JP2001112493A - Production of carbonyl compound by microorganism - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a production process for carbonyl compounds in high yield starting from corresponding nitrile compounds, provide a production process for a carbonyl compound bearing the carbonyl group converted from a specific cyano group in a polynitrile compound in high yield and provide a microorganism that catalyzes these reactions. SOLUTION: This production process for a carbonyl compound is characterized by using a microorganism in Burkholderia as a catalyst to convert a cyano group (CN) in a nitrile compound to a carbonyl group (COY) (Y is NH2 or OH). The microorganism in Burkholderia is also a new catalyst to be used in the reaction.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

[0001] The present invention relates to a nitrile compound in which a cyano group (CN) is converted to a COY group (where Y is NH ₂ or O ₂ ).
H The present invention relates to a method for obtaining a carbonyl compound from a nitrile compound as a raw material using a microorganism having the ability to convert the compound into a carbonyl compound, and a microorganism applied to the method.

[0002]

The carbonyl compound obtained by the method of the present invention is useful as a raw material for synthesizing pharmaceuticals, agricultural chemicals, dyes and other chemicals.

[0003]

2. Description of the Related Art As a method for producing a carbonyl compound from various nitrile compounds as raw materials, a method in which a cyano group is hydrolyzed with a strong alkali at a high temperature in the presence of a metal catalyst has been used. However, these methods have problems in that it is difficult to treat waste generated in the course of the reaction and to remove generated by-products.

Further, in order to chemically hydrolyze only a specific cyano group of a compound having a plurality of cyano groups in a molecule to selectively obtain a carbonyl compound having a cyano group, the compound is left unreacted. It is necessary to protect the cyano group to be protected or to isolate a substance produced by hydrolysis of only a specific cyano group from the mixture, but in each case, the process becomes multi-step and complicated, and the method to be put to practical use Has not been established.

[0005] In order to solve these problems, a method for producing a carbonyl compound utilizing the selective cyano group hydrolysis ability of a microorganism is known. For example, in U.S. Pat.No.4,629,700,
A method for producing cyanobenzoic acid from phthalonitrile using a microorganism of the genus Rhodococcus and a method for producing a carbonyl compound having a cyano group from various aliphatic dinitrile are disclosed. JP-A-58-209987 discloses a process for producing an aromatic carboxylic acid amide from an aromatic nitrile using a microorganism of the genus Corynebacterium or Nocardia. Further, EP 178,106 discloses a method for producing a carbonyl compound having a cyano group by selective hydrolysis of polynitrile from polynitrile using Gram-positive bacteria belonging to four genera including Rhodococcus. However, the reaction yield and reaction selectivity of these methods using a conventionally known series of microorganisms are not always sufficient for industrial implementation and have not been put to practical use.

[0006]

DISCLOSURE OF THE INVENTION The present invention is to provide a process for producing a carbonyl compound starting from a nitrile compound in a high yield by utilizing the selectivity of a microbial reaction. It is an object of the present invention to provide an industrially practicable method for producing a carbonyl compound having a cyano group by converting only a cyano group into a carbonyl group. Another object of the present invention is to search for and provide a microorganism genus capable of giving a high yield in the reaction.

[0007]

Means for Solving the Problems The present inventors have conducted extensive studies on the above-mentioned problems with respect to existing known microorganisms and a wide variety of microorganisms obtained from the environment such as soil, and as a result, have found that The inventors have found that a microorganism belonging to the genus Delia has the ability to hydrolyze the cyano group of a nitrile compound, and completed the present invention.

That is, the present invention relates to the following matters. [1] A method for producing a carbonyl compound, comprising converting a cyano group (CN) of a nitrile compound into a COY group (where Y represents NH ₂ or OH) using a microorganism of the genus Burkholderia. . [2] The method for producing a carbonyl compound according to [1], wherein the carbonyl compound is a carboxylic acid. [3] The method for producing a carbonyl compound according to claim 1, wherein the carbonyl compound is a carboxylic acid amide. [4] The method for producing a carbonyl compound according to [1] to [3], wherein the nitrile compound is an aromatic mononitrile compound or a heterocyclic mononitrile compound. [5] The nitrile compound is a polynitrile compound having two or more cyano groups in the molecule, and the carbonyl compound is a carbonyl compound having a cyano group [1] to [3].
The method for producing a carbonyl compound according to the above. [6] The polynitrile compound is o-phthalonitrile, isophthalonitrile or terephthalonitrile, and the carbonyl compound having a cyano group is a compound of the corresponding o-, m-
-Or a method for producing a carbonyl compound according to [5], which is p-cyanobenzoic acid. [7] The microorganism used is Burkholderia cepacia SD
814 (FERM P-17597) [1]-
The method for producing a carbonyl compound according to [6]. [8] Burke Holdelia Sephacia SD814 (FERM P
-17597).

[0009]

DESCRIPTION OF THE PREFERRED EMBODIMENTS The present invention will be described below in detail.
According to the present invention, a generally known microorganism of the genus Burkholderia is cultured by a conventional method, and the obtained cells are
By suspending and acting on an aqueous solution or suspension containing a nitrile compound, the desired carbonyl compound can be easily obtained at a high conversion rate.

The term "carbonyl compound" used in the present specification is defined as an aliphatic, aromatic or heterocyclic C
A compound having an OY group (Y represents NH ₂ or OH) or a mixture of the compounds.

Hitherto, a method for producing a carbonyl compound from a nitrile compound using a microorganism of the genus Burkholderia has not been known, and has been newly found by the present inventors in the present invention.

[0012] The microorganism of the genus Burkholderia used in the present invention is, for example, isolated from the soil in Japan by using a medium containing n-paraffin as the sole carbon source and energy source. Burke Holdelia
Sepacia SD814 (FERMP-17597). The mycological properties of the microorganism are as follows.

Bacteriological properties of Burkholderia cepacia SD814 (FERM P-17597) strain Form: Bacillus Size: 1 to 3 μm in major axis, 0.5 to 1 μm in minor axis Gram staining: negative Sporulation: not forming Motility: Yes Flagella: Polar hair Attitude to oxygen: Aerobic oxidase: Positive Catalase: Positive Color of colony: Light yellow Fluorescent dye formation: Negative Water-soluble dye formation: Negative PHB accumulation: Positive Growth at 40 ° C: Positive Human peroxidase: Negative Denitrification reaction: Negative Gelatin liquefaction: Positive Starch degradation: Negative Utilization of various compounds Assimilate: Glucose, D-xylose, D-ribose, levulinic acid, 2,3-butylene glycol, tryptamine Not assimilated: L-rhamnose, mesaconic acid, D-tartaric acid Quinone system: Q-8 GC content of intracellular DNA: 66 mol% The strain was identified by Bergey's Manual o f Systematic
Bacteriology Vol.1 (1984), Bergey's Manual of Deter
minative Bacteriology ninth Edition (1994), and Microbiology and Immunology, 36, 1251 (1992, Yabuuc
hi et.al).

The strain is a non-spore-forming gram-negative spore-forming bacterium, is aerobic, accumulates PHB, has a quinone Q-8, and is assimilating to various organic substances. From these and other factors, it was presumed to be Burkholderia Sepacia. However, it differs from existing Burkholderia cepacia in that it does not form a water-soluble dye.

As a result of these mycological examinations, the strain was concluded to be a new strain of Burkholderia cepacia, and was named Burkholderia cepacia SD814. Deposit No.FERM P-
Deposited as 17597.

The reaction of the present invention is most conveniently carried out, for example, by adding Burkholderia cepacia SD814 to an LB liquid medium.
% Peptone or other nutrient medium at 20 ° C to 40 ° C, preferably
Cultured at a temperature of 25 ° C. to 30 ° C. for about 24 hours, and the obtained culture solution contains a nitrile compound in an amount of 1 ppm to 50%, preferably 10 ppm to 1%.
This can be achieved by adding 0% and continuing to stir at the same temperature for about 1 to 200 hours. The nitrile compound to be added may not necessarily be completely dissolved, but a solvent, a surfactant or the like that improves the solubility or dispersibility in the reaction solution may be added. The nitrile compound may be added continuously or intermittently depending on the consumption of the nitrile compound as the reaction proceeds. In this case, the concentration of the nitrile compound in the reaction solution is not limited to the above.

As a medium carbon source for culturing microorganisms, sugars such as glucose, sucrose and fructose, organic substances such as ethanol, acetic acid, citric acid, succinic acid, lactic acid, benzoic acid and fatty acids and alkali metal salts thereof are used. , Aliphatic hydrocarbons such as n-paraffins, aromatic hydrocarbons, and natural organic substances such as peptone, meat extract, fish extract, soy flour, and bran, alone or in combination, usually from 0.01% to 30%, preferably 0.
It can be used at a concentration of about 1% to 10%.

Examples of a medium nitrogen source for culturing microorganisms include inorganic nitrogen compounds such as ammonium sulfate, ammonium phosphate, sodium nitrate and potassium nitrate; nitrogen-containing organic substances such as urea and uric acid; peptone; meat extract; fish extract; A natural organic substance such as soybean flour can be used alone or in combination thereof at a concentration of usually about 0.01% to 30%, preferably about 0.1% to 10%.
In addition, the cyano group is hydrolyzed by these strains and COY
The reaction raw material which becomes a group (however, Y represents NH ₂ or OH) is added in advance during the culture so that ammonium ions released by hydrolysis with the progress of the reaction become a nitrogen source of the microorganism. It is useful because it becomes.

If necessary, phosphates such as potassium dihydrogen phosphate, and metal salts such as magnesium sulfate, ferrous sulfate, calcium acetate, manganese chloride, copper sulfate, zinc sulfate, cobalt sulfate, and nickel sulfate may be used. It is added to improve the growth of fungi. The concentration varies depending on the culture conditions.
Usually 0.01% to 5% for phosphate, 10ppm to 1% for magnesium salt, 0.1ppm to 1000% for other compounds
It is about ppm. Depending on the selected medium, the growth of bacteria can be improved by adding about 1 ppm to 100 ppm of, for example, yeast extract, casamino acid, and yeast nucleic acid as a source of vitamins, amino acids, nucleic acids, and the like.

In order to improve the reactivity of bacteria with nitrile, nitrile compounds such as benzonitrile, tolunitrile, terephthalonitrile, isophthalonitrile, glucononitrile, adiponitrile, and n-butyronitrile, isobutyronitrile, and derivatives thereof in 10 ppm to 10 ppm
It is useful to add%. It is also useful to add a nitrile compound as a reaction raw material from the start of cultivation, also as a source of nitrile hydrolase.
In addition, the use of a nitrile compound as a carbon source and / or nitrogen source for microorganisms in culture instead of or in combination with the various carbon sources and nitrogen sources described above is also useful for inducing nitrile hydrolase activity. It is.

When any of the components is used, the pH of the medium is
It is desirable to adjust to 5-9, preferably 6-8. Microbial cells previously cultured in the medium as described above are separated from the culture solution by a method such as centrifugation or membrane filtration, and water containing a nitrile compound as a reaction material, physiological saline, or the pH of the culture. Re-suspending and reacting in a buffer solution consisting of phosphoric acid, acetic acid, boric acid, tris (hydroxymethyl) aminomethane, etc. and their salts adjusted to the same pH and reacting reduces the impurities in the reaction solution However, it is useful for simplifying the subsequent product separation. The pH during the reaction is
When a buffer having a sufficient concentration is used, it can be usually maintained, but when the above-mentioned pH is deviated due to the progress of the reaction, it is desirable to appropriately adjust the pH using sodium hydroxide, ammonia or the like.

The carbonyl compound formed in the reaction solution is
Depending on the properties in the reaction solution, it can be obtained by various commonly used methods such as centrifugation, membrane filtration, drying under reduced pressure, distillation, solvent extraction, salting out, ion exchange, and various types of chromatography.

When the reaction product is obtained as an aqueous solution, it is preferable to remove the microbial cells by a method such as centrifugation or membrane filtration under the conditions in which the product is dissolved. When the reaction product is obtained as a solid and the crystals are sufficiently large, the product can be fractionated using a stainless mesh or the like. If the crystals are so small that it is difficult to separate them from microorganisms,
For example, in the case of a carboxylic acid, a method is preferably used in which the cells are once removed as an aqueous solution under alkaline conditions or the like, and the cells are removed by a method such as centrifugation, membrane filtration, and the like, and then the conditions are recovered, and the precipitate is collected again for fractionation. However, this is not always the case when a method which can be clearly used to remove microorganisms, such as direct distillation of the reaction solution, can be used.

[0024] Depending on the nature of the reaction product, the accumulation of the product in the reaction solution may reduce the reaction rate. In such a case, a method in which water, physiological saline, and a reaction buffer are added to the reaction solution to dilute it continuously according to the concentration of the product is preferable. Further, at the time when the reaction rate is reduced, the bacteria are collected, and the supernatant is recovered as a product solution,
The reaction rate can be recovered by returning the separated bacteria to a solution or suspension containing the reaction raw materials again. These methods can be performed repeatedly or continuously as long as the nitrile hydrolysis activity of the microorganism is maintained.

The present invention is similarly carried out using a cell-free extract of a microorganism applicable to the present invention, and a product obtained by concentrating and extracting a component catalyzing the above reaction from the cell-free extract. be able to. Furthermore, the present invention can also be achieved by immobilizing a microorganism applicable to this reaction or an extract or extract thereof, on a poorly soluble carrier, and bringing the immobilized product into contact with a raw material solution. Examples of such an immobilization carrier include polyacrylamide, polyvinyl alcohol, poly-N-vinylformamide, polyallylamine, polyethyleneimine, methylcellulose, glucomannan,
Alginate, carrageenan, chitosan, polyurethanes, polyesters, and polymerization or cross-linked products thereof, such as a compound that forms a hardly water-soluble solid content containing a microorganism or an extractable component thereof, alone or as a mixture. be able to. In addition, activated carbon, porous ceramics, porous polymer moldings, nitrocellulose membranes, etc., a microorganism or its extract, an extraction component held on an object previously formed as a solid material, or a microorganism on the object. A grown solid / microbial cell complex can also be used.

As the nitrile compound used as a raw material of the present invention, for example, aliphatic nitrile, aromatic nitrile, heterocyclic nitrile, etc. can be preferably used.

Examples of the aliphatic nitrile include acetonitrile, propionitrile, n-butyronitrile, isobutyronitrile, n-valeronitrile, isovaleronitrile,
Capronitrile, malononitrile, succinonitrile,
Adiponitrile, glucononitrile, acrylonitrile, methacrylonitrile and the like can be mentioned.

Examples of aromatic nitriles include benzonitrile, tolunitrile, orthophthalonitrile, terephthalonitrile, isophthalonitrile, and substituted products of these aromatic nitrile compounds, such as chlorinated compounds, fluorinated compounds, nitrated compounds, aminated compounds. Preferred are orthophthalonitrile, terephthalonitrile and isophthalonitrile.

Examples of the heterocyclic nitrile include 3-cyanopyridine, 4-cyanopyridine, cyanoindoles and the like.

The carbonyl compound obtained by the hydrolysis reaction of the cyano group by the method of the present invention, that is, the production ratio of carboxylic acid and carboxylic acid amide varies depending on the kind of the nitrile compound selected as the raw material.

When the carbonyl compound is formed as a mixture of a carboxylic acid and a carboxylic acid amide, the composition ratio of the carboxylic acid and the carboxylic acid amide can be analyzed by a commonly known method such as HPLC or gas chromatography. When it is desired to obtain as a mixture of carboxylic acid and carboxylic acid amide, it can be obtained by simply separating the reaction solution as it is by the above various separation methods.

On the other hand, depending on the nitrile compound, only a carboxylic acid or a carboxylic acid amide may be produced. In such a case, the method of the present invention can be a method for producing a carboxylic acid or a carboxylic acid amide with high purity. For example, as an example of a nitrile compound when almost only a carboxylic acid is formed, phthalonitriles are mentioned, and terephthalonitrile and isophthalonitrile are preferable. The carbonyl compounds formed in this case are p-cyanobenzoic acid and m-cyanobenzoic acid, respectively, and both have a conversion of almost 100%.

As examples of the nitrile compound which forms only a carboxylic acid amide, 3-cyanopyridine and 4-cyanopyridine can be preferably mentioned. The carbonyl compounds formed in this case are nicotinamide and isonicotinamide, respectively, and both have a conversion of almost 100%.

Further, according to the present invention, only a specific cyano group of a polynitrile compound having a plurality of cyano groups can be hydrolyzed to be converted into a carbonyl compound having a cyano group. For example, Burkholderia Sepacia
In the conversion reaction of isophthalonitrile using SD814, selectivity for one of the two cyano groups of isophthalonitrile is high, and almost 100% of the isophthalonitrile consumed in the reaction and only one of the cyano groups are consumed. Is hydrolyzed to produce 3-cyanobenzoic acid. Compounds in which both of the two cyano groups are hydrolyzed, such as phthalic acid, are not formed. That is, the present invention, a plurality of chemically equivalent cyano groups, for example, only a specific cyano group among a plurality of cyano groups on the aromatic ring is highly selectively hydrolyzed,
This is a production method in which only a carbonyl compound having a cyano group can be obtained with high selectivity.

The method of the present invention can also be a method of producing each compound with a high purity when the carbonyl compound produced is either a carboxylic acid or a carboxylic amide.

[0036]

The present invention will be described below in more detail with reference to examples. These examples do not define the scope of the invention.

[Example 1] Burkholderia cepacia
SD814 was streaked on LB agar medium and placed in a thermostat at 30 ° C for 24 hours.
Cultured for hours. One platinum loop was scraped from the formed colony and suspended in 100 mL of LB liquid medium in a 500 mL baffled flask. The flask was placed in a 30 ° C. constant temperature rotary shaking incubator and cultured at 120 rotations per minute for 24 hours. The obtained microbial cells were collected by centrifugation at 10,000 g, and suspended in a 50 mM sodium / potassium phosphate buffer (pH = 7) in the same volume as the culture solution. Isophthalonitrile in bacterial suspension
0.2% (mass / volume) was added, and the mixture was placed in a thermostatic rotary shaking incubator at 30 ° C., and reacted at 120 rpm for 72 hours. The obtained reaction solution was adjusted to pH 2 with 2 mol / l hydrochloric acid, and an equal volume of ethyl acetate was added thereto, followed by stirring and extraction. The obtained ethyl acetate layer is appropriately diluted, and reverse-phase HPLC (column: Shodex DS-
613, eluent: 50% acetonitrile / 5 mM potassium phosphate buffer pH 3.0, flow rate 1 mL / min, detection: UV 210 nm), the retention time of the 3-cyanobenzoic acid sample in the reaction solution was the same. A peak component was observed. The peak components were separated and subjected to GC-mass spectrum analysis, and it was confirmed that each gave a fragment pattern suggesting the same structure as the standard. From the results of the relative comparison of the absorbance with the standard in HPLC, the concentration of 3-cyanobenzoic acid in the reaction solution was 1260 pp.
m. At this time, 3-cyanobenzamide and isophthalic acid were not detected in the reaction solution. H
When the consumption of isophthalonitrile was calculated under PLC conditions, the yield of 3-cyanobenzoic acid based on the consumed isophthalonitrile was 99.5%.

Example 2 Burkholderia cepacia
SD814 was cultured in the same manner as in Example 1 to prepare a cell suspension. Terephthalonitrile in bacterial suspension
0.2% (w / v) was added, and the mixture was placed in a 30 ° C. constant temperature rotary shaking incubator and reacted at 120 rotations per minute for 72 hours. The obtained reaction solution was adjusted to pH 2 with 2 mol / l hydrochloric acid, and an equal volume of ethyl acetate was added thereto, followed by stirring and extraction. The obtained ethyl acetate layer is appropriately diluted and subjected to reverse phase HPLC (column: Shodex DS-61).
3. Eluent: 50% acetonitrile / 5mM potassium phosphate buffer pH3.0, flow rate 1mL / min, detection: UV 240nm), the retention time of 4-cyanobenzoic acid standard in the reaction solution was the same. A peak component was observed. The peak components were separated and subjected to GC-mass spectrum analysis, and it was confirmed that each gave a fragment pattern suggesting the same structure as the standard. From the results of the relative comparison of the absorbance with the sample in HPLC, the concentration of 4-cyanobenzoic acid in the reaction solution was 1720 p.
pm. At this time, 4-cyanobenzamide and terephthalic acid were not detected in the reaction solution. H
When the consumption of terephthalonitrile was calculated under PLC conditions, the yield of 4-cyanobenzoic acid based on the consumed terephthalonitrile was 99.2%.

[Example 3] Burkholderia cepacia
SD814 was streaked on LB agar medium and placed in a thermostat at 30 ° C for 24 hours.
Cultured for hours. One platinum loop was scraped from the formed colony and inoculated into 100 mL of an inorganic salt medium in a 500 mL baffled flask. The composition of the inorganic salt medium is shown below. (Mineral salts _{medium) KH 2 PO 4 1.5 g Na} 2 HPO 4 1.5 g MgSO 4 7aq 0.2 g CaSO 4 2aq 10 mg FeSO 4 7aq 5 mg Yeast extract 20 mg / L after inoculation, immediately glucose 0.5%, Isofutaro Nitrile was added in an amount of 0.2%, and the flask was placed in a thermostatic rotary shaking incubator at 30 ° C., and cultured at 120 rpm for 144 hours. The obtained culture solution was adjusted to pH 2 with 2 mol / l hydrochloric acid, and an equal volume of ethyl acetate was added to the reaction solution, followed by stirring and extraction. The obtained ethyl acetate layer was appropriately diluted, and reverse phase HPL was used in the same manner as in Example 1.
When analyzed by C, a peak component having the same retention time as that of the 3-cyanobenzoic acid sample was observed. The peak components were separated and subjected to GC-mass spectrum analysis, and it was confirmed that each gave a fragment pattern suggesting the same structure as the standard. From the results of the relative comparison of absorbance with the sample in HPLC, the concentration of 3-cyanobenzoic acid in the reaction solution was 710 ppm.
It was calculated.

Example 4 Burkholderia cepacia
SD814 was cultured in the same manner as in Example 3 and 100 ml of an inorganic salt medium.
Immediately after inoculating L, 0.5% of glucose and 0.2% (mass / volume) of terephthalonitrile were added, and the flask was placed in a thermostatic rotary shaking incubator at 30 ° C. and cultured at 120 rotations per minute for 144 hours. The obtained culture solution was extracted in the same manner as in Example 3,
Analysis by reverse phase HPLC as in Example 2 revealed that 4-
A peak component having the same retention time as the cyanobenzoic acid sample was observed. The peak component was separated and subjected to GC-mass spectrum analysis, and it was confirmed that a fragment pattern suggesting the same structure as that of the sample was given. From the result of the relative comparison of the absorbance with the sample in HPLC, the concentration of 4-cyanobenzoic acid in the reaction solution was calculated to be 1210 ppm.

[Example 5] Burkholderia cepacia
SD814 was streaked on LB agar medium and placed in a thermostat at 30 ° C for 24 hours.
Cultured for hours. One platinum loop was scraped from the formed colony and suspended in 100 mL of LB liquid medium in a 500 mL baffled flask. The flask was placed in a 30 ° C. constant temperature rotary shaking incubator and cultured at 120 rotations per minute for 24 hours. The obtained microbial cells were collected by centrifugation at 10,000 g, and suspended in a 50 mM sodium / potassium phosphate buffer (pH = 7) in the same volume as the culture solution. Add 3-cyanopyridine to the bacterial suspension at 0.
2% (mass / volume) was added, and the mixture was placed in a thermostatic rotary shaking incubator at 30 ° C., and reacted at 120 rpm for 72 hours. The obtained reaction solution was centrifuged at 10,000 g, and the supernatant was subjected to gas chromatography (column carrier: Thermo-3000 / SHINCARBON A, manufactured by Shimadzu Corporation,
Analysis by vaporization chamber temperature: 300 ° C, column temperature: 250 ° C, constant temperature: nitrogen: 60 mL / min, detection: FID) revealed a peak component in the reaction mixture having the same retention time as the nicotinamide standard. The same reaction solution was subjected to GC-mass spectrum analysis, and it was confirmed that a fragment pattern suggesting the same structure as the nicotinamide standard was given. From the result of the relative comparison of the FID intensity with the sample in the gas chromatograph, the concentration of nicotinamide in the reaction solution was 420 ppm. At this time, nicotinic acid was not detected in the reaction solution.

[Example 6] Burkholderia cepacia
SD814 was cultured and harvested in the same manner as in Example 5, 0.2% (mass / volume) of 4-cyanopyridine was added to the prepared bacterial suspension, and the suspension was placed in a thermostated rotary shaking incubator at 30 ° C. The reaction was performed at 120 rotations for 72 hours. The obtained reaction solution was centrifuged at 10,000 g, and the supernatant was analyzed by gas chromatography under the same conditions as in Example 6.A peak component having a retention time consistent with the isonicotinamide sample was found in the reaction solution. Was. The same reaction solution was subjected to GC-mass spectrum analysis, and it was confirmed that it gave a fragment pattern suggesting the same structure as the isonicotinamide standard. From the result of the relative comparison of the absorbance with the sample in HPLC, the concentration of isonicotinamide in the reaction solution was 520 ppm. At this time, no isonicotinic acid was detected in the reaction solution.

[0043]

According to the present invention, a carbonyl compound can be obtained simply and efficiently from a nitrile compound as a raw material. Further, a benzoic acid compound having a cyano group, which is useful in various industrial fields, can be easily and efficiently obtained from a polynitrile compound, particularly an aromatic polynitrile compound as a raw material.

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme coat ゛ (Reference) (C12N 1/20 (C12N 1/20 C12R 1:01) C12R 1:01) (C12P 13/02 (C12P 13/02 C12R 1:01) C12R 1:01)

Claims (8)

[Claims] 1. The method according to claim 1, wherein the cyano group (CN) of the nitrile compound is COY-treated using a microorganism of the genus Burkholderia.
A method for producing a carbonyl compound, which comprises converting into a group (wherein, Y represents NH ₂ or OH). 2. A method in which a carbonyl compound is a carboxylic acid (Y is O
H The method for producing a carbonyl compound according to claim 1, wherein 3. The method for producing a carbonyl compound according to claim 1, wherein the carbonyl compound is a carboxylic acid amide (Y represents NH ₂ ). 4. The method for producing a carbonyl compound according to claim 1, wherein the nitrile compound is an aromatic mononitrile compound or a heterocyclic mononitrile compound. 5. The method for producing a carbonyl compound according to claim 1, wherein the nitrile compound is a polynitrile compound having a plurality of cyano groups in a molecule, and the carbonyl compound is a carbonyl compound having a cyano group. . 6. The polynitrile compound is o-phthalonitrile, isophthalonitrile or terephthalonitrile, and the carbonyl compound having a cyano group is
The method for producing a carbonyl compound according to claim 5, which is-, m-, or p-cyanobenzoic acid. 7. The method for producing a carbonyl compound according to claim 1, wherein the microorganism to be used is Burkholderia cepacia SD814 (FERM P-17597). 8. Burkholderia Sepasia SD814
(FERM P-17597).