JP2017136048A - Amide compound aqueous solution, method for producing amide compound aqueous solution, amide polymer and method for producing amide polymer - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide an amide compound aqueous solution obtained by hydration of a nitrile compound and a method for efficiently producing an amide compound aqueous solution, and an amide polymer and a method for producing the polymer.SOLUTION: The present invention provides an amide compound aqueous solution that comprises a bacterial cell comprising Pseudonocardia-derived nitrile-hydratase and/or a bacterial cell processed product thereof by 1-5,000 wtppm and a method for producing an amide compound aqueous solution, and an amide polymer obtained by polymerization of the amide compound and a method for producing the polymer.SELECTED DRAWING: None

Description

  The present invention relates to a method for producing an aqueous amide compound solution and an aqueous amide compound solution obtained by hydrating a nitrile compound using a bacterial cell and / or a treated product thereof as a catalyst, and an amide polymer and a polymer thereof. It relates to a manufacturing method.

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

  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.

 Amide compounds having an unsaturated bond are mainly used as raw materials for amide polymers. In recent years, acrylamide polymers obtained from acrylamide obtained by hydrating acrylonitrile as a raw material have been required to have higher quality. For example, there are flocculants in the use of acrylamide polymers, but acrylamide polymers used as flocculants have recently been required to have higher molecular weight while maintaining water solubility in accordance with the demand for improved performance. It has been. In addition, acrylamide polymers have applications such as papermaking additives. However, in order to further improve the quality of the obtained paper, a polymer having a better hue is required as the papermaking additive. Yes.

  In the method of using a microbial cell and / or a treated product thereof as a catalyst, an aqueous solution of an amide compound is generally obtained by reacting a nitrile compound with water in the presence of the microbial cell and / or treated product thereof. The resulting amide compound aqueous solution contains bacterial cells and / or treated products thereof.

  In order to remove the microbial compound contained in the amide compound aqueous solution and / or the treated product thereof as an impurity adversely affects the quality of the amide compound and the quality of the amide polymer, for example, Patent Document 1 and In patent document 2, the processing method by activated carbon is proposed. Patent Document 3 proposes a purification method using a porous hollow fiber membrane.

  However, the purification method using activated carbon and the purification method using a porous hollow fiber membrane inevitably have economic disadvantages such as requiring equipment dedicated to purification treatment. The methods of Patent Documents 1, 2, and 3 are insufficient as an efficient and high-quality amide compound production method, and there is still room for investigation.

Japanese Patent Publication No. Sho 61/115495 JP 2001/270857 pamphlet Japanese Patent Publication No. Sho 61/115495

  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. Also from the viewpoint of improving the quality of the amide polymer, a new method for producing an amide compound is desired.

  An object of the present invention is to provide an amide compound aqueous solution obtained by hydrating a nitrile compound, a method for efficiently producing an amide compound aqueous solution, an amide polymer, and a method for producing the polymer.

  The present inventors have conducted intensive research to solve the above problems. As a result, when an amide compound is produced using a specific microorganism, the quality of the amide compound and the quality of the amide polymer can be improved even when the microbial compound is less than a certain concentration and / or the treated product of the microbial cell. The inventors have found that there is no adverse effect and have arrived at the present invention.

That is, the present invention is as follows.
[1] An amide compound aqueous solution containing 1 to 5,000 wtppm of a microbial compound containing nitrile hydratase derived from Pseudonocardia genus and / or a treated product thereof [2] An amide compound in which the amide compound has an unsaturated bond Aqueous amide compound solution according to [1] above [3] Obtained by hydrating a nitrile compound using a microbial cell containing nitrile hydratase derived from Pseudonocardia and / or a treated product thereof as a catalyst Process for producing amide compound aqueous solution without removing bacterial cells and / or treated cells thereof from amide compound aqueous solution [4] At least one capable of homopolymerizing or copolymerizing with amide compound aqueous solution of amide compound according to [2] above Amide-based polymer obtained by copolymerization with various unsaturated monomers [5] The amide compound aqueous solution according to [2] A method for producing an amide polymer, characterized in that the liquid is homopolymerized or copolymerized with at least one unsaturated monomer copolymerizable with an amide compound

 In the present invention, an amide compound can be efficiently produced in the hydration reaction of a nitrile compound using a bacterial cell and / or a treated product thereof. In addition, by producing an amide polymer using the amide compound obtained by the above-described method, it is possible to obtain an excellent amide polymer having excellent hue and compatibility between water solubility and high molecular weight. .

  Hereinafter, the amide compound aqueous solution, the method for producing the amide compound aqueous solution, the amide polymer and the method for producing the polymer will be described.

[Amide compound aqueous solution]
The amide compound aqueous solution of the present invention is an aqueous solution containing an amide compound, and contains 1 to 5,000 wtppm of a microbial cell containing nitrile hydratase derived from Pseudonocardia and / or a treated product thereof. is there.

  The concentration of the amide compound in the amide compound aqueous solution is not particularly limited, and is preferably 1 to 99 wt%, more preferably 5 to 90 wt%, and still more preferably 10 to 60 wt%.

<Amide compound>
Examples of the amide compound include aliphatic amide compounds having 2 to 20 carbon atoms and aromatic amide compounds having 6 to 20 carbon atoms.

  Aliphatic amide compounds include, for example, saturated or unsaturated amides having 2 to 6 carbon atoms; specific examples include acetamide, propylamide, butyramide, isobutylamide, pentylamide, isopentylamide, hexylamide and the like. Aliphatic saturated monoamides; aliphatic saturated diamides such as malonamide, succinamide, and adipamide; and aliphatic unsaturated amides such as acrylamide, methacrylamide, and crotonamide.

  Examples of the aromatic amide compound include benzamide, o-, m- or p-chlorobenzamide, o-, m- or p-fluorobenzamide, o-, m- or p-nitrobenzamide, o-, m- or Examples include p-toluamide, o-, m- or p-pyridine amide.

  Of the amide compounds, aliphatic unsaturated amide compounds are preferable, and acrylamide and methacrylamide are more preferable.

<Bacteria and / or treated cells containing nitrile hydratase>
In the present invention, a microbial compound containing a nitrile hydratase derived from Pseudonocardia in an aqueous amide compound solution and / or a treated product thereof (hereinafter also simply referred to as “bacterial catalyst”) 1 Contains ˜5,000 wtppm.

  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”).

  As a microbial cell containing a nitrile hydratase derived from Pseudonocardia, a bacterium that produces a nitrile hydratase derived from Pseudonocardia and retains nitrile hydratase activity in an aqueous solution of a nitrile compound and an amide compound If it is a body, it will not specifically limit. 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 is exemplified as a representative example as described later, but is not particularly limited to Escherichia coli, and is not limited to Escherichia coli, and may belong to the genus Bacillus such as Bacillus subtilis. 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. ].

  The recombinant vector according to the present invention contains a gene encoding a nitrile hydratase derived from Pseudonocardia, and is obtained by linking a gene encoding a nitrile hydratase derived from Pseudonocardia to the vector. be able to. The vector is not particularly limited, and for example, commercially available expression plasmids typified by pET-21a (+), pKK223-3, pUC19, pBluescriptKS (+) and pBR322, nitriles derived from the genus Pseudonocardia By incorporating a gene encoding hydratase, an expression plasmid for the 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.

  The microbial cells producing nitrile hydratase derived from the genus Pseudonocardia as described above may be appropriately cultured and grown by a known method 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 above-mentioned Pseudonocardia-derived nitrile hydratase-producing microbial cells with a nitrile compound, various treatments such as collection by centrifugation or crushing to produce crushed cell bodies The cells subjected to any of these treatments are collectively referred to as a treated cell.

  The form of the disrupted cell is not particularly limited as long as it includes a cell producing nitrile hydratase derived from the genus Pseudonocardia. For example, the culture solution containing the cell itself, and centrifuging the culture solution Examples of the collected bacterial body separated and recovered in this manner, and those obtained by washing the collected bacterial body with physiological saline or the like.

 The amide compound aqueous solution of the present invention contains 1 to 5,000 wtppm of a microbial cell containing a nitrile hydratase derived from Pseudonocardia and / or a processed product thereof.

  The concentration of the fungus body containing nitrile hydratase derived from Pseudonocardia in the amide compound aqueous solution and / or the treated product thereof is preferably 1 to 3000 wtppm, more preferably 1 to 1,000 wtppm, More preferably, it is 1-500 wtppm.

[Method for producing amide compound aqueous solution]
The method for producing an aqueous solution of an amide compound according to the present invention comprises using a microbial compound containing a nitrile hydratase derived from Pseudonocardia and / or a treated product of the microbial compound from an aqueous amide compound solution obtained by hydrating the nitrile compound. It is a method for producing an aqueous amide compound solution characterized by not removing the body and / or the treated product thereof.

<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.

  Among the nitrile compounds, aliphatic unsaturated nitriles are preferable, and acrylonitrile and methacrylonitrile are more 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 multi-tubular cylindrical, spiral tube, spiral plate, plate, and double tube types.

  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, for example, a multi-tubular cylindrical type, a spiral tube type, a spiral plate type, a plate type, a double pipe type or the like is preferably installed on the external circulation line. The thing of the form directly installed in reactors, such as a formula and 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 0-50 degreeC, Preferably it sets to 10-40 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 may or may not have a temperature control device, but from the viewpoint of controlling the liquid temperature of the suspension of cell catalyst, It is preferable to have it.

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

 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 And 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 or more raw water supply pipes for supplying raw water to the reaction tank per 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 vessel, a stirring blade having an arbitrary shape can be selected. For example, 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, a full zone blade, etc. Can be mentioned. 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 to the reaction tank, the supply pipe for supplying the mixture of the raw water and the nitrile compound to the reaction tank may be one per reactor. Also good. 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 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 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 per reactor or a plurality of tubes. 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.

 The aqueous medium in which the bacterial cell catalyst is mixed when the bacterial cell catalyst 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 bacterial cell catalyst is mixed include a mixture of raw water and a nitrile compound, a bacterial cell catalyst, an unreacted raw material, a bacterial cell catalyst, a generated amide compound, and the like. .

  As a method of mixing the bacterial cell catalyst and the raw water when mixing the bacterial cell catalyst with the raw material water and supplying it to the reaction tank, 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 a method for supplying the mixture of the bacterial cell catalyst and the raw material water when the bacterial cell catalyst is mixed with the raw material water and then supplied to the reaction vessel. The method illustrated with the supply method of the raw material water in the case of supplying to a reaction tank independently can be used.

 The bacterial cell catalyst to be supplied to the reaction tank and the aqueous medium and / or the mixture containing the aqueous medium may be mixed at an arbitrary ratio. Preferably, the bacterial cell catalyst: 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 cell catalyst, the raw water, and the nitrile compound mixture when the cell catalyst is mixed with the raw material 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.

  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. 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 combining the external circulation line and the bacterial cell catalyst supply pipe. Examples include a method in which a part of the reaction mixture taken out of the tank 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 amount of the bacterial cell catalyst used varies depending on the reaction conditions, the type of catalyst and the form thereof, but is usually 1 to 5,000 wtppm, preferably 1 to 5 wt. It is 3,000 wtppm, More preferably, it is 1-1000 wtppm, More preferably, it is 1-500 wtppm.

  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.

  In the method for producing an aqueous amide compound solution of the present invention, it is not necessary to remove the microbial cells and / or treated products thereof from the aqueous amide compound solution obtained by the reaction. An aqueous compound solution can be obtained as a product.

[Amide polymer]
The amide polymer of the present invention is obtained by homopolymerizing the amide compound having an unsaturated bond obtained as described above or copolymerizing with at least one unsaturated monomer copolymerizable with the amide compound. Can be manufactured.

<Unsaturated monomer>
Examples of the unsaturated monomer copolymerizable with the amide compound having an unsaturated bond include unsaturated carboxylic acids such as acrylic acid, methacrylic acid, itaconic acid, maleic acid, fumaric acid, and salts thereof;
Sulfonic acids such as vinyl sulfonic acid, styrene sulfonic acid, acrylamidomethylpropane sulfonic acid and their salts;
Alkylaminoalkyl esters of (meth) acrylic acid such as N, N-dimethylaminoethyl methacrylate, N, N-diethylaminoethyl methacrylate, N, N-dimethylaminoethyl acrylate, or quaternary ammonium derivatives thereof;
N, N-dialkylaminoalkyl (meth) acrylamides such as N, N-dimethylaminopropylmethacrylamide, N, N-dimethylaminopropylacrylamide, or quaternary ammonium derivatives thereof;
Hydrophilic acrylamides such as acetone acrylamide, N, N-dimethylacrylamide, N, N-dimethylmethacrylamide, N-ethylmethacrylamide, N-ethylacrylamide, N, N-diethylacrylamide, N-propylacrylamide;
N-acryloylpyrrolidine, N-acryloylpiperidine, N-acryloylmorpholine;
Hydroxyethyl methacrylate, hydroxyethyl acrylate, hydroxypropyl methacrylate, hydroxypropyl acrylate;
Methoxypolyethylene glycol (meth) acrylate, N-vinyl-2-pyrrolidone;
Acrylamide;
Methacrylamide;
N, N-di-n-propylacrylamide, Nn-butylacrylamide, Nn-hexylacrylamide, Nn-hexylmethacrylamide, Nn-octylacrylamide, Nn-octylmethacrylamide, N- N-alkyl (meth) acrylamide derivatives such as tert-octylacrylamide, Nn-dodecylacrylamide, Nn-dodecylmethacrylamide;
N, N-diglycidylacrylamide, N, N-diglycidylmethacrylamide, N- (4-glycidoxybutyl) acrylamide, N- (4-glycidoxybutyl) methacrylamide, N- (5-glycidoxy N- (ω-glycidoxyalkyl) (meth) acrylamide derivatives such as pentyl) acrylamide, N- (6-glycidoxyhexyl) acrylamide;
(Meth) acrylate derivatives such as methyl (meth) acrylate, ethyl (meth) acrylate, butyl (meth) acrylate, lauryl (meth) acrylate, 2-ethylhexyl (meth) acrylate, glycidyl (meth) acrylate;
Examples include olefins such as acrylonitrile, methacrylonitrile, vinyl acetate, vinyl chloride, vinylidene chloride, ethylene, propylene, and butene, styrene, α-methylstyrene, butadiene, and isoprene.

  These monomers may be used alone or in combination of two or more.

<Polymerization initiator>
As the polymerization initiator, for example, a radical polymerization initiator can be used.

Examples of radical polymerization initiators include peroxides such as potassium persulfate, ammonium persulfate, hydrogen peroxide, and benzoyl peroxide; azobisisobutyronitrile, 2,2′-azobis (4-amidinopropane) dihydrochloride, Azo free radical initiators such as 4,4'-azobis (sodium 4-cyanovalerate);
Examples thereof include so-called redox catalysts in which the peroxide is used in combination with a reducing agent such as sodium bisulfite, triethanolamine, and ferrous ammonium sulfate.

  The above polymerization initiators may be used alone or in combination of two or more. The amount of the polymerization initiator is usually in the range of 0.001 to 5 wt% with respect to the total weight of the monomers.

  In the case of a single polymerization initiator, the polymerization temperature is usually in the range of 0 to 120 ° C, more preferably in the range of 5 to 90 ° C. In addition, the polymerization temperature does not always need to be maintained at a constant temperature, and may be appropriately changed as the polymerization proceeds. Usually, however, the polymerization temperature tends to increase as the polymerization proceeds. Therefore, it may be cooled as necessary.

  The molecular weight of the amide polymer of the present invention is not particularly limited, but is usually in the range of 100,000 to 50 million, and preferably in the range of 500,000 to 30 million.

  The amide polymer thus obtained is a polymer that is compatible with water solubility and high molecular weight, and is excellent in hue, such as a flocculant, a papermaking additive, and a petroleum recovery agent. Can be suitably used.

[Method for producing amide polymer]
As a method for producing the amide polymer of the present invention, for example, aqueous solution polymerization, emulsion polymerization and the like can be used.

  Among these, in the case of aqueous solution polymerization, the total concentration of the amide compound having an unsaturated bond and the unsaturated monomer added as necessary is usually 5 to 90% by weight.

The atmosphere during the polymerization is not particularly limited, but from the viewpoint of rapidly proceeding the polymerization, it is preferable to carry out the polymerization in an inert gas atmosphere such as nitrogen gas.
The polymerization time is not particularly limited, but is usually in the range of 1 to 20 hours.

The pH of the aqueous solution at the time of polymerization is not particularly limited, but polymerization may be carried out by adjusting the pH as necessary. In this case, usable pH adjusters include alkalis such as sodium hydroxide, potassium hydroxide and ammonia;
Mineral acids such as phosphoric acid, sulfuric acid, hydrochloric acid;
Examples include organic acids such as formic acid and acetic acid.

  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 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 raw water supply pipe, the pH regulator supply pipe, the acrylonitrile supply pipe, and the cell catalyst supply pipe are each directly connected to the first reactor.

 The first reactor was charged with 400 kg of water in advance. The wet cells obtained by the above culture method were suspended in pure water to obtain a cell suspension with a dry cell concentration of 0.3 wt%. This suspension was continuously fed at a rate of 10 kg / h while stirring in the first reactor. Further, acrylonitrile having a purity of 99.8% was continuously fed to the first reactor through an acrylonitrile feed pipe at a rate of 32 kg / h. Pure water was continuously supplied to the first reactor via a pure water supply pipe at a rate of 38 kg / h. The temperature of the reaction solution during the reaction was controlled by flowing 5 ° C. cooling water through the jacket type heat exchanger of the first reactor and the double tube of the second reactor so that the temperature of the reaction solution was 20 ° C. Using 0.1 M NaOH aqueous solution as a pH regulator, adjusting the supply amount so that the pH of the reaction solution is 7.5 to 8.5, and continuously to the first reactor through the pH regulator supply pipe Supplied. The pH of the reaction solution was measured at the outlet of the first reactor using the glass electrode method.

 The reaction solution is continuously withdrawn from the first reactor at a rate of 80 kg / h so that the reaction solution is 1 m above the bottom of the tank, and continuously supplied to the second reactor. The reaction was allowed to proceed further in the second reactor.

  Analysis was conducted 200 hours after the start of the reaction under the following HPLC conditions. The conversion to acrylamide at the first reactor outlet was 95%, and the acrylonitrile concentration at the second reactor outlet 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.

  The obtained reaction solution was neutralized with an aqueous acrylic acid solution to obtain an aqueous acrylamide solution having a concentration of 50 wt%.

 A part of the acrylamide aqueous solution was extracted and filtered using a filter, and the insoluble bacterial cell catalyst was separated by filtration. The bacterial cell catalyst separated by filtration was washed with pure water and then dried for 10 hours with a dryer at 120 ° C. The weight was measured after drying, and the concentration of the bacterial cell catalyst contained in the aqueous amide compound solution was calculated. The concentration was 360 wtppm.

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

[Production of acrylamide polymer]
Water was added to the acrylamide aqueous solution obtained by the above method to prepare an acrylamide sample for polymerization having a concentration of 20 wt%. 500 g of this acrylamide sample for 20 wt% polymerization was placed in a 1 L polyethylene container, and while maintaining the temperature at 18 ° C., dissolved oxygen in the liquid was removed through nitrogen, and immediately placed in a heat insulation block made of polystyrene foam.

Then, 200 × 10 ⁻⁶ mpm (molar ratio to acrylamide) of 4,4′-azobis (sodium 4-cyanovalerate), 200 × 10 ⁻⁶ mpm dimethylaminopropionitrile, and 80 × 10 ⁻⁶ mpm Of ammonium persulfate were each dissolved in a small amount of water and quickly poured into a 1 L polyethylene container in this order. Nitrogen gas was previously passed through these reagents, and before and after injection, a small amount of nitrogen gas was also passed through the polyethylene container to prevent oxygen gas from being mixed.

  When the reagent was injected, after the induction period of several minutes, the temperature inside the polyethylene container was observed to rise, so the supply of nitrogen gas was stopped. When the polyethylene container was held in the heat insulation block for about 100 minutes, the temperature inside the polyethylene container reached about 70 ° C. Therefore, the polyethylene container was taken out from the heat insulation block and immersed in 97 ° C. water for 2 hours to further proceed the polymerization reaction. Thereafter, it was immersed in cold water and cooled to stop the polymerization reaction.

  The water-containing gel of acrylamide polymer thus obtained was taken out from the polyethylene container, divided into small chunks and ground with a meat grinder. The ground hydrogel containing acrylamide polymer was dried with hot air at 100 ° C. for 2 hours and then pulverized with a high-speed rotary blade pulverizer to obtain a dry powdery acrylamide polymer. The obtained dry powdery acrylamide polymer was passed through a sieve, and a 32-42 mesh sample was collected and used as a polymer sample for subsequent tests.

[Example 2]
In the production of acrylamide of Example 1, instead of using a cell suspension with a dry cell concentration of 0.3 wt%, the procedure was carried out except that a cell suspension with a dry cell concentration of 0.5 wt% was used. In the same manner as in Example 1, acrylamide was produced. That is, the same reactor as in Example 1 was used, and 400 kg of water was charged in the first reactor in advance. The wet cells obtained by the culture method described in Example 1 of JP-A-2001-340091 were suspended in pure water to obtain a cell suspension with a dry cell concentration of 0.5 wt%. This suspension was continuously fed at a rate of 10 kg / h while stirring in the first reactor. Further, acrylonitrile having a purity of 99.8% was continuously fed to the first reactor through an acrylonitrile feed pipe at a rate of 32 kg / h. Pure water was continuously supplied to the first reactor via a pure water supply pipe at a rate of 38 kg / h. The temperature of the reaction solution during the reaction was controlled by flowing 5 ° C. cooling water through the jacket type heat exchanger of the first reactor and the double tube of the second reactor so that the temperature of the reaction solution was 20 ° C. Using 0.1 M NaOH aqueous solution as a pH regulator, adjusting the supply amount so that the pH of the reaction solution is 7.5 to 8.5, and continuously to the first reactor through the pH regulator supply pipe Supplied. The pH of the reaction solution was measured at the outlet of the first reactor using the glass electrode method.

 The reaction solution is continuously withdrawn from the first reactor at a rate of 80 kg / h so that the reaction solution is 1 m above the bottom of the tank, and continuously supplied to the second reactor. The reaction was allowed to proceed further in the second reactor.

  Analysis was conducted under the following HPLC conditions 200 hours after the start of the reaction. The conversion to acrylamide at the first reactor outlet was 98%, and the acrylonitrile concentration at the second reactor outlet 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.

  The obtained reaction solution was neutralized with an aqueous acrylic acid solution to obtain an aqueous acrylamide solution having a concentration of 50 wt%.

 A part of the acrylamide aqueous solution was extracted and filtered using a filter, and the insoluble bacterial cell catalyst was separated by filtration. The bacterial cell catalyst separated by filtration was washed with pure water and then dried for 10 hours with a dryer at 120 ° C. The weight was measured after drying, and the concentration of the bacterial cell catalyst contained in the amide compound aqueous solution was calculated. The concentration was 620 wtppm.

 The acrylamide polymer was produced in the same manner as in the production of the acrylamide polymer of Example 1. The obtained water-containing gel of acrylamide polymer was treated in the same manner as in Example 1 to obtain a polymer sample for use in the subsequent tests.

[Example 3]
In the production of acrylamide of Example 1, instead of using a cell suspension with a dry cell concentration of 0.3 wt%, the procedure was carried out except that a cell suspension with a dry cell concentration of 1.0 wt% was used. In the same manner as in Example 1, acrylamide was produced. That is, the same reactor as in Example 1 was used, and 400 kg of water was charged in the first reactor in advance. The wet cells obtained by the culture method described in Example 1 of JP-A-2001-340091 were suspended in pure water to obtain a cell suspension with a dry cell concentration of 1.0 wt%. This suspension was continuously fed at a rate of 10 kg / h while stirring in the first reactor. Further, acrylonitrile having a purity of 99.8% was continuously fed to the first reactor through an acrylonitrile feed pipe at a rate of 32 kg / h. Pure water was continuously supplied to the first reactor via a pure water supply pipe at a rate of 38 kg / h. The temperature of the reaction solution during the reaction was controlled by flowing 5 ° C. cooling water through the jacket type heat exchanger of the first reactor and the double tube of the second reactor so that the temperature of the reaction solution was 20 ° C. Using 0.1 M NaOH aqueous solution as a pH regulator, adjusting the supply amount so that the pH of the reaction solution is 7.5 to 8.5, and continuously to the first reactor through the pH regulator supply pipe Supplied. The pH of the reaction solution was measured at the outlet of the first reactor using the glass electrode method.

 The reaction solution is continuously withdrawn from the first reactor at a rate of 80 kg / h so that the reaction solution is 1 m above the bottom of the tank, and continuously supplied to the second reactor. The reaction was allowed to proceed further in the second reactor.

  Analysis was conducted under the following HPLC conditions 200 hours after the start of the reaction. As a result, the conversion rate to acrylamide at the outlet of the first reactor was 99.9% or more, and the acrylonitrile concentration at the outlet of the second reactor was the detection limit. The following (10 ppm by weight or less). The acrylamide concentration at the outlet of the second reactor was 53.1% by weight.

  The obtained reaction solution was neutralized with an aqueous acrylic acid solution to obtain an aqueous acrylamide solution having a concentration of 50 wt%.

 A part of the acrylamide aqueous solution was extracted and filtered using a filter, and the insoluble bacterial cell catalyst was separated by filtration. The bacterial cell catalyst separated by filtration was washed with pure water and then dried for 10 hours with a dryer at 120 ° C. When the weight was measured after drying and the concentration of the bacterial cell catalyst contained in the aqueous amide compound solution was calculated, the concentration was 1,200 wtppm.

 The acrylamide polymer was produced in the same manner as in the production of the acrylamide polymer of Example 1. The obtained water-containing gel of acrylamide polymer was treated in the same manner as in Example 1 to obtain a polymer sample for use in the subsequent tests.

[Example 4]
In the production of acrylamide of Example 1, instead of using a cell suspension with a dry cell concentration of 0.3 wt%, the procedure was carried out except that a cell suspension with a dry cell concentration of 3.0 wt% was used. In the same manner as in Example 1, acrylamide was produced. That is, the same reactor as in Example 1 was used, and 400 kg of water was charged in the first reactor in advance. The wet cell obtained by the culture method described in Example 1 of JP-A-2001-340091 was suspended in pure water to obtain a cell suspension with a dry cell concentration of 3.0 wt%. This suspension was continuously fed at a rate of 10 kg / h while stirring in the first reactor. Further, acrylonitrile having a purity of 99.8% was continuously fed to the first reactor through an acrylonitrile feed pipe at a rate of 32 kg / h. Pure water was continuously supplied to the first reactor via a pure water supply pipe at a rate of 38 kg / h. The temperature of the reaction solution during the reaction was controlled by flowing 5 ° C. cooling water through the jacket type heat exchanger of the first reactor and the double tube of the second reactor so that the temperature of the reaction solution was 20 ° C. Using 0.1 M NaOH aqueous solution as a pH regulator, adjusting the supply amount so that the pH of the reaction solution is 7.5 to 8.5, and continuously to the first reactor through the pH regulator supply pipe Supplied. The pH of the reaction solution was measured at the outlet of the first reactor using the glass electrode method.

 The reaction solution is continuously withdrawn from the first reactor at a rate of 80 kg / h so that the reaction solution is 1 m above the bottom of the tank, and continuously supplied to the second reactor. The reaction was allowed to proceed further in the second reactor.

  Analysis was conducted under the following HPLC conditions 200 hours after the start of the reaction. As a result, the conversion rate to acrylamide at the outlet of the first reactor was 99.9% or more, and the acrylonitrile concentration at the outlet of the second reactor was the detection limit. The following (10 ppm by weight or less). The acrylamide concentration at the outlet of the second reactor was 53.1% by weight.

  The obtained reaction solution was neutralized with an aqueous acrylic acid solution to obtain an aqueous acrylamide solution having a concentration of 50 wt%.

 A part of the acrylamide aqueous solution was extracted and filtered using a filter, and the insoluble bacterial cell catalyst was separated by filtration. The bacterial cell catalyst separated by filtration was washed with pure water and then dried for 10 hours with a dryer at 120 ° C. When the weight was measured after drying and the concentration of the bacterial cell catalyst contained in the aqueous amide compound solution was calculated, the concentration was 3,700 wtppm.

 The acrylamide polymer was produced in the same manner as in the production of the acrylamide polymer of Example 1. The obtained water-containing gel of acrylamide polymer was treated in the same manner as in Example 1 to obtain a polymer sample for use in the subsequent tests.

[Comparative Example 1]
In the production of acrylamide of Example 1, instead of using a cell suspension with a dry cell concentration of 0.3 wt%, the procedure was carried out except that a cell suspension with a dry cell concentration of 5.0 wt% was used. In the same manner as in Example 1, acrylamide was produced. That is, the same reactor as in Example 1 was used, and 400 kg of water was charged in the first reactor in advance. The wet cells obtained by the culture method described in Example 1 of JP 2001-340091 A were suspended in pure water to obtain a cell suspension with a dry cell concentration of 5.0 wt%. This suspension was continuously fed at a rate of 10 kg / h while stirring in the first reactor. Further, acrylonitrile having a purity of 99.8% was continuously fed to the first reactor through an acrylonitrile feed pipe at a rate of 32 kg / h. Pure water was continuously supplied to the first reactor via a pure water supply pipe at a rate of 38 kg / h. The temperature of the reaction solution during the reaction was controlled by flowing 5 ° C. cooling water through the jacket type heat exchanger of the first reactor and the double tube of the second reactor so that the temperature of the reaction solution was 20 ° C. Using 0.1 M NaOH aqueous solution as a pH regulator, adjusting the supply amount so that the pH of the reaction solution is 7.5 to 8.5, and continuously to the first reactor through the pH regulator supply pipe Supplied. The pH of the reaction solution was measured at the outlet of the first reactor using the glass electrode method.

 The reaction solution is continuously withdrawn from the first reactor at a rate of 80 kg / h so that the reaction solution is 1 m above the bottom of the tank, and continuously supplied to the second reactor. The reaction was allowed to proceed further in the second reactor.

  Analysis was conducted under the following HPLC conditions 200 hours after the start of the reaction. As a result, the conversion rate to acrylamide at the outlet of the first reactor was 99.9% or more, and the acrylonitrile concentration at the outlet of the second reactor was the detection limit. The following (10 ppm by weight or less). The acrylamide concentration at the outlet of the second reactor was 53.1% by weight.

  The obtained reaction solution was neutralized with an aqueous acrylic acid solution to obtain an aqueous acrylamide solution having a concentration of 50 wt%.

 A part of the acrylamide aqueous solution was extracted and filtered using a filter, and the insoluble bacterial cell catalyst was separated by filtration. The bacterial cell catalyst separated by filtration was washed with pure water and then dried for 10 hours with a dryer at 120 ° C. When the weight was measured after drying and the concentration of the bacterial cell catalyst contained in the aqueous amide compound solution was calculated, the concentration was 6,300 wtppm.

 The acrylamide polymer was produced in the same manner as in the production of the acrylamide polymer of Example 1. The obtained water-containing gel of acrylamide polymer was treated in the same manner as in Example 1 to obtain a polymer sample for use in the subsequent tests.

[Comparative Example 2]
In the production of acrylamide of Example 1, instead of using a cell suspension with a dry cell concentration of 0.3 wt%, the procedure was carried out except that a cell suspension with a dry cell concentration of 10.0 wt% was used. In the same manner as in Example 1, acrylamide was produced. That is, the same reactor as in Example 1 was used, and 400 kg of water was charged in the first reactor in advance. The wet cells obtained by the culture method described in Example 1 of JP-A-2001-340091 were suspended in pure water to obtain a cell suspension with a dry cell concentration of 10.0 wt%. This suspension was continuously fed at a rate of 10 kg / h while stirring in the first reactor. Further, acrylonitrile having a purity of 99.8% was continuously fed to the first reactor through an acrylonitrile feed pipe at a rate of 32 kg / h. Pure water was continuously supplied to the first reactor via a pure water supply pipe at a rate of 38 kg / h. The temperature of the reaction solution during the reaction was controlled by flowing 5 ° C. cooling water through the jacket type heat exchanger of the first reactor and the double tube of the second reactor so that the temperature of the reaction solution was 20 ° C. Using 0.1 M NaOH aqueous solution as a pH regulator, adjusting the supply amount so that the pH of the reaction solution is 7.5 to 8.5, and continuously to the first reactor through the pH regulator supply pipe Supplied. The pH of the reaction solution was measured at the outlet of the first reactor using the glass electrode method.

 The reaction solution is continuously withdrawn from the first reactor at a rate of 80 kg / h so that the reaction solution is 1 m above the bottom of the tank, and continuously supplied to the second reactor. The reaction was allowed to proceed further in the second reactor.

  Analysis was conducted under the following HPLC conditions 200 hours after the start of the reaction. As a result, the conversion rate to acrylamide at the outlet of the first reactor was 99.9% or more, and the acrylonitrile concentration at the outlet of the second reactor was the detection limit. The following (10 ppm by weight or less). The acrylamide concentration at the outlet of the second reactor was 53.1% by weight.

  The obtained reaction solution was neutralized with an aqueous acrylic acid solution to obtain an aqueous acrylamide solution having a concentration of 50 wt%.

 A part of the acrylamide aqueous solution was extracted and filtered using a filter, and the insoluble bacterial cell catalyst was separated by filtration. The bacterial cell catalyst separated by filtration was washed with pure water and then dried for 10 hours with a dryer at 120 ° C. When the weight was measured after drying and the concentration of the cell catalyst contained in the aqueous amide compound solution was calculated, the concentration was 12,300 wtppm.

 The acrylamide polymer was produced in the same manner as in the production of the acrylamide polymer of Example 1. The obtained water-containing gel of acrylamide polymer was treated in the same manner as in Example 1 to obtain a polymer sample for use in the subsequent tests.

<Testing method for acrylamide polymer>
The polymer samples obtained in Examples 1 to 4 and Comparative Examples 1 and 2 were evaluated for water solubility and color tone by the following methods.

Water solubility: In water solubility, 600 ml of water is put into a 1 L beaker, and 0.66 g (pure content 0.6 g) of polymer sample is added while stirring at 25 ° C. using a stirring blade having a predetermined shape. Stirring was performed for a period of time, and the resulting solution was filtered through a 150-mesh wire mesh, and water solubility was judged from the amount of insoluble matter and filterability. That is, ◎ is completely dissolved, ◯ is close to completely dissolved, there is an insoluble matter, △ is what can be filtered, △, the passage of the filtrate is slow, the insoluble matter is effectively filtered The thing which cannot be made was set as x.
Color tone: Regarding the color tone of the polymer, the polymer powder was visually evaluated.

  The evaluation results are shown in Table 1.

Claims (5)

Amide compound aqueous solution containing 1 to 5,000 wtppm of microbial cells containing nitrile hydratase derived from Pseudonocardia genus and / or processed cells thereof The amide compound aqueous solution according to claim 1, wherein the amide compound is an amide compound having an unsaturated bond. A microbial compound and / or a treated product thereof from an amide compound aqueous solution obtained by hydrating a nitrile compound using a nitrile hydratase derived from Pseudonocardia genus and / or a treated product thereof as a catalyst. For producing aqueous solution of amide compound without removing water An amide polymer obtained by homopolymerizing the amide compound aqueous solution according to claim 2 or copolymerizing with at least one unsaturated monomer copolymerizable with the amide compound. A method for producing an amide polymer, characterized in that the aqueous amide compound solution according to claim 2 is homopolymerized or copolymerized with at least one unsaturated monomer copolymerizable with the amide compound.