JP2007116916A - Method for producing optically active amino compound - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a method for efficiently producing an optically active amino compound in a mining and industrial manner by using a microbial cell of a microorganism. <P>SOLUTION: The method for production is carried out as follows. The microbial cell of the microorganism or a treated material thereof having ability to produce 1-benzyl-3-aminopyrrolidine from 1-benzyl-3-pyrrolidinone is made to act on a ketone compound represented by general formula (1) (wherein, R<SP>1</SP>and R<SP>2</SP>represent each independently any of a 6-10C aryl group which may have a substituent, a 6-10C aryl-1-6C alkyl group which may have a substituent, a 5- to a 10-membered heteroaryl group which may have a substituent or a 1-6C alkyl group which may have a substituent; and R<SP>1</SP>is a group having a higher priority sequence than that of R<SP>2</SP>by the sequence rule) in the presence of an amino group donor. Thereby, the resultant optically active amino compound represented by general formula (2) is collected. <P>COPYRIGHT: (C)2007,JPO&INPIT

Description

  The present invention relates to a method for producing an optically active amino compound using a microorganism.

  In general, optically active amines are synthesized into racemic forms by organic chemical synthesis methods to form diastereomeric salts with optically active acids such as tartaric acid, tartaric acid derivatives, and optically active mandelic acid, and optically resolved by crystallization. The method is performed (nonpatent literature 1). In addition, a method of deriving from a natural chiral pool such as an optically active amino acid such as L-aspartic acid is also known (Patent Documents 1, 2, and 3). Furthermore, in recent years, a method for producing a corresponding optically active amine from ketones using a microbial cell or an enzyme taken out has attracted attention. Examples of the method for producing amines by an enzymatic method include a method using a microorganism belonging to the genus Brevibacterium (Patent Document 4) and a transaminase method (Patent Documents 5 to 9). It is done.

  However, there are few references on biochemical production of 3-aminopyrrolidine or its derivatives from the corresponding ketone compounds. In Stirling et al. (Non-patent Document 2), there is an example of aminopyrrolidine in a compound that can be synthesized using ω-amino acid transaminase, but what kind of enzyme, reaction conditions, and reaction results are specifically described. The substance is not clear. In addition, in McCham et al. (Non-patent Document 3), there is a description of 1-benzyl-3-aminopyrrolidine as an example of an optically active amine that can be synthesized using ω-amino acid transaminase. Enzymes, reaction conditions, reaction results, etc. are not disclosed at all, and technical information is not disclosed sufficiently as in the above-mentioned literature.

  According to Kim et al. (Non-patent Document 4), when the ω-amino acid transaminase of Vibrio fluvialis JS17 strain accepts acetophenone and pyruvic acid, 3-aminopyrrolidine is weak, but an amino group donor. However, examples of using 1-benzyl-3-aminopyrrolidine as an amino group donor and examples of using 3-pyrrolidinone and 1-benzyl-3-pyrrolidinone as an acceptor are described. It is unclear whether 3-aminopyrrolidine, 1-benzyl-3-aminopyrrolidine can be synthesized or whether it acts on 1-benzyl-3-aminopyrrolidine.

As described above, 3-pyrrolidinone or a derivative thereof is used as an amino group acceptor to transfer an amino group from an amine or amino acid, and (S) -3-aminopyrrolidine or a derivative thereof is synthesized using a microbial cell or an enzyme. There is no literature describing a production method or a production method in which a microbial cell or enzyme is allowed to act on an enantiomer mixture of 3-aminopyrrolidine and a derivative to leave optically active 3-aminopyrrolidine or a derivative thereof.
JP 2002-212155 A JP 2002-316977 JP 63-273486 A Japanese Unexamined Patent Publication No. 3-103192 WO 97/15682 WO 00/26351 Japanese Patent Laid-Open No. 2001-8696 JP 2002-142793 A JP 2000-342276 A Organic Synthesis Handbook (Organic Synthesis Association, Maruzen, 761-766 (1990).) Stirling DL, "Chirality in Industry" (ed. Collins AN, Sheldrake GN & Crosby J.), 209-222, J. Wiley & Sons, NY (1992). Matcham GW & Bowen ARSTG., CHIMICA / chemistry today, 20-24 (1996). Shin JS & Kim BG, Biotechnol. Bioengi., 65 (2), 206-211 (1999).

  An object of the present invention is to provide a method for producing an optically active amino compound efficiently and industrially using microbial cells. In particular, it is an object to provide a method for efficiently producing specific (S) -form or (R) -form optically active amines, respectively.

As a result of intensive studies in view of these points, the present inventors have used a microbial cell having the ability to produce 1-benzyl-3-aminopyrrolidine from 1-benzyl-3-piperidinone or a treated product thereof. It has been found that (S) -form or (R) -form optically active amines having a specific structure can be efficiently produced from the corresponding ketone or the corresponding racemate, respectively, and the present invention has been completed.
Such an enzymatic method for synthesizing amines is a useful means for synthesizing amines for reasons such as few synthesis byproducts, normal temperature and normal pressure, safety and simplicity. Such a reaction cannot be carried out with the ω-amino acid transaminase, and such a technique has not been known at all.

（式中、Ｒ¹及びＲ²は、それぞれ独立して、下記Ａ群から選ばれる置換基を有していてもよいＣ_６−１０アリール基、下記Ａ群から選ばれる置換基を有していてもよいＣ_６−１０アリールＣ_１−６アルキル基、下記Ａ群から選ばれる置換基を有していてもよい５〜１０員ヘテロアリール基又は下記Ｂ群から選ばれる置換基を有していてもよいＣ_１−６アルキル基のいずれかを示し、Ｒ¹は順位則による優先順位がＲ²よりも高い基である；
Ａ群：ハロゲン原子、水酸基、Ｃ_１−４アルキル基、Ｃ_１−６アルコキシ基、ハロＣ_１−６アルキル基、−Ｏ（ＣＨ₂）_nＯ−（ｎは１または２）；
Ｂ群：ハロゲン原子、水酸基、Ｃ_１−６アルコキシ基；
または、Ｒ¹とＲ²とは互いに結合し、隣接する炭素原子と一緒になって５〜８員炭素環又は５〜８員ヘテロ環を形成していてもよく、該５〜８員炭素環又は５〜８員ヘテロ環は、下記置換基Ｃ群から選ばれる置換基を有していてもよい；
Ｃ群：ハロゲン原子、水酸基、Ｃ_１−４アルキル基、Ｃ_１−６アルコキシ基、Ｃ_６−１０アリール基、Ｃ_６−１０アリールＣ_１−６アルキル基）；

（式中、Ｒ¹及びＲ²は、それぞれ前記一般式（１）と同義である。）。
〔２〕
前記微生物がシュードモナス（Pseudomonas）属に属する微生物、セラチア（Serratia）属に属する微生物および配列番号：３で示される16S rDNA配列を含むキサントモナダセアエ(Xanthomonadaceae)科に属する微生物からなる群から選ばれる少なくとも１種の微生物である、〔１〕に記載の方法。
〔３〕
前記シュードモナス（Pseudomonas）属に属する微生物が、シュードモナス・コルガータ（Pseudomonas corrugata）10F6株であることを特徴とする〔２〕に記載の方法。
〔４〕
前記セラチア（Serratia.）属に属する微生物が、セラチア・エスピー（Serratia sp.）11E11株であることを特徴とする〔２〕に記載の方法。
〔５〕
前記配列番号：３で示される16S rDNA配列を含むキサントモナダセアエ(Xanthomonadaceae)科に属する微生物が、12A4株であることを特徴とする〔２〕に記載の方法。
〔６〕
前記ケトン化合物が、アセトフェノン、4-クロロアセトフェノン、4-メチルアセトフェノン、2-メトキシアセトフェノン、3-メトキシアセトフェノン、4-メトキシアセトフェノン、4-ヒドロキシアセトフェノン、1-(3,4-ジメトキシフェニル)プロパン-2-オン、1-(4-メトキシフェニル)プロパン-2-オン、1-（4-クロロフェニル）プロパン-2-オン、1-（4-ヒドロキシフェニル）プロパン-2-オン、1-（4-メチルフェニル）プロパン-2-オン、1-（3,4-メチレンジオキシフェニル）プロパン-2-オン、ベンジルアセトン、α-アセトナフトン、β-アセトナフトン、1-インダノン、α-テトラロン、β-テトラロン、7-メトキシ-2-テトラロン、1-ベンジル-3-ピロリジノン、1-メチル-3-ピロリジノン、1-ベンジル-3-ピペリドンおよび1-メチル-3-ピペリドンからなる群から選ばれる少なくとも１種の化合物であることを特徴とする〔１〕〜〔５〕のいずれかに記載の方法。
〔７〕
前記一般式（１）で表されるケトン化合物が下記一般式（３）で表されるケトン化合物であり、前記一般式（２）で表される化合物が下記一般式（４）で表される光学活性アミノ化合物であることを特徴とする〔１〕〜〔５〕のいずれかに記載の方法；

（式中、ＹはＨまたはアミノ基の保護基を示し、ｐは２〜３の整数、ｑは１〜２の整数を示し、ｐ>ｑである。）。
〔８〕
前記アミノ基の保護基が、Ｃ_１−６アルキル基、ベンジル基、ホルミル基またはｔ−ブトキシカルボニル基である、〔７〕に記載の方法。
〔９〕
前記ケトン化合物が、1-ベンジル-3-ピロリジノンであることを特徴とする〔７〕に記載の方法。
〔１０〕
前記アミノ基供与体が、
Ｌ−アラニン、イソプロピルアミン、1-ブチルアミン、sec-ブチルアミン、1,3-ジメチル-1-ブチルアミン、2-メチルブチルアミン、1-ペンチルアミン、2-ペンチルアミン、3-ペンチルアミン、1-ヘキシルアミン、2-ヘキシルアミン、1-ヘプチルアミンおよび2-ヘプチルアミンから選ばれる脂肪族アミン；
１−フェニルエチルアミン、2-フェニルエチルアミン、ベンジルアミン、3-アミノ-1-フェニルブタン、4-アミノ-１-フェニルペンタン、α−エチルベンジルアミンおよび1-(1-ナフチル)エチルアミンから選ばれる芳香族アミン；
2-アミノ-１-フェニルエタノールおよび2-フェニルグリシノールから選ばれるアミノアルコール類；および
１−アミノインダンからなる群から選ばれる１種である、〔１〕〜〔９〕のいずれかに記載の方法。
〔１１〕
前記作用を、メタノールまたはアセトニトリルを５〜５０％（全溶媒に対する割合（体積／体積））含む溶媒中で行うことを特徴とする〔１〕〜〔１０〕のいずれかに記載の方法。
〔１２〕
アミノ基受容体の存在下、1-ベンジル-3-ピロリジノンから1-ベンジル-3-アミノピロリジンを生成する能力を有する微生物の菌体あるいはその処理物を、下記一般式（５）で表されるアミノ化合物のエナンチオマー混合物に作用させ、残存する下記一般式（６）で表される光学活性アミノ化合物を採取することを特徴とするアミノ化合物の製法；

（式中、Ｒ¹及びＲ²は、それぞれ独立して、下記Ａ群から選ばれる置換基を有していてもよいＣ_６−１０アリール基、下記Ａ群から選ばれる置換基を有していてもよいＣ_６−１０アリールＣ_１−６アルキル基、下記Ａ群から選ばれる置換基を有していてもよい５〜１０員ヘテロアリール基又は下記Ｂ群から選ばれる置換基を有していてもよいＣ_１−６アルキル基のいずれかを示し、Ｒ¹は順位則による優先順位がＲ²よりも高い基である；
Ａ群：ハロゲン原子、水酸基、Ｃ_１−４アルキル基、Ｃ_１−６アルコキシ基、ハロＣ_１−６アルキル基、−Ｏ（ＣＨ₂）_nＯ−（ｎは１または２）；
Ｂ群：ハロゲン原子、水酸基、Ｃ_１−６アルコキシ基；
または、Ｒ¹とＲ²とは互いに結合し、隣接する炭素原子と一緒になって５〜８員炭素環又は５〜８員ヘテロ環を形成していてもよく、該５〜８員炭素環又は５〜８員ヘテロ環は、下記置換基Ｃ群から選ばれる置換基を有していてもよい；
Ｃ群：ハロゲン原子、水酸基、Ｃ_１−４アルキル基、Ｃ_１−６アルコキシ基、Ｃ_６−１０アリール基、Ｃ_６−１０アリールＣ_１−６アルキル基）；

（式中、Ｒ¹及びＲ²は、それぞれ前記一般式（５）と同義である。）。
〔１３〕
前記微生物がシュードモナス（Pseudomonas）属に属する微生物、セラチア（Serratia）属に属する微生物および配列番号：３で示される16S rDNA配列を有するキサントモナダセアエ(Xanthomonadaceae)科に属する微生物からなる群から選ばれる少なくとも１種の微生物である、〔１２〕に記載の方法。
〔１４〕
前記シュードモナス（Pseudomonas）属に属する微生物が、シュードモナス・コルガータ（Pseudomonas corrugata）10F6株であることを特徴とする〔１３〕に記載の方法。
〔１５〕
前記セラチア（Serratia.）属に属する微生物が、セラチア・エスピー（Serratia sp.）11E11株であることを特徴とする〔１３〕に記載の方法。
〔１６〕
前記配列番号：３で示される16S rDNA配列を含むキサントモナダセアエ(Xanthomonadaceae)科に属する微生物が、12A4株であることを特徴とする〔１３〕に記載の方法。
〔１７〕
前記一般式（５）で表されるアミノ化合物が、1-フェニルエチルアミン、α-エチルベンジルアミン、1-フェニル-2-アミノプロパン、1-(3,4-ジメトキシフェニル)-2-アミノプロパン、1-アミノインダン、1-アミノテトラリン、2-アミノテトラリン、1-フェニル-3-アミノブタン、3-アミノピロリジン、1-ベンジル-3-アミノピロリジン、3-アミノピペリジンおよび1-ベンジル-3-アミノピペリジンからなる群から選ばれる少なくとも１種であることを特徴とする〔１２〕〜〔１６〕のいずれかに記載の方法。
〔１８〕
前記一般式（５）で示される化合物が下記一般式（７）で表されるアミノ化合物であり、前記一般式（６）で示される化合物が下記一般式（８）で表される光学活性アミノ化合物であることを特徴とする〔１２〕〜〔１６〕のいずれかに記載の方法；

（式中、ＹはＨまたはアミノ基の保護基を示し、ｐは２〜３の整数、ｑは１〜２の整数を示し、ｐ>ｑである。）。
〔１９〕
前記アミノ基の保護基が、Ｃ_１−６アルキル基、ベンジル基、ホルミル基またはｔ−ブトキシカルボニル基である、〔１８〕に記載の方法。
〔２０〕
前記一般式（７）で表されるアミノ化合物が、1-ベンジル-3-アミノピロリジンであることを特徴とする〔１８〕に記載の方法。
〔２１〕
前記アミノ基受容体が、ピルビン酸、グリオキシル酸、2-ケトグルタル酸、アセトフェノン、プロピオフェノン、ベンジルアセトン、プロピオンアルデヒド、1-ブチルアルデヒド、ベンズアルデヒドおよびフェニルアセトアルデヒドからなる群から選ばれる少なくとも１種であることを特徴とする〔１２〕〜〔２０〕のいずれかに記載の方法。 That is, the present invention includes the following.
[1]
A microbial cell having the ability to produce 1-benzyl-3-aminopyrrolidine from 1-benzyl-3-pyrrolidinone in the presence of an amino group donor or a treated product thereof is represented by the following general formula (1). A process for producing an amino compound characterized by collecting an optically active amino compound represented by the following general formula (2) produced by acting on a ketone compound;

(In the formula, R ¹ and R ² each independently has a C _6-10 aryl group which may have a substituent selected from the following group A, and a substituent selected from the following group A. It may have a C _6-10 aryl C _1-6 alkyl group, a 5-10 membered heteroaryl group which may have a substituent selected from the following group A, or a substituent selected from the following group B. Any one of the _{optionally selected} C _1-6 alkyl groups, and R ¹ is a group having a higher priority than R ² according to the order rule;
Group A: halogen atom, hydroxyl group, C _1-4 alkyl group, C _1-6 alkoxy group, halo C _1-6 alkyl group, —O (CH ₂ ) _n O— (n is 1 or 2);
Group B: halogen atom, hydroxyl group, C _1-6 alkoxy group;
Alternatively, R ¹ and R ² may be bonded to each other and may form a 5- to 8-membered carbocyclic ring or a 5- to 8-membered heterocyclic ring together with adjacent carbon atoms, Alternatively, the 5- to 8-membered heterocycle may have a substituent selected from the following substituent group C;
C group: a halogen atom, a hydroxyl group, a C _1-4 alkyl group, a C _1-6 alkoxy group, a C _6-10 aryl group, a C _6-10 aryl C _1-6 alkyl group);

(In the formula, R ¹ and R ² have the same meanings as the general formula (1), respectively).
[2]
The microorganism is selected from the group consisting of a microorganism belonging to the genus Pseudomonas, a microorganism belonging to the genus Serratia, and a microorganism belonging to the family Xanthomonadaceae containing the 16S rDNA sequence represented by SEQ ID NO: 3. The method according to [1], wherein the method is at least one microorganism.
[3]
The method according to [2], wherein the microorganism belonging to the genus Pseudomonas is Pseudomonas corrugata 10F6 strain.
[4]
The method according to [2], wherein the microorganism belonging to the genus Serratia is Serratia sp. 11E11 strain.
[5]
The method according to [2], wherein the microorganism belonging to the family Xanthomonadaceae comprising the 16S rDNA sequence represented by SEQ ID NO: 3 is the 12A4 strain.
[6]
The ketone compound is acetophenone, 4-chloroacetophenone, 4-methylacetophenone, 2-methoxyacetophenone, 3-methoxyacetophenone, 4-methoxyacetophenone, 4-hydroxyacetophenone, 1- (3,4-dimethoxyphenyl) propane-2 -One, 1- (4-methoxyphenyl) propan-2-one, 1- (4-chlorophenyl) propan-2-one, 1- (4-hydroxyphenyl) propan-2-one, 1- (4-methyl Phenyl) propan-2-one, 1- (3,4-methylenedioxyphenyl) propan-2-one, benzylacetone, α-acetonaphthone, β-acetonaphthone, 1-indanone, α-tetralone, β-tetralone, 7 At least selected from the group consisting of -methoxy-2-tetralone, 1-benzyl-3-pyrrolidinone, 1-methyl-3-pyrrolidinone, 1-benzyl-3-piperidone and 1-methyl-3-piperidone The method according to any of characterized in that also a kind of the compound [1] to [5].
[7]
The ketone compound represented by the general formula (1) is a ketone compound represented by the following general formula (3), and the compound represented by the general formula (2) is represented by the following general formula (4). The method according to any one of [1] to [5], which is an optically active amino compound;

(In the formula, Y represents H or an amino-protecting group, p represents an integer of 2 to 3, q represents an integer of 1 to 2, and p> q).
[8]
The method according to [7], wherein the amino-protecting group is a C _1-6 alkyl group, a benzyl group, a formyl group, or a t-butoxycarbonyl group.
[9]
[7] The method according to [7], wherein the ketone compound is 1-benzyl-3-pyrrolidinone.
[10]
The amino group donor is
L-alanine, isopropylamine, 1-butylamine, sec-butylamine, 1,3-dimethyl-1-butylamine, 2-methylbutylamine, 1-pentylamine, 2-pentylamine, 3-pentylamine, 1-hexylamine, An aliphatic amine selected from 2-hexylamine, 1-heptylamine and 2-heptylamine;
Aromatics selected from 1-phenylethylamine, 2-phenylethylamine, benzylamine, 3-amino-1-phenylbutane, 4-amino-1-phenylpentane, α-ethylbenzylamine and 1- (1-naphthyl) ethylamine Amines;
The amino alcohol selected from 2-amino-1-phenylethanol and 2-phenylglycinol; and one selected from the group consisting of 1-aminoindane, according to any one of [1] to [9] Method.
[11]
The method according to any one of [1] to [10], wherein the action is performed in a solvent containing methanol or acetonitrile in an amount of 5 to 50% (ratio to the total solvent (volume / volume)).
[12]
A microbial cell having the ability to produce 1-benzyl-3-aminopyrrolidine from 1-benzyl-3-pyrrolidinone in the presence of an amino group receptor or a treated product thereof is represented by the following general formula (5). A method for producing an amino compound, which comprises reacting an enantiomeric mixture of an amino compound and collecting the remaining optically active amino compound represented by the following general formula (6);

(In the formula, R ¹ and R ² each independently has a C _6-10 aryl group which may have a substituent selected from the following group A, and a substituent selected from the following group A. It may have a C _6-10 aryl C _1-6 alkyl group, a 5-10 membered heteroaryl group which may have a substituent selected from the following group A, or a substituent selected from the following group B. Any one of the _{optionally selected} C _1-6 alkyl groups, and R ¹ is a group having a higher priority than R ² according to the order rule;
Group A: halogen atom, hydroxyl group, C _1-4 alkyl group, C _1-6 alkoxy group, halo C _1-6 alkyl group, —O (CH ₂ ) _n O— (n is 1 or 2);
Group B: halogen atom, hydroxyl group, C _1-6 alkoxy group;
Alternatively, R ¹ and R ² may be bonded to each other and may form a 5- to 8-membered carbocyclic ring or a 5- to 8-membered heterocyclic ring together with adjacent carbon atoms, Alternatively, the 5- to 8-membered heterocycle may have a substituent selected from the following substituent group C;
C group: a halogen atom, a hydroxyl group, a C _1-4 alkyl group, a C _1-6 alkoxy group, a C _6-10 aryl group, a C _6-10 aryl C _1-6 alkyl group);

(In the formula, R ¹ and R ² have the same meanings as in the general formula (5), respectively).
[13]
The microorganism is selected from the group consisting of a microorganism belonging to the genus Pseudomonas, a microorganism belonging to the genus Serratia, and a microorganism belonging to the family Xanthomonadaceae having the 16S rDNA sequence represented by SEQ ID NO: 3. [12] The method according to [12], which is at least one microorganism.
[14]
The method according to [13], wherein the microorganism belonging to the genus Pseudomonas is Pseudomonas corrugata 10F6 strain.
[15]
[13] The method according to [13], wherein the microorganism belonging to the genus Serratia. Is Serratia sp. 11E11 strain.
[16]
The method according to [13], wherein the microorganism belonging to the family Xanthomonadaceae containing the 16S rDNA sequence represented by SEQ ID NO: 3 is the 12A4 strain.
[17]
The amino compound represented by the general formula (5) is 1-phenylethylamine, α-ethylbenzylamine, 1-phenyl-2-aminopropane, 1- (3,4-dimethoxyphenyl) -2-aminopropane, 1-aminoindane, 1-aminotetralin, 2-aminotetralin, 1-phenyl-3-aminobutane, 3-aminopyrrolidine, 1-benzyl-3-aminopyrrolidine, 3-aminopiperidine and 1-benzyl-3-aminopiperidine The method according to any one of [12] to [16], wherein the method is at least one selected from the group consisting of:
[18]
The compound represented by the general formula (5) is an amino compound represented by the following general formula (7), and the compound represented by the general formula (6) is represented by the following general formula (8). A method according to any one of [12] to [16], which is a compound;

(In the formula, Y represents H or an amino-protecting group, p represents an integer of 2 to 3, q represents an integer of 1 to 2, and p> q).
[19]
[18] The method according to [18], wherein the amino-protecting group is a C _1-6 alkyl group, a benzyl group, a formyl group, or a t-butoxycarbonyl group.
[20]
The method according to [18], wherein the amino compound represented by the general formula (7) is 1-benzyl-3-aminopyrrolidine.
[21]
The amino group acceptor is at least one selected from the group consisting of pyruvic acid, glyoxylic acid, 2-ketoglutaric acid, acetophenone, propiophenone, benzylacetone, propionaldehyde, 1-butyraldehyde, benzaldehyde and phenylacetaldehyde. The method according to any one of [12] to [20], wherein

  Hereinafter, the meaning of terms, symbols, and the like described in the present specification will be described, and the present invention will be described in detail.

The “C _1-6 alkyl group” in the present specification is a monovalent group derived by removing any one hydrogen atom from an aliphatic hydrocarbon having 1 to 6 carbon atoms. Six straight-chain or branched alkyl groups are meant.
Specifically, for example, methyl group, ethyl group, 1-propyl group, 2-propyl group, 2-methyl-1-propyl group, 2-methyl-2-propyl group, 1-butyl group, 2-butyl group, 1-pentyl group, 2-pentyl group, 3-pentyl group, 2-methyl-1-butyl group, 3-methyl-1-butyl group, 2-methyl-2-butyl group, 3-methyl-2-butyl group 2,2-dimethyl-1-propyl group, 1-hexyl group, 2-hexyl group, 3-hexyl group, 2-methyl-1-pentyl group, 3-methyl-1-pentyl group, 4- Methyl-1-pentyl group, 2-methyl-2-pentyl group, 3-methyl-2-pentyl group, 4-methyl-2-pentyl group, 2-methyl-3-pentyl group, 3-methyl-3-pentyl Group, 2,3-dimethyl-1-butyl group, 3,3-dimethyl-1- Butyl group, 2,2-dimethyl-1-butyl, 2-ethyl-1-butyl group, 3,3-dimethyl-2-butyl group, 2,3-dimethyl-2-butyl group, and the like.

The “C _1-6 alkoxy group” in the present specification means an oxy group to which the above-defined “C _1-6 alkyl group” is bonded.
Specifically, for example, methoxy group, ethoxy group, 1-propyloxy group, 2-propyloxy group, 2-methyl-1-propyloxy group, 2-methyl-2-propyloxy group, 1-butyloxy group, 2 -Butyloxy group, 1-pentyloxy group, 2-pentyloxy group, 3-pentyloxy group, 2-methyl-1-butyloxy group, 3-methyl-1-butyloxy group, 2-methyl-2-butyloxy group, 3 -Methyl-2-butyloxy group, 2,2-dimethyl-1-propyloxy group, 1-hexyloxy group, 2-hexyloxy group, 3-hexyloxy group, 2-methyl-1-pentyloxy group, 3- Methyl-1-pentyloxy group, 4-methyl-1-pentyloxy group, 2-methyl-2-pentyloxy group, 3-methyl-2-pentyloxy group, 4 Methyl-2-pentyloxy group, 2-methyl-3-pentyloxy group, 3-methyl-3-pentyloxy group, 2,3-dimethyl-1-butyloxy group, 3,3-dimethyl-1-butyloxy group, Examples include 2,2-dimethyl-1-butyloxy group, 2-ethyl-1-butyloxy group, 3,3-dimethyl-2-butyloxy group, 2,3-dimethyl-2-butyloxy group and the like.

In the present specification, the “C _6-10 aryl group” refers to an aromatic hydrocarbon cyclic group having 6 to 10 carbon atoms, and specifically includes, for example, a phenyl group, a 1-naphthyl group, 2- And a naphthyl group.

  The “heteroatom” in the present specification means a sulfur atom, an oxygen atom or a nitrogen atom.

The “heteroaryl group” in the present specification means an aromatic ring group containing one or more heteroatoms in the atoms constituting the ring, and may be partially saturated. The ring may be a monocycle or a bicyclic heteroaryl group fused with a benzene ring or monoheteroaryl ring. The number of atoms constituting the ring is preferably 5 to 10 (C _5-10 heteroaryl group).
Specific examples of the heteroaryl group include a pyridyl group, a pyrazinyl group, a thienyl group, a furyl group, and a thiazolyl group.

  The “halogen atom” in the present specification means a fluorine atom, a chlorine atom, a bromine atom, or an iodine atom.

In the present specification, the “C _6-10 aryl C _1-6 alkyl group” is obtained by substituting any hydrogen atom in the above-mentioned definition “C _1-6 alkyl group” with the above-mentioned definition “C _6-10 aryl group”. Means group. The arylalkyl group is preferably a C _6-10 aryl C _1-4 alkyl group.
Specific examples include a benzyl group, a phenethyl group, and a 3-phenyl-1-propyl group.

The “halo C _1-6 alkyl group” represented in the present specification means a group obtained by substituting any hydrogen atom in the above-defined “C _1-6 alkyl group” with the above-mentioned “halogen atom”.

Substrate, product, amino group donor, amino group acceptor A method for producing an amino compound according to the present invention comprises converting 1-benzyl-3-aminopyrrolidine from 1-benzyl-3-pyrrolidinone in the presence of an amino group donor. A microbial cell having the ability to be produced or a processed product thereof is allowed to act on a ketone compound represented by the following general formula (1), and an optically active amino compound represented by the following general formula (2) is collected. It is characterized by that.

Although described in detail ketone compound, and R ^1, R ² of the ketone compound (1), and R ^1, R ² amino compound obtained (2) is as defined.
In the formula, R ¹ and R ² each independently have a C _6-10 aryl group which may have a substituent selected from the following group A, and a substituent selected from the following group A. Or a C _6-10 aryl C _1-6 alkyl group, a 5- to 10-membered heteroaryl group which may have a substituent selected from the following group A, or a substituent selected from the following group B indicates either be a C _1-6 alkyl group, R ¹ is the priority by sequence rule is higher group than R ^2.

The substituents of Group A and Group B are not limited as long as they are inert to the reaction, and the following substituents are preferable.
Group A: halogen atom, hydroxyl group, C _1-4 alkyl group, C _1-6 alkoxy group, halo C _1-6 alkyl group, —O (CH ₂ ) n O— (n is 1 or 2);
Group B: halogen atom, hydroxyl group, C _1-6 alkoxy group.

R ¹ and R ² may be bonded to each other and may form a 5- to 8-membered carbon ring or a 5- to 8-membered heterocycle together with adjacent carbon atoms. The 5- to 8-membered carbocycle or 5- to 8-membered heterocycle may have a substituent selected from the following substituent group C. The substituents of group C are not limited as long as they are inert to the reaction, but the following substituents are preferable.
Group C: C _1-4 alkyl group, C _1-6 alkoxy group, C _6-10 aryl C _1-6 alkyl group.

Among these, (i) any ^one of R ¹ and R ^{2 is} a C _1-6 alkyl group _optionally having a substituent selected from Group B, and any one is a substituent selected from Group A or a combination of good _{C 6-10} aryl _{C 1-6} alkyl group optionally having a substituent which is selected from optionally _{C 6-10} aryl group or group a have, or, (ii) R ¹ and R ² are bonded to each other, and examples thereof include a 5- to 8-membered carbocycle or a 5- to 8-membered heterocycle combined with an adjacent carbon atom. The ring may have a substituent selected from the following substituent group C.

Of the C _6-10 aryl groups that may have a substituent selected from Group A, preferably an unsubstituted C _6-10 aryl group, or a halogen atom, a hydroxyl group, C _1- C _6-10 aryl groups having ₄ alkyl groups or C _1-6 alkoxy groups can be mentioned.
Among C _6-10 aryl groups which may have a substituent, a phenyl group, a naphthyl group, a halogenated phenyl group, a hydroxyphenyl group, a C _1-4 alkylphenyl group, a C _1-4 alkoxy are more preferable. A phenyl group is mentioned, Especially preferably, a phenyl group, a naphthyl group, a chlorophenyl group, a hydroxyphenyl group, a toluyl group, and a methoxyphenyl group are mentioned.

Among C _6-10 aryl C _1-6 alkyl groups which may have a substituent selected from the following group A, preferably an unsubstituted C _6-10 aryl group C _1-6 alkyl group, or And a C _6-10 aryl C _1-6 having a halogen atom, a hydroxyl group, a C _1-4 alkyl group, a C _1-6 alkoxy group or —O (CH ₂ ) _n O— (n is 1 or 2) as a substituent. An alkyl group is mentioned.

Among the C _6-10 aryl C _1-6 alkyl groups which may have a substituent, more preferably a benzyl group, a phenethyl group, a halogenated benzyl group, a hydroxybenzyl group, a C _1-4 alkylbenzyl group, C _1-4 alkoxybenzyl group, di- _C1-4 alkoxybenzyl group, methylenedioxybenzyl group can be mentioned, particularly preferably benzyl group, phenethyl group, chlorobenzyl group, hydroxybenzyl group, methylbenzyl group, methoxybenzyl group. , Dimethoxybenzyl group and methylenedioxybenzyl group.

Among 5 to 10-membered heteroaryl groups which may have a substituent selected from Group A, preferably an unsubstituted 5 to 10-membered heteroaryl group or C _1-4 alkyl group is used as a substituent. And a 5- to 10-membered heteroaryl group (C _1-4 alkyl 5- to 10-membered heteroaryl group).
More preferable examples of the 5- to 10-membered heteroaryl group which may have a substituent include a pyridyl group, a piperazyl group, a thienyl group, a furyl group and a thiazolyl group.

Among the C _1-6 alkyl groups which may have a substituent selected from the group B, an unsubstituted C _1-6 alkyl group is preferable, and a C _1-4 alkyl group is more preferable.
Among the C _1-6 alkyl groups which may have a substituent, a methyl group, an ethyl group, a propyl group, an isopropyl group, a butyl group, an isobutyl group, and a t-butyl group are particularly preferable.

R ¹ and R ² may be bonded to each other and may form a 5- to 8-membered carbocyclic ring or a 5- to 8-membered heterocyclic ring together with adjacent carbon atoms. Alternatively, the 5- to 8-membered heterocyclic ring may have a substituent inert to the reaction, and may specifically have a substituent selected from the following substituent group C.
Group C: halogen atom, hydroxyl group, C _1-4 alkyl group, C _1-6 alkoxy group, C _6-10 aryl group, C _6-10 aryl C _1-6 alkyl group.

As the 5- to 8-membered carbocycle, the number of carbon atoms constituting the ring is preferably 5 to 8 (5- to 8-membered carbocycle), and the ring may have a double bond. It may be a bicyclic ring in which a benzene ring and a non-aromatic ring are condensed.
Specific examples include a cyclopentyl ring, a cyclohexyl ring, a cycloheptyl ring, a cyclooctyl ring, an indane ring, and a 1,2,3,4-tetrahydronaphthalene (tetralin) ring.
As the substituent, a C _1-6 alkoxy group is preferable.

As the 5- to 8-membered heterocycle, the number of atoms constituting the ring is preferably 5 to 8 (5- to 8-membered heterocycle), and 1 to 3 heteroatoms are contained in the atoms constituting the ring. A non-aromatic or monovalent ring which may have a double bond therein and is monocyclic. The hetero atom is preferably a nitrogen atom.
Specific examples of the hetero ring include a pyrrolidine ring, a piperidine ring, an imidazolidine ring, an imidazoline ring, a pyrazolidine ring, a pyrazoline ring, and a piperazine ring.
Preferred examples of the substituent include a C _1-4 alkyl group and a C _6-10 aryl C _1-6 alkyl group.

In the compound represented by the general formula (1), more preferably, a compound represented by the following general formula (3) is exemplified, and in the compound represented by the general formula (2), more preferably, The compound represented by General formula (4) is mentioned.

  In the formula, Y represents a protecting group for H or an amino group, p represents an integer of 2 to 3, q represents an integer of 1 to 2, and p> q.

Preferred examples of the protecting group for an amino group include C _1-6 alkyl groups such as a methyl group and an ethyl group, a formyl group, a benzyl group, and a t-butoxycarbonyl group (BOC group).

  Of these, a benzyl group and a t-butoxycarbonyl group (BOC group) are more preferable.

  p is preferably 2 or 3, and q is preferably 1.

  Specific examples of the compound represented by the general formula (1) and (3) include acetophenone, 4-chloroacetophenone, 4-methylacetophenone, 2-methoxyacetophenone, 3-methoxyacetophenone, 4-methoxy Methoxyacetophenone, 4-hydroxyacetophenone, 1- (3,4-dimethoxyphenyl) propan-2-one, 1- (4-methoxyphenyl) propan-2-one, 1- (4-chlorophenyl) propan-2-one 1- (4-hydroxyphenyl) propan-2-one, 1- (4-methylphenyl) propan-2-one, 1- (3,4-methylenedioxyphenyl) propan-2-one, benzylacetone, α-acetonaphthone, β-acetonaphthone, 1-indanone, α-tetralone, β-tetralone, 7-methoxy-2-tetralone, 1-benzyl-3-pyrrolidinone, 1-methyl-3-pyrrolidinone, 1-benzyl-3- Piperi Emissions include a 1-methyl-3-piperidone or the like.

  Examples of the amino group donor that can be used in the method of the present invention include general amino acids and amines. Specifically, for example, L-alanine, isopropylamine, 1-butylamine, sec-butylamine, 1,3-dimethyl-1-butylamine, 2-methylbutylamine, 1-pentylamine, 2-pentylamine, 3-pentylamine Aliphatic amines such as 1-hexylamine, 2-hexylamine, 1-heptylamine, 2-heptylamine; 1-phenylethylamine, 2-phenylethylamine, benzylamine, 3-amino-1-phenylbutane, 4- Aromatic amines such as amino-1-phenylpentane, 1-phenylpropylamine (α-ethylbenzylamine), 1- (1-naphthyl) ethylamine; amino such as 2-amino-1-phenylethanol and 2-phenylglycinol Examples of alcohols include cyclic amines such as 1-aminoindane.

Of these, L-alanine, isopropylamine, 1-butylamine, sec-butylamine, 1-pentylamine, 2-pentylamine, 1-hexylamine, 2-hexylamine, benzylamine, 1-phenylethylamine, 2-phenylethylamine, 2-phenylglycinol, 1-aminoindane, 3-amino-phenylbutane, and α-ethylbenzylamine Note that, in the amino group-containing compound, the compound having an asymmetric carbon atom is the optically active compound. Any one or a mixture of two or more thereof can be appropriately selected and used.

  The amount of such an amino group donor to be used is preferably 1.0 to 5 equivalents relative to the substrate. In addition, a substrate means the compound represented by (1), (3), (5), (7) used as a raw material.

Another embodiment of the process for producing an amino compound according to the present invention is a microbial cell having the ability to produce 1-benzyl-3-aminopyrrolidine from 1-benzyl-3-pyrrolidinone in the presence of an amino group receptor, or a microbial cell thereof The treated product is allowed to act on an enantiomeric mixture of an amino compound represented by the following general formula (5), and the remaining optically active amino compound represented by the following general formula (6) is collected.

In the formula, R ¹ and R ² have the same meanings as R ¹ and R ² in the general formula (1), respectively.

In the compound represented by the general formula (5), more preferably, a compound represented by the following general formula (7) is exemplified, and in the compound represented by the general formula (6), more preferably, The compound represented by General formula (8) is mentioned.

In the formula, Y represents a protecting group for H or an amino group, p represents an integer of 2 to 3, q represents an integer of 1 to 2, and p> q.
The amino group has the same meaning as the amino group in Y represented by the general formula (3).

  More specifically, the amino compounds represented by the general formulas (5) and (7) include 1-phenylethylamine, α-ethylbenzylamine, 1-phenyl-2-aminopropane, 1- (3, 4-dimethoxyphenyl) -2-aminopropane, 1-aminoindane, 1-aminotetralin, 2-aminotetralin, 1-phenyl-3-aminobutane, 3-aminopyrrolidine, 1-benzyl-3-aminopyrrolidine, 3- Examples include aminopiperidine, 1-benzyl-3-aminopiperidine and the like.

  Examples of the amino group receptor in the method of allowing a microbial cell to act on an enantiomeric mixture of the amino compound include general α-keto acids, ketones, and aldehyde compounds. Specifically, preferably, pyruvic acid, glyoxylic acid, 2-ketoglutaric acid, acetophenone, propiophenone, benzylacetone, propionaldehyde, 1-butyraldehyde, benzaldehyde, phenylacetaldehyde and the like can be mentioned.

  Of these, pyruvic acid, glyoxylic acid, 2-ketoglutaric acid, acetophenone, butyraldehyde, propionaldehyde, and benzaldehyde are more preferable.

  The amount of amino group receptor used is preferably 1.0 to 5 equivalents relative to the substrate.

Microorganism The microorganism that can be used in the present invention only needs to have the ability to produce 1-benzyl-3-aminopyrrolidine from 1-benzyl-3-pyrrolidinone. The reaction of the present invention can be carried out using such microbial cells or treated products thereof.

  Specific examples of such microorganisms include, for example, Pseudomonas corrugata 10F6 strain, Serratia sp. 11E11 strain, and Xanthomonasadaae (containing 16S rDNA of SEQ ID NO: 3). Examples include the microorganism 12A4 strain belonging to the family Xanthomonadaceae). These are newly isolated from soil in Japan as microorganisms having high transamination activity by the present inventors. Table 1 shows these morphological and physiological characteristics.

The 10F6 strain is a gram-negative gonococci of motility because it is gram-stained negative, Neisseria gonorrhoeae, no spore, motility, catalase positive, oxidase positive, non-fermentative.
The 11E11 strain is a kind of intestinal bacteria because of Gram staining negative, Neisseria gonorrhoeae, no spore, motility, oxidase reaction negative, and glucose fermentability.
The 12A4 strain is a gram-negative gonococci with gram staining, gonococcus, abspore, motility, positive catalase reaction, positive oxidase reaction, and non-fermentative.

  Furthermore, 16S ribosomal RNA genomic DNA was analyzed as a genetic method. InstaGene Matrix (BIORAD) was used for the extraction of genomic DNA, and the operation followed the protocol of BIORAD. Using the extracted genomic DNA as a template, a region of about 500 bp on the 5 'end side of the 16S ribosomal RNA gene (16S rDNA) was amplified by PCR. As the primers, 5F and 531R were used.

  Thereafter, the amplified base sequence was sequenced to obtain a 16S rDNA sequence. For PCR, MicroSeq 500 16S rDNA Bacterial Identification PCR Kit (Applied Biosystems) was used. MicroSeq 500 16S rDNA Bacterial Identification Sequencing Kit (Applied Biosystems) was used for the cycle sequence.

  GeneAmp PCR System 9600 (Applied Biosystems) was used as the thermal cycler, and ABI PRISM 3100 DNA Sequencer (Applied Biosystems) was used as the DNA sequencer. The basic operation from PCR to cycle sequence was in accordance with Applied Biosystems protocol (P / N 4346296 Rev. B).

  A homology search was performed using the base sequence of the obtained 16S rDNA. MicroSeq Microbial Identification System Software V.1.4.1 was used for homology search and molecular phylogenetic tree generation, and MicroSeq Bacterial 500 Library v.0023 (Applied Biosystems) was used as the database for homology search. The molecular phylogenetic tree was performed by the neighbor-joining method.

  As a result, the 16S rDNA sequence was sequence 1 for the 10F6 strain, sequence 2 for the 11E11 strain, and sequence 3 for the 12A4 strain. The phylogenetic tree is shown in FIGS.

  Since sequence 1 of 10F6 strain was 100% homologous to that of Pseudomonas corrugata, and was located in the same location as the 16S rDNA of Pseudomoans corrugata in the molecular phylogenetic tree (Fig. 1), 10F6 strain was identified as Pseudomoans corrugata. Identified. This result is also consistent with the previous morphological and physiological characteristics.

  The 16S rDNA sequence 2 of 11E11 strain showed the highest homology with that of Serratia fonticola with a homology of 97.72%. In addition, it was included in the cluster formed by Serratia 16S rDNA and formed a cluster with 16S rDNA of Serratia proteamaculans subsp. Proteamculance (Fig. 2). Since there was no sequence that completely matched sequence 2, it was not possible to identify to the species level at this time, and the 11E11 strain was identified as Serratia sp. This result is also consistent with the previous morphological and physiological characteristics.

  16S rDNA sequence 3 of strain 12A4 showed the highest homology with 96.79% homology with that of Xantomonas campestris subsp. Betilicola and Xanthomonas axonopodis subsp. Glycines. Moreover, it formed a cluster with 16S rDNA of Pseudomonas pictorum on the molecular phylogenetic tree (FIG. 3). Since no highly homologous sequence was found for SEQ ID NO: 3, it could not be identified to the genus level at this time, and the 12A4 strain was identified as a strain belonging to the family Xanthomonadaceae including Xanthomonas.

  As for Pseudomonas pictorum, as a result of 16S rRNA base sequence classification, it is inappropriate to be Pseudomonas genus, it is appropriate to transfer to Xanthomonadaceae family, genus level identification Has been reported to require further research (Anzai Y. et al., Int. J. Syst. Evol. Microbiol., 50, 1563-1589 (2000).).

  Pseudomoans corrugata 10F6 strain, Serratia sp. 11E11 strain, strain XA 12 Deposited at the National Institute of Technology and Evaluation (NPMD), which is an independent administrative agency located at 5-8 (〒 292-0818), dated September 21, 2005, with accession numbers NITE P-139, NITE P-140 and NITE P-141 are assigned. The present invention includes not only the deposited microorganism itself but also its mutant strain having the above-mentioned ability and a strain that has been genetically engineered.

  The medium for culturing the microorganism used in the present invention is not particularly limited as long as the microorganism can grow.

  For example, any of the above microorganisms can be used as the carbon source. Specifically, sugars such as glucose, fructose, sucrose, and dextrin, alcohols such as sorbitol and glycerol, fumaric acid, citric acid, Organic acids such as acetic acid and propionic acid and salts thereof, hydrocarbons such as paraffin, toluene, cresol, benzoic acid and the like, or a mixture thereof can be used.

  Examples of nitrogen sources include ammonium salts of inorganic acids such as ammonium chloride, ammonium sulfate, and ammonium phosphate, ammonium salts of organic acids such as ammonium fumarate and ammonium citrate, meat extract, yeast extract, corn steep liquor, and casein hydrolysis. Products, inorganic organic nitrogen-containing compounds such as urea, or mixtures thereof.

  In addition, nutrient sources used in normal culture, such as inorganic salts, trace metal salts, vitamins, and the like can be appropriately mixed and used.

Further, if necessary, factors that promote the growth of microorganisms, factors that increase the ability to produce the target compound of the present invention, specifically 1-propylamine, 2-propylamine, 1-butylamine, sec-butylamine, 1- Aliphatic amines such as pentylamine, 2-pentylamine, 1-hexylamine, 2-hexylamine, 1-heptylamine, 2-heptylamine, 1-octylamine, 2-octylamine, cyclohexylamine, 1-benzyl- 0.01 g of cyclic amines such as 3-aminopyrrolidine and 3-aminopyrrolidine, aromatic amines such as 1-phenylethylamine, 2-phenylethylamine, 1-aminoindane, 1-aminotetralin and 1- (1-naphthyl) ethylamine / L to 10 g / L An appropriate concentration can also be added. Alternatively, a substance such as CaCO ₃ effective for maintaining the pH of the medium can be added.

  As the culture method, the medium pH is 3 to 11, preferably 4 to 8, the culture temperature is 15 to 60 ° C., preferably 20 to 45 ° C., and anaerobic or aerobic conditions suitable for the growth of the microorganism. The culture is performed for about 5 to 240 hours, preferably about 12 to 120 hours.

Method for Producing Amino Compound A method for producing an amino compound according to the present invention comprises a microbial cell having the ability to produce 1-benzyl-3-aminopyrrolidine from 1-benzyl-3-pyrrolidinone in the presence of an amino group donor or The treated product is allowed to act on the ketone compound represented by the general formula (1) or (3), and the optically active amino compound represented by the general formula (2) or (4) to be produced is collected. It is a feature.

  In addition, the method for producing an amino compound according to the present invention includes a microbial cell having the ability to produce 1-benzyl-3-aminopyrrolidine from 1-benzyl-3-pyrrolidinone in the presence of an amino group receptor, or a processed product thereof. Is allowed to act on the enantiomeric mixture of the amino compound represented by the general formula (5) or (7) to collect the remaining optically active amino compound represented by the general formula (6) or (8). It is a feature.

  The substrate is a ketone compound represented by the general formula (1) or (3) or an amino compound represented by the general formula (5) or (7), in a concentration range in which substrate inhibition of the enzyme does not occur, What is necessary is just to add to reaction systems, such as a culture medium intermittently or continuously, Usually, 0.01 to 20% (wt / wt) (ratio of the mass of the substrate with respect to the mass of the reaction liquid) is added.

  The substrate can be added as it is dissolved or dispersed in water as it is, dissolved in water or dissolved in an organic solvent such as DMSO that does not affect the reaction, or dispersed in a surfactant or the like. Good.

  The microorganisms used in the present invention can be used in the following manner: 1) a method in which a substrate, an amino group donor or an amino group acceptor is added to the medium from the beginning and cultured, and 2) the culture solution is used as it is. A method of adding an amino group donor or amino group acceptor, a ketone compound or an amino compound as a substrate to the solution, 3) separating the cells by centrifugation, etc., and washing the buffer as it is or after washing, There is a method in which an amino group donor or amino group acceptor or a substrate is added to a product resuspended in water or the like and reacted.

  The microbial cell may be a living microbial cell, or may be a microbial cell crushed material, a treated product that has been subjected to a treatment such as acetone treatment, toluene treatment, or lyophilization.

  The cells of the microorganisms can be immobilized on carrageenan gel, alginic acid gel, polyacrylamide gel, cellulose, agar, etc. by a known method, and reacted in a reactor using an ultrafiltration membrane or the like. You can also.

  A coenzyme such as pyridoxal-5'-phosphate (PLP) necessary for the reaction can also be added.

  The reaction temperature is preferably 5 to 80 ° C, more preferably 15 to 70 ° C. The reaction pH may be appropriately selected within the range in which the enzyme reacts, but it is usually preferably pH 4 to 12, more preferably pH 6 to 11, and a buffer solution or a pH stat can be used.

  The reaction can be performed by standing, shaking, or stirring.

  The solvent used in the reaction is usually water, but an organic solvent such as alcohol can be added within a range that does not affect the reaction. In particular, in the reaction for producing 1-benzyl-3-aminopyrrolidine, methanol and acetonitrile are added in an amount of 5 to 50% by volume with respect to 1-benzyl-3-pyrrolidinone, thereby producing 1-benzyl-3-aminopyrrolidine. Optical purity can be improved.

  In the case of ketone compounds that are unstable in air, the reaction yield can be obtained by replacing the gas phase of the reaction solution with an inert gas such as nitrogen, helium, or argon, or by carrying out the reaction in a gas stream. Can be improved.

  The produced amino compound represented by the general formula (2), (4), (6) or (8) or the remaining amino compound represented by the general formula (5) or (7) is subjected to ultrafiltration, It can be recovered and purified by combining ordinary methods such as concentration, column chromatography, extraction, activated carbon treatment and distillation.

As described above, in the present invention, by using the ketone compound represented by the general formula (1) or (3) as a substrate, the optically active (S) amino acid represented by the general formula (2) or (4) is used. The compound can be obtained with high purity.
In particular, by using a 3-pyrrolidinone derivative as a substrate, an (S) -3-aminopyrrolidine derivative can be efficiently produced.

  In particular, optically active 1-benzyl-3-aminopyrrolidine is important as a raw material for pharmaceuticals such as antibiotics, antibacterial agents, dopamine D3 receptor agonists, and CCR3 antagonists.

In the present invention, an optically active (R) amino compound represented by the general formula (6) or (8) is used by using the amino compound represented by the general formula (5) or (7) as a substrate. Can be obtained with high purity.
In particular, the (R) -3-aminopyrrolidine derivative can be selectively left from the enantiomeric mixture of the 3-aminopyrrolidine derivative.

EXAMPLES Next, although an Example demonstrates this invention concretely, this invention is not limited only to these Examples. Hereinafter, in the examples, 1-benzyl-3-pyrrolidinone is abbreviated as PNB, and 1-benzyl-3-aminopyrrolidine is abbreviated as APB.
In the present invention, quantitative determination of each amine, amino acid, and ketone and measurement of optical purity were performed by high performance liquid chromatography.

Determination of PNB, APB, 1-phenylethylamine Column: Luna 5u C18, φ2.0mm x 150mm (Phenomenex)
Mobile phase: 40 mM phosphate buffer pH 2.5 (including 5 mM sodium hexanesulfonate) / acetonitrile [9: 1, v / v]
Column temperature: 40 ° C
Flow rate: 0.2mL / min
Detection: 254nm
Retention time: PNB 11.1 minutes, 1-phenylethylamine 14.3 minutes, APB 17.8 minutes

Determination of acetophenone <br/> Column: Luna 5u C18, φ2.0mm × 150mm (phenomenex)
Mobile phase: 0.1% trifluoroacetic acid aqueous solution / 0.1% trifluoroacetic acid acetonitrile solution [65:35, v / v]
Column temperature: 40 ° C
Flow rate: 0.2mL / min
Detection: 254nm
Retention time: 15.8 minutes

Determination of 3-amino-1-phenylbutane <br/> Column: Luna 5u C18, φ2.0mm × 150mm (Phenomenex)
Mobile phase: 40 mM phosphate buffer pH 2.5 (including 5 mM sodium hexanesulfonate) / acetonitrile [0 to 5 minutes [9: 1, v / v] constant, 5 to 20 minutes [9: 1, v / v] → [6: 4, v / v] linear concentration gradient elution)
Column temperature: 40 ° C
Flow rate: 0.2mL / min
Detection: 254nm
Retention time: 10.5 minutes

Determination of alanine and glutamic acid (derivatization with dub silk chloride)
The reaction solution is centrifuged at 15000 rpm for 5 minutes to obtain a supernatant. Take 20 μL of this supernatant and add 80 μL of 0.2 M NaHCO ₃ —Na ₂ CO ₃ buffer, pH 9.0. To this, add 200 μL of a 10 mM doubsyl chloride solution in acetone, seal tightly, and heat at 70 ° C. for 15 minutes. After cooling to room temperature, 100 μL of 1 M HCl is added and this is used as the HPLC sample.

Column: Luna 5u C18, φ2.0mm x 150mm (Phenomenex)
Mobile phase: 45 mM potassium acetate buffer, pH 4.0 / acetonitrile [65:35, v / v]
Column temperature: 40 ° C
Flow rate: 0.2mL / min
Detection: 436nm
Retention time: Glutamic acid 11.0 minutes, Alanine 19.2 minutes

APB optical purity measurement (preparation of measurement sample from reaction solution)
Take 100 μL of the reaction solution in a 2 mL centrifuge tube, and add 150 μL of pure water. Add 1N NaOH to pH 12 or higher. Add 0.5mL of 1-butanol, shake and mix gently, and remove 1-butanol layer as much as possible. 1-Butanol is distilled off under reduced pressure to obtain a residue.

(GITC derivatization)
Add 100 μL of acetonitrile to the residue and dissolve. Take 50 μL of this, add 100 μL of 0.4% GITC acetonitrile solution, and let stand at room temperature for 10 minutes. To this, 50 μL of 0.2% ethanolamine in acetonitrile is added and left at room temperature for 3 minutes. This is appropriately diluted with an HPLC mobile phase to obtain an optical purity measurement HPLC sample.
GITC: 2,3,4,6-tetra-O-acetyl-β-D-glucopyranosyl isothiocyanate

Column: Luna 5u C18, φ2.0mm x 150mm (Phenomenex)
Mobile phase: 400 mM ammonium acetate buffer, pH 4.0 / methanol [58:38, v / v]
Column temperature: 30 ° C
Flow rate: 0.2mL / min
Detection: 254nm
Retention time: R body 23.5 minutes, S body 26.3 minutes

Measurement of optical purity of 1-phenylethylamine <br/> Column: CROWNPAK CR (+), φ4.6 mm × 150mm (Daicel Chemical Industries)
Mobile phase: Perchloric acid aqueous solution, pH 1.5
Column temperature: 30 ° C
Flow rate: 0.6mL / min
Detection: 254nm
Retention time: S body 16.0 minutes, R body 20.4 minutes

Measurement of optical purity of 3-amino-1-phenylbutane
GITC was performed in the same manner as APB, and an HPLC sample was prepared.
Column: Luna 5u C18, φ2.0mm x 150mm (Phenomenex)
Mobile phase: 400 mM ammonium acetate buffer, pH 4.0 / methanol [50:50, v / v]
Column temperature: 50 ° C
Flow rate: 0.2mL / min
Detection: 254nm
Retention time: R body 63.6 minutes, S body 67.3 minutes

In addition, the composition of “bacteria preparation medium 1” and “bacteria preparation medium 2” used in the following examples is as follows.
(A medium)
Glucose 3g, yeast extract (Kyokuto Pharmaceutical) 0.3g, KH ₂ PO ₄ 1g, K ₂ HPO ₄ 3g, MgSO ₄ · 7H ₂ O 0.5g, 1-aminopentane 1g, 10mL of mineral solution of Table 2 are mixed and removed Add ionic water to dissolve and adjust the pH to 7.0 with a total volume of 1000 mL.

調製方法：脱イオン水900mLにニトリロ三酢酸を溶かし、
1N KOHでpH6.5とする。次いで各金属塩を溶解させる。
最後に1N KOHでpH7.0とする。

Preparation method : Dissolve nitrilotriacetic acid in 900 mL of deionized water,
Adjust to pH 6.5 with 1N KOH. Each metal salt is then dissolved.
Finally, the pH is adjusted to 7.0 with 1N KOH.

(Medium B)
Medium with yeast extract of medium A 2g / L

(C medium)
Medium containing 0.3 g / L of Adecanol LG-294 added to medium B

(D medium)
Bouillon medium: 10 g of meat extract (Kyokuto Pharmaceutical), 10 g of polypeptone (Nippon Pharmaceutical), and 5 g of NaCl were dissolved in deionized water to a total volume of 1000 mL and adjusted to pH 7.3.

(Comparative Example 1)
Reaction with a known ω-amino acid transaminase-producing bacterium Pseudomonas putida IFO 14796 Pseudomonas putida IFO 14796 (F-126 strain) is prepared by a known method (Yonaha K. et al., Agric. Biol. Chem). , 47 (10), 2257-2265 (1983).). Specifically, a medium consisting of 1 g of glycerin, 2 g of peptone, 1 g of KH ₂ PO _4, 1 g of K ₂ HPO ₄ , 0.5 g of yeast extract, 2 g / L of alanine, pH 7.2, 50 mL in a 500 mL conical flask with a fold After sterilization at 121 ° C. for 15 minutes, one platinum ear was inoculated from a preserved slant, and cultured with shaking at 30 ° C. for 24 hours.

The cells collected from the culture by centrifugation were further washed once with 10 mM potassium phosphate buffer (pH 7.0). The cells were suspended in 10 mM potassium phosphate buffer (pH 7.0). Cell concentration OD600 = 24 at final concentration, amino group donor 100 mM (sec butylamine only 200 mM), acceptor 50 mM, 0.05 mM PLP, 100 mM potassium phosphate buffer, pH 8.0 (adjusted with HCl or NaOH), total The mixture was mixed so that the liquid volume became 0.50 mL, left at 25 ° C., and the reaction was carried out for 24 hours with N ₂ substitution of the gas phase.

  After completion of the reaction, the produced amine and amino acid were quantified by HPLC and shown in Table 3. Similarly, the cell concentration OD600 = 24 at the final concentration, racemic APB 50 mM as the amino group donor, pyruvic acid 30 mM, 0.05 mM PLP, 100 mM potassium phosphate buffer, pH 8.0 (in HCl or NaOH) The mixture was mixed so that the total liquid volume became 0.50 mL, and the shaking reaction was performed at 25 ° C. for 24 hours. The results are shown in Table 4.

  PWB does not produce any APB from PNB even when reacted with Pseudomonas putida IFO 14796 with ω-amino acid transaminase activity, and no PNB is produced even when racemic APB is allowed to act on pyruvate as a receptor. Thus, the novel action of the strain of the present invention was clarified.

Example 1
5 mL of each medium A was put in a φ21 mm test tube and sterilized at 121 ° C. for 15 minutes. Then, one strain of the strains shown in Table 5 was inoculated from a preserved slant and cultured with shaking at 25 ° C. for 48 hours.

The bacterial cells centrifuged from the total amount of each culture solution were washed once with 10 mM potassium phosphate buffer (pH 7.0). This cell was suspended in 100 mM potassium phosphate buffer (pH 7.0) to make 0.2 mL. To this was added 0.3 mL of the following reaction solution, and the gas phase was replaced with N ₂ gas, followed by a stationary reaction at 25 ° C. for 24 hours. After completion of the reaction, the reaction solution was diluted 10-fold with 0.1 M HCl, the supernatant was separated by centrifugation, the supernatant was analyzed by high performance liquid chromatography, and APB was quantified. APB was converted to GITC and the optical purity was measured by high performance liquid chromatography. The results are shown in Table 5.

Substrate solution (1)
PNB 50mM
(S) -1-Phenylethylamine 50 mM
PLP 0.083mM
In 100 mM potassium phosphate buffer, pH 7.0

Example 2
Three strains shown in Table 6 were cultured, collected, washed and suspended in the same manner as in Example 1, and reacted by mixing 0.3 mL of the following substrate solution and 0.2 mL of the cell suspension. After substituting the gas phase with N ₂ gas, a stationary reaction was performed at 25 ° C. for 24 hours, and quantitative determination and optical purity measurement were performed in the same manner as in the Examples. The results are shown in Table 6.

Substrate solution (2)
PNB 50mM
Racemic sec-Butylamine 200mM
PLP 0.083mM
In 100 mM potassium phosphate buffer, pH 7.0

Example 3
Pre-culture was performed by shaking for 24 hours at 25 ° C. in a conical Erlenmeyer flask containing 50 mL of D medium sterilized with Pseudomonas corrugata 10F6. C culture medium 1200mL each in 4 Maruryo Bioengine MDL-200 2.0L mini jars, sterilized at 121 ° C for 20 minutes, cooled, then inoculated 12mL each of the above culture solution, 580 rpm, 25 ° C The cells were cultured for 48 hours with aeration at 0.5 vvm.

After completion of the culture, 2 L of the culture broth was collected, and the cells were collected by centrifugation and suspended in deionized water to 120 mL. To this was added 6 mL of 5 mM PLP, 6.54 g (54 mmol) (S) -1-phenylethylamine, 6.31 g (36 mmol) g of PNB, and the whole was made up to 300 mL with 100 mM potassium phosphate buffer, pH 7.0. I put it in a 1L vessel of the miniger. The reaction was carried out at 25 ° C. for 48 hours at 200 rpm while N ₂ gas (100 mL / min) was passed through the gas phase.

  After the reaction was completed, 19.6 g / L of APB was formed in the reaction solution, and 1.02 g / L of PNB remained. The reaction yield after correcting for the decrease in the liquid volume was 87.9%. The cells were removed by centrifugation and the supernatant was separated. Next, the pH was adjusted to 3 with sulfuric acid, and PNB was extracted with 280 mL of methyl isobutyl ketone. Next, NaOH was added to the aqueous layer to adjust the pH to 12.5, and extracted twice with 270 mL of 1-butanol. The 1-butanol layer was collected and washed with 70 mL of saturated saline. The 1-butanol layer was concentrated under reduced pressure to obtain 6.43 g of crude (S) -APB. The chemical purity was 70.4%, the optical purity was 87.9% ee, and the consistent yield was 71.3%.

Example 4 Comparison of amino group donors

In the same manner as in Example 3, Pseudomonas corrugata 10F6 strain was cultured. After centrifugation, the cells were washed once with 10 mM potassium phosphate buffer pH 7.0, and then the cells were suspended in 100 mM potassium phosphate buffer pH 7.0 so that OD600nm was 60.

After mixing 0.40 mL of this bacterial cell suspension with 0.60 mL of the substrate solution or suspension containing each amine shown in Table 7, the gas phase was replaced with N ₂ gas, and then allowed to stand at 25 ° C. for 24 hours. And quantified in the same manner as in Example 1. The composition of the substrate solution or suspension is as follows.
Donor amine 100 mM (for (S) -1-phenylethylamine), 200 mM (for other racemic amines)
PLP 0.083mM
PNB 50mM
100 mM potassium phosphate buffer, pH 7.0 (pH adjusted with HCl)
The results are shown in Table 7.

Example 5
Place 50 mL of each medium A in a 500 mL Erlenmeyer flask with folds, sterilize at 121 ° C for 15 minutes, and then store Pseudomonas corrugata 10F6 strain and Serratia sp. 11E11 strain from a preserved slant. Inoculated and cultured with shaking at 140 rpm for 48 hours at 25 ° C. The culture solution is collected, collected by centrifugation, washed once with 10 mM potassium phosphate buffer pH 7.0, and the cells are suspended in 100 mM potassium phosphate buffer pH 7.0 so that the OD 600 nm is 60. did. After mixing 0.40 mL of this bacterial cell suspension and 0.60 mL of the substrate suspension containing each amine shown in Table 8, the gas phase was replaced with N ₂ gas, and then the mixture was allowed to stand at 25 ° C. for 24 hours. The amount was determined in the same manner as in Example 1. The results are shown in Table 8.

Substrate suspension Donor amine 167 mM (for optically active or asymmetric carbon-free amines), 333 mM (for racemic amines)
PLP 0.083mM
PNB 83mM
100 mM potassium phosphate buffer, pH 7.0 (pH adjusted with HCl)

Example 6
Comparison of amino group acceptors with (S) -1-phenylethylamine as donor
Place 50 mL of medium A in a 500 mL fritted Erlenmeyer flask, sterilize at 121 ° C for 15 minutes, and then store Pseudomonas corrugata 10F6 strain, Serratia sp. 11E11 and 12A4 strains from the preserved slant. Platinum ears were inoculated and cultured with shaking at 25 ° C. for 48 hours. The bacterial cells centrifuged from each culture solution were washed once with 10 mM potassium phosphate buffer (pH 7.0). This cell was suspended in 100 mM potassium phosphate buffer (pH 7.0), and a cell suspension concentrated 15 times with respect to the culture solution was prepared. 50 mM (S) -1-phenylethylamine, 50 mM amino group receptor shown in Table 9, 100 mM potassium phosphate buffer solution containing 0.167 mM PLP 0.3 mL of each substrate solution of pH 7.0 and cell suspension 0.2 mL was mixed, allowed to stand at 25 ° C., and the gas phase was replaced with N ₂ and reacted for 24 hours. After completion of the reaction, acetophenone produced by HPLC was quantified, and the relative values with pyruvate as 100 are shown in Table 9.

  In all three strains, glyoxylic acid was a better receptor than pyruvic acid. Especially in 10F6 strain, PNB was the best receptor.

Example 7
Synthesis of various amines In the same manner as in Example 6, Pseudomonas corrugata 10F6 strain and Serratia sp. 11E11 strain were cultured, collected and washed. Next, the cells were suspended in 100 mM potassium phosphate buffer (pH 7.0), and the amount of the solution was adjusted so that the OD was 60. 167 mM L-alanine or 333 mM sec butylamine, 83 mM each ketone shown in Table 10, 100 mM potassium phosphate buffer pH 0.1 (adjusted with HCl) containing 0.167 mM PLP and each substrate suspension 0.3 mL 0.2 mL of the cell suspension was mixed, allowed to stand at 25 ° C., and the gas phase was substituted with N ₂ for 24 hours.

  After completion of the reaction, the amine corresponding to the produced ketone was quantified by HPLC, and its optical purity was measured. The results are shown in Table 10. As the amino group donor, sec-butylamine was better than L-alanine. (S) -1-phenylethylamine is produced from acetophenone and (S) -3-amino-1-phenylbutane is produced from benzylacetone with high optical purity. By using the strain of the present invention, not only APB but also It has been found that a wide range of other useful (S) -amino compounds can be synthesized.

(Example 8)
Preparation of (R) -APB from racemic APB
Place 50 mL of B medium in a 500 mL fritted Erlenmeyer flask and sterilize at 121 ° C for 15 minutes. Then inoculate one Pseudomonas corrugata strain (Pseudomonas corrugata) 10F6 from a preserved slant and shake at 25 ° C for 48 hours. Culture was performed. After centrifugation and washing once with 10 mM potassium phosphate buffer (pH 7.0), the cells were suspended in 100 mM potassium phosphate buffer (pH 8.0). Final concentration of 50mM racemic APB, 50mM pyruvate, 0.05mM PLP, 100mM potassium phosphate buffer pH8.0 (adjusted with HCl), cell concentration OD600 = 12,6, total solution volume 0.50mL Were mixed, and shaken at 25 ° C. for 24 hours. After the reaction was completed, the remaining APB concentration and optical purity were measured. The results are shown in Table 11.

本発明の菌株をAPBのエナンチオマー混合物に作用させることにより、(R)-APBを調製できた。

(R) -APB could be prepared by allowing the strain of the present invention to act on an enantiomeric mixture of APB.

Example 9
Effect of methanol and acetonitrile on APB synthesis reaction Pseudomonas corrugata 10F6 strain was cultured in the same manner as in Example 8. The cells collected from the culture by centrifugation were further washed once with 10 mM potassium phosphate buffer (pH 7.0). The cells were suspended in 10 mM potassium phosphate buffer (pH 7.0), and the amount of the solution was adjusted so that OD600 was 96. A predetermined amount of 0.075 mL of the bacterial cells, 0.075 mL of the substrate solution, methanol or acetonitrile was added, and the liquid volume was adjusted to 0.30 mL with pure water. These were allowed to stand at 25 ° C., and the gas phase was substituted with N ₂ for 24 hours.

  After completion of the reaction, the produced APB was quantified by HPLC, and its optical purity was further measured. The results are shown in Table 12. It was found that the optical purity of the APB produced was improved by adding 5 to 35% by volume of methanol and acetonitrile.

基質溶液組成：120mM PNB、480mM secブチルアミン、0.2mM PLP（100mM カリウムリン酸緩衝液, pH7.0中）

Substrate solution composition: 120 mM PNB, 480 mM sec butylamine, 0.2 mM PLP (in 100 mM potassium phosphate buffer, pH 7.0)

According to the present invention, an (S) isomer having a specific structure can be obtained by using a microbial cell having a capability of producing 1-benzyl-3-aminopyrrolidine from 1-benzyl-3-piperidinone or a processed product thereof. The (R) -form optically active amines can be efficiently produced from the corresponding ketone or the corresponding racemate.
Such a method for synthesizing amines by an enzymatic method can be performed at room temperature and normal pressure with few synthesis byproducts.

FIG. 1 is a phylogenetic tree of the 10F6 strain and related strains by the neighbor joining method. FIG. 2 is a phylogenetic tree of the 11E11 strain and related strains by the neighbor joining method. FIG. 3 is a phylogenetic tree of the 12A4 strain and related strains by the neighbor joining method.

Claims (21)

A microbial cell having the ability to produce 1-benzyl-3-aminopyrrolidine from 1-benzyl-3-pyrrolidinone in the presence of an amino group donor or a treated product thereof is represented by the following general formula (1). A process for producing an amino compound characterized by collecting an optically active amino compound represented by the following general formula (2) produced by acting on a ketone compound;

(In the formula, R ¹ and R ² each independently has a C _6-10 aryl group which may have a substituent selected from the following group A, and a substituent selected from the following group A. It may have a C _6-10 aryl C _1-6 alkyl group, a 5-10 membered heteroaryl group which may have a substituent selected from the following group A, or a substituent selected from the following group B. Any one of the _{optionally selected} C _1-6 alkyl groups, and R ¹ is a group having a higher priority than R ² according to the order rule;
Group A: halogen atom, hydroxyl group, C _1-4 alkyl group, C _1-6 alkoxy group, halo C _1-6 alkyl group, —O (CH ₂ ) _n O— (n is 1 or 2);
Group B: halogen atom, hydroxyl group, C _1-6 alkoxy group;
Alternatively, R ¹ and R ² may be bonded to each other and may form a 5- to 8-membered carbocyclic ring or a 5- to 8-membered heterocyclic ring together with adjacent carbon atoms, Alternatively, the 5- to 8-membered heterocycle may have a substituent selected from the following substituent group C;
C group: a halogen atom, a hydroxyl group, a C _1-4 alkyl group, a C _1-6 alkoxy group, a C _6-10 aryl group, a C _6-10 aryl C _1-6 alkyl group);

(In the formula, R ¹ and R ² have the same meanings as the general formula (1), respectively).   The microorganism is selected from the group consisting of microorganisms belonging to the genus Pseudomonas, microorganisms belonging to the genus Serratia and microorganisms belonging to the family Xanthomonadaceae containing the 16S rDNA sequence represented by SEQ ID NO: 3. The method of claim 1, wherein the method is at least one microorganism.   3. The method according to claim 2, wherein the microorganism belonging to the genus Pseudomonas is a Pseudomonas corrugata 10F6 strain.   The method according to claim 2, wherein the microorganism belonging to the genus Serratia. Is a Serratia sp. 11E11 strain.   The method according to claim 2, wherein the microorganism belonging to the family Xanthomonadaceae containing the 16S rDNA sequence represented by SEQ ID NO: 3 is the 12A4 strain.   The ketone compound is acetophenone, 4-chloroacetophenone, 4-methylacetophenone, 2-methoxyacetophenone, 3-methoxyacetophenone, 4-methoxyacetophenone, 4-hydroxyacetophenone, 1- (3,4-dimethoxyphenyl) propane-2 -One, 1- (4-methoxyphenyl) propan-2-one, 1- (4-chlorophenyl) propan-2-one, 1- (4-hydroxyphenyl) propan-2-one, 1- (4-methyl Phenyl) propan-2-one, 1- (3,4-methylenedioxyphenyl) propan-2-one, benzylacetone, α-acetonaphthone, β-acetonaphthone, 1-indanone, α-tetralone, β-tetralone, 7 At least selected from the group consisting of -methoxy-2-tetralone, 1-benzyl-3-pyrrolidinone, 1-methyl-3-pyrrolidinone, 1-benzyl-3-piperidone and 1-methyl-3-piperidone The method according to any one of claims 1 to 5, wherein it is one compound. The ketone compound represented by the general formula (1) is a ketone compound represented by the following general formula (3), and the compound represented by the general formula (2) is represented by the following general formula (4). The method according to any one of claims 1 to 5, which is an optically active amino compound;

(In the formula, Y represents H or an amino-protecting group, p represents an integer of 2 to 3, q represents an integer of 1 to 2, and p> q). The method according to claim 7, wherein the amino-protecting group is a C _1-6 alkyl group, a benzyl group, a formyl group, or a t-butoxycarbonyl group.   The method according to claim 7, wherein the ketone compound is 1-benzyl-3-pyrrolidinone. The amino group donor is
L-alanine, isopropylamine, 1-butylamine, sec-butylamine, 1,3-dimethyl-1-butylamine, 2-methylbutylamine, 1-pentylamine, 2-pentylamine, 3-pentylamine, 1-hexylamine, An aliphatic amine selected from 2-hexylamine, 1-heptylamine and 2-heptylamine;
Aromatics selected from 1-phenylethylamine, 2-phenylethylamine, benzylamine, 3-amino-1-phenylbutane, 4-amino-1-phenylpentane, α-ethylbenzylamine and 1- (1-naphthyl) ethylamine Amines;
The method according to any one of claims 1 to 9, which is an aminoalcohol selected from 2-amino-1-phenylethanol and 2-phenylglycinol; and one selected from the group consisting of 1-aminoindan.   The method according to any one of claims 1 to 10, wherein the action is performed in a solvent containing 5 to 50% of methanol or acetonitrile (ratio (volume / volume) to the total solvent). A microbial cell having the ability to produce 1-benzyl-3-aminopyrrolidine from 1-benzyl-3-pyrrolidinone in the presence of an amino group receptor or a treated product thereof is represented by the following general formula (5). A method for producing an amino compound, which comprises reacting an enantiomeric mixture of an amino compound and collecting the remaining optically active amino compound represented by the following general formula (6);

(In the formula, R ¹ and R ² each independently has a C _6-10 aryl group which may have a substituent selected from the following group A, and a substituent selected from the following group A. It may have a C _6-10 aryl C _1-6 alkyl group, a 5-10 membered heteroaryl group which may have a substituent selected from the following group A, or a substituent selected from the following group B. Any one of the _{optionally selected} C _1-6 alkyl groups, and R ¹ is a group having a higher priority than R ² according to the order rule;
Group A: halogen atom, hydroxyl group, C _1-4 alkyl group, C _1-6 alkoxy group, halo C _1-6 alkyl group, —O (CH ₂ ) _n O— (n is 1 or 2);
Group B: halogen atom, hydroxyl group, C _1-6 alkoxy group;
Alternatively, R ¹ and R ² may be bonded to each other and may form a 5- to 8-membered carbocyclic ring or a 5- to 8-membered heterocyclic ring together with adjacent carbon atoms, Alternatively, the 5- to 8-membered heterocycle may have a substituent selected from the following substituent group C;
C group: a halogen atom, a hydroxyl group, a C _1-4 alkyl group, a C _1-6 alkoxy group, a C _6-10 aryl group, a C _6-10 aryl C _1-6 alkyl group);

(In the formula, R ¹ and R ² have the same meanings as in the general formula (5), respectively).   The microorganism is selected from the group consisting of a microorganism belonging to the genus Pseudomonas, a microorganism belonging to the genus Serratia, and a microorganism belonging to the family Xanthomonadaceae having the 16S rDNA sequence represented by SEQ ID NO: 3. 13. A method according to claim 12, wherein the method is at least one microorganism.   The method according to claim 13, wherein the microorganism belonging to the genus Pseudomonas is Pseudomonas corrugata 10F6 strain.   The method according to claim 13, wherein the microorganism belonging to the genus Serratia. Is a Serratia sp. 11E11 strain.   The method according to claim 13, wherein the microorganism belonging to the family Xanthomonadaceae containing the 16S rDNA sequence represented by SEQ ID NO: 3 is the 12A4 strain.   The amino compound represented by the general formula (5) is 1-phenylethylamine, α-ethylbenzylamine, 1-phenyl-2-aminopropane, 1- (3,4-dimethoxyphenyl) -2-aminopropane, 1-aminoindane, 1-aminotetralin, 2-aminotetralin, 1-phenyl-3-aminobutane, 3-aminopyrrolidine, 1-benzyl-3-aminopyrrolidine, 3-aminopiperidine and 1-benzyl-3-aminopiperidine The method according to claim 12, wherein the method is at least one selected from the group consisting of: The compound represented by the general formula (5) is an amino compound represented by the following general formula (7), and the compound represented by the general formula (6) is represented by the following general formula (8). The method according to any one of claims 12 to 16, which is a compound;

(In the formula, Y represents H or an amino-protecting group, p represents an integer of 2 to 3, q represents an integer of 1 to 2, and p> q). The method according to claim 18, wherein the amino-protecting group is a C _1-6 alkyl group, a benzyl group, a formyl group, or a t-butoxycarbonyl group.   The method according to claim 18, wherein the amino compound represented by the general formula (7) is 1-benzyl-3-aminopyrrolidine.   The amino group acceptor is at least one selected from the group consisting of pyruvic acid, glyoxylic acid, 2-ketoglutaric acid, acetophenone, propiophenone, benzylacetone, propionaldehyde, 1-butyraldehyde, benzaldehyde and phenylacetaldehyde. 21. The method according to any one of claims 12 to 20, wherein:

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