JP2017136049A - Process for producing amide compound and apparatus for producing amide compound - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide an efficient process for producing an amide compound that is industrially very important and an apparatus for producing an amide compound suitably used in the process.SOLUTION: Provided is a process for producing an amide compound by using microbial cells containing nitrile hydratase and/or a cell-treated product thereof, in which the microbial cell and/or the processed product thereof are supplied to a reaction tank, in which a liquid is kept at 10 to 40°C, at a temperature of -20°C or higher from and less than the temperature of a liquid in the tank. Also provided is an apparatus for producing an amide compound comprising: a reaction tank for producing an amide compound using bacterial cells containing nitrile hydratase and/or a cell-treated product thereof; and a microbial cell catalyst-storing tank for supplying the microbial cells and/or the processed product thereof at a temperature of -20°C or higher and less than the temperature of a liquid in the reaction tank.SELECTED DRAWING: None

Description

  The present invention relates to a method for producing an amide compound and an apparatus for producing an amide compound, which use a microbial cell and / or a treated product thereof as a catalyst and hydrate a nitrile compound to obtain an amide compound.

  As one of the main production methods of amide compounds, a hydration method using a nitrile compound as a raw material is used in many cases, and as an industrial production method of an amide compound such as acrylamide, for example, a metal such as Raney copper A method of hydrating a nitrile compound such as acrylonitrile using copper as a catalyst, or a method of hydrating a nitrile compound using a microbial cell containing nitrile hydratase and / or a treated product thereof as a catalyst in recent years. Are known.

  The method of using bacterial cells and / or treated products thereof as a catalyst has attracted industrial attention because of high conversion and selectivity of nitrile compounds such as acrylonitrile.

 Since the cell catalyst may cause a decrease in activity at a temperature exceeding the heat resistance of the cell, the reaction vessel is usually operated at a temperature at which the activity is maintained. For example, in Patent Document 1, in a method for producing acrylamide from acrylonitrile using a biocatalyst that is easily deactivated by heat, the reaction temperature and the reaction temperature are efficiently removed while being an industrial reaction scale. A method has been proposed in which the temperature difference from the temperature of the cooling water introduced into the heat exchanger is maintained within a certain range. Patent Document 2 discloses a method for producing acrylamide from acrylonitrile using a biocatalyst having a nitrile hydratase, and a method for producing a high-quality acrylamide having a higher concentration with a smaller amount of catalyst, and storing at less than 30 ° C. A method of using the prepared acrylonitrile has been proposed.

 However, in the methods described in Patent Document 1 and Patent Document 2, temperature control is performed only in the reaction tank and the supplied raw materials, and it is insufficient to efficiently use the bacterial cell catalyst. There is still room for consideration.

International Publication No. 2010/038832 Pamphlet International Publication No. 2011/138966 Pamphlet

  In the hydration reaction of a nitrile compound using a microbial cell and / or a treated product thereof, it is required to produce an amide compound more efficiently.

  An object of the present invention is to provide a method for efficiently producing an amide compound and an apparatus for producing an amide compound suitably used in the method.

  The present inventors have conducted intensive research to solve the above problems. As a result, the inventors have found that the above problems can be solved by a method for producing an amide compound having the following steps, and have reached the present invention.

That is, the present invention is as follows.
[1] In a method for producing an amide compound from a nitrile compound using a microbial cell containing nitrile hydratase and / or a treated product thereof, the temperature of the liquid containing at least the nitrile compound in the reaction vessel is 10 to 10%. An amide characterized by supplying a bacterial cell and / or a treated product thereof to a reaction vessel maintained at 40 ° C. at a liquid temperature in the reaction vessel of −20 ° C. or higher and lower than the liquid temperature in the reaction vessel. Compound production method [2] The amide compound is acrylamide or methacrylamide The production method according to [1] [3] A microbial compound containing nitrile hydratase and / or a treated product thereof, A reaction tank to be produced, and a cell catalyst storage tank for supplying cells and / or treated cells thereof at a temperature not lower than −20 ° C. and not higher than the temperature in the reaction tank. Apparatus for producing an amide compound

 In the present invention, a method for efficiently producing an amide compound without reducing the activity of the bacterial cell catalyst by supplying the bacterial cell and / or the treated product thereof to the reaction vessel at a specific temperature, and the method It is possible to provide an apparatus for producing an amide compound that is suitably used for the above.

  Hereinafter, the manufacturing method and manufacturing apparatus of the amide compound of this invention are demonstrated.

[Method for producing amide compound]
The method for producing an amide compound of the present invention is a method for producing an amide compound using a microbial cell containing nitrile hydratase and / or a treated product thereof, and a microbial cell maintained at a specific temperature and / or its bacterium. Supplying a body treatment product to the reaction vessel.

<Bacteria and / or treated cells containing nitrile hydratase>
In the present invention, a microbial cell containing nitrile hydratase and / or a treated product thereof (hereinafter, also simply referred to as “bacterial cell catalyst”) is used as a hydration catalyst of a nitrile compound to an amide compound.

  Nitrile hydratase refers to an enzyme (protein) having the ability to hydrolyze a nitrile compound to produce a corresponding amide compound (hereinafter also referred to as “nitrile hydratase activity”).

  The microbial cell containing nitrile hydratase is not particularly limited as long as it produces nitrile hydratase and retains nitrile hydratase activity in an aqueous solution of a nitrile compound and an amide compound. The nitrile hydratase-producing cells include Nocardia, Corynebacterium, Bacillus, thermophilic Bacillus, Pseudomonas, and Micrococcus. , Rhodococcus genus represented by Rhodochrous species, Acinetobacter genus, Xanthobacter genus, Streptomyces genus, Rhizobium genus, Klebsiella genus, Klebsiella Representative of the genus Enterobacter, Erwinia, Aeromonas, Citrobacter, Achromobacter, Agrobacterium or thermophila Genus Pseudonocardia, Bacteridium, Brevibak Bacteria such as belonging to the helium (Brevibacterium) genus can be mentioned. These may be used alone or in combination of two or more.

 In addition, a transformant in which a nitrile hydratase gene cloned from these bacterial cells is highly expressed in an arbitrary host, and one or more of the constituent amino acids of the nitrile hydratase using other DNA by using recombinant DNA technology And a transformant in which a mutant nitrile hydratase having further improved amide compound resistance, nitrile compound resistance, and temperature resistance is expressed by substitution, deletion, deletion or insertion. In addition, as an arbitrary host mentioned here, Escherichia coli can be mentioned as a representative example as in Examples described later, but is not particularly limited to Escherichia coli, and Bacillus subtilis and other Bacillus genera. Other strains such as fungi, yeasts and actinomycetes are also included. As an example of such a case, MT-10822 [This strain is the Institute of Biotechnology, Institute of Industrial Technology, Ministry of International Trade and Industry, 1-3 1-3 Higashi, Tsukuba, Ibaraki, on February 7, 1996 (currently Tsukuba, Ibaraki) City East 1-1-1 Tsukuba Center Chuo No. 6 (National Institute of Technology and Evaluation, Patent Biological Deposit Center) under the accession number FERMBP-5785, based on the Budapest Treaty on the international recognition of deposits of bacterial cells under patent procedures It has been deposited. ].

  Among these bacterial cells, in terms of having a highly active and highly stable nitrile hydratase, cells belonging to the genus Pseudonocardia and the nitrile hydratase gene cloned from the bacterial cells can be used in any host. A highly expressed transformant and a transformant expressing a mutant nitrile hydratase are preferred. In addition, the said transformant is preferable at the point which raises the stability of nitrile hydratase more and the activity per microbial cell is higher.

  Also preferred are Rhodococcus rhodochrous J-1, which can highly express nitrile hydratase in the cells, and transformants in which the nitrile hydratase gene cloned from the cells is highly expressed in any host. . The microbial cells producing the nitrile hydratase can be prepared by a general method known in the fields of molecular biology, biotechnology, and genetic engineering.

  The recombinant vector according to the present invention contains a gene encoding nitrile hydratase, and can be obtained by linking a gene encoding nitrile hydratase to the vector. The vector is not particularly limited. For example, a gene encoding nitrile hydratase in a commercially available expression plasmid represented by pET-21a (+), pKK223-3, pUC19, pBluescriptKS (+), pBR322, etc. Thus, the expression plasmid of nitrile hydratase can be constructed. The host organism to be used for transformation is not limited as long as the recombinant vector is stable and capable of self-propagation and can express foreign DNA traits. For example, E. coli is a good example. The transformant having the ability to produce nitrile hydratase can be obtained by introducing it into Bacillus subtilis, yeast or the like.

  Bacteria that produce nitrile hydratase as described above may be appropriately cultured and grown by known methods to produce nitrile hydratase. As a medium used in this case, either a synthetic medium or a natural medium can be used as long as it contains a suitable amount of carbon source, nitrogen source, inorganic salts and other nutrients. For example, after inoculating the cells in a normal liquid medium such as LB medium or M9 medium, an appropriate culture temperature (generally 20 ° C. to 50 ° C., but in the case of thermophilic bacteria, 50 ° C. or higher) However, it can also be prepared by culturing. Culturing can be performed using a conventional culture method such as shaking culture, aeration and agitation culture, continuous culture, or fed-batch culture in a liquid medium containing the culture components. The culture temperature of the transformant is preferably 15 to 37 ° C. The culture conditions are not particularly limited, and may be appropriately selected depending on the type of culture and the culture method, and it is sufficient that the strain can grow and produce nitrile hydratase.

  In the present invention, in order to react the bacterial body producing the above-mentioned nitrile hydratase with a nitrile compound, various treatments such as collection of bacteria by centrifugation, etc., and crushing to produce a crushed bacterial body, The microbial cells that have been subjected to any of these treatments are collectively referred to as processed microbial cells.

  The form of the disrupted cell is not particularly limited as long as it contains a nitrile hydratase-producing cell. For example, the culture solution containing the cell itself, the culture solution is centrifuged and separated and recovered. And those obtained by washing the collected cells with physiological saline or the like.

<Nitrile compound>
Examples of the nitrile compound include an aliphatic nitrile compound having 2 to 20 carbon atoms and an aromatic nitrile compound having 6 to 20 carbon atoms, which may be used alone or in combination of two or more.

  Examples of the aliphatic nitrile compound include saturated or unsaturated nitriles having 2 to 6 carbon atoms; specifically, acetonitrile, propionitrile, butyronitrile, isobutyronitrile, valeronitrile, isovaleronitrile, capronitrile Aliphatic saturated mononitriles such as malononitrile, succinonitrile, adiponitrile, and the like; and aliphatic unsaturated nitriles such as acrylonitrile, methacrylonitrile, and crotonnitrile.

Examples of the aromatic nitrile compound include benzonitrile, o-, m- or p-chlorobenzonitrile, o-, m- or p-fluorobenzonitrile, o-, m- or p-nitrobenzonitrile, o- , M- or p-tolunitrile, o-, m- or p-cyanopyridine, benzyl cyanide.
Among the nitrile compounds, acrylonitrile and methacrylonitrile are preferable.

<Water (raw water)>
The raw material water is not particularly limited, and purified water such as distilled water or ion exchange water can be used.

<PH regulator>
The pH adjuster is used to adjust the pH of the reaction mixture to a suitable range for keeping the activity of the bacterial cell catalyst good.
When the pH suitable for the reaction is less than 7, an acid can be used as a pH adjuster.

As the acid used as the pH adjuster, either an inorganic acid or an organic acid can be used. Examples of inorganic acids include hydrohalic acids such as hydrogen chloride, hydrogen bromide, and hydrogen iodide, hypochlorous acid, chlorous acid, chloric acid, perchloric acid, hypobromite, bromite, bromine Examples thereof include halogenated oxoacids such as acid, perbromic acid, hypoiodic acid, iodic acid, iodic acid, and periodic acid, sulfuric acid, nitric acid, phosphoric acid, and boric acid. Examples of organic acids include formic acid, acetic acid, propionic acid, acrylic acid, methacrylic acid, crotonic acid, succinic acid, malonic acid, fumaric acid, maleic acid, citric acid, lactic acid, benzoic acid and other carboxylic acids such as methanesulfonic acid, Examples include sulfonic acids such as ethanesulfonic acid, benzenesulfonic acid, and p-toluenesulfonic acid. The acid used as the pH adjusting agent can be used in any state of gas, solid and liquid, but considering the ease of supply to the reaction vessel, it is preferable to use the acid in the liquid state. The acid in the state is more preferably used as an aqueous solution. In consideration of controllability of pH adjustment of the reaction tank, it is more preferable to use a liquid acid as an aqueous solution. The concentration of the acid when used as an aqueous solution is not particularly limited, but it is difficult to adjust pH when a high concentration aqueous solution is used. Hereinafter, it is more preferably 1 wt% or more and 50 wt% or less.
When the pH suitable for the reaction is higher than 7, a base can be used as a pH regulator.

  As the base used as the pH adjuster, either an inorganic base or an organic base can be used. Examples of the inorganic base include alkali metal hydroxides such as lithium hydroxide, sodium hydroxide, and potassium hydroxide, alkaline earth metal hydroxides such as magnesium hydroxide and calcium hydroxide, lithium carbonate, and sodium carbonate. And alkali metal carbonates such as potassium carbonate, alkali metal hydrogen carbonates such as lithium hydrogen carbonate, sodium hydrogen carbonate and potassium hydrogen carbonate, and ammonia. Examples of the organic base include trimethylamine, triethylamine, aniline, and pyridine. The base used as the pH adjusting agent can be used in any state of gas, solid, and liquid, but considering the ease of supply to the reaction vessel, it is preferable to use the base in the liquid state. The base in a state is more preferably used as an aqueous solution. In consideration of controllability of pH adjustment of the reaction tank, it is more preferable to use a liquid base as an aqueous solution. The concentration of the base when used as an aqueous solution is not particularly limited, but it is difficult to adjust the pH when a high concentration aqueous solution is used. Hereinafter, it is more preferably 1 wt% or more and 50 wt% or less.

<Reaction tank>
As the reaction tank, a single-stage reaction tank composed of one reactor may be used, or a multi-stage reaction tank composed of a plurality of reactors may be used. As the reactor, a tank reactor or a tube reactor may be used. As the tank reactor, a reactor equipped with a stirrer is preferable. When the tank reactor has a stirrer, the stirrer blade of the stirrer can be of any shape, for example, propeller blade, flat paddle blade, pitched paddle blade, flat turbine blade, pitched turbine blade, ribbon blade, anchor blade , Full zone wings. One stirring blade may be provided, or a plurality of stirring blades may be provided.

 An external circulation line equipped with a circulation pump may be installed in the reaction tank. One external circulation line provided with a circulation pump may be installed in the reaction tank, or a plurality of external circulation lines may be installed.

  When an external circulation line equipped with a pump is installed in the reaction tank, the pump provided in the external circulation line is not particularly limited. For example, a turbo pump such as a centrifugal pump, a tilt pump or an axial pump And positive displacement pumps such as reciprocating pumps and rotary pumps.

 When installing an external circulation line equipped with a pump in the reaction tank, a part of the reaction mixture is taken out from the reaction tank using the pump provided in the external circulation line and reacted via the external circulation line. Returned to the tank. It is preferable that a temperature control device is installed in the external circulation line installed in the reaction tank. As the temperature control device, a heat exchanger is preferably used. Examples of the heat exchanger include a multi-tube cylindrical type, a spiral tube type, a spiral plate type, a plate type, and a double pipe type.

 When installing an external circulation line equipped with a pump in a multistage reaction tank composed of a plurality of reactors, all of the external circulation lines equipped with a pump may be installed in each reactor. It may be installed in only one reactor. In the case of multiple reactors, if the external circulation line equipped with a pump is installed only in one of the reactors, it is installed in the first reactor (the most upstream reactor). It is preferred that

  Using a reaction tank composed of a first-stage tank reactor and a second-stage tube reactor, the reaction liquid taken out from the tank reactor is further reacted in the tube reactor, and the tank reaction A reaction tank in which an external circulation line equipped with a pump is installed in the vessel is more preferable because the conversion rate can be improved.

  The tank reactor and the tube reactor may or may not be equipped with a heat exchanger as long as the nitrile hydratase activity of the bacterial cell catalyst is maintained. In order to control, the reactor preferably comprises a heat exchanger. As the heat exchanger, it is preferable to install a multi-tubular cylindrical type, a spiral tube type, a spiral plate type, a plate type, a double pipe type, etc. on the external circulation line. The thing of the form directly installed in reactors, such as a coil type, is mentioned. When the reactor is a tubular reactor, the reactor itself can be composed of a multi-tubular cylindrical or double-tube heat exchanger.

  Examples of the reaction method include (1) a method in which a reaction is carried out after all of the cell catalyst, reaction raw material (nitrile compound and raw water) and pH regulator are charged in a reaction tank at once (batch reaction), (2) fungus After a part of the body catalyst, reaction raw material (nitrile compound and raw water) and pH regulator are charged into the reaction tank, the remaining cell catalyst, reaction raw material (nitrile compound and raw water) and pH are continuously or intermittently added. A method of performing a reaction by supplying a regulator (semi-batch reaction), (3) a continuous or intermittent supply of a cell catalyst, a reaction raw material (nitrile compound and raw water) and a pH regulator, and a reaction solution (bacteria) Including continuous catalyst, unreacted raw material, pH adjuster and generated amide compound, etc.), while continuously removing the reaction solution in the reaction vessel without taking out the whole reaction solution. Continuous reaction) and the like. Among these, a continuous reaction is preferable because it is easy to industrially produce an amide compound in large quantities and efficiently.

  The reaction is performed in the presence of a cell catalyst. The use form of the cell catalyst is preferably a suspension bed. For example, in the case of continuous reaction, a suspension of the cell catalyst may be prepared and the suspension supplied to the reaction vessel.

  In addition, when using the multistage reaction tank comprised from several reactors as a reaction tank, as the structure, (a) Cell body catalyst, reaction raw material (nitrile compound and raw material water), and pH regulator are the upper reaction. A serial type mode in which a reaction liquid (including a bacterial cell catalyst, an unreacted raw material, a pH regulator, and a generated amide compound) is supplied to the lower reactor and is taken out from the upper reactor, (B) A parallel mode in which a cell catalyst, a reaction raw material (nitrile compound and raw water) and a pH adjuster are directly supplied to two or more reactors (without going through other reactors) can be mentioned.

  For example, in the case of performing a continuous reaction using a multistage reaction tank, the supply destination of the cell catalyst, the reaction raw material (nitrile compound and raw water) and the pH adjuster is the first-stage reactor (the most upstream reaction). It is not limited only to the reactor), but may be a reactor after the second stage (reactor located downstream).

  Although the reaction tank temperature which is the liquid temperature in a reaction tank is based also on the heat resistance of a microbial cell catalyst, it is normally set to 10-40 degreeC, Preferably it sets to 10-30 degreeC. When the reaction vessel temperature is in the above range, it is preferable in that the nitrile hydratase activity of the bacterial cell catalyst can be maintained well.

  The reaction tank temperature refers to the liquid temperature in the reactor when the reaction tank is composed of only one reactor, and in the case where the reaction tank is composed of a plurality of reactors. Refers to the liquid temperature. The reaction vessel temperature can be measured, for example, by a thermocouple method (eg, K type). The reaction vessel temperature can be measured at an arbitrary location in the reaction vessel, and specifically can be measured at the reaction vessel outlet (reaction liquid outlet).

Volume of the reaction vessel is not particularly limited, considering the industrial production, usually 0.1 m ³ or more, preferably from 1 to 100 m ^3, more preferably 5 to 50 m ^3. When the reaction vessel is composed of a plurality of reactors, the volume refers to the volume of each reactor.

  The reaction is generally carried out under normal pressure, but can also be carried out under pressure in order to increase the solubility of the nitrile compound. The pH in the reaction vessel is not particularly limited as long as it is a suitable range for maintaining the activity of the bacterial cell catalyst, but is preferably in the range of pH 5 to pH 10. When the pH is within the above range, it is preferable in that the nitrile hydratase activity can be favorably maintained.

  When the reaction tank is composed of only one reactor, the pH of the reaction tank refers to the pH within the reactor, and when the reaction tank is composed of a plurality of reactors, The pH of the The pH of the reaction vessel can be measured by, for example, an indicator method, a hydrogen electrode method, a quinhydrone electrode method, an antimony electrode method, or a glass electrode method. The pH of the reaction vessel can be measured at any place in the reaction vessel, and specifically, can be measured at the reaction vessel outlet (reaction liquid outlet).

<Cell catalyst storage tank>
The cell catalyst storage tank has a catalyst supply pipe that holds a cell catalyst suspension and supplies the cell catalyst suspension to the reaction tank. The cell catalyst storage tank has a temperature control device from the viewpoint of controlling the liquid temperature of the cell catalyst suspension.

 The temperature control device in the fungus body storage tank may be provided on an external circulation line that the fungus body storage tank may have, or may be provided in the fungus body storage tank itself. Examples of the temperature control device in the bacterial cell storage tank include a heat exchanger. As the heat exchanger, for example, an external circulation line of a bacterial cell storage tank such as a multi-tubular heat exchanger, a spiral tube heat exchanger, a spiral plate heat exchanger, a plate heat exchanger, a double tube heat exchanger, etc. Or a type installed directly in a cell catalyst storage tank such as a jacket type heat exchanger or a coil type heat exchanger.

 The bacterial cell catalyst storage tank may have a stirrer or may not have a stirrer, but it is preferable to have a stirrer from the viewpoint of controlling the liquid temperature of the bacterial cell catalyst. When the cell catalyst storage tank has a stirrer, the stirring blade of the stirrer can be of any shape, for example, propeller blade, flat paddle blade, pitched paddle blade, flat turbine blade, pitched turbine blade, ribbon blade, anchor blade , Full zone wings. One stirring blade may be provided, or a plurality of stirring blades may be provided.

<Supply of reaction raw materials>
The raw water supplied to the reaction tank may be supplied alone to the reaction tank, or may be supplied to the reaction tank after being mixed with the nitrile compound. When supplying raw material water to the reaction tank alone, there is no particular restriction on the position of the supply port to the reaction liquid in the reaction tank of the raw water supply pipe, and even if it is installed above the reaction liquid, May be installed.

  The method for supplying raw water when supplying raw water alone to the reaction vessel is not particularly limited, and a known method can be used. As a method of supplying the raw water to the reaction tank, a device having a liquid transport function can be used. For example, a centrifugal pump, a tilting pump, an axial flow pump, or a turbo pump, a reciprocating pump, a rotary pump, Pumps such as positive displacement pumps and conveyors such as screw conveyors can be used. As a method for supplying the raw water to the reaction tank, the raw water can be supplied without using the equipment having the liquid transport function. When equipment with a liquid transport function is not used, for example, a raw water storage tank for storing raw water is installed at the top of the reaction tank, the raw water storage tank and the reaction tank are connected by a raw water supply pipe, and gravity is used. There is a method of supplying by dropping. Or the method of supplying using the pressure difference of the raw material water storage tank and reaction tank which arise by pressurizing a raw material water storage tank is mentioned.

  There may be one raw water supply pipe for supplying raw water to the reaction tank, or a plurality of raw water supply pipes may be provided for one reactor. There may be one supply port for the raw water supply pipe per supply pipe, or a plurality of supply ports may be provided. There is no restriction | limiting in particular in the shape of the supply port of a raw material water supply pipe, Any can be used suitably if it is a shape normally used.

 As a method of mixing the raw water and the nitrile compound in the case of supplying the raw water and the nitrile compound to the reaction tank, a known method can be used, for example, a method of stirring and mixing using a stirring tank, a raw material The method of mixing water and a nitrile compound in piping is mentioned. In the case of using a stirring tank, the shape of the stirring tank is not particularly limited, but a cylindrical stirring tank is generally used, and it can be used in any case of a vertical cylindrical shape and a horizontal cylindrical shape. it can. The stirring tank may be provided with a baffle plate or may not be provided with a baffle plate.

  In the case of using a stirring tank, a stirring blade having an arbitrary shape can be selected, and examples thereof include a propeller blade, a flat paddle blade, a pitched paddle blade, a flat turbine blade, a pitched turbine blade, a ribbon blade, an anchor blade, and a full zone blade. It is done. One stirring blade may be provided, or a plurality of stirring blades may be provided.

 In order to mix the raw material water and the nitrile compound in the pipe, the raw water and the nitrile compound can be directly mixed by combining the raw water supply pipe and the nitrile compound supply pipe. A method of actively mixing by installing a line mixer can be mentioned.

  When the raw water and nitrile compound are mixed and then supplied to the reaction vessel, the nitrile compound may be completely dissolved in water after mixing with water, but it is not necessarily required to be completely dissolved in water. What is necessary is just to mix by arbitrary ratios. Preferably, the ratio of water: nitrile compound is 100: 1 to 1: 100, more preferably 50: 1 to 1:50, and even more preferably 10: 1 to 1:10 by volume.

  When the raw water and the nitrile compound are mixed and then supplied to the reaction vessel, a known method can be used for supplying the raw water and the nitrile compound mixture. Specifically, the above raw water is used alone. The method illustrated by the supply method of raw material water in the case of supplying to a reaction tank can be used.

  When mixing the raw water and the nitrile compound and then supplying them to the reaction vessel, there may be one supply pipe for supplying a mixture of the raw water and the nitrile compound to the reaction vessel. May be. The supply port of the supply pipe may be one for each supply pipe, or a plurality of supply ports may be provided. There is no restriction | limiting in particular in the shape of the supply port of a supply pipe | tube, Any can be used suitably if it is a shape normally used.

 The nitrile compound supplied to the reaction vessel may be supplied to the reaction vessel after being mixed with the raw water, or may be supplied alone to the reaction vessel. When supplying the nitrile compound alone to the reaction vessel, there is no particular restriction on the position of the supply port to the reaction solution in the reaction vessel of the nitrile compound supply pipe, and even if it is installed above the reaction solution, May be installed.

 The supply method of the nitrile compound when supplying the nitrile compound alone to the reaction vessel is not particularly limited, and a known method can be used. Specifically, the raw material water is supplied to the reaction vessel. The method exemplified in the method can be used.

 There may be one nitrile compound supply pipe for supplying the nitrile compound to the reaction tank, or a plurality of nitrile compound supply pipes may be provided for one reactor. The supply port of the nitrile compound supply pipe may be one for each supply pipe, or a plurality of supply ports may be provided. The shape of the supply port of the nitrile compound supply pipe is not particularly limited, and any shape that is usually used can be suitably used.

<Supply of pH regulator>
The pH adjuster supplied to the reaction tank may be supplied alone to the reaction tank, or may be supplied to the reaction tank after being mixed with the aqueous medium and / or the mixture containing the aqueous medium supplied to the reaction tank. When supplying the pH regulator alone to the reaction tank, the position of the supply port to the reaction liquid in the reaction tank of the pH regulator supply pipe is not particularly limited. You may install in a reaction liquid.

 The supply method of the pH adjuster when supplying the pH adjuster alone to the reaction vessel is not particularly limited, and a known method can be used. Specifically, the above raw water is supplied to the reaction vessel. The method exemplified in the supplying method can be used.

 The number of pH adjusting agent supply pipes for supplying the pH adjusting agent to the reaction vessel may be one per reactor or plural. The supply port of the supply pipe for the pH adjusting agent may be one for each supply pipe, or a plurality of supply ports may be provided. The shape of the supply port of the pH adjusting agent supply pipe is not particularly limited, and any shape that is usually used can be suitably used.

 The aqueous medium in which the pH adjusting agent is mixed when the pH adjusting agent and the aqueous medium and / or the mixture containing the aqueous medium are mixed and then supplied to the reaction vessel is preferably raw water. Examples of the mixture containing an aqueous medium in which the pH adjuster is mixed include a reaction liquid containing a mixture of raw water and a nitrile compound, a cell catalyst, an unreacted raw material, a pH adjuster, and a generated amide compound. .

  As a method of mixing the pH regulator and the raw water when the pH regulator is mixed with the raw water and then fed to the reaction vessel, a known method can be used. The method illustrated by the method of mixing a nitrile compound can be used.

 A known method can be used as the method for supplying the mixture of the pH regulator and the raw water when the pH regulator is mixed with the raw water and then supplied to the reaction tank. The method illustrated by the supply method of raw material water in the case of supplying to a reaction tank independently can be used.

 The pH regulator and the aqueous medium and / or the mixture containing the aqueous medium to be supplied to the reaction tank may be mixed in any ratio, and preferably the pH regulator: the aqueous medium and / or the mixture containing the aqueous medium. The ratio is 1: 1 to 1: 100,000,000, more preferably 1:10 to 1: 10,000,000 in volume ratio.

  A known method can be used as a method of mixing the mixture of the pH regulator, the raw water and the nitrile compound when the pH regulator is mixed with the raw water and the nitrile compound and then supplied to the reaction vessel. The method exemplified in the method of mixing the raw material water and the nitrile compound can be used.

  When the pH regulator is mixed with the reaction solution containing the bacterial cell catalyst, unreacted raw material, pH regulator and the generated amide compound and then supplied to the reaction vessel, the pH regulator and bacterial cell catalyst, unreacted raw material, pH As a method of mixing with the reaction liquid containing the regulator and the generated amide compound, for example, the following method can be used. An external circulation line equipped with a pump for returning a part of the reaction mixture to the reaction tank is installed in the reaction tank, and the reaction is performed as a mixture containing an aqueous medium by coupling the external circulation line and the pH regulator supply pipe. Examples include a method in which a part of the reaction mixture taken out from the tank is used and a part of the reaction mixture and the pH adjuster are directly mixed in an external circulation line. A part of the reaction mixture directly mixed in the external circulation line and the pH adjusting agent are supplied to the reaction tank as they are using the external circulation line.

<Supply of bacterial cell catalyst>
The bacterial cell catalyst is supplied to a reaction tank in which the liquid temperature in the tank is kept at 10 to 40 ° C. at a temperature not lower than the liquid temperature in the reaction tank −20 ° C. and lower than the liquid temperature in the reaction tank. When the supply temperature of the bacterial cell catalyst is lower than −20 ° C. than the liquid temperature in the reaction vessel, the activity of the bacterial cell catalyst is lowered due to a rapid temperature rise when supplied to the reaction vessel, and the amide compound is efficiently produced. Is difficult to manufacture. When the supply temperature of the bacterial cell catalyst is higher than the temperature in the reaction tank, the activity of the bacterial cell catalyst is lowered while it is held in the bacterial cell storage tank, and it is difficult to efficiently produce the amide compound.

  The bacterial cell catalyst supplied to the reaction tank may be supplied alone to the reaction tank, or may be supplied to the reaction tank after being mixed with a mixture containing an aqueous medium supplied to the reaction tank. When supplying the cell catalyst alone to the reaction vessel, the installation position of the supply port to the reaction solution in the reaction vessel of the cell catalyst supply pipe is not particularly limited. You may install in a reaction liquid.

 The supply method of the bacterial cell catalyst when supplying the bacterial cell catalyst alone to the reaction vessel is not particularly limited, and a known method can be used. Specifically, the above raw water is supplied to the reaction vessel. The method exemplified in the supplying method can be used.

 The cell catalyst supply pipe for supplying the cell catalyst to the reaction vessel may be one for each reactor, or a plurality of supply tubes may be provided. There may be one supply port of the supply pipe for the bacterial cell catalyst, or a plurality of supply ports may be provided for each supply pipe. The shape of the supply port of the fungus body catalyst supply pipe is not particularly limited, and any shape having a shape normally used can be used.

  When a mixture containing a cell catalyst and an aqueous medium is mixed and then supplied to the reaction vessel, the mixture containing the aqueous medium mixed with the cell catalyst is a cell catalyst, unreacted raw material, cell catalyst and production A reaction mixture containing the amide compound prepared.

  As a method of mixing with a reaction mixture containing a cell catalyst and a cell catalyst, an unreacted raw material, a pH regulator, a generated amide compound, and the like, for example, the following method can be used. As a mixture containing an aqueous medium, an external circulation line equipped with a pump for returning a part of the reaction mixture to the reaction tank is installed in the reaction tank, and the external circulation line and the cell catalyst supply pipe are combined. There is a method in which a part of the reaction mixture taken out from the reaction vessel is used and a part of the reaction mixture and the cell catalyst are directly mixed in an external circulation line. A part of the reaction mixture and the cell catalyst directly mixed in the external circulation line are supplied as they are to the reaction tank using the external circulation line.

  The reaction time (retention time of the reaction solution) is usually 0.5 to 50 hours, preferably 2 to 25 hours. The reaction time refers to the total reaction time (retention time of the reaction liquid) in all reactors.

[Amide compound production equipment]
The apparatus for producing an amide compound of the present invention comprises a reaction vessel for producing an amide compound using a microbial cell containing nitrile hydratase and / or a treated product thereof, and a liquid temperature in the reaction vessel of -20 ° C or higher. A cell catalyst storage tank is provided for supplying cells and / or treated cells thereof at a temperature equal to or lower than the liquid temperature in the tank.
The configuration of the reaction tank is as described above.

  Next, examples of the present invention will be specifically described, but the present invention is not limited to these examples.

[Example 1]
[Preparation of microbial cells containing nitrile hydratase]
In accordance with the method described in Example 1 of JP-A-2001-340091, no. Three clone cells were obtained and similarly cultured by the method of Example 1 to obtain wet cells containing nitrile hydratase.

[Production of acrylamide]
In order to obtain a final product having an acrylamide concentration in the aqueous solution of 50% by weight, the reaction was performed under the following conditions.

A tank-type stirring tank with a SUS jacket type heat exchanger (volume 2 m ³ ) having a tank inner diameter of 1.3 m and a straight body length of 1.6 m was prepared as a bacterial cell catalyst storage tank. The wet cells obtained by the above culture method were suspended in pure water in a cell catalyst storage tank to prepare a 2 m ³ cell catalyst suspension. Cooling water at 10 ° C. was circulated through the jacket of the cell catalyst storage tank, and stirred at a rotational speed of 60 rpm using a three-blade propeller blade having a blade diameter of 65 cm and a blade width of 6.5 cm. The temperature of the suspension of the cell catalyst in the cell catalyst storage tank was 10 ° C.

A tank reactor with a SUS jacket heat exchanger (volume: 1 m ³ ) having a tank inner diameter of 1 m and a straight body length of 1.36 m, equipped with a stirrer as the first reactor, and a volume of 0.5 m as the second reactor. ³ SUS double tube reactors were prepared. The first reactor was charged with 400 kg of water in advance. While the inside of the first reactor was agitated, the cell catalyst suspension prepared above was continuously fed at a rate of 11 kg / h. Further, acrylonitrile having a purity of 99.8% was continuously supplied to the first reactor through an acrylonitrile supply pipe at a rate of 32 kg / h. Pure water was continuously supplied through a pure water supply pipe at a rate of 37 kg / h. The temperature of the reaction solution during the reaction was controlled to 25 ° C. by flowing 5 ° C. cooling water through the jacket of the first reactor and the double tube of the second reactor. A 0.1 M NaOH aqueous solution was used as a pH adjuster, and the supply amount was adjusted so that the reaction pH was 7.5 to 8.5. The reaction solution is continuously withdrawn from the first reactor at a rate of 80 kg / h so that the liquid level of the reaction solution during the reaction is 1 m from the bottom of the tank, and continuously supplied to the second reactor. The reaction was allowed to proceed further in the second reactor.

  When analysis was performed under the following HPLC conditions 160 hours after the start of the reaction, the conversion to acrylamide at the outlet of the first reactor was 95%, and the acrylonitrile concentration at the outlet of the second reactor was below the detection limit (10 Weight ppm or less). The acrylamide concentration at the outlet of the second reactor was 53.2% by weight.

Here, the analysis conditions were as follows.
・ Acrylamide analysis conditions:
High-performance liquid chromatograph: LC-10A system (manufactured by Shimadzu Corporation)
(UV detector wavelength 250 nm, column temperature 40 ° C.)
Separation column: SCR-101H (manufactured by Shimadzu Corporation)
Eluent: 0.05% (volume basis) -phosphoric acid aqueous solution / acrylonitrile analysis conditions:
High-performance liquid chromatograph: LC-10A system (manufactured by Shimadzu Corporation)
(UV detector wavelength 200 nm, column temperature 40 ° C.)
Separation column: Wakosil-II 5C18HG (manufactured by Wako Pure Chemical Industries)
Eluent: 7% (volume basis) -acetonitrile, 0.1 mM-acetic acid,
An aqueous solution containing 0.2 mM sodium acetate at various concentrations

[Example 2]
In the production of acrylamide of Example 1, the reaction was carried out in the same manner as in Example 1 except that 5 ° C cooling water was circulated instead of circulating 10 ° C cooling water through the jacket of the cell catalyst storage tank. The temperature of the suspension of the cell catalyst in the cell catalyst storage tank was 5 ° C.

  When the above HPLC analysis was performed 160 hours after the start of the reaction, the conversion to acrylamide at the outlet of the first reactor was 94%, and the acrylonitrile concentration at the outlet of the second reactor was below the detection limit (less than 10 ppm by weight) It became. The acrylamide concentration at the outlet of the second reactor was 53.1% by weight.

[Comparative Example 1]
In the production of acrylamide of Example 1, the reaction was carried out in the same manner as in Example 1 except that 2 ° C. cooling water was circulated instead of circulating 10 ° C. cooling water through the jacket of the cell catalyst storage tank. The temperature of the cell catalyst suspension in the cell catalyst storage tank was 2 ° C.

  160 hours after the start of the reaction, analysis was performed under the following HPLC conditions. As a result, the conversion rate to acrylamide at the first reactor outlet was 90%, and the acrylonitrile concentration at the second reactor outlet was 40 ppm by weight. It was. The acrylamide concentration at the outlet of the second reactor was 52.2% by weight.

[Example 3]
[Preparation of microbial cells containing nitrile hydratase]
In accordance with the method described in Example 1 of JP-A-2001-340091, no. Three clone cells were obtained and similarly cultured by the method of Example 1 to obtain wet cells containing nitrile hydratase.

[Production of acrylamide]
In order to obtain a final product having an acrylamide concentration in the aqueous solution of 50% by weight, the reaction was performed under the following conditions.

A tank-type stirring tank with a SUS jacket type heat exchanger (volume 2 m ³ ) having a tank inner diameter of 1.3 m and a straight body length of 1.6 m was prepared as a bacterial cell catalyst storage tank. The wet cells obtained by the above culture method were suspended in pure water in a cell catalyst storage tank to prepare a 2 m ³ cell catalyst suspension. Cooling water at 10 ° C. was circulated through the jacket of the cell catalyst storage tank, and stirred at a rotational speed of 60 rpm using a three-blade propeller blade having a blade diameter of 65 cm and a blade width of 6.5 cm. The temperature of the suspension of the cell catalyst in the cell catalyst storage tank was 10 ° C.

A tank reactor with a SUS jacket heat exchanger (volume: 1 m ³ ) having a tank inner diameter of 1 m and a straight body length of 1.36 m, equipped with a stirrer as the first reactor, and a volume of 0.5 m as the second reactor. ³ SUS double tube reactors were prepared. The first reactor was charged with 400 kg of water in advance. While the inside of the first reactor was agitated, the cell catalyst suspension prepared above was continuously fed at a rate of 11 kg / h. Further, acrylonitrile having a purity of 99.8% was continuously supplied to the first reactor through an acrylonitrile supply pipe at a rate of 32 kg / h. Pure water was continuously supplied through a pure water supply pipe at a rate of 37 kg / h. The temperature of the reaction solution during the reaction was controlled to be 20 ° C. by flowing 5 ° C. cooling water through the jacket of the first reactor and the double tube of the second reactor. A 0.1 M NaOH aqueous solution was used as a pH adjuster, and the supply amount was adjusted so that the reaction pH was 7.5 to 8.5. The reaction solution is continuously withdrawn from the first reactor at a rate of 80 kg / h so that the liquid level of the reaction solution during the reaction is 1 m from the bottom of the tank, and continuously supplied to the second reactor. The reaction was allowed to proceed further in the second reactor.

  When analysis was performed under the following HPLC conditions 160 hours after the start of the reaction, the conversion to acrylamide at the outlet of the first reactor was 95%, and the acrylonitrile concentration at the outlet of the second reactor was below the detection limit (10 Weight ppm or less). The acrylamide concentration at the outlet of the second reactor was 53.1% by weight.

[Comparative Example 2]
In the production of acrylamide of Example 3, the reaction was performed in the same manner as in Example 3 except that 20 ° C. cooling water was circulated instead of circulating 10 ° C. cooling water through the jacket of the cell catalyst storage tank. The temperature of the cell catalyst suspension in the cell catalyst storage tank was 20 ° C.

  When analysis was performed under the following HPLC conditions 160 hours after the start of the reaction, the conversion to acrylamide at the outlet of the first reactor was 60%, and the acrylonitrile concentration at the outlet of the second reactor was 3.9% by weight. It became. The acrylamide concentration at the outlet of the second reactor was 48.7% by weight.

[Comparative Example 3]
In the production of acrylamide of Example 3, 5 ° C. cooling water was passed through the jacket of the first reactor and the double tube of the second reactor so that the temperature of the reaction solution during the reaction was 20 ° C. Instead of temperature control, 5 ° C cooling water is circulated through the jacket of the first reactor and the double tube of the second reactor so that the temperature of the reaction liquid during the reaction is 10 ° C. The reaction was carried out in the same manner as in Example 3 except that.

  160 hours after the start of the reaction, analysis was performed under the following HPLC conditions. As a result, the conversion rate to acrylamide at the first reactor outlet was 88%, and the acrylonitrile concentration at the second reactor outlet was 200 ppm by weight. It was. The acrylamide concentration at the outlet of the second reactor was 51.0% by weight.

Claims (3)

In the method for producing an amide compound from a nitrile compound using a bacterial cell containing nitrile hydratase and / or a treated product thereof, the liquid temperature of the liquid containing at least the nitrile compound in the reaction vessel is 10 to 40 ° C. Production of an amide compound, characterized in that a cell and / or a treated product thereof is supplied to a maintained reaction vessel at a temperature of a liquid temperature in the reaction vessel of −20 ° C. or higher and lower than the liquid temperature in the reaction vessel. Method The production method according to claim 1, wherein the amide compound is acrylamide or methacrylamide. A reaction vessel for producing an amide compound produced using a bacterial cell containing nitrile hydratase and / or a treated product thereof, and a liquid temperature in the reaction vessel of −20 ° C. or higher and lower than the liquid temperature in the reaction vessel Device for producing an amide compound comprising a cell catalyst storage tank for supplying cells and / or treated cells thereof at a temperature