System and method for solid phase aerobic fermentation

Abstract

The invention provides a system for solid phase fermentation, comprising: - a fermentation device configured to supply a substrate and substrate-treating microorganisms and / or enzymes therein and also to discharge a resulting product and waste substances; and - one or more hydraulic systems configured to provide, in the fermentation device, a liquid or a mixture of liquids which or at least one of which is saturated with bubbles of aerating gas of micro-, and / or nano-, and / or smaller size and / or dissolved oxygen; - an aeration device configured to provide saturation of the liquid with bubbles of aerating gas of micro-, and / or nano-, and / or smaller size, and / or dissolved oxygen; - a filtration device configured to separate the solid phase from a circulating liquid. A method for solid-phase aerobic fermentation and a method for producing a biotechnological product using said system are also provided. The invention provides uniform aeration of the substrate, regulation of its temperature and humidity, promotes acceleration of enzymatic processes, prevents bonding and drying of the substrate, reduces the size of the fermentation device and production areas, increases the number of possible products, and improves performance of the fermentation device and the entire system as a whole.

Description

[0001] SYSTEM AND METHOD FOR SOLID PHASE AEROBIC FERMENTATION

[0002] Field of the invention

[0003] The invention relates to biotechnology, in particular to a system and a method for solidphase aerobic fermentation, as well as a method for producing a biotechnological product using, for aeration purposes, one or more hydraulic systems configured to provide in the fermentation device presence of a liquid or a mixture of liquids, which or at least one of which is saturated with bubbles of aerating gas of micro-, and / or nano-, and / or smaller size, and / or dissolved oxygen.

[0004] Background

[0005] A solid-phase fermentation is a biotechnological process for obtaining useful products from solid substrates using microorganisms and / or enzymes. This method produces antibiotics, growth hormones, biofuels, enzymes, organic acids, food additives, biopesticides, and other products.

[0006] Rice, wheat, barley, legumes, as well as various wastes from the production and / or processing of agricultural products, such as cake, oilseed meal, bran, waste from the pulp and paper industry, renewable plant biomass and others, are usually used as substrates.

[0007] Unlike cultivation in liquid media, solid-phase fermentation usually allows to obtain higher product concentrations, however, working with solid substrates has a number of features and problems that need to be taken into account when planning the technological process:

[0008] - it is necessary to maintain a certain level of aeration and humidity throughout the thickness of the substrate, while due to high concentration of solid particles, uniform aeration and humidification are often difficult;

[0009] - constant aeration with air dries the substrate, as a result, air channels appear in the substrate, and the substrate can dry out;

[0010] - during the fermentation process, the substrates can bond together, which makes it difficult for the fermentation process to proceed evenly, so they usually try to distribute the substrate in the thinnest possible layers, which increases the size of the fermentation devices and the production area as a whole;

[0011] - some difficulties arise in removing the heat generated in the process of metabolism and temperature regulation in the reactor;

[0012] - when using mixing devices, it is important not to damage the solid substrate in order to provide conditions for the growth of microorganisms close to natural; - due to the large number of holes in the fermenter, contamination with unwanted microorganisms is possible;

[0013] - taking into account these aspects, it is important to maintain the performance of the fermenter when scaling the process.

[0014] Various designs of systems for solid-phase aerobic fermentation are known.

[0015] Most of them comprise:

[0016] - a fermentation device configured to supply a substrate and substrate-treating microorganisms and / or enzymes therein, as well as to discharge a resulting product and waste substances used in the fermentation process;

[0017] - a condition regulation system in the fermentation device, including a humidity regulation system in the fermentation device, an aeration system to provide aeration in the fermentation device, a temperature regulation system in the fermentation device, etc.

[0018] The humidity regulation system typically supplies a humidity control liquid, typically water, to the fermentation device. The aeration is usually performed by supplying a flow of aerating gas, in particular air, and the aeration system is usually a system for supplying such gas. Said liquid and gas are usually supplied to the fermentation device through separate manifolds, often with a time gap. The temperature regulation system typically includes circulation of a separate temperature regulation liquid. However, known solutions have a number of disadvantages.

[0019] As noted above, in order to ensure uniform aeration, uniform humidification and uniform temperature regulation, the substrate is arranged in layers as thin as possible, which increases the size of the fermentation devices and the production area as a whole or reduces the performance of the existing fermentation device.

[0020] Also, due to the difficulties associated with uneven humidification, the speed of redox processes and the entire fermentation as a whole decreases.

[0021] The distribution of liquid and gas is usually carried out through the use of special mixing devices. For example, CN102199521A provides a solid-phase fermentation system in which, to ensure a uniform distribution of substances during fermentation, a fermentation device is provided with a double-helix mixing device. However, the disadvantage of such structures is the possibility of destruction of the substrate during rotation of the mixing device. Also, the aerating gas cannot penetrate into the deep layers of the substrate and, as a result, it is necessary to reduce the thickness of the substrate, which increases the size of the fermentation device and production areas or reduces the performance of the existing fermentation device. In CN101496574A, supplying a humidified air to the fermentation device is proposed, and an aeration system includes an air supply device, as well as an air humidifier. Thus, aeration is also carried out by a gas flow (humidified air). It is assumed that such approach will solve the problem of humidification and temperature control, however, even in the case of humidification, uniform aeration and uniform distribution of temperature and humidity are still not achieved upon contact of air with the substrate, because the majority of moisture and the main part of air remain on the surface of the substrate. In addition, this approach creates a problem of bonding or, conversely, drying of the substrate, which makes it difficult for further uniform passage of air through it and, as a result, does not allow the fermentation process to proceed evenly. In this regard, it is necessary to reduce the thickness of the substrate, which increases the size of the fermentation device and the required production areas in general or reduces the performance of the existing fermentation device.

[0022] A system according to patent RU 2235767 C2 is also known from the prior art. Said system comprises a fermentation device containing inside a plurality of modular bases in the form of plates, wherein said plates are permeable to air and water and are connected to the wall of the tank so that neither air nor water can pass from the side. The fermentation device is once filled with water for the purpose of supplying microorganisms, after feeding microorganisms, the water is drained. The aeration system includes an air supply system, the air is supplied through a separate manifold located on the bottom of the fermentation device. The air is supplied after the water has been drained from the fermentation device. The temperature regulation system includes a system for supplying a separate cooling liquid under each of the modular bases.

[0023] The disadvantage of this solution is also insufficiently uniform aeration, which leads to formation of zones with unevenly distributed temperature and humidity, and this in turn reduces the performance of the fermentation device. During such aeration, large air bubbles are formed, which, upon collision with the substrate, cannot penetrate inside and provide sufficient aeration. Also, due to the difficulties associated with uneven humidification, the speed of redox processes and the entire fermentation as a whole decreases. Bonding or, conversely, drying the substrate is also possible, which makes it difficult for further uniform passage of air therethrough and, as a result, does not allow the fermentation process to proceed evenly. In this regard, it is necessary to reduce the thickness of the substrate, which increases the size of the fermentation device and the required production areas in general or reduces the performance of the existing fermentation device.

[0024] WO2018165411 discloses an aerobic fermentation system including a reservoir, an aeration system comprising a gas sparger in fluid connection with a reservoir for supplying compressed gas to an internal volume of the reservoir, and a recirculation circuit in fluid connection with an outlet of the reservoir. The recirculation circuit includes an eductor in fluid communication with an oxygen-containing gas source, a static mixer downstream of the eductor, a heat exchanger downstream of the ejector, and a distributor downstream of the heat exchanger. The aeration system provides mixing and transfer of oxygen mass to the fermentation composition in the tank. The fermentation composition passes through the eductor, the static mixer, the heat exchanger and the recirculation circuit distributor and back to the tank. Oxygen is transferred from the oxygen-containing gas to the fermentation composition, and heat from the fermentation composition is removed in the recirculation circuit.

[0025] However, this system also has a number of disadvantages. In particular, circulation of the entire substrate occurs in the system without separating the liquid and solid phases, which will obviously cause circulation problems in the solid-phase cultivation of insoluble or poorly soluble substrates, which, in the opinion of the present inventors, reduces the fermentation efficiency.

[0026] WO202 1097794 discloses a system and a method for aerobic fermentation. The system includes a fermentation reservoir, application of a certain strain for material fermentation; a separation reservoir; at least two microinterface generators and a return pipeline. Compared to the traditional method, due to the destruction of air with the formation of micrometer-sized microbubbles and mixing of microbubbles with raw materials with the formation of a gas-liquid emulsion, the gas-liquid interface area increases, thereby increasing the efficiency of the system.

[0027] However, in this case, to saturate the raw material with oxygen, an air supply is provided to form a gas-liquid mixture in the upper part of the fermentation device, and the air supply is also separately provided to the lower part of the fermentation device, directly to the raw material supply zone, ensuring the mass exchange of raw material with air. According to the inventors of the present invention, deep and uniform saturation of raw materials with air under such conditions is difficult, which reduces efficiency of fermentation, as well as the proposed design with separation of the fermentation zone complicates the maintenance of this device.

[0028] Effective use only for small volumes and complexity of scaling for industrial purposes are also disadvantages of known fermentation devices and methods.

[0029] Thus, there is a need to improve solid-phase aerobic fermentation systems to ensure more uniform aeration of the substrate, uniform regulation of its temperature and humidity, acceleration of oxidative processes, prevention of bonding and drying of the substrate, eliminating the need to reduce the thickness of the substrate, and therefore reduce the size of the fermentation device and production areas and increase the performance of the fermentation device as a whole. It would also be advisable to increase the amount of possible products obtained during the fermentation process. Summary of the invention

[0030] The invention provides a system for solid phase aerobic fermentation, the system comprising:

[0031] - a fermentation device configured to supply a substrate and substrate-treating microorganisms and / or enzymes therein and to discharge a resulting product and waste substances;

[0032] - one or more hydraulic systems configured to provide, in the fermentation device, presence of a liquid or a mixture of liquids, which or at least one of which is saturated with bubbles of aerating gas of micro-, and / or nano-, and / or smaller size, and / or dissolved oxygen, wherein said one or more hydraulic systems comprise:

[0033] - an aeration device configured to provide saturation of the liquid with bubbles of the aerating gas of micro-, and / or nano-, and / or smaller size, and / or dissolved oxygen before feeding it into the substrate for fermentation;

[0034] - a filtration device configured to separate a solid phase from a circulating liquid supplied to said aeration device, and then back to the substrate.

[0035] According to one of the embodiments of the invention, the aeration device is arranged outside the fermentation device. According to another embodiment of the invention, the aeration device is arranged inside the fermentation device.

[0036] In one of the embodiments of the present invention the liquid is water. Also, according to one of the embodiments of the invention, the liquid further comprises one or more additives selected from the group consisting of salts, derivatives of salts, alcohols, acids and alkalis, esters, polyesters, ammonia and organic solvents.

[0037] According to one of the embodiments of the invention, the aerating gas is a gas selected from the group consisting of oxygen, air, or a gas mixture comprising oxygen.

[0038] According to one of the embodiments, the system for solid phase aerobic fermentation is configured to supply said microorganisms and / or enzymes together with said liquid or mixture of liquids to the fermentation device.

[0039] According to one of the embodiments, the system for solid phase aerobic fermentation further comprises a temperature regulation system.

[0040] According to one of the embodiments, a cooler and / or a heater is additionally integrated into said one or more hydraulic systems.

[0041] According to one of the embodiments, the fermentation device is a fermenter selected from the group consisting of a plate fermenter, a packed fermenter, a multi-tier fermenter of any design, a tray-type fermenter, a compacted bed fermenter, a rotating drum-type fermenter, a rocking solidphase fermenter, a container-type fermenter provided with a stirring device, and a fluidized bed fermenter.

[0042] According to one of the embodiments, the resulting product is extracted and discharged using said liquid or mixture of liquids.

[0043] According to one of the embodiments, the system for solid phase aerobic fermentation further comprises a system for supplementing said liquid or mixture of liquids after discharging the resulting product.

[0044] According to one of the embodiments, a discharge device is connected to the fermentation device to discharge the resulting product.

[0045] According to one of the embodiments, the system for solid phase aerobic fermentation further comprises a drainage device for discharging the circulating liquid. Also, according to one of the embodiments, said drainage device is connected to said hydraulic system to circulate said liquid in the system for solid-phase aerobic fermentation.

[0046] According to one of the embodiments, the filtration device is located outside the fermentation device and coupled to the fermentation device.

[0047] According to one of the embodiments, the system for solid phase aerobic fermentation further comprises a circulating liquid storage tank coupled to said fermentation device via the hydraulic system.

[0048] According to one of the embodiments, the system for solid phase aerobic fermentation further comprises an exhaust gas discharge system to discharge exhaust gas from the fermentation device.

[0049] The present invention also proposes a method for solid phase aerobic fermentation, the metjod including performing solid phase aerobic fermentation using the system to saturate a liquid or a mixture of liquids in a fermentation device with bubbles of aerating gas of micro, and / or nano- , and / or smaller size, and / or dissolved oxygen.

[0050] Moreover, the invention proposes a method for producing a biotechnological product, the method including:

[0051] - feeding to a fermentation device included in the claimed system for solid-phase aerobic fermentation a substrate for producing said product and substrate-treating microorganisms and / or enzymes;

[0052] - performing a solid-phase aerobic fermentation using said system, providing saturation of a liquid or a mixture of liquids in the fermentation device with bubbles of aerating gas of micro-, and / or nano-, and / or smaller size, and / or dissolved oxygen; - extracting and discharging a resulting product, and al discharging waste substances.

[0053] According to one of the embodiments, the resulting product is extracted and discharged by said liquid or mixture of liquids.

[0054] The distinctive feature of the present invention is an aeration device (saturation of a liquid with bubbles of aerating gas of micro-, and / or nano-, and / or smaller size, and / or dissolved oxygen), allowing aeration not from a gas stream, but from a liquid stream provided by one or more hydraulic systems. This liquid is saturated with bubbles of aerating gas of micro, and / or nano-, and / or smaller size. The aerating gas contains oxygen, i.e. the aerating liquid contains oxygen in the aerating gas bubbles. In addition to the aerating gas bubbles, the liquid may also contain oxygen dissolved therein. Thus, the oxygen of the aerating gas enters the fermentation device in sufficient quantity and, accordingly, deeply and uniformly saturates the substrate with said liquid or mixture of liquids.

[0055] The aeration device is optionally arranged outside the fermentation device, which allows to more efficiently saturate the raw material with oxygen.

[0056] In the presence of such aeration system, a need for a separate supply of moisture-regulating liquid is eliminated and the mass transfer of gas and liquid is improved. Due to the reduced size of the bubbles, a greater supply of oxygen is provided, which improves the process of solid-phase aerobic fermentation. The use of a liquid as a carrier allows the gas to be more evenly distributed in the substrate as opposed to its supply with a gas flow.

[0057] The invention thus provides a more uniform aeration of the substrate due to a deeper penetration of the aerating gas bubbles and / or dissolved oxygen. At the same time, the humidity and temperature of the substrate are regulated. When the temperature of said liquid or mixture of liquids is suitably regulated, said liquid or mixture of liquids immediately provides the desired temperature. More uniform aeration and regulation of humidity and temperature create more comfortable conditions for the growth and metabolism of microorganisms and thus provide an increase in the performance of the system for solid-phase aerobic fermentation and the system as a whole.

[0058] Thus, due to the use of said liquid or mixture of liquids, which or at least one of which is saturated with bubbles of aerating gas of micro-, and / or nano-, and / or smaller size (hereinafter referred to as aerating liquid for brevity), and / or dissolved oxygen, a separate humidity regulation system and a temperature regulation system may be absent in the claimed system for solid-phase aerobic fermentation. In one of the embodiments of the present invention, said system for solid phase aerobic fermentation can further comprise an additional temperature regulation system providing temperature regulation in the fermentation device by circulating the temperature regulation liquid.

[0059] The saturation of the liquid with aerating gas bubbles and / or oxygen and the supply of such liquid to the substrate improves the oxidative enzymatic processes in the solid and liquid phases. The liquid saturated with oxygen and / or aerating gas bubbles accelerates the oxidation processes occurring during fermentation, and, accordingly, the entire fermentation process and increases the performance of the fermentation device and the entire system as a whole.

[0060] In addition, the supply of aerating gas in the liquid stream also prevents the above- mentioned bonding and drying of substrates. Thus, a need to divide the substrate into thinner layers is eliminated. This allows to reduce the required dimensions of the fermentation device and the required production areas for the process at the same capacity or, conversely, to increase the capacity of existing fermentation devices and the entire system as a whole.

[0061] Moreover, in some cases, use of the claimed system for solid-phase aerobic fermentation allows to increase the amount of products obtained as a result of fermentation. Said aerating liquid or mixture of liquids simultaneously performs the role of an extractant, and allows the products in the liquid phase to be immediately extracted from the substrate. In conventional processes, the product remains in the solid phase-substrate, which is extracted from the fermentation device and then sent to separate the product from it. By using the claimed system using an aerating liquid, the present invention allows to simultaneously obtain a product in the solid phase and, additionally, a product in the liquid phase. For example, the resulting soluble single-cell protein fractions can be immediately extracted from the substrate with an aerating liquid.

[0062] According to one of the embodiments of the invention, the aeration device is arranged outside the fermentation device, thereby achieving the additional advantage that the supply of the liquid saturated with dissolved oxygen to the fermentation device can be carried out both in the upper substrate layers and in the lower substrate layers, or combined.

[0063] A further distinctive feature of the present invention is the presence of the filtration device that separates the solid phase from the circulating liquid, which allows effective fermentation in the solid phase. In particular, the filtration device, which allows to effectively separate the liquid and solid phases, provides preparation of the liquid for further circulation in the system. The filtration system is used for the cyclic use of the waste liquid and to enable its purification, followed by saturation of the liquid with gas bubbles and feeding it back to the fermentation device.

[0064] The filtration device can be arranged both outside the fermentation device and inside the fermentation device. In one of the embodiments of the present invention, said liquid can be water, in particular any water, for example, commonly used water, unprepared water, specially prepared water, mineralized, mineralized, ionized, deionized, distilled water or any other water, or a mixture of such waters, or any other mixtures with such waters, including emulsions.

[0065] In another embodiment of the present invention, the liquid can further comprise one or more additives, for example, selected from the group consisting of salts, salt derivatives, alcohols, acids, and alkalis, esters, polyesters, ammonia, and organic solvents. The composition of the additives may vary during the fermentation process and depends on the resulting product.

[0066] In an additional embodiment, a mixture of these liquids of different compositions can be used.

[0067] In one of the embodiments of the present invention, the aerating gas can be oxygen, air, or any gas mixture containing oxygen.

[0068] In one of the embodiments of the present invention, the claimed system for solid phase aerobic fermentation can be configured to supply said microorganisms and / or enzymes in conjunction with said liquid or mixture of liquids to the fermentation device. This provides another technical effect - simplified supply of microorganisms and / or enzymes and their more uniform distribution over the substrate.

[0069] In one of the embodiments of the present invention, the temperature regulation system and the aeration system are combined and the temperature regulation is carried out by said aerating liquid or mixture of liquids which or at least one of which is saturated with bubbles of aerating gas of micro-, and / or nano-, and / or smaller size, and / or dissolved oxygen.

[0070] That is, the temperature in the fermentation device can be regulated by the same aerating liquid.

[0071] In one of the embodiments of the present invention, a cooler and / or a heater can be integrated into the one or more hydraulic systems. Thus, uniform temperature regulation in the fermentation device can be accomplished by said one or more hydraulic systems using said liquid or mixture of liquids saturated with aerating gas bubbles and / or dissolved oxygen. Temperature regulation can be carried out, for example, by turning the cooler and / or the heater on / off, by regulating the flow rate and / or composition of said liquid or mixture of liquids, by a combination of said methods, or by any other suitable methods.

[0072] This approach saves the liquid used, as well as reducing the number of necessary technological holes in the fermentation device, which reduces the likelihood of contamination. The amount of process equipment and necessary liquids is also reduced, which simplifies the design of the system and the entire fermentation process while maintaining or increasing the performance of the fermentation device and the entire system as a whole.

[0073] In one of the embodiments, any suitable fermenter can be used as the fermentation device, for example, selected from the group consisting of a plate fermenter, a packed fermenter, a tower fermenter with or without shelves, a tray-type fermenter, a compacted bed fermenter, a rotating drum-type fermenter, a rocking solid-phase fermenter, a container-type fermenter provided with a stirring device, a fluidized bed fermenter, and any other. Those skilled in the art, on the basis of general knowledge of biotechnology, will be able to easily select the necessary structure of the fermenter, depending on the type of substrate, the microorganisms and / or enzymes used, and other factors. The present invention is not limited to the type and design of the fermenter, and can be applied to any types and designs of fermenters.

[0074] The supply of substrate-treating microorganisms and / or enzymes and / or other desired components can be carried out either separately or with the supply of substrate (during, before, after the supply). The supply of substrate-treating microorganisms and / or enzymes and / or other desired components can be accomplished by any method using suitable devices known by those skilled in the art.

[0075] The substrate can be selected and fed to the fermentation device also by any method known by those skilled in the art. Thus, the substrate can be pre-arranged in a fermentation device, or introduced with microorganisms and / or enzymes.

[0076] In one of the embodiments of the invention, the substrate and / or microorganisms and / or enzymes and / or other desired components can be supplied to the fermentation device by the hydraulic system included in the aeration system. In one of the embodiments of the invention, the system for solid phase aerobic fermentation is configured to supply a substrate and / or microorganisms and / or enzymes and / or other desired components in conjunction with an aerating liquid.

[0077] In one of the embodiments of the present invention, the claimed system for solid phase aerobic fermentation can be configured to extract and discharge the resulting product with said liquid or mixture of liquids. In one of the embodiments of the claimed methods, the resulting product is also extracted and discharged by said liquid or mixture of liquids. Thus, discharging of the resulting product can be carried out not only in the solid phase, but also in the liquid phase. Therefore, said system for solid-phase aerobic fermentation can be configured to discharge the resulting product with an aerating liquid. In one embodiment of the present invention, the aerating liquid is used to extract the product from the substrate, and discharge the product with the waste liquid. That is, the claimed system for solid-phase aerobic fermentation and the claimed methods allow to increase the amount of products obtained: not only products in the solid phase can be obtained, but also liquid-soluble products discharged in the liquid phase.

[0078] In one of the embodiments of the present invention, said system for solid phase aerobic fermentation can further comprise a system for supplementing of aerating liquid after the resulting product is discharged, since the product is discharged in the liquid phase and the aerating liquid supply shall be compensated. Such system can be any device for supplementing and replenishing sufficient amount of aerating liquid to perform solid phase aerobic fermentation.

[0079] Thus, the same liquid (aerating liquid) is both an oxygen carrier for aeration, and a source of moisturizing liquid, and a temperature regulator, and accelerates oxidation processes in the substrate with acceleration of the entire fermentation process as a whole, and is a carrier for supplying microorganisms and / or enzymes and / or substrate and / or other necessary components, and also an extractant for extraction and discharge of products, it also prevents bonding and drying of the substrate, reduces the size of the fermentation device and production areas, increases the performance of the fermentation device and the entire system as a whole, and also reduces the amount of necessary process liquids and simplifies the fermentation process.

[0080] In one of the embodiments of the present invention, a discharge device can be connected to the fermentation device to discharge the resulting product. In one of the embodiments of the present invention, the resulting product can be discharged, for example, from the side of the fermentation device opposite to the supply of the substrate by means of a suitable discharge device, for example a screw pump, screw or other device, or under its own weight by opening the shut-off valves, or by means of a combination of said discharging methods.

[0081] In one of the embodiments of the present invention, the system for solid-phase aerobic fermentation can further comprise one or more drainage devices to remove waste materials from the fermentation process. In one of the embodiments of the present invention, the system for solid phase aerobic fermentation can further comprise one or more drainage devices to remove the waste liquid from the fermentation process. In this case, the discharged liquid, e.g. water, can be either discharged into the various water intake device(s) or system(s) or used cyclically. The waste liquid can further comprise fermentation products, that is, the discharge of the waste liquid and the discharge of the resulting product can be combined.

[0082] In one of the embodiments, said drainage device is connected to said hydraulic system to circulate said liquid in the system for solid phase aerobic fermentation. This approach allows reducing the flow rate of the liquid used. In one of the embodiments of the present invention, the system for solid phase aerobic fermentation can further comprise a waste liquid storage tank wasted during the fermentation process. The liquid storage tank can be used to collect drainage and accumulate liquid before mixing this liquid with the gas.

[0083] In one of the embodiments of the present invention, the system for solid phase aerobic fermentation can further comprise an exhaust gas discharge system to discharge exhaust gas from the fermentation device. Exhaust gas discharging provides more uniform aeration with fresh gas, creates a more uniform temperature regime, prevents drying of the substrate, and also creates more comfortable conditions for the growth of microorganisms, which contributes to an increase in the productivity of the system for solid-phase aerobic fermentation and the entire system as a whole.

[0084] The present invention therefore can optionally comprise one or a combination of the following features:

[0085] - feeding into a fermentation device said microorganisms and / or enzymes together with said liquid or mixture of liquids;

[0086] - additional temperature regulation system;

[0087] - a cooler and / or a heater is integrated into said one or more hydraulic systems;

[0088] - extracting and discharging the resulting product using said liquid or mixture of liquids;

[0089] - a system for supplementing said liquid or mixture of liquids after discharging of the resulting product;

[0090] - discharging the resulting product by a discharge device connected to the fermentation device;

[0091] - drainage to discharge waste liquid;

[0092] - the drainage device is connected to said hydraulic system to circulate said liquid in the system for solid-phase aerobic fermentation;

[0093] - waste liquid storage tank;

[0094] - a exhaust gas discharge system to discharge exhaust gas from the fermentation device.

[0095] After and / or during the fermentation process, in the case of cyclic use of the liquid, the latter can be saturated with useful soluble products, which can be withdrawn together with the liquid for further extraction or drying, and the withdrawn volume can be supplemented with the necessary composition, for example, with the initial one.

[0096] The proposed system can be provided and additionally contain other additional elements for solid-phase aerobic fermentation.

[0097] Thus, the system can include an additional humidity regulation liquid supply system or temperature and humidity sensors and regulators. The claimed method for solid-phase aerobic fermentation and a method for producing a biotechnological product include performing solid-phase aerobic fermentation using the claimed system in accordance with all possible variants thereof specified above, obtaining all these results.

[0098] In the implementation of the claimed methods, aeration in the fermentation device is provided by an aerating liquid or a mixture of liquids, which or at least one of which is saturated with bubbles of aerating gas of micro-, and / or nano-, and / or smaller size, and / or contains oxygen dissolved therein.

[0099] Thus, all of these embodiments and optional additional features of the system for solidphase aerobic fermentation, as well as the described technical effects, fully apply to the claimed methods.

[0100] Brief Description of the Drawings

[0101] Fig. l is a schematic diagram of solid-phase aerobic fermentation according to the present invention, which reflects the variants and principles of operation of the claimed system for solidphase aerobic fermentation with mandatory and optional elements, as well as embodiments of the claimed method for solid-phase aerobic fermentation and a method for producing a biotechnological product.

[0102] Fig. 2 is a schematic diagram of solid-phase aerobic fermentation according to the present invention with shown capabilities for drainage and cycling use of liquid shown.

[0103] Fig. 3 illustrates one non-limiting example of a system for solid phase aerobic fermentation.

[0104] Fig. 4 illustrates another non-limiting example of a system for solid phase aerobic fermentation.

[0105] The Applicant considers it necessary to emphasize that the claimed invention does not depend on the type and design of the fermentation device and can be applied to any types and designs of such devices.

[0106] The arrangement of the elements and the direction of the flows in the presented drawings are indicated schematically and can be any such that solid-phase aerobic fermentation is possible. Those skilled in the art of biotechnology will appreciate possible options for location and direction of such flows, and such options are encompassed within the scope of protection of the present disclosure.

[0107] Detailed Description The present invention will now be discussed in more detail, indicating examples of embodiments of the invention that do not limit the present invention.

[0108] The terms “comprising” or “including” are interchangeable and imply that, in addition to said system and method elements, said system and methods may include other elements.

[0109] The term “aeration” in the context of the present invention means the process of saturating a liquid with bubbles of aerating gas of micro-, and / or nano-, and / or smaller size, and / or dissolved oxygen to provide oxygen during aerobic fermentation.

[0110] The term “aerating liquid” means any liquid used in the fermentation system that is preliminarily or during the fermentation process saturated with aerating gas bubbles and / or dissolved oxygen. This liquid may initially be a separately supplied liquid for aeration purposes, or a liquid used for temperature regulation, supply of substrate, microorganisms, enzymes, and other purposes.

[0111] The term “saturated with bubbles of aerating gas” means that the aerating liquid is fully or partially saturated with the bubbles of the specified size. This may be achieved, for example, by using special saturating devices or by mixing liquid and gas with a certain force provided by the design of a hydraulic system, a fermentation device or other additional devices.

[0112] The terms “bubbles of micro-, and / or nano-, and / or smaller size” are intended to mean that the liquid comprises bubbles having micro-bubble sizes (ranging from 1 pm to 1 mm) or nanobubble sizes (ranging up to 1 pm). Also, the liquid, in addition to these bubbles, contains a gas dissolved therein. For example, the size of the bubbles can be less than 1 mm, or less than 0.1 mm, or less than 1 pm. These ranges are inclusive of any values included therein. Measurement error of at least ±10% should also be taken into account.

[0113] The term “providing, in a fermentation device, presence of a liquid or a mixture of liquids which or at least one of which is saturated with bubbles of aerating gas of micro-, and / or nano-, and / or smaller size, and / or dissolved oxygen” means that said one or more hydraulic systems supply such liquid or mixture of liquids to the fermentation device. Such provision is created due to the aeration device, which can be arranged both outside and inside the fermentation device. In this case, a contact of said liquid or mixture of liquids saturated with aerating gas bubbles and / or dissolved oxygen with the substrate and / or a pass of said liquid or mixture of liquids through the substrate are provided.

[0114] The amount of aerating liquid is determined individually and, for example, is sufficient and / or effective to perform solid-phase aerobic fermentation. The terms "a", "an" and "the" may include plural forms. So, for example, some embodiments of the present invention include one or more substrates, one or more microorganisms, one or more enzymes, one or more products, etc.

[0115] Any microorganisms suitable for aerobic fermentation purposes can be used as microorganisms. Examples are fungi, yeasts, bacteria, their consortia, and any other variants. The microorganisms can be selected from obligate aerobes, facultative anaerobes, microaerophiles, etc. Those skilled in the art, based on general knowledge of biotechnology, will be able to easily select the necessary microorganism to obtain the desired product. The microorganisms can be supplied as subculture in any suitable way using any suitable subculture supply systems.

[0116] Separate enzymes such as cellulases, laccases, phenol oxidases, peroxidases, pectinases, hemicellulases, amylases, lipases, chitinases and others can also be used for solid-phase aerobic fermentation.

[0117] As a substrate, rice, wheat, barley, legumes, various wastes of production and / or processing of agricultural products, such as cake, oilseed meal, bran, pulp and paper industry wastes, renewable plant biomass and others can be used. Below are examples of using specific substrates to produce specific products. For example, without being limited thereto, straw bale, wheat bran, or fruit processing wastes can be used.

[0118] Any suitable fermenter can be used as the fermentation device. Examples are plate fermenters, packed fermenters, tower fermenters with or without shelves, tray-type fermenters, compacted-bed fermenters, rotating-drum-type fermenters, rocking solid-phase fermenters, container-type fermenters provided with a stirring device, fluidized-bed fermenters, and any others, including multi-tier constructs. Multi-tier systems in solid-phase fermenters allow the substrate to be divided into several levels, reducing pressure on the lower layers and preventing them from compacting. Such systems are a structure consisting of several trays or shelves, where a substrate is placed on each tier, wherein the possible shape of the trays includes, but is not limited to: flat trays, perforated trays, inclined trays, trays with sides, concave or U-shaped trays, vibrating trays with an inclined bottom, cone-shaped or pyramidal trays, lattice trays, spherical trays, cylindrical trays, a cone-shaped cover, a spherical cover, and any combinations thereof.

[0119] Thus, when using multi-tier systems for fermentation, a number of advantages are achieved:

[0120] - uniform load distribution: the substrate is distributed in trays, and the pressure on each tier is minimal, which preserves the porous structure of the substrate; - improvement of aeration: multi-tier structures facilitate air access to each level of the substrate, which contributes to a better exchange of gases;

[0121] - process monitoring convenience: processes occurring at different levels are easier to monitor and adjust. Temperature, humidity and aeration can be regulated separately for each tier.

[0122] - flexibility in control: it is easier to maintain a uniform distribution of microorganisms and nutrients throughout the mass of the substrate.

[0123] Such fermenters are often used in the production of enzymes and other bioproducts, where it is important to maintain uniform aeration and access to oxygen for microorganisms.

[0124] To organize continuous fermentation in multi-tier systems of solid-phase fermenters, mechanisms can be used to move the substrate from the upper tiers to the lower tiers. This ensures a continuous treatment cycle of the substrate and maintains the high efficiency of the process. The following solutions are possible for moving the substrate from the upper tiers to the lower tiers:

[0125] 1. Gravitational displacement

[0126] During gravitational movement, the substrate on the upper tier moves under the influence of gravity through special holes or inclined trays to the lower levels. For this purpose, adjustable valves or hatches can be provided on each tier, opening at the right time.

[0127] The advantage of this method is the simplicity of design and minimal maintenance costs, since there are no moving mechanical parts.

[0128] 2. Mechanical conveyors or scraper systems

[0129] Here, special conveyor systems, such as belt or scraper conveyors, can be used to move the substrate. With their help, the substrate is collected from the upper tiers and moved to the lower tiers.

[0130] The advantage of this method is to ensure uniform movement of the substrate, accurate control over the speed of movement and the amount of substrate.

[0131] 3. Screw (auger) conveyors

[0132] An auger conveyor (or multiple conveyors) can move a substrate from one tier to another. The system can be configured to operate automatically by taking a certain amount of substrate to the next level.

[0133] The advantage of this option is the ability to work with dense substrates and reliability during continuous operation.

[0134] 4. Pneumatic displacement systems

[0135] Pneumatic movement systems are systems that use airflow to move the substrate. The air flow can lift the light substrate and carry it to the desired areas of the fermenter, moving from the upper tiers to the lower tiers. The advantage of this method is good aeration of the substrate during movement and minimal mechanical impact on the substrate.

[0136] 5. Vibrating trays

[0137] In this case, vibrating trays with a perforated bottom are used, through which the substrate can gradually move down, vibrating at the desired frequency. Vibration facilitates the movement of the substrate, preventing it from bonding and compacting.

[0138] The advantage of such approach is the gradual and uniform movement of the substrate, which improves process control.

[0139] 6. Rotary systems with blades

[0140] In this case, rotary blades or augers are installed on each tier, which move the substrate into the holes for its transfer to the lower level.

[0141] The advantage of the described method is the automation of the process with the possibility of adjusting the amount of substrate moved.

[0142] Each of these systems can be adapted to specific production requirements, depending on the type of substrate, the volume of fermentation and the necessary conditions for microorganisms.

[0143] Those skilled in the art, on the basis of general knowledge of biotechnology, will be able to easily select the necessary structure of the fermenter, depending on the type of substrate, the microorganisms and / or enzymes used, and other factors. The present invention is not limited to the type and design of the fermenter, and may be applied to any types and designs of fermenters.

[0144] Said liquid saturated with dissolved oxygen and / or aerating gas bubbles can be water, in particular any water, for example commonly used water, unprepared water, specially prepared water, mineralized, mineralized, ionized, deionized, distilled water or any other water, or a mixture of such waters, or any other mixtures with such waters, including emulsions.

[0145] In another embodiment of the present invention, the liquid can further comprise additives such as, for example, salts, salt derivatives, alcohols, acids and alkalis, esters, polyesters, ammonia and organic solvents. The composition of the additives may vary during the fermentation process and depends on the resulting product.

[0146] In one embodiment, a mixture of these liquids of different compositions can be used.

[0147] The aerating gas is oxygen, air, or any gas mixture containing oxygen.

[0148] According to another embodiment of the present invention, the fermentation device and the system for solid-phase aerobic fermentation are further configured to remove (extract) and discharge the resulting product from the substrate and, accordingly, from the fermentation device, wherein the obtained product is extracted and discharged using said liquid or mixture of liquids, which is or at least one of which is saturated with bubbles of aerating gas of micro-, and / or nano- , and / or smaller size, and / or contains oxygen dissolved therein.

[0149] Fig. l is a schematic diagram of solid-phase aerobic fermentation according to the present invention, according to which the claimed system for solid-phase aerobic fermentation operates and the claimed method for solid-phase aerobic fermentation and a method for producing a biotechnological product are carried out.

[0150] The arrangement and direction of the flows in Fig. 1 and the other drawings are schematic, and can be any such that solid-phase aerobic fermentation is possible. Those skilled in the art of biotechnology will appreciate the possible options for the location and direction of such flows, and such options are encompassed within the scope of protection of the present disclosure.

[0151] A substrate 12, as well as microorganisms (subculture) and / or enzymes 11, are supplied into the fermentation device 1.

[0152] The supply of substrate-treating microorganisms and / or enzymes and / or other desired components can be carried out both separately and with the supply of substrate (during, before, after the supply of substrate). The supply of substrate-treating microorganisms and / or enzymes and / or other desired components can be accomplished by any method using suitable devices known by those skilled in the art. The substrate can be selected and fed to the fermentation device also by any method known by those skilled in the art. Thus, the substrate can be pre-arranged in a fermentation device, or introduced with microorganisms and / or enzymes.

[0153] A liquid or mixture of liquids 55 is supplied to the system and provide a presence of a liquid or a mixture of liquids in the fermentation device by a hydraulic system 3 (which there may be one or more), which or at least one of which is saturated with bubbles of aerating gas 22 of micro-, and / or nano-, and / or smaller size. Accordingly, said liquid or mixture of liquids also comprises oxygen dissolved therein. As noted above, said one or more hydraulic systems either supply such liquid to the fermentation device or create it in the fermentation device.

[0154] By way of example, Fig. 1 shows a hydraulic system 3, which is a system for providing a presence of liquid or a mixture of liquids in a fermentation device, which or at least one of which is saturated with bubbles of aerating gas 22 of micro-, and / or nano-, and / or smaller size, and / or dissolved oxygen. Said hydraulic system comprises a device 2 for saturating a liquid or a mixture of liquids with gas bubbles, into which aerating gas 22 is supplied, and a pipeline through which a liquid or a mixture of liquids saturated with bubbles of aerating gas of micro-, and / or nano-, and / or smaller size, and / or dissolved oxygen is supplied to the fermentation device. A liquid-gas mixing device or a gas bubble generator can be used. The hydraulic system can be adapted to use devices to saturate liquid or mixture of liquids with gas bubbles, using, for example, the properties of the turbine of the vortex pump and effectively absorbing the gas with the liquid or several liquids and adding it under pressure to the process. All said devices can be arranged both inside and outside the fermentation device.

[0155] In one of the embodiments of the invention, the substrate and / or microorganisms and / or enzymes and / or other desired components can be fed to the fermentation device by the hydraulic system 3.

[0156] Said liquid or mixture of liquids, which or at least one of which is saturated with bubbles of aerating gas of micro-, and / or nano-, and / or smaller size, and / or dissolved oxygen, passes through the substrate, creates and regulates the necessary conditions for the implementation of solid-phase aerobic fermentation.

[0157] Due to this, the substrate is properly converted into a product under the action of microorganisms and / or enzymes, ensuring the required amount of oxygen contained in the aerating gas. Thus, the system for solid-phase aerobic fermentation comprises an aeration system comprising one or more hydraulic systems configured to provide, in the fermentation device, presence of a liquid or a mixture of liquids, which or at least one of which is saturated with bubbles of aerating gas of micro-, and / or nano-, and / or smaller size, and / or dissolved oxygen.

[0158] Due to the fact that the oxygen of the aerating gas passes through the substrate in the liquid, deep penetration of oxygen into the substrate, uniform aeration and simultaneous uniform regulation of temperature and humidity are ensured, since the temperature of the supplied liquid can be regulated accordingly. The saturation of the liquid with aerating gas bubbles and the supply of such liquid to the substrate improves the oxidative enzymatic processes in the solid and liquid phases. The liquid saturated with aerating gas bubbles accelerates the oxidation processes that occur during fermentation, which speeds up the entire fermentation process and increases the performance of the fermentation device and the entire system as a whole. Regular passage of liquid through the substrate prevents bonding and drying of the substrate, so it is not necessary to reduce the thickness of its layer, which in turn allows to reduce or not to increase the size of the fermentation device and production areas or increases the performance of existing fermentation devices and the entire system.

[0159] The substrate is converted into product(s), which can be in either a solid phase or a liquid phase (in the exiting waste liquid). That is, by applying a hydraulic system and said liquid or mixture of liquids, the amount of products obtained can be increased. By way of example, the resulting product is discharged from the fermentation device together with the substrate (position 10 in Fig. 1) and / or with the waste liquid (position 10’ in Fig. 1).

[0160] The waste substances are discharged, for example, as follows: the liquid is discharged through the system 4, the gas - through the gas discharge system 8. The waste substrate is also discharged from the fermentation device in any suitable manner.

[0161] The discharge of the resulting product and waste substances from the fermentation device 1 is also carried out using any suitable systems and methods.

[0162] In this case, drainage through a drainage storage tank 5 and a filtration system 6 are used to discharge the waste liquid.

[0163] Further, optionally, said liquid can be directed back into the fermentation device. For this purpose, it is further purified, if necessary, its temperature is optionally regulated by means of a temperature controller 7, and then the liquid is fed into the system 2 for saturation with gas, and then returned as an aerating liquid to the fermentation device through the hydraulic system 3.

[0164] The invention can make use of one or more hydraulic systems and one aerating liquid or mixture of liquids. For example, in case of a large fermentation device, several hydraulic systems can be used: one or more aerating, one or more temperature regulation systems (cooling systems). Two or more aerating systems and two or more temperature regulation systems, etc. may be used. Accordingly, a single liquid or a mixture of liquids may be used for these purposes.

[0165] In one of the embodiments of the present invention, said system for solid phase aerobic fermentation can further comprise a system for supplementing of aerating liquid after the resulting product is discharged, since the product is discharged in the liquid phase and the aerating liquid supply shall be compensated. Such system can be any device for supplementing and replenishing sufficient amount of aerating liquid to perform solid phase aerobic fermentation.

[0166] The temperature regulation system can be implemented as shown in the diagram or as a separate feed to the cooling / heating agent fermentation device. For example, the cooling / heating agent can be fed through the jacket of the fermentation device. A heat exchanger can also be arranged within the fermentation device, that is, the fermentation device can further include a heat exchanger. An aerating liquid or any other agent can be used, for example, as the cooling / heating agent.

[0167] In one of the embodiments, the temperature regulation system can include cooling / heating assembly(ies) within the fermentation device connected by a connection to pipes to discharge and supply coolant / heating liquid that are arranged outside the fermentation device. In one of the embodiments, the temperature regulation system can be a cooling and / or heating system and can be combined with a hydraulic system providing a liquid saturated with an aerating gas, in which case the saturated liquid is supplied via a temperature regulation system, which can have a system of droppers or openings to discharge the liquid into the fermentation device.

[0168] After and / or during the fermentation process, in the case of cyclic use of the liquid, the latter can be saturated with useful soluble products, which can be withdrawn together with the liquid for further extraction or drying, and the withdrawn volume can be supplemented with the necessary composition, for example, the initial one.

[0169] Specific embodiments of the invention illustrated in Figs. 2-4 are described below. These examples are provided solely for the purpose of better understanding the invention, and are not intended to limit the scope of protection.

[0170] Fig. 2 shows an example of a system for solid-phase aerobic fermentation according to the present invention with shown capabilities for drainage and cyclic use of liquid.

[0171] The substrate 12, for example a bale of flax fiber, after steam pretreatment, is placed in the fermentation device 1. Further, the hydraulic system 3 is filled with liquid 55, for example water, then in the aeration device 2 the water is saturated with an aerating gas 22, for example air, to form a liquid saturated with dissolved oxygen and / or aerating gas bubbles of micro-, and / or nano-, and / or smaller size, and it is fed into the fermentation device 1, where the substrate is irrigated from above, increasing or maintaining humidity and simultaneously aerating the substrate due to the air contained in such aerating liquid. An inoculant, e.g., one of or a consortium of basidiomycetes, e.g., white rot fungi selected from those having a selective ability to degrade lignin without substantially degrading cellulose, is supplied through the microorganisms supply system 11. Water saturated with aerating gas and / or dissolved oxygen, passing through the substrate from top to bottom, drains through the drainage system 4, then enters the storage tank 5. After that, passing through the filtration system 6, it reaches the temperature controller 7, then it is saturated in cyclic manner with air in said aeration device 2. The fermentation device 1 is equipped with a gas discharge system 8. As a result of the required biological time cycle, the resulting product is obtained in the form of delignified flax biomass enriched with cellulose. Such process does not harm the environment and is environmentally friendly. A heater / cooler can be integrated into the hydraulic system. Thus, this liquid with gas bubbles performs the role of not only of uniform aeration and humidification, but also provides temperature regulation.

[0172] The proposed system creates comfortable conditions for the growth of microorganisms and, as indicated above, increases the performance of the fermentation device and the entire system. In addition, performing aeration using a liquid flow prevents the substrate from bonding and drying. Thus, a need to divide the substrate into thinner layers is eliminated. This allows to reduce the required dimensions of the fermentation device and the required production areas for the process, as well as to increase the performance of the device with the same dimensions.

[0173] Fig. 3 relates to an example of operation of the claimed system for solid-phase aerobic fermentation and the claimed methods in a case where the fermentation device is a fermenter with a shelf system.

[0174] The substrate 12, for example wheat bran, after steam pretreatment is placed through the substrate supply system 15 to the fermentation device 1. Further, the hydraulic system 3 is filled with liquid 55, e.g. water. Further, in the device 2 for liquid saturation with gas, water is saturated with an aerating gas 22, for example air, to form a liquid saturated with dissolved oxygen and / or aerating gas bubbles of micro-, and / or nano-, and / or smaller size, and it is supplied into the fermentation device 1, so that irrigation occurs evenly from the top of the substrate, thereby increasing or maintaining humidity and simultaneously aerating the substrate due to dissolved air. An inoculant, for example a producer of a single-cell protein, is supplied through the subculture supply system 11. Water, passing through the substrate from top to bottom, drains through the drainage system 4, then enters the storage tank 5. After that, passing through the filtration system 6, it reaches the temperature controller 7, then it is saturated in cyclic manner with air in said aeration device 2. The fermentation device 1 is equipped with a gas discharge system 8. The substrate is moved along the shelves 13 in a serpentine course from top to bottom due to their inclination and / or vibration of the shelves 13 and / or vibration of the fermentation device 1. As a result of the required biological time cycle, a resulting product is obtained in the form of a singlecell protein, which is discharged through the product discharge system from the fermentation device 10.

[0175] Such fermentation process ensures uniform aeration of the substrate due to deeper penetration of the bubbles of aerating gas and / or dissolved oxygen, regulation of the temperature of the substrate and increase in the performance of the fermentation device and the entire system as a whole. In this case, it is not required to supply a separate humidity regulating liquid, since the aerating liquid performs the function of humidification of the substrate. Also, the saturation of the liquid with aerating gas bubbles and / or dissolved oxygen and the supply of such liquid to the substrate accelerates the oxidative enzymatic processes in the solid and liquid phases and the entire fermentation process as a whole. In addition, performing aeration using a liquid flow prevents the substrate from bonding and drying. Thus, a need to divide the substrate into thinner layers is eliminated. This allows to reduce the required dimensions of the fermentation device and the required production areas for performing the process, and also increases the productivity of the fermentation device and the entire system as a whole.

[0176] Fig. 4 shows an example of operation of the claimed system for solid-phase aerobic fermentation when the filtration device is arranged outside the fermentation device, wherein the fermentation device is a fermenter with a stirring device.

[0177] The fermenter (1) is prepared for operation by washing with 1% baking soda solution, rinsing with tap water to remove soda residues and steam sterilization at 121°C for 40 min using a steam generator (not shown in the diagram). A similar preparation procedure is used for the other elements of the process flow diagram. Steam generator (not shown in the diagram) for automatic sterilization for all sections of the system (vessel equipment, pipeline sections) by supplying steam through tap-ins to pipelines, preferably installed near the shut-off valves. The recommended treatment mode when using both a steam generator and a laboratory autoclave is 121 °C for 40 minutes.

[0178] The substrate is prepared in an amount sufficient to fill the fermenter (1) to 70-75% of its full volume and sterilized by autoclaving at 121°C for 40 min.

[0179] For the technological process, water is used that meets the requirements for drinking water. The water supplied to the fermenter (1), the storage tank (5) and for direct (feeding from above) and reverse (feeding from below) washing of the bacterial filter (6) with the return of solid particles to the fermenter is prepared by autoclaving at 121°C for 20 minutes and cooling, oxygen saturation in the aeration device (2).

[0180] The subculture (inoculum) is prepared in a microbiological laboratory in the form of a suspension of bacterial cells and endospores obtained as a result of deep cultivation of the producer cells in a suitable liquid medium or as a result of washing the culture obtained on an agarized medium with sterile water.

[0181] Filling of the fermenter and fermentation are carried out in compliance with the aseptic conditions.

[0182] Fermentation can be realized in a periodic and continuous mode. The sterile substrate is loaded from above into the sterile fermenter (1) and filled to 70- 75% of the total volume. Further, sterile oxygen-saturated water is supplied to the fermenter (1) in an amount at least providing free sliding of the raw material particles relative to each other, but not more than the content level in the fermenter of 75% of the total volume. The content of the fermenter is stirred for 1-5 minutes using a stirring device (stirrer), then the subculture is introduced. During cultivation, the temperature is maintained in the range of 25-65 °C, pH 4.0- 11.0, the stirring speed is 5-60 rpm, the water saturated with oxygen and / or aerating gas bubbles (from above or below) is supplied in an amount sufficient to provide the producer cells with oxygen, and an equivalent amount of liquid is taken from the apparatus (from below). Sterile acid or alkali solutions can be added to adjust the pH during cultivation.

[0183] The extracted liquid contains particles of raw materials colonized by producer cells and free producer cells. Further, this liquid enters the filter (6) with a pore diameter of 100 pm (=0.1 mm), on which the particles of raw materials are separated in the form of a precipitate and returned to the fermenter (1), and the filtrate flows into the filtrate storage tank (5). To maintain the producer cells in the active state, a water saturated with dissolved oxygen in an amount sufficient to provide the cells with oxygen is allowed to be supplied in this tank (5). To increase the protein content in the producer's cells, sources of nutrients in the form of mineral salts are introduced into the culture medium. Preferably, the application is in the form of a concentrated solution (1 : 100).

[0184] The sterile water tank (9) serves to supplement, if necessary, the sufficient liquid level of the storage tank (5). At a distance of 14 from the bottom of the storage tank (5), an outlet is provided to the suction line of the pump, for heating or cooling to the heat exchanger-heater (16) or the heat exchanger-cooler (17) and further to the gas saturation device (2).

[0185] Atmospheric air is supplied to the gas saturation device (2) by the compressor (18) through the fine filter (19). Optionally, oxygen from the compressed oxygen cylinder (20) can be introduced into the air flow upstream of the fine filter (19).

[0186] The liquid saturated with dissolved oxygen then enters the fermenter (1) and is used to supply the producer culture with air oxygen.

[0187] During fermentation, the following parameters are monitored in the fermenter (1) and the filtrate storage tank (5):

[0188] 1 - temperature,

[0189] 2 - pH,

[0190] 3 - oxygen at the inlet to the fermenter (1) and at the outlet / carbon dioxide dissolved and in the gas phase,

[0191] 4 - optical density at a wavelength of 600 nm, 5 - redox potential;

[0192] 6 - humidity;

[0193] 7. - density;

[0194] 8. - liquid level in the storage tank (5);

[0195] In all examples presented, it is also possible that said aerating liquid or mixture of liquids acts as an extractant to extract the resulting product from the substrate and discharge it from the fermentation device. That is, the fermentation device and the system for solid-phase aerobic fermentation are further configured to remove (extract) and discharge the resulting product from the substrate and, accordingly, from the fermentation device, wherein the extraction and discharging the resulting product is carried out with said liquid or mixture of liquids, which is or at least one of which is saturated with bubbles of aerating gas of micro-, and / or nano-, and / or smaller size, and / or contains oxygen dissolved therein. The small bubbles contained in the liquid or mixture of liquids, in addition to the function of aeration and temperature regulation, also provide a simultaneous more complete extraction of the resulting product from the substrate. The product can be discharged together with the specified liquid and further, if necessary, purified by any available methods. If necessary, the amount of liquid in the system is supplemented to the required amount.

[0196] This approach allows to increase the amount of products obtained as a result of fermentation. For example, in traditional processes, the product remains in a solid phase-substrate, which is extracted from the fermentation device and further isolated. The present invention allows to simultaneously obtain a solid-phase product and additionally a liquid-phase product.

[0197] For example, soluble proteins, e.g., soluble fractions of a single-cell protein, can be produced and immediately extracted from the substrate by an aerating liquid. The resulting product is removed from the fermentation device together with the waste liquid. An example of the discharge is shown in Figs. 1-3 with position 10’. If necessary, the product is pasteurized and thus, for example, a fodder product for young animals is obtained. In this way, enzymes can also be obtained in liquid form, which can be used without further extraction. For example, these may be any soluble enzymes, in particular proteases, lipases, cellulases, mannases, laccases, phenol oxidases, peroxidases, pectinases, hemicellulases, amylases, chitinases, and others.

[0198] In a conventional method without the use of an aerating liquid, the product is only in the solid phase, and according to the present invention, it is possible to obtain products both in the solid and liquid phase, which increases the number of possible products, simplifies the processes for their production, reduces the cost and time for obtaining biotechnological products. To enhance the extraction (recover) of the product, the aerating liquid can further comprise salts or salt derivatives for the production of salt-soluble products, alcohols for the production of alcohol-soluble products, acid or alkali for the production of acid- and alkali-soluble products. The liquid can also additionally contain, for example, esters, polyesters, ammonia, organic solvents and other additives. A single liquid or a mixture of liquids can be used. The composition of said liquid or mixture of liquids may vary during the fermentation process and depends on the products obtained. The composition and amount of these additional additives are selected in such a way as to enable solid-phase aerobic fermentation.

[0199] As can be seen from the examples given, the achievement of the technical effect, which consists in:

[0200] - more uniform aeration (distribution of the aerating gas and, accordingly, oxygen dissolved in the liquid);

[0201] - uniform regulation of temperature and humidity;

[0202] - acceleration of oxidative processes in the fermentation process, which accelerates the entire fermentation process as a whole;

[0203] - preventing bonding and drying of the substrate;

[0204] - eliminating the need to reduce the thickness of substrates;

[0205] - and, accordingly, reducing the size of the fermentation device and production areas with the same performance or increasing the performance of the fermentation device and the system as a whole;

[0206] - the possibility of supplying microorganisms and / or enzymes together with an aerating liquid;

[0207] - increasing the number of possible products, the possibility of obtaining products not only in the solid but also in the liquid phase, does not depend on a type of fermentation device.

[0208] The claimed method of solid-phase aerobic fermentation includes performing solid-phase aerobic fermentation using the claimed system in accordance with all possible variants thereof.

[0209] Thus, for the implementation of the method for solid-phase aerobic fermentation:

[0210] - a substrate and necessary microorganisms and / or enzymes are introduced into the fermentation device;

[0211] - fermentation is carried out to obtain a product, providing aeration in the fermentation device using the claimed aeration system using one or more hydraulic systems; - product and waste substances (liquid, gas, etc.) are discharged, wherein the aeration is performed by one or more hydraulic systems, at least one of which provides, in the fermentation device, presence of a liquid or a mixture of liquids, which or at least one of which is saturated with dissolved oxygen and / or bubbles of aerating gas of micro-, and / or nano-, and / or smaller size.

[0212] That is, when the method is implemented in the fermentation device, the aeration in the fermentation device is provided with an aerating liquid or a mixture of liquids, which or at least one of which is saturated with bubbles of aerating gas of micro-, and / or nano-, and / or smaller size, and / or contains oxygen dissolved therein.

[0213] As can be seen from the examples presented, the implementation of the claimed method ensures the achievement of all the above effects. That is, the following is provided:

[0214] - more uniform aeration (distribution of aerating gas);

[0215] - uniform regulation of temperature and humidity;

[0216] - acceleration of oxidative processes in the fermentation process, which accelerates the entire fermentation process as a whole;

[0217] - preventing bonding and drying of the substrate;

[0218] - elimination of the need to reduce the thickness of substrates;

[0219] - accordingly, a reduction in the size of the fermentation device and production areas with the same performance or an increase in the performance of the fermentation device and the system as a whole are achieved;

[0220] - the possibility of supplying microorganisms and / or enzymes together with an aerating liquid;

[0221] - an increase in the number of possible products, the possibility of obtaining products not only in the solid but also in the liquid phase.

[0222] Various biotechnological products can be obtained using the claimed system and method for solid-phase aerobic fermentation, for example, but not limited to single-cell protein, polyhydroxyalkanoates, including polyhydroxybutyrate, cellulose, fermentable sugars, biofuels, antibiotics, polyunsaturated fatty acids (PUFAs), enzymes, organic acids, fermented foods, alcoholic beverages, biopesticides, microbial polysaccharides, flavors, biosurfactants, phenolic compounds, biofertilizers, and others.

[0223] Thus, the present invention also relates to a process for producing a biotechnological product, comprising: - feeding into a fermentation device included in the claimed system for solid-phase aerobic fermentation a substrate for producing said product and substrate-treating microorganisms and / or enzymes;

[0224] - performing solid-phase aerobic fermentation in the claimed system for solid-phase aerobic fermentation providing of regulation of conditions in the fermentation device, including by aeration and temperature regulation;

[0225] - extraction and discharge of the resulting product, as well as discharge of waste substances; wherein the aeration in the fermentation device is provided by a liquid or a mixture of liquids, which or at least one of which is saturated with dissolved oxygen and / or bubbles of aerating gas of micro-, and / or nano-, and / or smaller size.

[0226] In one of the embodiments, the extraction and / or discharge of the resulting product is optionally carried out by said liquid or mixture of liquids.

[0227] Embodiments of said method include those described above relating to a system for solidphase aerobic fermentation. Exemplary embodiments of the method are described in detail above when describing the operation of a system for solid-phase aerobic fermentation.

[0228] As can be seen from the examples presented, the implementation of the claimed method ensures the achievement of all the above effects. That is, the following is provided:

[0229] - more uniform aeration (distribution of aerating gas);

[0230] - uniform regulation of temperature and humidity;

[0231] - acceleration of oxidative processes in the fermentation process, which accelerates the entire fermentation process as a whole;

[0232] - preventing bonding and drying of the substrate;

[0233] - elimination of the need to reduce the thickness of substrates;

[0234] - accordingly, a reduction in the size of the fermentation device and production areas with the same performance or an increase in the performance of the fermentation device and the system as a whole are achieved;

[0235] - the possibility of supplying microorganisms and / or enzymes together with an aerating liquid;

[0236] - an increase in the number of possible products, the possibility of obtaining products not only in the solid but also in the liquid phase. It should be noted that from the examples provided, it is understood that the system and methods of the invention can optionally comprise one or more of the additional features recited herein.

[0237] In addition, it should be noted that those skilled in the art, after familiarizing himself with the present invention, can assume possible modifications and / or changes in the claimed system and method of solid-phase aerobic aeration, as well as the method for producing a biotechnological product. Such modifications and / or alterations are encompassed herein.

Claims

AMENDED CLAIMS received by the International Bureau on 27 April 20261. A system for solid phase aerobic fermentation, the system comprising:- a fermentation device configured to supply a solid substrate and substrate-treating microorganisms and / or enzymes therein and to discharge a resulting product and waste substances;- one or more hydraulic systems configured to provide, in the fermentation device, presence of a liquid or a mixture of liquids, which or at least one of which is saturated with bubbles of aerating gas of micro-, and / or nano-, and / or smaller size, and / or dissolved oxygen, wherein said one or more hydraulic systems comprise:- an aeration device configured to provide saturation of the liquid with bubbles of the aerating gas of micro-, and / or nano-, and / or smaller size, and / or dissolved oxygen before feeding it into the solid substrate for fermentation;- a filtration device configured to separate a solid phase from a circulating liquid supplied to said aeration device and then back to the substrate.

2. The system of claim 1, wherein said aeration device is arranged outside the fermentation device.

3. The system of claim 1, wherein said aeration device is arranged inside the fermentation device.

4. The system as claimed in any one of claims 1 to 3, wherein said liquid is water.

5. The system of claim 4, wherein said liquid further comprises one or more additives selected from the group consisting of salts, derivatives of salts, alcohols, acids and alkalis, esters, polyesters, ammonia and organic solvents.

6. The system as claimed in any one of claims 1-5, wherein the aerating gas is a gas selected from the group consisting of oxygen, air, or a gas mixture comprising oxygen.

7. The system as claimed in any one of claims 1-6, wherein the system is configured to supply said microorganisms and / or enzymes together with said liquid or mixture of liquids to the fermentation device.

8. The system as claimed in any one of claims 1-7, further comprising a temperature regulation system.

9. The system as claimed in any one of claims 1-8, wherein a cooler and / or a heater is additionally integrated into said one or more hydraulic systems.

10. The system as claimed in any one of claims 1-9, wherein the fermentation device is a fermenter selected from the group consisting of a. plate fermenter, a packed fermenter, a multilevel fermenter of any design, a tray-type fermenter, a compacted bed fermenter, a rotating drum-type fermenter, a rocking solid-phase fermenter, a container-type fermenter provided with a stirring device, and a fluidized bed fermenter.

11. The system as claimed in any one of claims 1-10, wherein the resulting product is extracted and discharged using said liquid or mixture of liquids.

12. The system of claim 11, further comprising a system for supplementing said liquid or mixture of liquids after discharging the resulting product.

13. The system as claimed in any one of claims 1-12, wherein a discharge device is connected to the fermentation device to discharge the resulting product.

14. The system as claimed in any one of claims 1-13, further comprising a drainage device for discharging the circulating liquid.

15. The system of claim 14, wherein said drainage device is connected to said hydraulic system to circulate said liquid in the system for solid-phase aerobic fermentation.

16. The system as claimed in any of claims 1-15, wherein the filtration device is disposed outside the fermentation device and coupled to the fermentation device.

17. The system as claimed in any one of claims 1-16, further comprising a circulating liquid storage tank coupled to said fermentation device via the hydraulic system.

18. The system as claimed in any one of claims 1-17, further comprising an exhaust gas discharge system to discharge exhaust gas from the fermentation device.

19. A method for solid-phase aerobic fermentation, the method including performing solidphase aerobic fermentation using the system of any one of claims 1-18 to saturate a liquid or a mixture of liquids in a fermentation device with bubbles of aerating gas of micro, and / or nano-, and / or smaller size and / or dissolved oxygen.

20. A method for producinga biotechnological product, the method including:- feeding into a fermentation device included in the system of any of claims 1-18 a solid substrate for producing said product and substrate-treating microorganisms and / or enzymes;- performing solid-phase aerobic fermentation using the system of any one of claims 1 to 18, providing saturation of a liquid or a mixture of liquids in a fermentation device with bubbles of aerating gas of micro-, and / or nano-, and / or smaller size, and / or dissolved oxygen;- extracting and discharging a resulting product, and discharging waste substances.

21. The method of claim 20, wherein the resulting product is extracted and discharged by said liquid or mixture of liquids.

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