 A with a voltage of 0.45 V and a transformer power of 0.63 kW. At the same time, for 30 s, the shell 7 heats up to a temperature of 500-550 ^o C, providing heating of the air in the chamber closed on both sides (“deaf”) to 300-325 ^o C, which causes sterilization of the chamber. When there is liquid in the chamber cavity (for example, when the dispenser-pipeline or flow circuit is disconnected while the pump is running), two consecutive 30-second heating is carried out, the first to eject and evaporate the liquid (1.5 cm ³ ) from the chamber through the valve, the second - for final sterilization of the chamber cavity. With these short-term heating of the connector, the valve body is heated by heat transfer from the shell to a temperature of not more than 90 ^o C, provided that the initial temperature of the case does not exceed 50 ^o C. Quick cooling of the connectors, after heating, is carried out in the MFP box by pouring them with a solution and then blowing air from the nozzle.

 Ensuring sufficient aseptic conditions during operation of the installation is achieved as follows.

 For washing, checking the tightness of the fermenter and its flow path, samplers (main and backup sets), special workplaces are equipped in the working room (the creation of automated places is possible).

 Fermenter filter cartridges and box filters are checked for permeability at special stands.

 The insulating efficiency of the box is checked during the commissioning of the installation and during its periodic maintenance according to existing proven methods. Sterilization of the fermenter, flow path and samplers in the autoclave and chemical sterilization of the box is carried out according to the modes worked out during the testing of this equipment and indicated in the installation passport.

Aerosol treatments of the box with an inactivating solution from the nozzle are carried out before sterile connections are made and after, as well as after the introduction of replaceable removable samplers, flow path and any other elements into the box from the working room. Before removing the listed equipment from the box, it is processed in prechambers. At the end of the work, the box is completely processed, including, if necessary, processing and aseptic replacement of primary exhaust filters. For this, the valve on the bubbler line is opened (by connecting the supply filter directly to the air cavity of the fermenter), a rubber bulb is pressed to the nozzle of this filter, which is compressed under vacuum - 0.2 kgf • cm ² , and a rubber tube with a clamp and a return fitting is connected to the exhaust filter nozzle from a valve self-locking connector through which the fermenter is evacuated to a predetermined value by turning on the aeration inducer.

 Similarly, in a sterile (ready-to-use) interchangeable fermenter, a slight excess pressure is created with the use of a special elastic rubber can swell to a predetermined volume instead of a pear.

 Thus, transportation of the fermenter before and after can be carried out with an indication of its tightness, and sealed canisters can be used to transport the flow path, metering vessels and sampler.

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Claims (3)

1. A small fermentation unit containing a biological insulating box with a fermenter in it with a mechanical mixing device and sensors located in the lid, providing control of temperature, intensity of heat generation, stirrer speed, redox potential, hydrogen index, optical concentration of cells, pressure in fermenter, levels of foam and culture fluid, as well as with equipment and automated systems located outside the isolation box moms providing control of the cultivation process in flow regimes "q ^T -STAT", "eH-STAT", "pH-STAT", "OK-STAT", characterized in that it is made in the form of a single prefabricated stand with an area of not more than 0, 5 m ² and a height of no more than 2 m, inside of which there are removable, autoclavable fermenters with a flow path, a sampler, the stand additionally contains a control panel and a base table on which the module modules of automated systems and connectors for connecting sensors are fixed, between remote control and table located isol a box, the volume of which is not more than 0.1 m ³ , but not less than three volumes of the fermenter, having a lower chamber equipped with a sealed door and a window, made in such a way that when the fermenter is placed in it, its cover and the bottom of the upper chamber are located at the same level with the formation of a countertop, for viewing, lighting and access to which the box has one frontal and two side windows, as well as two glove openings in the front panel and two prechambers designed to transmit the flow path and the sampler and located and at the corners of the rear wall of the box, while the internal doors of the prechamber, sampler, fermenter lid with a flow path and their connecting nodes are located between the glove openings at a distance of not more than 0.3 m from them, removable fermenter with a flow path and a sampler are connected in the box between themselves and with the stand by means of connectors, sealed valve self-locking connectors and sealed electrical connectors.  2. A small fermentation unit containing a removable, autoclavable fermenter equipped with a stirrer, a mechanical defoamer, a heat insulating jacket, a tubular heat exchanger, a bubbler and a siphon built in through the cover, as well as connection points for dispensers, a sampler and sensors, characterized in that the fermenter is additionally contains inside the system of sterilizing filtration of gases entering and leaving during aeration, including a supply filter with a bellows valve or with a solenoid m valve, connected to the bubbler through the through hole above the seat in the valve or valve body, as well as an exhaust filter, hermetically connected to the upper cavity of the fermenter through its jacket and a radial hole in the cover, each filter having a glass-like housing with a hermetically screwed cover containing a cylindrical filter cartridge, filter housings, and a valve or valve housing are vertically mounted in the fermenter cap and located under the cap in its upper cavity, and the filter cap and the valve handwheel Whether the solenoid valve disposed outside the lid of the fermenter, and the filter cover and protruding from the lid of the fermenter heat exchanger tubes have similar choke self-closing of the valve connector, whereby the fermenter is connected with an aeration and temperature control installation systems. 3. A small fermentation unit, characterized in that it contains a device for aseptic connection-disconnection of pipelines, including an electrically sterilizable connector, which consists of a hermetically connected to each other through a heat-resistant gasket with a union nut or using flanges of a bellows valve having a connecting chamber with an electric heating element, and a spring valve made of stainless steel, a welding transformer with connectors, while the valve has a lock outside the seat and stem with a spacer ring and pusher for sequential opening of the seat and the spring valve connected to the valve, a cylindrical shell of the connecting chamber located under the seat and made as a unit with the valve body and containing contact rings from two ends is used as an electric heating element to which, through flexible insulated cables with spring-loaded contact clamps or via busbars with clamps, the secondary winding of the transformer is connected and short-circuited an informator, moreover, the spring valve, geometric, thermal and electrical parameters of the connector sterilized by electric heating, and the transformer with connectors are selected in such a way that when the power is turned on for a specified time, but not more than 30 s, the cavity of the connecting closed chamber of the connector is heated from the shell to temperature of at least 300 ^o With the release of excess pressure through the valve, and the valve and valve bodies are heated with no more than 90 ^o C.

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