JPH0793874B2 - Bioreactor - Google Patents

Description

TECHNICAL FIELD The present invention relates to a bioreactor which is a reaction device using a microorganism as a catalyst.

(Prior Art) Biochemical reactions such as decomposition, synthesis and chemical conversion of substances catalyzed by microorganisms are used in a wide range of fields such as organic synthesis, food industry and analytical chemistry. The reaction apparatus used for this type of reaction generally comprises a reaction tank using a microorganism as a catalyst, and the reaction liquid obtained in such a reaction tank is filtered by various filters to separate it from the microorganism and other mixtures. It

(Problems to be Solved by the Invention) By the way, in the above case, it is possible to use a cross-flow filter as a filter, and the filter and the reaction tank are communicated with each other through a circulation path, and the reaction solution is discharged from the reaction tank. Is continuously flowed along one side of the filter of the filter, while a part of the reaction liquid is filtered while the reaction liquid flowing keeps the cake accumulated on one side of the filter at a minimum thickness, and is stable. The filtration is maintained for a long time. Further, in order to increase the filtration rate, it is preferable to increase the circulation flow rate of the reaction solution to further reduce the thickness of the cake to be deposited, but increasing the circulation flow rate causes damage to microorganisms and deactivates the microorganisms. Therefore, the circulation flow rate is 2 m, for example.
The filtration rate could not be increased because it could not be increased more than / sec.

Therefore, an object of the present invention is to increase the filtration rate of a reaction solution in a bioreactor, which is a reaction means using a microorganism as a catalyst.

(Means for Solving the Problem) The present invention is a bioreactor which is a reaction device using a microorganism as a catalyst, and the first of the bioreactors is a cross-flow filtration containing a reaction tank using a microorganism as a catalyst and a tubular filter. Equipped with a vessel, a circulation passage for allowing the two to communicate with each other in the longitudinal direction along the outer circumference or the inner circumference of the cylindrical filter from the reaction tank, and the cylindrical filter in the circumferential direction. It is characterized by being rotatable.

A second bioreactor according to the present invention is provided with a storage tank for storing a reaction liquid and a cross-flow filter containing a tubular filter to which microorganisms for chemically reacting the reaction liquid are fixed, and The tubular filter is capable of communicating with each other so as to flow the liquid to be reacted from the storage tank in the longitudinal direction along the outer circumference or the inner circumference of the tubular filter, and the tubular filter is rotatable in the circumferential direction. It is characterized by.

Thus, the microorganisms used in the present invention are bacteria, radioactive fungi, molds, yeasts and the like, and these microorganisms are glycoamylase, aminoacidase, glugose isomerase, β-galactosidase, cellulase, invertase, asparaginase, aspal. Enzymes such as tase, catalase, protease, ribase, lysine decarboxylase, hexokinase, tryptophan synthase, glycerol dehydrogenase and the like are included.

When the reaction using a microorganism as a catalyst is carried out in a reaction tank, the microorganism is suspended and cultured in various pre-cultured areas, and this is stored in the reaction solution together with the reaction liquid having the same composition as the pre-cultured area. .
Further, when performing such a reaction in the filter, the outer peripheral side or the inner peripheral side of the cylindrical filter described below is immersed in the suspension described above, and the microorganism is cultured and immobilized in the state where the filter is left stationary. To do. The details of the immobilizing means are described in the specification of Japanese Patent Application No. 63-72228, which is a prior application of the present applicant.

The cross-flow filter contains a cylindrical filter, and the reaction liquid or the liquid to be reacted flows longitudinally along the outer or inner circumference of the filter, during which part of the liquid flows through the filter. The filtrate then flows out of the vessel. In addition, the cylindrical filter is rotatable in the circumferential direction,
The fluid is rotated during filtration. A filter of this type is shown as a rotary crossflow filter in the specification of Japanese Patent Application No. 63-77070 filed by the present applicant. The tubular filter is made of an inorganic material such as alumina, silica, silica-alumina, ceramics such as zirconia, porous glass, carbon, etc., and an appropriate tubular shape such as a pipe shape, a monolith shape, or a honeycomb shape is adopted. .

(Operation / Effect of Invention) In the bioreactor having such a configuration, the biochemical reaction using the microorganism as a catalyst is performed in the reaction tank or the filter, and the reaction liquid in the reaction tank or the reaction liquid in the storage tank is It flows in the longitudinal direction along the outer circumference or the inner circumference of the cylindrical filter. The flowing liquid applies a shearing force in the longitudinal direction to the cake deposited on the outer or inner circumference of the filter, and also applies a shearing force in the circumferential direction to the cake due to the rotation of the filter in the circumferential direction. Therefore, the cakes deposited on the outer or inner circumference of the filter are separated by the action of these two kinds of shearing forces and their growth is suppressed, and a large filtration rate is maintained for a long time without increasing the circulation flow velocity of the fluid. be able to. In addition, since it is not necessary to increase the circulation flow side, damage to the microorganisms is small and there is no risk of deactivating the microorganisms.

(Example) Production of lactase in a mold broth, purification of lactase, and sterilization test were carried out.

(1) Bioreactor The bioreactor used for the test consisted of a reaction tank 11, a cross flow filter 12, and a circulation system path 13 communicating these both 11 and 12 as shown in the drawing. After the culture is completed, the culture solution is circulated by driving the circulation pump 14 after the culture is completed. A cylindrical ceramic filter 15 is housed in the cross flow filter 12. The filter 15 has an outer diameter of 50 mm, an inner diameter of 36 mm, and a length of 300 mm and is formed by coating the outer periphery with a separation membrane, and the average pore diameter of the separation membrane is 0.
It has a total filtration area of 0.047 m ² . The cross-flow filter is of the same type as the rotary cross-flow filter shown in the specification of Japanese Patent Application No. 63-77070 filed by the present applicant, and the filter 15 is rotatable in the circumferential direction. In the filter 12, the culture fluid flows in the longitudinal direction along the outer periphery of the filter 15 while the culture fluid is circulating, and the filter 15 rotates in the circumferential direction during this period.

(2) Test conditions At the end of culturing, the mold broth has a mold concentration of 0.2 wt% and a molecular weight of 1
It contains 30,000 lactase as a suspension enzyme at a temperature of 3
It has a temperature of 0 ° C, a viscosity of 5 cp, a specific gravity of 1 and a PH5. The conditions and results of the purification and sterilization (filtration) of the culture broth are shown in a separate table, and the results when the standard cross-flow filtration method is adopted are also shown in the table as a comparative example. The cross-flow filter used as a comparative example is a fixed type in which the filter unit does not rotate, and the filter unit has 4 units.
This filter is arranged in parallel. Each filter has an outer diameter of 10 mm, an inner diameter of 7 mm, and a length of 500 mm, and is formed by coating a separation membrane on the inner periphery. The average pore diameter and the total filtration area of the separation membrane are 0.1 μm as in the case of the filter 15 of the embodiment. 0.044
m ² .

In the attached table, the filtration rate, lactase permeation rate, and mold inactivation rate are average values from the start of filtration to 60 minutes, and during this period, the culture solution was reduced from 4 l to 2 l. In addition, lactase permeability and mold inactivation rate in the attached table were measured by the following methods.

O Lactase permeability The enzyme activity of the stock solution and the filtrate is measured based on the β-galactosidase specific activity measuring method *, and calculated by the following formula.

(Note) *: Enzyme activity measurement methods, p. 53 (published by Boehringer Mannheim Yamanouchi Co., Ltd.) ○ Mold inactivation rate Dilute the stock solution and the filtrate, incubate the mold for a certain period, measure the number of colonies, and calculate by the following formula To do.

However, A is the number of colonies in 1 ml of the stock solution (cfu / ml) B is the number of colonies in 1 ml of the filtrate (cfu / ml) (3) Results As is clear from the above table, when the filtration rate in the present example and the comparative example were constant, the lactase permeability was higher and the mold inactivation rate was about 10% in the present example than in the comparative example. 1 /
It is understood that 10 is extremely useful. Also,
In order to increase the lactase permeability and decrease the mold deactivation rate in Comparative Examples, it is necessary to reduce the circulation flow rate and the filtration rate.

[Brief description of drawings]

The drawing is a schematic configuration diagram of a bioreactor according to an embodiment of the present invention. Explanation of code 11 …… Reaction tank, 12 …… Cross flow filter, 13 …… Circulation path, 15 …… Filter.

Claims (2)

[Claims] 1. A cross-flow filter containing a reaction tank using a microorganism as a catalyst and a tubular filter,
The tubular filter is provided with a circulation passage that allows the two to communicate with each other in the longitudinal direction along the outer or inner circumference of the tubular filter, and the tubular filter is rotatable in the circumferential direction. A bioreactor characterized by. 2. A storage tank for storing a liquid to be reacted and a cross-flow filter containing a cylindrical filter to which microorganisms for chemically reacting the liquid to be reacted are fixed, and the storage is made by communicating these two with each other. A bioreactor comprising a circulation system passage for causing a liquid to be reacted from a tank to flow in the longitudinal direction along the outer circumference or the inner circumference of the tubular filter, wherein the tubular filter is rotatable in the circumferential direction.