JPS62198383A - Bioreactor element and its production - Google Patents

Abstract

PURPOSE:The microorganism immobilization layer in which microorganism containing enzymes are immobilized in many pores and can pass through liquid and the microorganism separation layer which can selectively pass through only the liquid in the microorganism suspension are integrated to increase productivity of bioreactions. CONSTITUTION:The objective bioreactor element is integrally composed of the microorganism immobilization layer E1 in which a microorganism such as a bacterium M containing enzymes such as glucamylase are immobilized in its fine pores and the pores can pass through liquid, and of the microorganism separation layer E2 which can selectively pass through only the liquid in the microorganism suspension. In the biochemical reactions by means of the elements, the reaction mixture is allowed to pass through in the arrow direction from the layer E1 to the layer E2, while the microorganism M effects the biochemical reactions to produce the reaction mixture containing the objective compound. The mixture already passed through the layer E2 and is free from the cell bodies of the microorganism. Accordingly, the process with the bioreactor elements according to the present invention is in no need of the step of microorganism separation to increase the production efficiency of the objective product.

Description

DETAILED DESCRIPTION OF THE INVENTION [Industrial Application Field] The present invention relates to a bioreactor element in which microorganisms containing enzymes used in biochemical reactions are immobilized, and a method for producing the same.

[Prior art]

Biochemical reactions using this type of bioreactor element are used in a wide range of fields such as organic synthesis, food industry, and analytical chemistry. Various methods are known as a means of immobilizing (sometimes).

One of the various immobilization methods is the physical adsorption method in which microorganisms are physically adsorbed onto the element substrate, but in such methods, the interaction between the microorganisms and the element substrate is weak, so the microorganisms may be released from the element substrate during the reaction. The problem is that it is easy. Other immobilization means include water-insoluble beads.

Glutaraldehyde is a covalent bonding method that covalently bonds microorganisms to various element substrates in the form of pellets.

There are cross-linking methods in which microorganisms are cross-linked to the element substrate using a cross-linking agent having two or more functional groups such as bisdiazobenzidine, and ionic bonding methods in which microorganisms are bonded to the element substrate through ionic bonding. The problem is that immobilization changes the properties of the microorganisms and significantly reduces their activity.

Furthermore, other immobilization methods include an entrapment method in which microorganisms are wrapped in a polymeric gel lattice such as agar or carrageenan, or covered with a semipermeable polymer film; However, there is a problem that the activity of microorganisms decreases during comprehensive adjustment, and another problem is that the activity per unit microorganism is low because it shows activity only on the gel surface.

In order to deal with these problems, the present applicant has filed a patent application filed in
In the application No. 50322 (Japanese Unexamined Patent Publication No. 60-43382), we proposed a bioreactor element to solve the problems of the comprehensive method mentioned above.
A bioreactor element for solving the above-mentioned difficulties of the physical adsorption method is proposed in a patent application dated 2009 (title of the invention: Bioreactor element and method for producing the same). Therefore, in all of these bioreactor elements, a ceramic honeycomb structure is adopted as the element base, and the characteristics of such a structure are effectively used to enhance the immobilization of microorganisms and increase the contact of the reaction liquid with the microorganisms. It is something.

[Problem that the invention seeks to solve]

By the way, in these bioreactor elements, the reaction liquid flows in along the partition walls of the honeycomb structure that constitutes the element and flows out as a product liquid, so that microorganisms that have grown on the walls of the partition walls of the structure and the partition walls Since the microorganisms that have been slightly detached from the inside are mixed in the product solution in a suspended state, a subsequent microorganism separation step is essential.

Furthermore, even in these bioreactor elements, it is not possible to bring the reaction solution into contact with many of the immobilized microorganisms efficiently in a short period of time, and there is room for improvement in terms of reaction efficiency.

Therefore, an object of the present invention is to eliminate the need for microorganism separation steps because no microorganisms are mixed in the product solution, and to enable the reaction solution to effectively contact a large number of immobilized microorganisms in a short period of time. An object of the present invention is to provide a bioreactive element with high reaction efficiency and a method for producing the same.

[Means for solving problems]

In order to achieve this object, the first invention of the present invention is as follows:
It is integrally equipped with a microorganism immobilization layer that has microorganisms containing enzymes immobilized in a large number of pores and has a liquid permeability, and a microorganism separation layer that has a selective permeability that allows only the liquid in the microorganism suspension to permeate. It is located in the bioreactor element.

Further, the second invention of the present invention has pores capable of immobilizing microorganisms in a microorganism suspension obtained by suspending microorganisms containing an enzyme in an aqueous solution containing nutrients, and has liquid permeability. An element substrate integrally provided with a first layer and a second layer having a selective permeability that allows only the liquid in the suspension to pass therethrough is immersed, and the first F! A microorganism immobilization layer characterized in that the gas in the pores of the first layer is replaced with the suspension, and then the microorganisms in the pores of the first layer are cultured, multiplied, and immobilized in the pores. and a microbial separation layer.

Accordingly, in the present invention, the element substrate means a bioreactor component component before immobilizing microorganisms, and as shown in FIG. The first layer becomes
7'! No. 27' which becomes a microbial separation layer with El and itself!
It is integrally equipped with E2.

The element base is alumina and silica.

Silica-alumina, zirconia, porous glass.

It is made of an inorganic material such as carbon, which is usually a constituent material of ceramics, and is formed into an appropriate shape in relation to the reaction vessel, such as a plate shape, a pipe shape, or a honeycomb shape. Since the first layer ε1 of the element substrate becomes a microorganism immobilization layer, it is important that it has a large number of pores E3 for immobilizing microorganisms M, and the reaction liquid is not allowed to pass through the microorganism immobilization layer. It is important that the material has liquid permeability because it causes a reaction. In addition, the average pore diameter of the large number of pores E3 is related to the size of the microorganism M to be immobilized, and is preferably equal to or 40 times the size of the microorganism M. If the pore diameter is smaller than the size of the microorganism M, the microorganism If the pore size exceeds 40 times, microorganisms that have been cultured and proliferated within the pores E3 are likely to be easily detached from the pores. Note that all of the large number of pores E3 do not need to have liquid permeability, but many of them need to have liquid permeability. Therefore, pore E3
Most of the things are the 1st JL! It is a communicating hole that opens on both sides of El, and the remaining part may be a non-communicating hole that opens only on either side of the first FjE1g). In the present invention, the porosity is a ratio based on the pores of the pores E3 including all communicating pores and non-communicating pores in the first layer ε1, and the porosity is largely related to the amount of immobilized microorganisms M, and is from 30% to 8
0%, preferably 40% to 70%. Porosity is 30
If it is less than 80%, the amount of immobilized microorganisms is insufficient and
If it exceeds %, problems will arise regarding the strength of the immobilized layer.

Also, the second J'! of the element base! E2 itself is a microorganism separation layer, and it functions to separate microorganisms M that become cloudy in the product liquid that has passed through the microorganism immobilization layer.
The pore E4 is smaller than the size of the microorganism M by 0.01
It is preferable to have an average pore diameter of μ or more and liquid permeability. More preferably, the pore size of the pore E4 is such that the microorganism M
The size is 0.5 to 0.1 times that of . When the pore size is less than 0.01μ, the flow resistance in the microorganism separation layer becomes large, and it is necessary to increase the supply pressure of the reaction liquid. Further, the thickness of the second layer ε2 (microorganism separation N) is 0.
01μ to toooμ, preferably 0.1μ to 10μ
It is 0μ. If the thickness of the second layer ε2 is less than 0.01μ, it is difficult to adjust the diameter of the pores E4 in the same layer, and if it exceeds 1000μ, the pressure loss during permeation of the product liquid in the same layer becomes large. . Each layer of the bioreactor element according to the present invention is formed by each layer El of the element substrate.
, it has the same structural features as E2.

In such an element base, the first r&! El is obtained by forming, firing and heat-treating under the same conditions as those for forming ordinary ceramic porous bodies, and the second part ε2 is the first part.
It is formed on one side of the layer El, on the inner or outer circumferential surface, on the inner wall surface or on the outer wall surface, etc. by means such as adhering a gel film by the sol-gel method, high-pressure bonding of fine powder, or adhesion of porous glass. . Note that the average pore diameter, porosity, etc. of pores E3 and E4 in each layer El and ε2 are determined as appropriate based on the composition of the material used, mechanical treatment conditions such as pulverization of the material, heat treatment conditions, firing conditions, and other phase separation treatment conditions. This allows adjustment to a desired value.

In the present invention, the microorganisms M used for immobilization are not particularly limited, but include, for example, bacteria, actinobacteria, molds, yeasts, etc. Enzymes include glucoamylase, aminoacidase, glucose isomerase, β-galactosidanise, and cellulase. .

invertase, amataparaginase, aspartase,
Examples include catalase, protease, lipase, lysine decarboxylase, hexokinase, tryptophan synthase, glycerol dehydrogenase, and the like. In addition, the size of each of the above-mentioned microorganisms is 0.1μ to 1
50μ (general bacteria 0.5μ~1μ), fungi 2μ~1μ
0μ, yeast fungi 5# to 10μ. In the present invention, a microorganism M containing any of these enzymes is suspended in an aqueous solution containing nutrients to prepare a microorganism suspension, and the element substrate described above is immersed in the suspension. Next, in this state, the first layer E1 of the element base is removed by degassing under reduced pressure.
The gas in the pores E3 of the first layer El is replaced with a microorganism suspension, and then the microorganisms M in the pores E3 of the first layer El are cultured, grown, and immobilized in the same pores E3. Thereby, the first layer E1 of the element base becomes a microorganism immobilization layer, and a bioreactor element integrally provided with the microorganism immobilization layer and the second layer E2, which is a microorganism isolation layer, is obtained.

[Action/effect of the invention]

In the biochemical reaction using the bioreactor element according to the present invention, the reaction solution is permeated from the microorganism immobilization layer side of the bioreactor element to the microorganism a layer side as shown by the arrow in FIG. During this time, a biochemical reaction is caused by the microorganism M to obtain a product liquid containing the target product.

Since the product liquid has already passed through the microbial separation layer, it does not contain free microorganisms rVI, and therefore the conventional microbial separation process is completely unnecessary, increasing the production efficiency of the desired product liquid, and increasing the separation process. By omitting this, the space of the reactor can be narrowed and the cost of the device can be reduced.

In addition, since the reaction solution permeates through the microorganism immobilization layer, it is possible to contact a much larger number of immobilized microorganisms in a shorter time and more efficiently than when the reaction solution flows along the microorganism immobilization layer. The reaction efficiency can be significantly improved in a short time. Therefore, if this bioreactor element is used in series with a conventional bioreactor element and a reaction liquid in which a biochemical reaction is occurring to some extent is permeated through the bioreactor element, the reaction liquid will be absorbed at the point of permeation. The reaction proceeds to further increase the production efficiency of the desired product liquid.

[Examples and comparative examples]

(1) Preparation of element substrate Each partition has a two-layer structure of the 1N layer and the 2nd layer 171
A similar honeycomb element substrate al-a17 was prepared by the following method, and honeycomb element substrates bl, b2 each having a single layer structure in which each partition wall was made of cordierite were prepared.
(Outer diameter 5cm, length 30c+n, cell opening length 1mm)
and b3 (outer diameter 6 marks, length 10 cyn, cell opening length 1
mm) was produced by a conventionally known method.

In addition, a vibrator-like element base a18 having a two-layer structure, an inner and outer layer with a first layer on the outer periphery and a second layer on the inner periphery, was produced by the method described below.

(la) Various types of first rfi single-layer honeycomb structures made of cordierite (outer diameter 5 cm, length 15 cm, cell opening length 1 mm) were manufactured by a conventionally known method, and each honeycomb structure was Close both the upper and lower openings of the through hole by pasting tape or the like. On the other hand, α-alumina powder with an average particle size of 1 μm was wet-pulverized to obtain a supported slurry, each of the honeycomb structures described above was immersed in the slurry to support the slurry, and after drying, the slurry was fired at 500°C for about 3 hours. Thus, a honeycomb-like element base AL-WA17 in which the first N and second layers were integrated was obtained.

(lb) First layer single-layer vibrator structure made of cordierite, outer diameter 12 mm, inner diameter 6 mm, length 10 cm
) is prepared by a conventionally known method, and the outer circumferential surface of the structure is covered by pasting it with a tape or the like on the outer circumference of the structure. Such a structure was immersed in a slurry similar to the supporting slurry shown in item (Ia) above to support the same slurry, and after drying, it was heated at 500°C for about 30 minutes.
By firing for a period of time, a vib-like element base a18 integrally provided with the second layer on the inner periphery was obtained.

(2) Immobilization of m-organisms (2a) Alcohol-fermented yeast Satucharomyces cerevisiae (about 5μ in size) was added to a culture solution (yeast extract 0.15%,
Nll+CI 0.25%, Niyu IIPO, 0.55
%.

Mg5O+-711jL00.025%, NaC,1
, O: 1%, CaCl, 0.001%.

Suspend in 0.3% citric acid (all weight %) at ρ115.4 at a rate of 10 pieces/m. Honeycomb-like element substrates al~al? are added to the yeast suspension. , bl, b2 are immersed;
While vacuum degassing using an aspirator for 15 minutes, remove the element substrate. , bl, and b2. In this case, each element substrate a1 to a1
In No. 7, one open end was closed to prevent the suspension from flowing into the longitudinal through hole surrounded by the second N.

Next, each of these element substrates al-al? ,bl
, b2 was set in a constant temperature bath at 30° C., and the yeast that had entered the pores was cultured once for 3 days to allow the yeast to proliferate. As a result, each element base al-a17. Yeast is immobilized on bl and b2, and each bioreactor element A
t~AI? , 81.82.

Each bioreactor element A1 to A17. Bl, 8
The configuration of No. 2 is shown in Table 1.

(2b) Acetobacter acetiform bacteria (size approximately 1μ)
Preculture medium C (if 10 g of glycose.

Polypeptone 10g, yeast extract 10g, ethanol 2
0B, ice vinegar MlOg) to IOke/III, e! g to make it cloudy. The honeycomb element base b3 is immersed in this suspension, and one open end of the vibrator element base alB is closed to prevent the suspension from flowing into the inner hole of the vibrator element base alB, as described in 9. Immerse it in the suspension and keep it at 30°C.

The acetic acid bacteria are left to stand still for 4 days at pH 3.3.

These elemental substrates a1B, b3 are then transferred to a preculture medium with the same components as preculture medium C, where each element substrate a18. Acetobacter in b3 at 30℃, pl+3
．． 3 and cultured for 36 hours. As a result, each element base a18. Bioreactor elements 818 and 83 in which acetic acid bacteria are immobilized on b3 are produced. The composition of each bioreactor element AI8,133 is shown in Table 2.

(2c) The above-mentioned acetic acid bacteria were comprehensively immobilized on beads with a diameter of 3III+ by a conventional method using 3% by weight of sodium alginate to produce a bead-shaped bioreactor element B4.

(3) Honeycomb-shaped bioreactor elements are housed in upper and lower two stages, with fB& on the outside of the reactor and lm11.12 on the outside, and fB & on the inside of the inner cylinder 1. The upper element 13 is one of each bioreactor element A1-A17, and the lower element 14 is similar to bioreactor element B2. As shown in FIGS. 2 to 5, the upper element 13 is provided with a microorganism separation layer 13b at a predetermined portion of the partition wall of the microorganism immobilization layer 13a, and a large number of first microorganisms surrounded by the immobilization layer 13a. The through holes 13c serve as flow paths for the reaction liquid, and the numerous second through holes 13d surrounded by the separation layer 13b serve as flow paths for the product liquid.

In this upper stage element 13, each second through hole 1
The lower ends of the cylindrical lid 15 are sealed as shown in FIG. It penetrates and opens in the attached body 15. The lid body 15 is equipped with a discharge conduit 15a for the produced liquid approximately in the center of its top, and has negative pressure conduits 1 connected to a vacuum pump on both sides thereof.
5b and a positive pressure conduit 15c connected to a compression pump, and a predetermined gap is maintained between the inner cylinder 11 and the upper end thereof.

In this state, the inner cylinder 11 and the lid 15 are arranged concentrically within the outer cylinder 12, and the lid 15 protrudes from the upper end of the outer cylinder 12 in an airtight manner. A reaction liquid supply pipe 12a is connected to the bottom of the outer cylinder 12.
The reaction liquid supplied from the first stage first flows through the through holes of the lower stage 14 along their partition walls, and during this time, the first stage reaction occurs. Next, the reaction solution is transferred from each first through hole 13c of the upper element 13 to the immobilized J! ! 13a and separation 7@13b, and flows into each second through hole 13d, during which a second stage reaction occurs, and flows out from the discharge conduit 15a through each second through hole 13d1. Each first through hole 13c
A part of the reaction liquid that has flowed into the chamber and gas components generated by the reaction are transferred from the gap between the inner cylinder 11 and the lid body 15 to both [11, 12
A part of the reaction liquid flows out to the annular passage P between the two, and a part of the reaction liquid flows back to the lower end side of the lower stage 14. Note that the outside *12 was kept at a desired temperature by a heat insulating means (not shown), and was kept at about 30° C. in the reaction test described below.

On the other hand, an exhaust pipe 12b is provided outside the top of the outer cylinder 12. The exhaust pipe 12b communicates with the annular passage P, and a float 16a and a solenoid valve 16b are disposed at the inner end thereof. flow) 16a is the solenoid 16 of the solenoid valve 16b
It controls the opening and closing of a switch 16e interposed between C and electric field Bd.When the liquid level in the outer cylinder 12 is above a predetermined height, the float 16a moves upward and closes the switch 16e, causing the liquid level to reach a predetermined level. If the height is less than the height, the flow 16a moves downward and closes the switch 16e. As a result, when the generated gas increases in the outer cylinder 12 and the liquid level falls below a predetermined height due to the gas pressure, the switch 16e closes, the solenoid 16c of the solenoid valve 16b is energized, and the solenoid valve 16b opens. . As a result, the generated gas in the outer cylinder 12 is discharged through the solenoid valve 16b, and then the liquid level in the outer cylinder 12 rises and the switch 16e
opens, energization to the solenoid 16c is stopped, and the solenoid valve 16b closes.

(3b) As shown in FIGS. 7 to 9, in the second reaction device 20, the upper element 23 is an assembly of bioreactor elements A1B according to the present invention, and the lower element 24 is a bioreactor element B3, It differs from the first reactor 10 in these points. Upper element 2
3 is made up of a large number of elements A18 assembled in a cylindrical shape, and is housed in a portion above the lower element 24 in the inner cylinder 21, and their upper ends penetrate the bottom of the lid body 25 in a liquid-tight manner. A through hole 23a opens inside the lid body 25. In the upper element 23, the gaps between the two elements A18 serve as flow passages 23b for the reaction liquid, and each flow passage 23b opens at the upper end of the inner cylinder 21 and communicates with the annular passage P.

Note that the lower end of each through hole 23a is sealed as shown in the filled part shown in FIG. 9, and the outer cylinder 22 is kept at a desired temperature by a heat insulating means (not shown).

Therefore, in such a second reactor 20, the "penetrations" of the elements 24 flow along their partition walls, during which time a first stage reaction occurs. Next, the reaction liquid flows from each flow path 23b of the upper stage element 23 through the immobilized J523c on the outer periphery of each element A1B and the inner layer 23d, and flows into each through hole 23a, during which time a second stage reaction occurs. and flows out from the discharge conduit 25a through each through hole 23a. Note that in this second reaction device 20 as well, the exhaust pipe 2
2b is equipped with a flow) 26g and a solenoid valve 26b, and the exhaust of generated gas and the like is performed in the same manner as in the first reactor 10. In addition, the other configurations are the same as the first reactor 10 and function in the same way as the first reactor IO, and members corresponding to those of the first reactor 10 are designated with similar numerals in the 20s. Therefore, the explanation will be omitted.

(4) Reaction test (4a) Using the first reaction apparatus 10 in which each of the bioreactor elements A1 to A17 is employed as the upper stage element 13 and the bioreactor element B2 is employed as the lower stage element 14, pH 5, An ethanol production reaction test was conducted using the 20% by weight glycose solution prepared in Example 4 as a reaction liquid.

In this test, the temperature was 30°C, the supply rate of the glycose solution was 40rrTR/br, and the ethanol concentration and microorganisms (free The bacterial cell concentration was measured. In addition, in this test, 1
Positive pressure is applied to the inside of the device 10 through the positive pressure conduit 15c for 2 seconds several times a day, and the separation layer 1 of each element ^1-A17 is
The bacterial cells clogged on the upstream side of layer 3b were allowed to flow out to the solidified layer 13a side. The results obtained are shown in the columns of test numbers 1 to 17 in Table 1. In these tests, negative pressure was supplied into the lid 15 through the negative pressure conduit 15b to reduce the pressure inside the lid 15.

In addition, in the first reaction apparatus 10, instead of the upper and lower elements 13.14, a bioreactor element with the same length as the total length of these elements 13.14)'81.
．． No. 82 was employed individually, and the above-mentioned ethanol production test was conducted using the apparatus. The results obtained are shown in the column of test number 18.19 in Table 1.

(4b) A second reaction device 20 in which an element formed by collecting a large number of bioreactor elements A18 is used as the upper stage element 23, and a bioreactor element 83 is employed as the lower stage element 24.
An acetic acid fermentation test was conducted using In this test,
A reaction solution having the same composition as the preculture medium C shown in section (2b) was prepared at a temperature of 30°C, ρh of 3.3°, and a supply rate of 50 m, i'
/hr, and at the same time when supplying the reaction solution, air was supplied at 250 hr.
It was fed at a rate of m/min. Fifteen days after the start of the reaction, the acetic acid concentration and the microorganism (free bacterial cell) concentration in the reaction product liquid discharged from the discharge conduit 25a in a steady state were measured. The obtained results are shown in the column of test number 20 in Table 2.

In addition, in the second reaction device 20, a bead-shaped bioreactor element B4 is adopted in place of the upper and lower stage elements 23.24, and the same element B4 is filled into the inner cylinder 21 in an amount equivalent to the both stage elements 23.24. The above acetic acid fermentation test was conducted. The obtained results are shown as test number 2 in Table 2.
Shown in column 1.

The pore diameters shown in the chart above are measured by the known mercury pressure method.

Table 2 below (5) Discussion Test results of test numbers 1 to 12 shown in Table 1 and test number 1
Comparison with the test results of 8.19 shows that in the former case, the amount of ethanol produced was large, and therefore the reaction efficiency was high, and there was no single bacterial cell. This indicates that the elements Al to A12 according to the present invention used in the former contribute. Furthermore, referring to test numbers 13 to 17, in the element according to the present invention, the pore diameter, porosity, etc. of the immobilization layer greatly affect the reaction efficiency, and the pore diameter of the separation layer greatly affects the separation efficiency of free bacterial cells. The significance of using Elmate and conventional Pi and Oriakdar elements in series is that the test results (Test No. 1, 11°, 12.1
9 etc.), and in this reaction test, the amount of ethanol produced is the highest when the length of both elements is 15 cm.

On the other hand, referring to the test results in Table 2, the element A18 according to the present invention has superior reaction efficiency compared to the bead-shaped Nilemene 84 based on the conventional entrapment method, and has almost complete ability to isolate S. It can be seen that it has

[Brief explanation of drawings]

Fig. 1 is a super-enlarged schematic partial sectional view of an element according to the present invention, Fig. 2 is a partially cutaway schematic perspective view of a first reaction device employing the same element, and Fig. 3 is the same as Fig. 2. A cross-sectional view in the direction of arrow ■-■, Figure 4 is the arrow IV-IV in Figure 2.
5 is an enlarged view of the arrow V portion in FIG. 3,
Fig. 6 is an electrical circuit of a solenoid valve, Fig. 7 is a partially cutaway schematic perspective view of a second reaction device employing an element according to the present invention, and Fig. 8 is a diagram showing the direction of arrows ■-■ in Fig. 7. Cross section, No. 9
The figure is a sectional view taken along arrow IX-IX in FIG. 7. Explanation of symbols 10.2.0... Reactor, 11.21... Inner cylinder,
12.22...Outer cylinder, 13.23...Upper stage element, 13a, 23c...φ fixing layer, 13b, 23d...
... Separation layer, 14.24 ... Lower element, 15.
25... Lid body, 16a. 26a...Float, 16b, 26b---Solenoid valve,
El...first layer, El...second layer, E3, E4
...Pore, M...Microorganism.

Claims (7)

[Claims] (1) A microorganism immobilization layer with liquid permeability in which microorganisms containing enzymes are immobilized in a large number of pores, and a microorganism separation layer with selective permeability that allows only the liquid in the microorganism suspension to pass through, are integrated. A bioreactor element prepared for the purpose. (2) The bioreactor element according to claim 1, wherein the microorganism immobilization layer and the microorganism separation layer are made of an inorganic material. (3) The microorganism immobilization layer is the same size as the microorganism or 40
A bioreactor element according to claim 1 or 2, comprising a large number of pores with twice the average pore diameter. (4) The bioreactor element according to claim 1, 2, or 3, wherein the microorganism immobilization layer has a porosity of 30% to 80%. (5) The microorganism separation layer is smaller than the microorganism size 0.0
The bioreactor element according to claim 1, 2, 3 or 4, comprising a large number of pores with an average pore diameter of 1 μ or more. (6) The bioreactor element according to claim 1, 2, 3, 4, or 5, wherein the microorganism separation layer has a thickness of 0.01μ to 1000μ. (7) In a microbial suspension obtained by suspending microorganisms containing enzymes in an aqueous solution containing nutrients, a first layer having pores capable of immobilizing the microorganisms and having liquid permeability, and the suspension; An element base integrally equipped with a second layer having a selective permeability that allows only the liquid inside to pass through is immersed, and the gas in the pores of the first layer is removed from the suspension by depressurization and degassing. replace,
Thereafter, the microorganisms in the pores of the first layer are cultured, multiplied, and immobilized in the pores. Manufacturing method.