WO2006003790A1

WO2006003790A1 - Reaction apparatus for hydrophobic compound and method of reaction therewith

Info

Publication number: WO2006003790A1
Application number: PCT/JP2005/010952
Authority: WO
Inventors: Kenichi Noda; Shiro Irisa; Tomonari Kogure; Minoru Sakata; Akihiko Tokida
Original assignee: Bussan Nanotech Research Institute, Inc.
Priority date: 2004-06-30
Filing date: 2005-06-15
Publication date: 2006-01-12
Also published as: JP2007275689A

Abstract

A reaction apparatus for hydrophobic compound for carrying out reaction of hydrophobic compounds with the use of a biocatalyst, characterized by having a microchannel chip comprising a first channel for circulation of a hydrophobic liquid material containing a hydrophobic compound, a second channel for circulation of a water base liquid material containing a biocatalyst and a third channel for circulation of the hydrophobic liquid material and water base liquid material which is connected to the first channel and second channel. By virtue of this reaction apparatus, the reaction efficiency of hydrophobic compound can be satisfactorily enhanced in a wide variety of reactions conducted in the presence of a biocatalyst. Accordingly, the provided reaction apparatus for hydrophobic compound and method of reaction therewith are applicable to, being important in organic chemistry, a hydrolysis reaction, redox reaction, transition reaction, substitution reaction, elimination reaction, condensation reaction, addition reaction, isomerization reaction, asymmetric synthesis reaction, etc., and future progress is promising in the field of biocatalytic reaction for hydrophobic compounds.

Description

Specification

Hydrophobic compound reactor and reaction method

Technical field

The present invention relates to a hydrophobic compound reaction apparatus and reaction method using a biocatalyst.

Background art

[0002] Processes in which chemical reactions are carried out under normal temperature and pressure using biocatalysts such as cells or enzymes are widely used in the production of various fermented foods and various pharmaceuticals including antibiotics. In the production of chemical products, the use of biocatalysts is progressing for the production of ethanol and acrylamide for fuels.

[0003] However, the targets of these reactions are all water-soluble compounds. When biocatalysts are used for hydrophobic compounds that are important in the chemical industry, hydrophobic reactions in the presence of hydrophilic biocatalysts are necessary. Progress has been delayed due to the low reaction efficiency of organic compounds. Currently, these measures include the ability to use hydrolyzing enzymes such as relatively high hydrophobicity and lipase (see, for example, Patent Document 1 below), and surfactants for dispersing and adding hydrophobic compounds in aqueous solutions. Has been attempted (for example, see Patent Document 2 below).

Patent Document 1: Japanese Patent Laid-Open No. 06-296499

Patent Document 2: Japanese Translation of Special Publication 2002-527073

Disclosure of the invention

Problems to be solved by the invention

[0004] However, the method for reacting a hydrophobic compound described in Patent Documents 1 and 2 described above has the following problems.

[0005] That is, lipase functions as a catalyst only in a relatively simple reaction such as hydrolysis, and in organically important reactions such as oxidation-reduction reaction, transfer reaction, substitution reaction, elimination reaction, etc. As a result, I got enough results.

[0006] The addition of a surfactant is not only expensive for the reaction but also causes a phenomenon such as foaming, which is undesirable for increasing the reaction efficiency.

[0007] The present invention has been made in view of the above circumstances, and has been made hydrophobic by a wide variety of reactions. It is an object of the present invention to propose a reaction apparatus and a reaction method for a hydrophobic compound that can sufficiently increase the reaction efficiency of the compound and reduce the cost required for the reaction.

Means for solving the problem

[0008] As a result of intensive studies to solve the above problems, the present inventors have thought that the reason why the reaction efficiency of the hydrophobic compound is not sufficient as described above is due to the following reason. That is, in general, when a hydrophobic compound is reacted using a biocatalyst as described above, the hydrophobic compound and the biocatalyst are in sufficient contact, and the reaction of the hydrophobic compound proceeds sufficiently in the presence of the biocatalyst. It is considered a thing. However, in reality, the reaction has not progressed sufficiently, and the reason is not clear. As a result of intensive studies, the present inventors have thought that the low reaction efficiency of the hydrophobic compound as described above may be caused by a reaction vessel such as a flask. That is, the present inventors considered that the reaction efficiency may be lowered in the above reaction vessel due to insufficient contact area between the hydrophobic compound and the biocatalyst. As a result of intensive studies, the present inventors have found that the above problems can be solved by the following invention, and have completed the present invention.

That is, the present invention is a reaction apparatus for a hydrophobic compound that reacts a hydrophobic compound with a biocatalyst, and includes a first channel that circulates a hydrophobic liquid substance containing the hydrophobic compound, and the biocatalyst. A microchannel chip having a second channel through which an aqueous liquid material flows, and a third channel connected to the first channel and the second channel and through which the hydrophobic liquid material and the aqueous liquid material flow is provided.

[0010] According to this hydrophobic compound reactor, a hydrophobic liquid substance containing a hydrophobic compound and an aqueous liquid substance containing a biocatalyst are transferred from the first channel and the second channel to the third channel in the microchannel chip. Since it is distributed, the distributed hydrophobic liquid substance and aqueous liquid substance come into contact in the third channel. Then, the surface area of the interface between the hydrophobic liquid substance and the aqueous liquid substance increases, so that the contact efficiency between the hydrophobic compound and the biocatalyst can be greatly improved. For this reason, in the presence of the biocatalyst, the reaction efficiency of the hydrophobic compound can be increased, and as a result, the reaction can be performed at a reaction rate several tens of times higher. In addition, the ability to react quickly and efficiently with hydrophobic fluids with poor compatibility and aqueous fluids allows the application to various variations of hydrophobic compounds and biocatalysts. You can. Further, since addition of an extra surfactant or the like is unnecessary, the reaction of the hydrophobic compound can be performed at a low cost, and the reaction product can be obtained with high purity.

[0011] Further, the channel width of the channel is preferably lmm or less. When the channel width is 1 mm or less, the effect of reducing the diffusion rate can be obtained, and the surface area of the interface between the hydrophobic fluid and the biocatalyst can be increased. Therefore, in the presence of the biocatalyst, the hydrophobic compound The reaction of can be dramatically promoted.

[0012] The reaction method of the present invention is a reaction method of a hydrophobic compound in which a hydrophobic compound is reacted using a biocatalyst, and is an aqueous liquid containing a hydrophobic liquid substance containing a hydrophobic compound and a biocatalyst. The substance is contacted in a channel having a channel width of 1 mm or less.

[0013] According to this reaction method, a hydrophobic liquid substance containing a hydrophobic compound and an aqueous liquid substance containing a biocatalyst are circulated in a channel having a channel width of 1 mm or less. When in contact with an aqueous liquid material, the surface area at each interface further increases. For this reason, it is possible to greatly improve the contact efficiency between the hydrophobic compound and the biocatalyst. In addition, the reaction efficiency of the hydrophobic compound can be increased in the presence of the biocatalyst, and as a result, the reaction can be performed at a reaction rate several tens of times higher. Furthermore, since a hydrophobic fluid having poor compatibility and an aqueous fluid can be reacted quickly and efficiently, it can be applied to hydrophobic compounds and biocatalysts of various types. Further, since addition of an extra surfactant or the like is unnecessary, the reaction of the hydrophobic compound can be performed at a low cost, and the reaction product can be obtained with high purity.

[0014] Preferably, the hydrophobic liquid substance and the aqueous liquid substance are circulated in the third channel, and the phases of the hydrophobic liquid substance and the aqueous liquid substance are brought into contact with each other in a separated state. As described above, when the hydrophobic liquid substance and the aqueous liquid substance are brought into contact with each other, the contact efficiency can be greatly improved.

[0015] Further, it is preferable that droplets of the hydrophobic liquid substance and aqueous liquid substance droplets are formed in the third channel, and are alternately circulated and brought into contact with each other. As described above, when the hydrophobic liquid substance and the aqueous liquid substance are brought into contact with each other, the contact efficiency can be greatly improved.

[0016] Furthermore, the third channel is composed of a hydrophobic liquid substance and an aqueous liquid substance. It is preferable that a hydrophobic liquid substance and an aqueous liquid substance are brought into contact with each other by forming a liquid. As described above, when the hydrophobic liquid substance and the aqueous liquid substance are brought into contact with each other, the contact efficiency can be greatly improved.

[0017] It is preferable that the biocatalyst is a microorganism having an ability to incorporate a hydrophobic compound into a cell. Microorganisms that have the ability to take up hydrophobic compounds into cells effectively take up hydrophobic substances into cells at the contact interface between hydrophobic liquid substances and aqueous liquid substances, increasing the efficiency of contact with enzymes in the microorganisms, Promote the reaction.

[0018] In addition, the microorganism may be a Pichia yeast, Candida yeast, Pseudomonas bacterium, Rhodococcus bacterium, Mycobacterium bacterium, Bacillus bacterium, Brevibacillus bacterium, or Diobacillus bacterium. I like it. Since these microorganisms have a high ability to take various hydrophobic substances into the body, the reaction is further promoted.

The invention's effect

[0019] According to the reaction apparatus and the reaction method of the hydrophobic compound of the present invention, the reaction efficiency of the hydrophobic compound can be sufficiently increased in a wide variety of reactions in the presence of the biocatalyst. Therefore, according to the reaction apparatus and reaction method of the hydrophobic compound of the present invention, hydrolysis reaction, oxidation reduction reaction, transfer reaction, substitution reaction, elimination reaction, condensation reaction, addition reaction, It can be applied to isomerization reactions, asymmetric synthesis reactions, etc., and progress can be expected in the field of biocatalytic reactions of hydrophobic compounds in the future.

Brief Description of Drawings

FIG. 1 is a plan view showing a microchannel chip according to this embodiment of a hydrophobic compound reaction apparatus according to the present invention.

2 is a partial cross-sectional view taken along the thickness direction of a main body portion constituting the microchannel chip of FIG.

FIG. 3 is a cross-sectional view showing first and second modes of a first channel, a second channel, and a third channel.

FIG. 4 is a partial plan view showing a microchannel chip according to another embodiment of the hydrophobic compound reactor according to the present invention.

[FIG. 5] FIG. FIG. 6 is a partial front view showing a dispersion unit 31.

FIG. 6 is a graph showing reaction rate measurement results according to Examples:! To 3 and Comparative Example 1.

FIG. 7 is a graph showing the reaction rate measurement results of Examples 4 and 5 and Comparative Example 2.

Explanation of symbols

[0021] 1 ... 1st channel, 2 ... 2nd channel, 3 ... 3rd channel, 4, 7 ... Receiver, 5 ... Substrate, 6 to 3rd tube, 8 ... 4th tube, 10 ... Microchannel Tip, 11… First tube, 12 ··· Second tube, 21 ··· First syringe, 22 ··· Second syringe, 31 ··· Dispersing rod, 32 ················· Channel width.

BEST MODE FOR CARRYING OUT THE INVENTION

Hereinafter, preferred embodiments of the present invention will be described in detail with reference to the drawings.

FIG. 1 is a plan view showing a microchannel chip according to this embodiment of the hydrophobic compound reactor according to the present invention. As shown in FIG. 1, the microchannel chip 10 has a substrate 5 made of an inert material, and a first channel 1 for allowing a hydrophobic liquid substance to flow through one surface 5a of the substrate 5; A second channel 2 is formed to distribute the aqueous liquid material. On the other hand, the third channel 3 is formed on the first surface 5a of the substrate 5 to distribute the hydrophobic liquid substance and the aqueous liquid material. The third channel 3 is formed by meandering on the first surface 5a of the substrate 5. Has been. One end of the third channel 3 is connected to the first channel 1 and the second channel 2, and the other end of the third channel 3 extends to the edge of the entire surface 5a.

For this reason, the hydrophobic liquid material circulated in the first channel 1 and the aqueous liquid material circulated in the second channel 2 come into contact in the third channel 3. The other end of the third channel 3 is connected to the receiver 4 via the third tube 6. A branch channel 3 a is connected to the third channel 3, and the branch channel 3 a is connected to the receiver 7 via the fourth tube 8. In this case, the reaction products are accommodated in separate receivers 4 and 7, and multiple analyzes necessary for identifying compounds can be simultaneously performed in each of them. Alternatively, in the receiver 4, a reaction product can be taken out as necessary, or a device for detecting the reaction product can be connected.

FIG. 2 is a partial cross-sectional view along the thickness direction of the substrate 5. In FIG. 2, the cross-sectional shape of the third channel 3 is a quadrangle, and the channel width d of the third channel 3 is preferably lmm or less. It is. Since the channel width d is in the above range, when the hydrophobic liquid substance and the aqueous liquid substance are in contact with each other in the third channel 3, the contact area at the interface increases, and thus the reaction is more fully promoted. be able to. At the same time, it is possible to react quickly by utilizing the effect of shortening the diffusion time of molecules, that is, the effect of minimizing diffusion rate control. On the other hand, when the channel width d exceeds lmm, the effect of increasing the interfacial surface area per unit volume and the effect of minimizing the diffusion rate can be sufficiently obtained as compared with the case where the channel width d is 1 mm or less. Tend to become impossible. The channel width d is more preferably 0.:! To 500 xm.

[0026] The length of the third channel 3 can be arbitrarily determined in accordance with the convenience of the apparatus and reaction. “Channel width” refers to the length in the direction perpendicular to the thickness direction of the substrate. When the cross-sectional shape of the third channel 3 is a square, it means the horizontal length, and when it is a trapezoid or a triangle, it means the longest length in the above direction. In the case of a circular or semi-circular shape, the length of the diameter and the length of the diameter in the case of an ellipse are the longest in the direction.

Although not shown, the cross-sectional shapes of the first channel 1 and the second channel 2 are the same as those of the third channel 3, and the channel width d is the same as that of the third channel 3.

The first channel 1 is connected to the first syringe 21 via the first tube 11, and the second channel 2 is connected to the second syringe 22 via the second tube 12.

Accordingly, the hydrophobic liquid substance is accommodated in the first syringe 21 and further the aqueous liquid substance is accommodated in the second syringe 22, so that the hydrophobic liquid substance passes through the first tube 11 and enters the first channel 1. The aqueous liquid substance can be injected into the second channel 2 via the second tube 12. Therefore, the hydrophobic liquid substance and the aqueous liquid substance are contacted in the third channel 3, and the reaction that is the object of the present invention is performed.

[0030] According to the above reaction apparatus, when the first syringe 21 is operated, the hydrophobic liquid substance containing the hydrophobic compound is injected from the first syringe 21 into the first channel 1 via the first tube 11. On the other hand, when the second syringe 22 is actuated, an aqueous liquid substance containing a biocatalyst is injected into the second channel 2 from the second syringe 22 through the second tube 12. The hydrophobic liquid material flows from the first channel 1 to the third channel 3, and the aqueous liquid material flows from the second channel 2 to the third channel 3. In the third channel 3, the hydrophobic liquid material and the aqueous liquid are circulated. Substance Contact. At this time, since the third channel 3 is a channel in the microchannel chip 10, when the circulated hydrophobic liquid substance and the aqueous liquid substance come into contact with each other, the surface area of each increases, and the hydrophobic compound and the biocatalyst are separated. Contact efficiency can be greatly improved. For this reason, the reaction efficiency of the hydrophobic compound can be sufficiently increased in the presence of the biocatalyst, and as a result, the reaction can be performed at a reaction rate several tens of times higher. In addition, since the reaction efficiency of the hydrophobic compound can be sufficiently increased as described above, the hydrolysis reaction, oxidation-reduction reaction, transfer reaction of the hydrophobic compound in the presence of the biocatalyst is organically important. Substitution reactions, elimination reactions, condensation reactions, addition reactions, isomerization reactions, asymmetric synthesis reactions, and the like can also be performed. Furthermore, since the reaction efficiency of the hydrophobic compound can be sufficiently increased without using a surfactant as described above, the hydrophobic compound can be reacted at a low cost.

[0031] Furthermore, according to the above reaction apparatus, the hydrophobic liquid substance and the aqueous liquid substance circulate separately in the first channel 1 and the second channel 2, so that the hydrophobic liquid substance and the aqueous liquid substance are separated from each other. Can be passed through the third channel 3 at a desired speed. That is, the amount of the hydrophobic compound flowing through the first channel 1 and the amount of the biocatalyst flowing through the second channel 2 can be freely controlled as necessary. Therefore, generation of unreacted hydrophobic compounds can be sufficiently suppressed.

[0032] It is preferable that the hydrophobic liquid substance and the aqueous liquid substance are brought into contact with each other by the following method because the contact efficiency is improved. For example, (A) a method of forming and flowing phases in the third channel and bringing them into contact with each other in a separated state, and (B) a method of forming droplets in the third channel and alternating the droplets. And (C) a method of forming an emulsion in the third channel and bringing it into contact.

[0033] In these methods (A) and (B), the shape of the third channel 3, the contact angle of each of the hydrophobic liquid substance and the aqueous liquid substance with respect to the material of the third channel 3, the viscosity, the amount of the flowing liquid, and Each can be realized by controlling the speed of distribution and the like. The method (C) can be realized by, for example, another embodiment described later.

[0034] The biocatalyst used for the reaction of the hydrophobic reactant can be reused. That is, it is injected from the second syringe 22 and passes through the second tube 12 and the third channel 3. After reaching the receiver 4, the biocatalyst may be removed from the receiver 4 and reinjected into the second syringe 22. Since this reaction is a biocatalytic reaction, it can be used repeatedly until the catalyst is deactivated.

[0035] The hydrophobic compound may be a solid or liquid or a volatile compound. However, if it is a solid, it must be dissolved or dispersed in some hydrophobic solvent. Examples of hydrophobic compounds include toluene, naphthalene, phenol, benzaldehyde, anisole, catechol, hydroxybenzoic acid and other aromatic compounds or derivatives thereof, thiophene, benzothiophene, dibenzothiophene, furan, imidazole, thiazole, Heterocyclic compounds such as pyridine, pyrazine, indole or derivatives thereof, C5-C20 aliphatic hydrocarbons such as pentane, hexane, pentadecane, octadecane, arachidonic acid, linoleic acid, or derivatives thereof, α-vinene, limonene Terpenes such as menthol, ketones, diketones, polyketones, epoxides, esters, latatones, amino acids and the like. Examples of the hydrophobic solvent include aromatic solvents such as toluene, aliphatic solvents such as heptane, hexane, cyclohexane, η tetradecane, and isooctane; dimethyl ether, tetrahydroxyfuran, t-butyl methyl ether, diisopropyl ether, and the like. Ether solvents, ester solvents such as ethyl acetate, and halogen solvents such as methylene chloride.

[0036] Examples of the biocatalyst include, for example, enzymes such as lipase, esterase, protease, amylase, isomerase, dehydrogenase, lyase, transferase, kinase, anoredolase, ligase, Schizosaccharomyces yeast, Pichia yeast , Candida yeast, Pseudomonas bacteria, Sphingomonas bacteria, Alkagenes bacteria, Burkholderia bacteria, Acinetopacter bacteria, Corynebacterium bacteria, Bacillus bacteria, Brevibacillus bacteria, Giobacillus bacteria, Nocardia Microbial power S such as genus bacteria, Rhodococcus bacteria, Gordonia bacteria, or Mycobacterium bacteria. These can be used by dissolving or dispersing in water or the like.

FIG. 3 is a cross-sectional view showing the second to third aspects of the third channel 3. The channel 21a of the second embodiment shown in (a) of FIG. 3 is a channel formed by laminating two base materials. In other words, grooves are provided in the lower part of one base material 23a and the upper part of the other base material 23b, Channel 21a is formed by shelling. The channel 21b of the third embodiment shown in (b) of FIG. 3 is formed by laminating three base materials. That is, a through groove is provided in the base material 24b which is an intermediate layer, and the base material 24a and 24c are laminated so as to sandwich the base material 24b between the upper part and the lower part, thereby forming the channel 21b. Note that channel 21a and channel 2 lb are equivalent to channel 3.

[0038] These base materials 23a, 23b, 24a to 24c can be arbitrarily determined, glass such as quartz glass or Pyrex (registered trademark) glass, polymers such as PDMS, polycarbonate or polyimide, iron, stainless steel, aluminum In addition, metals such as nickel or copper, silicon, or the like can be used, and the respective base materials in the case of stacking may be of the same type or different types.

FIG. 4 is a partial plan view showing a microchannel chip according to another embodiment of the hydrophobic compound reactor according to the present invention. As shown in FIG. 4, the microchannel chip 200 of the present embodiment is the first implementation except that the tip portion la of the first channel 1 is opposed to the end portion of the third channel 3 via the dispersion portion 31. The configuration is the same as that of the microchannel chip 10 of the embodiment.

[0040] Here, the dispersing portion 31 is, for example, plate-shaped and has a plurality of through holes 32 as shown in FIG. Therefore, the first channel 1 is connected to the third channel 3 through the through hole 32 of the dispersion part 31. FIG. 5 is a partial front view showing the dispersion portion 31 as viewed from the direction of the force from the tip end portion la of the first channel 1 to the end portion of the third channel 3.

[0041] According to the microchannel chip 200, when the hydrophobic liquid substance is circulated through the first channel 1 and the aqueous liquid substance is circulated through the second channel 2, the hydrophobic liquid substance flowing through the first channel 1 is The first channel 1 reaches the tip la. On the other hand, the aqueous liquid material flowing through the second channel 2 flows into the third channel 3. Then, the hydrophobic liquid material that has reached the tip portion la of the first channel 1 flows into the third channel 3 through the through hole 32 of the dispersion portion 31, and the hydrophobic liquid material and the aqueous liquid material are mixed. . At this time, the hydrophobic liquid substance has a microsphere shape. Therefore, the emulsion is formed by mixing the hydrophobic liquid substance and the aqueous liquid substance, and the hydrophobic liquid substance and the aqueous liquid substance come into contact with each other. When the hydrophobic liquid substance and the aqueous liquid substance come into contact with each other in this way, Tactile efficiency can be greatly improved.

[0042] The present invention is not limited to the above embodiment. For example, in the above embodiment, a diaphragm pump or the like may be used instead of the force syringes 21 and 22 that use the syringes 21 and 22 to distribute the hydrophobic liquid substance and the aqueous liquid substance.

In the above embodiment, it is also useful to provide check valves in the first channel 1, the second channel 2, and the third channel 3. If a check valve is provided, fluid backflow can be prevented, and problems due to backflow can be prevented.

Furthermore, it is also useful to provide a heating device or a cooling device for the microchannel chip 10 used in the above embodiment. When the reaction is carried out using the reaction apparatus of the present invention, the chemical reaction includes an exothermic reaction and an endothermic reaction. Therefore, if a device capable of adjusting the temperature is provided, the reaction can be promoted more reliably. In addition, when a temperature change is caused by a chemical reaction, the fluid force S expands, which may cause unnecessary bubbles, which may cause clogging of the channel. For this reason, it is useful to provide a device capable of adjusting the temperature from this point.

Furthermore, in the above-described embodiment, the force branch channel 3 a in which the branch channel 3 a is branched and connected to the fourth tube 8 on the other end side of the third channel 3 is not necessarily required. Or conversely, there may be two or more branch channels 3a.

Example

Hereinafter, the present invention will be described in more detail with reference to Examples and Comparative Examples, but the present invention is not limited to the following examples.

[Example 1]

First, the nocardiooff-ohmic bacterium Rhodococcus sp. Was cultured, and using this bacterium, a cell suspension A, an aqueous liquid, was prepared so that the cell concentration was OD = 10. While n

660

-Hydrophobic liquid A in which dibenzothiophene was dissolved in tetradecane at a ratio of 0.02 wt% (ImM) was prepared. Dibenzothiophene has the property of being oxidized to 2-hydroxybiphenyl and releasing sulfate ions into an aqueous solution in the presence of the nocardiochrome bacterium Rhodococcus sp.

[0048] Next, the reaction apparatus shown in Fig. 1 was prepared. Then, the cell suspension A is transferred to the second syringe 2 2 (opening diameter φ 4.5 mm), and the hydrophobic liquid A was injected into the first syringe 21 having the same opening diameter as the second syringe 22. Then, the hydrophobic liquid A is circulated from the first syringe 21 via the first tube 21 to the first channel 1 and the cell suspension A is passed from the second syringe 22 via the second tube 12 to the second channel 2. Circulated. Then, the hydrophobic liquid A and the cell suspension A were brought into contact in the third channel having a channel width of 100 zm, and the reaction of dibenzothiophene was performed. At this time, it was circulated so that the ratio of the hydrophobic liquid A and the cell suspension A was 1: 1 by volume. The reaction time was 30 seconds, and the concentration of 2-hydroxybiphenyl, the reaction product at this time, was measured. Figure 6 shows the result of converting this measured value per minute. In FIG. 6, the result of this example is indicated by “Reaction A”.

[0049] (Example 2)

The concentration of 2-hydroxybiphenyl as a reaction product was measured in the same manner as in Example 1 except that the reaction time was 1 minute. Figure 6 shows the conversion results per minute. In FIG. 6, the result of this example is indicated by “Reaction B”.

[Example 3]

The concentration of 2-hydroxybiphenyl as a reaction product was measured in the same manner as in Example 1 except that the reaction time was 3 minutes. Figure 6 shows the conversion results per minute. In FIG. 6, the result of this example is indicated by “Reaction C”.

[0051] (Example 4)

Mycobacterium sp. Having the ability to oxidize carbon-carbon double bonds was cultured in a culture vessel sealed with ethylene gas and sealed. Using this bacterium, a cell suspension B, which is an aqueous liquid, was prepared so that the cell concentration was OD = 17. Meanwhile, to n—

600

A hydrophobic liquid B in which styrene was dissolved in xadecane at a ratio of 2 wt% (174 mM) was prepared. Styrene, a raw material, has the property of being oxidized to R-styrene oxide (R_l, 2_epoxyethylbenzene) in the presence of Mycobacterium sp.

[0052] The cell suspension B is used instead of the cell suspension A, the hydrophobic liquid B is used instead of the hydrophobic liquid A, and the reaction time is 0.1 hour (6 minutes). Except for this, the production amount (/ ig) of R-styrene oxide as a reaction product was measured in the same manner as in Example 1. In this case, R— The optical purity of styrene oxide was 99% or more. Figure 7 shows the results of converting these measured values per hour. In FIG. 7, the result of this example is indicated by “Reaction D”.

[0053] (Example 5)

The production amount (z g) of R-styrene oxide as a reaction product was measured in the same manner as in Example 4 except that the reaction time was 0.2 hour (12 minutes). The optical purity at this time was 99% or more. Figure 7 shows the result of converting this measured value per hour. In FIG. 7, the result of this example is indicated by “Reaction E”.

[Comparative Example 1]

The concentration of 2-hydroxybiphenyl as a reaction product was measured in the same manner as in Example 1 except that a 100 ml Erlenmeyer flask was used without using a microchannel chip, and the reaction time was 1 hour. Figure 6 shows the conversion results per minute. In FIG. 6, the result of this comparative example is indicated by “control”.

[0055] (Comparative Example 2)

Production of R-styrene oxide as a reaction product in the same manner as in Example 4 except that a test tube (made of glass, φ21 mm X 200 mm) was used without using a microchannel chip, and the reaction time was 20 hours. The amount (/ ig) was measured. The optical purity at this time was 99% or more. Figure 7 shows the result of converting this measured value per hour. In FIG. 7, the result of this example is indicated by “control”.

[0056] From the results of the above Examples:! To 5 and Comparative Examples 1 and 2, it was found that the reaction efficiency of the hydrophobic compound according to the present invention was greatly improved. This confirms that the reaction efficiency of the hydrophobic compound according to the present invention can be sufficiently increased. In addition, since the reaction efficiency can be sufficiently increased as described above, it is considered that a hydrophobic compound can be reacted in the presence of a biocatalyst in a wide variety of reactions. Industrial applicability

[0057] According to the present invention, the reaction efficiency of the hydrophobic compound can be sufficiently increased in a wide variety of reactions. In other words, it is applied to hydrolysis reactions, oxidation-reduction reactions, transfer reactions, substitution reactions, elimination reactions, condensation reactions, addition reactions, isomerization reactions, asymmetric synthesis reactions, etc. that are important in organic chemistry. It is possible to make progress in the field of biocatalytic reactions of hydrophobic compounds in the future.

Claims

The scope of the claims

[1] A hydrophobic compound reaction apparatus for reacting a hydrophobic compound using a biocatalyst, and a first channel for circulating a hydrophobic liquid substance containing the hydrophobic compound,

A second channel for distributing an aqueous liquid substance containing a biocatalyst, and

An apparatus for reacting a hydrophobic compound, comprising: a microchannel chip connected to the first channel and the second channel and having a third channel through which the hydrophobic liquid substance and the aqueous liquid substance circulate.

[2] The hydrophobic compound reactor according to [1], wherein the channel width force of the third channel is not more than Slmm.

[3] A method for reacting a hydrophobic compound by using a biocatalyst to react the hydrophobic compound with a hydrophobic liquid substance containing the hydrophobic compound and an aqueous liquid substance containing the biocatalyst with a channel width of lm m or less. A method for reacting a hydrophobic compound, characterized in that the contact is made in a channel having a surface.

[4] The hydrophobic liquid substance and the aqueous liquid substance are circulated in the third channel, and the phases of the hydrophobic liquid substance and the aqueous liquid substance are brought into contact with each other in a separated state. Item 4. A method for reacting a hydrophobic compound according to Item 3.

5. The sparse structure according to claim 3, wherein droplets of the hydrophobic liquid material and droplets of the aqueous liquid material are respectively formed in the third channel, and are circulated and contacted alternately. Reaction method of aqueous compound.

[6] The emulsion consisting of the hydrophobic liquid substance and the aqueous liquid substance is formed in the third channel, and the hydrophobic liquid substance and the aqueous liquid substance are brought into contact with each other. The reaction method of the hydrophobic compound as described.

7. The method for reacting a hydrophobic compound according to any one of claims 3 to 6, wherein the biocatalyst is a microorganism having an ability to take up the hydrophobic compound into cells.

[8] The microorganism is Pichia yeast, Candida yeast, Pseudomonas bacteria, Rhodoco 8. The method for reacting a hydrophobic compound according to claim 7, wherein the method is a bacterium belonging to the genus Bacus, Mycobacterium, Bacillus, Brevibacillus, or Gibacillus.