HU203288B - Apparatus for carrying out biocatalytic processes by means of biocatalyzer of solid phase - Google Patents

Abstract

The reaction space (15, 75) of the apparatus where reaction between the liquid phase and solid granular biocatalyst takes place is separated from the space that serves for intensive contact between the gas - generally oxygen - and the liquid phase. The latter space is the interior of a circulation pipe (50, 85). One end of the circulation pipe (50) leads out from the reaction space (15, 75) and the other end leads into it. The circulation pipe (15, 75) is divided by a butt-in pump (38, 86) into suction branch (37, 87) and pressure branch (39, 88). In the direction of the medium-flow (g) proceeding from the pump (38, 86) towards the point of leading the pressure branch into the reaction space, successively first a flow-accelerating element (49, 89) is built into the pressure branch (39, 88), then a gas food pipe (48, 92) is leading in, and thereafter gas dispersing elements (45, 46) are built in. Filter 21 prevents loss of catalyst. <IMAGE>

Description

The present invention relates to an apparatus for carrying out biocatalytic processes using a solid phase biocatalyst.

One of the fastest growing areas of biotechnology is the use of solid phase biocatalysts. Solid phase biocatalysts are so-called. It is created by an immobilization (immobilization) technique, the essence of which is to locate catalytically active biological materials in the reactor space in order to prevent them from entering the flowing phase. (J. Chem. Techn. Biotechn., 1984, 34, 127) Solid phase biocatalysts can be prepared by immobilizing isolated enzymes, dead cells or cellular constituents, or living cells (microbes, plant or animal cells). In specific cases, the most appropriate of these options will be selected on the basis of economic and / or stability considerations.

The use of solid phase biocatalysts has significant economic advantages over conventional biocatalytic processes, namely:

- enabling continuous or quasi-continuous technology;

- increase in volumetric productivity;

- reduce operational (eg wastewater) and product recovery costs.

Another advantage of producing secondary metabolites by fixed cellular fermentation is that the continuous mode of operation reduces the viscosity of the fermentlc, thereby improving oxygen delivery; certain overgrowth problems can be avoided and the loss of a chemically unstable product is reduced. (Biotechnol, Bioeng. 1983.25,2399-2411.)

A variety of reactor types have been developed to perform solid phase biocatalytic operations. The most common in industrial size is the so-called. for example, glucose isomerization, amino acid resolution, or ethanol fermentation. (Chem. Eng. Sci. 1985.40, 1321-1354.) Industrial-size fluidization reactors are used in the treatment of wastewater with fixed biomass (J. Walter Pllut, Control Ced. 1977,49,816). There are also examples of industrial applications of membrane reactors (Chem. Ing. Tech. 1988, 60, 16-23).

There are many types of pilot plant and laboratory-sized reactors. The most commonly used fixed bed and fluidization reactors have been implemented in multi-unit form (horizontal and vertical cascade design). Among other things, in order to prevent process CO2 enrichment (J. Chem. Tech. Biotech. 1987, 39, 75-84) and to reduce axial substrate solution re-mixing (Proc. 4th Eur. Congr. Biotech. Vol. 1, 1987, 273276). The so-called. external and internal loop reactors are mainly used to implement fixed cellular fermentation processes. (Biotech. Bioerg. 1987, 30, 498-504; J. Chem. Tech. Biotechnol. 1986, 36, 415-426.)

Although many reactor types exist to perform solid phase biocatalysis, as noted above, quite a number of reactor engineering problems remain unresolved, primarily for processes where one of the substrates is a gas phase (usually oxygen). Gaseous-liquid transfer is of great importance in most conventional biocatalytic systems, for example, air-demanding microbes require the use of so-called gaseous effluents. oxygen transfer rate (OTR) to 150 mmol / dm ³ / h. In addition, to utilize the useful volume of the device, it is desirable that said OTR value be achieved at a moderately high ventilation rate (1-1.5 wp). Because the presence of carrier particles for immobilizing bioactive materials in most cases impairs gas-liquid transfer, the carrier particles enhance agglomeration of gas bubbles (Proc. 4th Eur. Congr. Biotech. 1987, Vol. 1.101-104) - with three conventional gas distribution fluidizers. in bubble column or loop systems, up to a fraction of the aforementioned CTR value is achieved (Chem. Eng. Sci. 1987, 42, 543-553; Biotech. Bioeng. 1988, 32,677-688). It is difficult to intensify gas distribution because of the relatively low mechanical stability of most carriers, thus avoiding the application of significant forces in the catalytic zone, e.g. it is also unable to withstand the forces of liquid or liquid-gas rays.

In some fermentation processes, which are carried out with microbial, plant, and animal cell cultures that are particularly susceptible to infection, the additional difficulty is to transfer the normally gel-encapsulated cells to the fermentor under sterile conditions. Namely, the structure of the fermenter generally does not allow, or only by a very complicated process, the formation of sterile catalyst particles in situ.

SUMMARY OF THE INVENTION It is an object of the present invention to provide an apparatus for carrying out biocatalytic processes using a solid phase biocatalyst that provides intense gas-liquid material transfer which does not adversely affect catalyst carrier particles of low mechanical stability and which combines gel-type carrier particles with specific sterility requirements. training equipment and bioreactor functions.

The present invention is based on the following findings:

When the part of the apparatus providing the high specific gas-liquid transfer surface is separated from the device part containing the solid phase biocatalyst, the turbulent forces promoting gas dispersion do not burden the catalyst particles and consequently do not damage them. It is further recognized that the gas-liquid phase mixing can be significantly intensified by substantially increasing the gas-liquid phase mixing by circulating in a dispersion element containing only a volume of the catalytic zone of the apparatus. a relatively small proportion, preferably not more than 10%. We have further discovered that the gas-liquid is transported

By selecting the reaction direction and velocity of the gas-liquid emulsion formed in the equipment part of the serving unit, and the structure (design) of the reaction space, the latter can ensure uniform circulation of the catalyst particles and increase the residence time of the gas bubbles. design, it can be adapted to coagulate the droplets formed by the drip forming unit on the device cover prior to biocatalysis containing a biocatalyst (e.g., a cell or spore suspension) in the reaction zone, thereby enabling the inoculation of the biocatalyst particles in situ. Based on these findings, the object of the present invention has been accomplished with an apparatus having a liner It has a reaction space, means for feeding the catalyst and a fluid and gas into the reaction space, means for discharging the product obtained by biocatalysis and the process gas, and a circulation pump, characterized in that the two ends of the reaction space an overflow recirculation line at the locations above which has a filter element for retaining solid catalyst supports in the reactor space prior to the exit from the reactor space for the discharge of the treated medium; the recirculation pump is integrated into the recirculation line, which divides into a pressure branch and a suction branch, and a flow accelerator element is inserted in the pressure branch after the pump, followed by a gas inlet pipe and then a pressure branch gas dispenser ).

In a preferred embodiment, a phase separation chamber having a gas outlet stump is connected to the area extending above the area, and the suction branch of the circulation line extends from the lower part of this chamber to which the liner in the reaction space extends.

According to another advantageous feature of the invention, the cylindrical reactor space is also enclosed at the bottom by a cylindrical substrate having a channel tangentially connected to the opening of the collector opening in the pressure branch of the circulation line. It is expedient that the upper surface of my support is provided with an oblique curve from the bottom of the pressure collector opening to the top of this opening, which is a sand-threaded surface, and that the outlet provided with a valve outside the support and covered by a filter element from the reactor compartment above, a conical flow modifying element extending downwardly into the inside of the liner is mounted.

It is also preferred that the nozzle for supplying the fluid to be treated is inserted into the pressure line of the circulation line between the pump and the flow accelerator and the nozzle for discharging the product obtained as a result of the biocatalytic process from the suction line of the circulation line.

A further feature of the invention is that the device has a nozzle in the phase separation chamber for feeding the solid biocatalyst material into the reaction space.

In a preferred embodiment of the apparatus, there is provided a drip-forming head for feeding into the phase separation chamber a droplet of a barrel containing biocatalyst material, such as bacteria, which is connected to a mixing vessel for receiving the solids. In this case, it is preferable to have a valve inserted between the dropper head and the container, and if the dropper head has drip forming elements directed towards the inside of the phase separation chamber.

If the density of the solid catalyst particles is lower than that of the liquid phase involved in the biocatalytic process, it is expedient to use an apparatus embodiment in which the bottom taper, the upper cylindrical reaction tube, has a larger diameter upper cylindrical portion and an intermediate taper. having a smaller diameter lower cylindrical portion and having a conical chamber downstream of the continuation of the conical lower portion of the reaction space to which the suction branch of the circulation line is connected and the pressure section of the circulation line being tangentially introduced into the lower region of the upper a filter element is provided in front of the suction branch opening out of the chamber. In this case, it is advantageous for the auxiliary gas flow nozzle to be guided through the wall defining the conical chamber towards the opening of the lower cylindrical portion of the liner extending into the chamber.

According to a further feature of the invention, the length of the pressure branch of the circulation pipe is several times greater than the length of the suction branch. As a result, the number of dispersants can be increased and thus the gas-liquid transfer rate can be increased.

In a further embodiment, the apparatus has a gas dispersion element (s) having a configurator, a diffuser and a gap between them. Such gas dispersing elements are primarily incorporated in the vertical or proximal section or sections of the pressure line of the circulation line. Gas dispersion elements having a rod and a spiral member surrounding it are introduced into the horizontal or near-horizontal section or sections of the ram.

The invention will now be described in more detail with reference to the accompanying drawings, which show some preferred embodiments and structural details of the apparatus. In the drawings la. FIG. 4A is a schematic, partially sectional view of an embodiment of the apparatus;

lb. 1a. Figure 6 is a sectional view taken along line A τA;

it's down. Fig. 2A is a vertical sectional view of a larger scale with a drop forming element;

see ld. Fig. 4A is a larger scale axonometric drawing of a dispersing element;

1 e. Fig. 2A is a front view of another embodiment of a dispersing element;

2a. FIG

203288Β is a schematic vertical sectional view;

2b. three 2a. FIG. 3B illustrates a way of connecting the apparatus of FIG.

3a. FIG. 4a is a vertical sectional view of a further embodiment of the apparatus;

3b. 3a. Fig. 6 is a sectional view taken along the line F-F in Fig. 4A.

1a to 1e. The apparatus shown in FIGS. 1 to 4 is a form suitable for encapsulating airborne microorganisms or their spores in a gel and for carrying out quasi-continuous fermentation with microorganisms (spores) encapsulated in the gel (e.g., fixing Brevibacterium flavum on Carrageenan and using it for L-tryptophan biosynthesis).

The main parts of the equipment are the boy drum entry unit I located below; an autorecirculation reactor unit H above this; a phase III separation unit; a sputtering unit IV attached to its lid and a gas-dispersing unit V for forced circulation, which interconnect the fluid introduction unit I and the phase separation unit III bypassing the reaction unit Π.

The fluid delivery unit I has a cylindrical base member 1 which is tangentially exterior to the male dumb feeding nipple 2 which, as shown in FIGS. As shown in FIGS. 1 to 4, it continues in the tangential channel 3 formed in my sub-base 1. The lower bounding surface 5 of the space 4 (Fig. 1a) (see also Fig. 1b) rises in a forehead with respect to the longitudinal vertical vertical axis of the device, starting from the lower boundary edge of the vertical entry plane ending.

A cone-shaped flow modifying element 8 is fixed at a distance, with its apex, on the vertical central geometric axis x above my support 1, which is secured to my support 1 by the screw 7. A cylindrical disk-like longitudinal filter element 10 is located between the base of the flow modifier 8 and the front face of the circular central portion 9 (see also Fig. 1b) of the base member 1. Preferably, the cone angle of the flow modifier 8 is between 50 and 70 ° and is proportional to the diameter dk of its base board as (l-1, 2): 2.

Incorporated into the cylindrical base 1 of the fluid delivery unit I is a two-pass vertical 11,12 truncated. All stubs are used to remove the liquid phase in the catalyst carrier-free form (as the filter element 10 will retain these particles as will be seen), while stub 12 is intended to completely remove the liquid-solid suspension from the apparatus. The stubs 11,12 can be opened and closed by means of the valves 13 and 14, respectively, which are located below (below) the base 1.

The autocirculation reaction unit van has a reaction space 15 bounded by a cylindrical housing 15a. The inside diameter DI of the housing 15a is the same as the diameter Da of my cylindrical base 1. The housing 15a is spaced apart by a spacer housing 164 so that there is a narrow annular thermostatic space 16 between the housing and this housing. Inside the reaction space 15, a liner 17 having a diameter d i is centrally located, in other words, the housing 15a, the bottom thermostatic jacket and the liner 17 are concentric to the vertical geometric center axis. Said diameter D i is suitably proportioned to d ι as 2: 1. The lower edge of the liner 17 extends below the tip of the cone-shaped flow modifier 8, i.e. the liner partially encircles the upper part of the flow modifier. The vertical distance Ai between the lower edge of the liner 17 and the circle of diameter dk cut out by the vertical projection of the liner is selected such that the surface of the imaginary cylindrical surface between this circle and the lower edge of the liner is close to the cross-sectional area of the liner be the same ·

The phase separation unit ΙΠ has a chamber 18 having an upwardly extending truncated conical portion 19 and a cylindrical upper portion 20 extending therethrough. The lower portion 19 is connected to the reaction space 15. The upper portion 20 preferably has a diameter >2 of about 1.5 to 2 times the Dt diameter of the reaction space 15. The height β2 of the phase III separation unit is about 1/3 to 1/4 of the height of the reaction space 15. The conical lower portion 19 of the phase separation unit AIII is preferably about one third of the height of the upper portion 20. The height L of the fluid introduction unit I, the reaction unit és and the phase separation unit vagy, i.e. the reactor itself, should preferably not exceed 5-6 times the diameter DI of the reaction space.

The mantle-shaped lower portion 19 is enclosed within a sheath enclosing a filter element 21, preferably made of mesh fabric, which is thus of a truncated cone shape. The aperture size of the filter element 21 is selected such that it impedes the gas from escaping from the liquid in the direction of the lower resistance inside the chamber. From the lower portion 19, a drainage outlet 22 and a sampling outlet 23 protrude. The vertical insertion tube 17 extends into the interior of the lower portion 19 of the phase separation unit III, about 0.01-0.02 m above the plane defined by the center lines of the stubs 23 and 22 (preferably the stubs 22 and 23 are horizontal and centered in the same horizontal plane). are located).

The upper 20 of the AIII phase separation unit is closed by 24 sheets (sheets). Inside the lid 24 is mounted symmetrically to the vertical geometric central axis x, the cylindrical drop forming head 25 of the spool forming unit IV. Thus, the dripping head and the lower part 19 and the upper part 20 of the phase separation unit ΙΠ are concentric with the central vertical axis x. The dripping head 25 may also be connected by a valve 26 and a nozzle 36, and the dripping head IV may be surrounded by a thermostatic jacket (not shown) and preferably have a volume of about one tenth the volume of the container 27 (tumbler).

the lower conical, top cylindrical container 27, which is concentric with the vertical axis geometric axis x, is closed by a lid 27a, and from the container through this lid 30 injection ports 31 filling ports (filling port),

-432 203288Β and 34 stub outlets with an overpressure line (not shown) connected to the latter. The casing of the container 27 is designated 27b; this mantle is spaced apart by a thermostatic mantle 27c which defines a narrow thermostatic space 27d with the mantle 27b. The volume of the container 27 is preferably about 50% of the volume of the reaction space 15 of the H unit. The mixing propeller 33 extends into the reservoir 27, and in the lower part of its frustoconical shape, the driving rod of which is in the center of the vertical axis geometry and driven by the motor A.

At the base of the drip-forming head 25 there are incorporated drip-forming elements 35, one of which is in section and in larger scale. FIG. A filter 36 is formed in the drop head 25 above the base plate, preferably of a mesh size of about 0.1 mm. The approximate number of drop forming elements 35 circularly distributed on the motherboard can be determined by the following empirical formula:

/ 100.Di [mA ² Irish

Depending on the nature of the sol, the dt hole diameter (dt) of the droplets 35 is 0.3-0.5 mm.

The forced gas dispersion unit V of the apparatus is provided with a circulating line, all of which has a 50 end, one end of which is connected to the 2 port of the fluid inlet unit I and the other end to the liquid outlet port 22 of the phase separation unit. The circulation line 50 (having a maximum diameter preferably about 1/4 to 1/6 of the diameter DI of the reaction space 15) has two sections: the suction branch 37 extends from the fluid outlet 22 to the variable flow circulation pump 38 and the discharge section 39 from pump 38 fluid inlet extends to 2 stumps. The location of the pump 38 in the circulation pipe 50 is selected so as to be as close as possible to the nozzle 22, i.e., the longitudinal extension 39 of the circulation pipe 50 is as long as possible. The suction branch and the pressure branch 39 can be short-circuited by means of a valve 41 in the by-pass 40.

The flow accelerator element 42, which is embedded in the ram 39, is preferably a configurator (shown in Fig. 1a by breaking the tube wall) to accelerate the flow of liquid at high speed. A nozzle 43 is provided for smoothing the gas flow, which is provided in the conduit 42, after the confluent 42, for the flow of vapor required for biochemical reaction in the downstream direction of the confluent 42 in order to introduce steam, generally oxygen (air) p located. Dispersing elements 45, also visible by bursting of the tube, are incorporated in the section of the ram 39 between the pipe 44 and the nipple 2, one of which is illustrated in larger scale in axonometric drawing. 1 to 3, which provide intensive gas-liquid mixing and dispersion of the gas phase into tiny bubbles. The la. and see. The dispersion element 45 shown in Fig. 1b has a centrally tapered slit having a confuser at its front end, a diffuser, and a diffuser 45a, as shown in Fig. 1d, on the one hand and a bifurcator 45 on the other hand. 45 c) and preferably can be installed in a vertical pipe section in a top-down flow of gas-liquid. That's down. The dispersion member 46 shown in FIG. 4B is formed by a rod 46a and a spiral member 46b which surrounds it 10; it is desirable to incorporate this dispersing element 46 in a horizontal tube section. The 45.46 dispersant elements are interchangeable and their typical dimensions are determined by the air requirement of the particular cell culture and the reactor size.

The forced-circulation gas dispersion unit V also includes a strainer 48 for inlet medium (arrow o, Fig. 1a) and a strainer 49 for product removal (/ arrow, fig. La), which, by means of their associated valves 47a, 47b zárhatóknyithatók.

The apparatus described above operates as follows:

The container 27 of the AIV droplet forming unit is filled with a suitable concentration of aqueous Na-alginate salt through the filling port 31, with the valve 26 closed, and the filling port is sealed and the entire reactor (bioreactor) sterilized by steam sterilization. In order to carry out this operation, steam is introduced through the gas dispersion unit V pipe 44, which is discharged from the system through the outlet port 28 from the chamber 18 of the phase separation unit. Also, the container 27 of the AIV droplet forming unit is sterilized with the 120 'C steam introduced into the thermostatic space 27d while stirring the solution in the container 27 to improve heat transfer. After completion of the sterilization operation, the reactor compartment and the circulation conduit 50 are dewatered through two inlets 12 of fluid inlet unit I, and the reactor is cooled to room temperature with cooling water circulating in thermostatic space 16a and cooling water circulating in thermostatic space 27d.

In the next step, the reaction space 15 and part of the chamber 18 are filled up to the height of the upper rim of the liner 17 by means of a pump 38 with sterile CaCl 2 precipitant solution fed into the system via the 48 port.

The solution of the solution in the tank 27 is inoculated with the Brevibacterium flavum cell suspension through the inoculum 30, and the inoculated solution is homogenized with the mixing propeller until uniform cell concentration is reached. Subsequently, by opening valve 26, the resulting ho55 mogenized sol-cell suspension is transferred from the container 27 to the cylindrical droplet forming head 25 of the solder forming unit ΙΠ. Here, the filter 36 captures coarse particles that may be present in the suspension, and the suspension itself flows into the droplets 35. The suspension flows down through the nozzle 34 under the action of sterile pressurized air fed into the container 27; the flow rate can be controlled by varying the air pressure.

The maximum of 25 drop-forming units

-5HU 203288Β w = 0.4. is suitable for dispensing a suspension of cell volume / volume (n = 35 dm / h) (number of droplets in the formula: na 35). According to the formula, for example, a 0.2 m diameter reactor 5 achieves a feed rate of ~ 20 dm ^3-1 h, which approximates the performance of the highest-performance known vibration word dropper used in a large laboratory size, but does not, in contrast, .

The droplets of the cell / sol suspension drop from the drop head 25 into the CaCl2 precipitator solution 15 in the reaction space and solidify therein and settle through the liner 17 to the bottom of the reactor. During the formation of the gel granules, the tube 44 and the nozzle 43 embedded therein have an approx. A sterile air stream of volume velocity corresponding to a linear velocity relative to a 1 cm / s blank plant cross-section is fed to the reactor via the circulation line 50, through port 20 and through channel 3 (Figure 1b). As a result of the air jet tangentially introduced and forced by a helical upward flow through the face-threaded surface 5 (Fig. B), the gel particles deposited on the substrate 1 together with the CaCl2 solution rise in the cylindrical space between the wall 15a and the outer surface of the liner 17. Upon reaching the top edge of the liner 17 and falling through it, they sink again, thereby circulating the gel particles in the reaction space 15. The flow generated by the sterile air stream 30 improves the solid-liquid transfer, i.e. the Ca ² * ions, into the gel particles so that they gel in their entire cross-section in a shorter time than the known processes and consequently the gel-sealed cells are less exposed. CaCl2 solution may cause adverse effects.

After completion of the gel particle formation, valve 26 is closed and the overpressure exerted through the nozzle 34 to the interior of the container 27 is opened, the valve 13 is opened, and the CaCl2 solution is removed from the device via the nozzle 11 and the autocirculation valve 41 is removed. with the pump open, the pump 38 begins to add sterile medium to the reactor. (If necessary, the gel particles are rinsed with sterile distilled water prior to the addition of the medium.) The medium is passed through the 48 nozzle into the circulation line 50. As soon as the level of the nutrient solution reaches the upper edge of the liner 17, the nutrient solution is discontinued and the valve 41 is closed, however, the pump 38 is used to circulate the nutrient solution further. A sterile filtered air stream is fed through tube 44 and nozzle 43 (the latter serving to balance the gas flow) to the pressure branch 39 (arrow p, Fig. 1a) of the circulation line 50 with a flow rate of 1 to 1.5 watt of bacterial culture. ventilation rate. In the case of air-demanding microorganisms, it is desirable that oxygen transfer rate (OTR) should be within the range of 150-250 mmol O2 / dm ³ h. This OTR interval in the apparatus can be achieved by adjusting the volume flow rate of the liquid stream circulated by the pump 38 to 30-80% of the actual volume flow rate of the gas stream (sterile air flow). In the dispersion elements 45 and 46 embedded in the circulation conduit 65 (see also Figures 1 and 2), a very fine gas-liquid dispersion is produced by the high turbulence gas-liquid flow and the material transfer surface is also intensively regenerated. The pressure drop in the dispersing elements 45,46 do not exceed .10 ⁵ Pa (0,81,0) overall.

At the fluid inlet 2 and tangential channel 3 (Fig. 1b), the gas-liquid dispersion enters the substrate 1 of the reactor and, due to the sand-like design of the surface 5, is forced upwardly in a spiral movement. The fluid containing the liquid-lifting gas phase flowing at a rate of 1020 m / s causes the fluid to move in a spiral path in the reaction space 15. The gas bubble, agglomerated during elevation, is more elongated than its liquid phase, but also moves in a helical path, increasing both the average residence time of the bubbles in the reaction space and the resulting shear stresses due to different directional forces that increase the agglomeration of the bubble. The liquid stream captures the catalyst carrier gel particles and rises into the phase III separator unit, above the rim of the liner 17, and through the liner to the back stream of the autocirculation fluid generated by the gas bubbles. The flow modifying element 8 makes the flow of gel particles (solid phase biocatalyst) and liquid phase smoother and also prevents dead space in the apparatus. The liquid flowing down through the liner 17 also entices finely divided gas bubbles,

The gas exits the liquid phase in the phase III separator and exits through the nozzle 28. The liquid stream leaves the phase separation unit via the nozzle 22, but can only flow into the nozzle 22 through the filter 21, which retains the biocatalyst particles (gel particles). From the nozzle 22, the liquid enters the inlet branch 37 of the circulating pipe 50 and from there through the pump 28 to the discharge branch 39.

The progress of the fermentation process and the state of the culture medium can be monitored by analyzing samples taken from the 23 stumps that flow out of the phase III separation unit. In the production stage of the fermentation, the feed medium and overflow (not shown) connected to the stubs 48 and / or 49, or the feed and take-off pumps can also be used for continuous feed and product feed.

When a medium change is required, opening the valve 13 (Fig. 1a) through the all nozzle will drain the fermentation broth and fill the reactor with fresh medium as described in detail above.

After completion of the complete fermentation cycle, the fermentation broth is lowered through the nozzle 12 by opening the valve 14 together with the catalyst carrier gel particles and the equipment is sterilized by steam.

2a, 2b. The embodiment of Figures 1 to 4 is, with the exception of the absence of a barrier forming unit, essentially the same as Ia. Figures

-32EN 203288Β, therefore, the previously used reference numerals are used mutatis mutandis to denote the same equipment parts. 2a, 2b. The apparatus of FIG. 1B is configured in a form suitable for quasi-continuous fermentation with microorganisms fixed on a preformed particulate carrier. A special feature of the process is that anaerobic or low-air gas, but also a by-product (such as carbon dioxide), inhibits fermentation, such as ethanol fermentation with Saccharomyces cells, and is therefore an inert gas (e.g. nitrogen) or inert gas to displace CO2 from the liquid phase. air mixture is used. A further feature of the process is that the product also has an inhibitory effect on fermentation, therefore, to reduce fluid re-mixing, several equipment is operated in a cascade arrangement.

As already indicated, FIG. The fluid inlet unit I of the apparatus of FIG. 5a, the thermostat 51 is provided with a jacket which defines the narrow thermostat space 51a. Above the phase separation unit nem, there is not a solder drop unit, but a pellet feeding strain 51 carrying the chamber 53 through its lid 53, a strain 54 for feeding the cell suspension and a gas (air) outlet 55. The chamber 18 is surrounded by a thermostat jacket 56 that defines the thermostat compartment 56a from the outside. Typical preferred geometric dimensions for this embodiment are: dk: Di-1: 1.7; Di '.di Z 1.8: 1. In this case, the pressure gas circulation gas dispersion unit also provides a connection between the individual members of the cascade arrangement, and the diameter of its 50 circulation lines is about one-fifth of the diameter DI of the reaction space. The flow accelerator 42, preferably a confuser, in the circulating pipe 50 is again used to greatly increase the velocity of the liquid stream, the nozzle 43 is used to smooth the gas flow through the pipe 44, and the dispersing elements 45,46 for intensive gas-liquid mixing and is intended to ensure the dispersion of the gas phase into small bubbles.

The stubs 49 and 48 connected to the valves 47a and 40b, respectively, are provided in FIGS. 4A to the individual members of the cascade arrangement shown in FIG. If the process of FIG. 2a. 2b. Referring to FIG. 2a, the second member of the cascade arrangement illustrates that the stub 48 serves to supply the feed fluid, which is a fluid withdrawal for the first member, preferably performed by the pump 57. Fluid is drawn through the port 49 (Fig. 2a), which is fed to the third member of the cascade, and pump 58 is provided for this operation (Fig. 2b). The pump 59 is the feed means for the entire cascade line. The volumetric velocity of the liquid streams carried by the pumps 57, 58, 59 is preferably adjusted by level control. The cascade elements can also be connected to a system of wires and valves, which allows changing their order during operation. . The Xi-X 3 members (reactors) of the cascade reactor (cascade series) are connected by lines 60, 61.

2a, 2b. The apparatus according to Figure 1B shall function as follows:

1-3 pounds of the cascade series (Fig. 2b) are filled with nutrient solution using pumps 57-59. The valves 47b are then closed and the cascade reactors are steam sterilized according to the generally known sterilization procedure for fermenters. After completion of the sterilization process, the reactors are cooled, the fermentation temperature adjusted, and biocatalyst particles containing fixed Saccharomyces cells, or sterile carrier particles through Saccharomyces cells 52 are introduced into the sterilized growth medium via the 52 the cell suspension is added and the medium is adjusted to the level of the upper rim of the liner 17 (Fig. 2a).

In the next step, a sterile inert gas or inert gas-air mixture is introduced through the nozzle 44 into the K1-K3 members of the cascade reactor at a flow rate sufficient to displace the CO2 generated during the fermentation and to autocirculate the liquid / solid suspension in the reactor space. In order to ensure intense gas-liquid material transfer, the circulation pumps 38 circulate the liquid through cascade members through the dispersing elements 45 and 46 in the circulation lines 50 (see also with reference to Figs. 1a-le). The O2-containing gas exits from the liquid phase in each of 18 cells (reactors) in the chamber 18 of the phase III separation unit and leaves the reactor through the nozzle 55 and the liquid leaves the reactor space 15 through the nozzle 21 (the filter 21 retains the catalyst particles) and returns to the pump 38 at the intake branch 37 of the circulation line 50 and from there to the discharge branch 39.

After reaching a predetermined cell count, pumps 57-59 (Fig. 2b) are used to initiate continuous sterile nutrient feeding and perform continuous product harvesting.

The device according to the invention is shown in FIG. and 3b. The embodiment of Figure 1B is a biochemical reaction catalyzed by a fixed enzyme, one substrate of the enzyme in a liquid phase and the other substrate in a gas phase, which is fixed on a low density, deformable, tiny particulate carrier. An example of such a reaction is the oxidation of glucose to gluconic acid in the presence of a glucose oxidase enzyme fixed to a polyacrylamide bead polymer support.

The main parts of the apparatus are provided with an upwardly expanding truncated cone chamber 63 of the fluid outlet unit VI, from which a fluid outlet section 64 extends, into which a flow section 65 for feeding auxiliary gas flow is inserted. A nozzle 69 provided with a valve 66 is provided for removing fluid in the apparatus in a state free of catalyst particles. The openings of the stumps 64,67 and 69 extending into the chamber 63 are delimited by the same truncated cone-shaped filter element 70, preferably made of mesh, from the interior of the chamber. The aperture size of the filter element 70 is in this case and such

-7H 203288Β is chosen to impart a resistance to the gas phase that increases upward in the reaction space towards a lower resistance. A nozzle 71 directed to the interior of the chamber 63 is connected to the auxiliary gas flow nozzle 65 for smoothing the gas flow. The taper angle of the chamber 63 is preferably 50-70 °, medium. The diameter Sa may be about one-third the diameter of the reactor, and the height b may be about one-fifth of the total height L of the reactor.

VI. A fluid circulation unit is connected from above with an autocirculation reaction unit, having an upwardly expanding truncated conical lower part 73 and a cylindrical upper part 74. The lower part of the A73 is considered to be an extension (expansion) of 63 chambers. The height of the lower part 73 is preferably proportional to the height of the upper part 74 at least 11: 3. The total height E of the compactor (formed by the fluid drainage unit VI and the reaction unit II) is preferably not greater than 1.8-2.5 times the diameter Βi of the reaction space 75 (cylindrical upper part 74). The reactor is provided with a reference numeral 76 in the vertical central geometric axis Xi, which is comprised of three parts: a lower cylindrical section 77 of smaller diameter E1, an upper cylindrical section 78 of larger diameter E3 and an upwardly extending truncated cone 79 The tapered part, the length of which is from bottom to top, is designated by the reference letters h2 and h3. Preferably, ci: c ₂ : c3 = 3: 1: 3 and ei: e ₂ = 1: 4, and e3'T? I - 1: 2. The height H1 of the reaction space 75 is equal to or nearly equal to the total height of the insert tube 76, but the lower end of the lower cylindrical portion 77 of the insert extends into the interior of the chamber 63 and is spaced approximately Ba / 3 above the tip of the nozzle 71. . The liquid discharge unit VI and the reaction unit Π are surrounded by a thermostatic jacket 80a defining a thermostatic space 80.

In the upper cylindrical portion 78 of the liner 76, about a lower quarter of its height C3 extends tangentially to the fluid-fed nozzle 81 which passes through the wall of the container 72 (and the thermostatic jacket 80a). The stub 81 is preferably horizontal and may have a diameter of about 1/10 of the diameter ¢ 3 of the upper cylindrical section 78.

The reaction space 75 is closed at the top by a lid 82 through which the gas outlet nozzle 83 protrudes from the reaction space and into which the catalyst supply nozzle 84 enters.

A recirculation line 85 is connected to one of the ends 64 of the fluid discharge unit VI and to the fluid supply nozzle 81 of the reactor space at which it is connected. and down. The dispersion elements 45, 46 of FIG. The circulation line 85 has a suction branch 87 from a nozzle 64 to a circulating pump 86 and a nozzle 88 from a pump 86 to a fluid-fed nozzle 81. The latter includes 89 configurators for greatly increasing the flow rate of the fluid. A nozzle 92 containing a nozzle 91 is provided for feeding the gas stream. Dispersing elements 45 (see Fig.) Are provided in the vertical section of the ram 88 between the flow accelerator 89 and the nozzle 81, and the dispersing elements 46 (Fig. 1a) are integrated in the horizontal sections; such a dispersing element 46 is also located in the stub 81 as shown in FIG. 3b. is clearly shown.

A nozzle 93 containing a valve 94 extends into the suction port 87, and a nozzle 95 for supplying the fresh substrate solution is connected to the discharge nozzle 88, comprising a flow accelerator 89, preferably a confluent valve 96.

3a. and 3b. The apparatus according to Figure 1B shall function as follows:

Preferably, the chemically sterilized equipment is filled with a polyacrylamide bead enzyme suspended in a glucose substrate through the catalyst feeder port 84. The volume of the fixed enzyme charge preferably amounts to about 3040% of the reactor volume. Subsequently, with the aid of pump 86, a glucose substrate solution is introduced into the outlet 88 of the circulating line 85 through the nozzle 95, opening the valve 96, and feeding it through the tube 81 to the reaction space 75. The feed is continued until the liquid level reaches the upper edge of the cartridge 76. The valve 94 is then closed.

After adjusting the reaction temperature with the liquid circulated in the thermostatic chamber 80, an auxiliary air stream providing autocirculation of the liquid phase and the catalyst particles is initiated through the auxiliary gas flow inlet 65 and the nozzle 71.

Preferably, the volumetric rate is adjusted to about 10% of the volumetric flow rate of the oxygen flow of the reaction (fed to the nozzle 92). Bubbles emerging from the nozzle 71, when introduced into the lower cylindrical portion 77 of the nozzle 76, exert a suction effect on the biocatalyst particles having a density less than or equal to that of the liquid suspended therein. In this way, upward fluid movement in the liner 76 is generated and circulation is closed through the space between the outer wall ι of the liner 76 and the inner surface of the container wall 72. The liner 76 has a high degree of agglomeration of the bubbles, so that the auxiliary air stream as an oxygen source is practically insignificant and serves only to regulate the particle circulation.

The required oxygen concentration in the liquid is provided by the forced circulation of the liquid through the circulation line 86 via the circulating pump 86: the pump 86 extracts the liquid while the filter element 70 retains the catalyst particles in the chamber 63. The pump 86 injects the fluid into the ram 88, which is mixed therein through the tube 92 and the nozzle 91 with the feed air stream supplied in accordance with the aperture p, and wherein the dispersing elements 45,46 are a-la. 1 to 4, provide the necessary oxygen transport from the gas phase to the liquid phase.

From the ram 88, the gas-liquid dispersion flows through the fluid feed tube 81 into the upper cylindrical portion 78 of the reactor liner 76, flowing through the rim and moving downwardly along the outer wall of the reservoir 72 with the catalyst particles -8EN 203288Β actuator bottom and gas exits on failure and leaves the reactor through the 83 port.

The significance of the tangential fluid introduction through the tube 81 is that the highly oxygenated substrate solution moving in the helical path contacts the catalyst particles very intensively and over a long period of time, thereby ensuring proper conversion.

The continuous mode of operation can be accomplished through a 95 nozzle, by adding an o-directional substrate solution with the valve 96 open, and by t-taking a product via the nozzle 93, or by serially switching several reactors.

In the reactor batch mode, the product can be discharged from the chamber 63 through the opening 67 by opening valve 66. Complete emptying of the reactor can be achieved by opening valve 68 through port 69.

Advantageous effects of the invention may be summarized as follows:

An essential advantage of the apparatus is that it enables high specific oxygen demand biocatalytic operations in the presence of a solid phase biocatalyst to be maximally exploited in the presence of a solid phase biocatalyst in a way that is economically feasible to maximize the utilization of the solid phase biocatalyst , the advantages mentioned in the introduction, while at the same time intensifying the gas-liquid contact and the smooth circulation of the biocatalyst particles fixed on the support, eliminate the problems of material transfer in conventional processes. It is also a significant advantage that, since the reaction space in the apparatus is separated from the field of intense vapor-liquid contact, the catalyst particles are not subjected to strong mechanical stress and thus are not damaged during the biocatalytic operation.

When using cell cultures that are particularly susceptible to infection as a biocatalyst, a preferred embodiment of the apparatus offers the possibility of encapsulating the cells in situ, i.e., making the particulate solid biocatalyst within the apparatus under more intense and secure conditions than those known in the art.

A further advantage is that the introduction of inert gas (or a mixture of inert gas and air), through its intensive distribution, enables the elimination of harmful gaseous by-products from the liquid phase of the fermentor, while economically using inert gas.

Finally, it is an important advantage that, when used as a member of a cascade series, it is also possible to perform highly efficient biocatalytic operations in which the product or / and the by-product gas (e.g. carbon dioxide) inhibits the biocatalytic process (e.g. fermentation).

Of course, the invention is not limited to the embodiments of the apparatus described in detail above, but may be practiced in many ways within the scope of the claims.

Claims (15)

PATENT CLAIMS An apparatus for carrying out a biocatalytic process by means of a solid phase biocatalyst, the apparatus comprising a reactor space comprising a cartridge tube, a catalyst for feeding the reaction medium, a feed medium and a gas, and a product and biogas produced by the biocatalysis. 10, and a circulation pump, characterized in that it has at its two ends a circulating line (50; 85) which overlaps the reaction space (15; 75) and out of the reaction space (15; 75) for discharging the medium treated therein. a filter element (21; 70) for retaining solid catalyst supports in the reaction space (15; 75) is provided prior to its outlet; the recirculation pump (38; 86) being integrated in the recirculation line (50; 85) which is connected to a pressure branch (39; 86); 88) and divides into a suction branch (37; 87), and a first flow accelerator (42; 89) is installed in the pressure branch (39; 88) after the pump, firstly downstream of the pump and then a gas inlet pipe (44). ; 92) it sticks out; and then the ram (39; 88) The dispersion element (s) 25 (45,46) comprises (1a, 2a and 3a. figure). Apparatus according to claim 1, characterized in that a phase separation chamber (18) having a gas outlet nozzle (28, 55) is connected to the reaction space (15) and the suction branch (37) of the circulating pipe (50) is They start from the lower part of the part (18), where the insert tube (17) located in the reaction space (15) extends (1a, Fig. 2a). Apparatus according to claim 1 or 2, 35, characterized in that the cylindrical reaction space (15) is closed at the bottom also by a cylindrical support (1), in which the pressure branch (39) of the circulation line (50) has a channel (3) . 40 Apparatus according to Claim 3, characterized in that the upper surface (5) of the base member (1) is formed as an oblique arc-shaped surface (la. And) extending from the bottom to the top of the opening of the pressure branch (39). lb. 45). Apparatus according to claim 3 or 4, characterized in that there is a manifold (11) with a valve (13) on the outside of the base member (1) and with a filter (10) on the top of the reaction space (15). 50, a conical flow modifier (8) extending upwardly from the filter element (10) and extending downwardly into the inside of the insert tube (17) (Figs. 1a and 2a). 6. An arrangement according to any one of claims 1 to 4, characterized in that the nozzle (48; 95) for supplying the fluid to be treated to the pressure branch (39; 88) of the circulation line (50; 85) and the flow accelerator (42; 89). ) and the biocatalytic process 60, the outlet for the resulting product 60 extends from the suction branch (37; 87) of the circulation line (50) (1a, Figs. 2a, 3a). 7. Apparatus according to any one of claims 1 to 3, characterized in that, in order to feed the solid biocatalyst lyser material into the reaction space (15), The 203288Β phase separation chamber (18) has a throat (52) (Fig. 2a). 8. Apparatus according to any one of claims 1 to 3, characterized in that a drop forming head (25) for feeding the droplets of the biocatalyst material, such as bacteria, into the phase separation chamber (18) is connected to the mixing means (1a) for receiving the solids. figure). Apparatus according to claim 8, characterized in that a valve (26) is inserted between the drop forming head (25) and the container (27) (Fig. 1a). Apparatus according to claim 8 or 9, characterized in that the drop forming head (25) has drop forming elements (Figs. 1a and le) directed towards the inside of the phase separation chamber (18). 11. Apparatus according to any one of claims 1 to 4, characterized in that the lower conical insertion tube extending in the upper cylindrical reaction space (75) has a larger diameter (e3) upper cylindrical portion (78) and a smaller diameter (ei) lower cylindrical portion connected thereto. (77) has; and the apparatus having a conical chamber (63) extending downwardly from the conical lower part (73) of the reaction space (75) to which the suction branch (87) of the circulating line (85) is connected; and a pressure branch (88) of the circulation conduit (85) is tangentially inserted in the lower region of the upper cylindrical portion (78) of the liner (76) and a filter member (70) is provided in front of the suction branch (87) extending from the chamber (63). (Figures 3a, 3b). Apparatus according to claim 11, characterized in that the auxiliary gas flow supply nozzle (71) is passed through a wall defining a conical chamber (63) which is connected to the lower cylindrical portion (77) of the insert tube (76) extending into the chamber (63). (Fig. 3a). 13. Apparatus according to any one of claims 1 to 6, characterized in that the length of the pressure branch (39; 88) of the circulation line (50; 85) is several times greater than the length of the suction branch (37; 88). 14. Apparatus according to any one of claims 1 to 4, characterized in that it has a gas dispersing element (45) having a confuser (45b), a diffuser (45c) and a connecting gap (45a) therein (see Figure). 15. Apparatus according to any one of claims 1 to 5, characterized in that it has a dispersing element (46) having a rod (46a) and a spiral member (46b) surrounding it (Fig. 1b).