公开(公告)号：US11591577B2

公开(公告)日：2023-02-28

申请号：US17177823

申请日：2021-02-17

Applicant: API CORPORATION

Inventor： Ryoma Miyake ,  Hiroshi Kawabata

IPC: C12N9/02 ,  C12P17/12 ,  C12N9/10 ,  C12N9/06 ,  C12N15/09

Abstract: A novel method of producing high-purity hydroxy-L-pipecolic acids in an efficient and inexpensive manner while suppressing the production of hydroxy-L-proline is provided. The method includes allowing an L-pipecolic acid hydroxylase, a microorganism or cell having the ability to produce the enzyme, a processed product of the microorganism or cell, and/or a culture liquid comprising the enzyme and obtained by culturing the microorganism or cell, to act on L-pipecolic acid as a substrate in the presence of 2-oxoglutaric acid and ferrous ion, wherein the L-pipecolic acid hydroxylase has the properties: (1) the enzyme can act on L-pipecolic acid in the presence of 2-oxoglutaric acid and ferrous ion to add a hydroxy group to the carbon atom at positions 3, 4, and/or 5 of L-pipecolic acid; and (2) the enzyme has a catalytic efficiency (kcat/Km) with L-proline that is equal to or less than 7 times the catalytic efficiency (kcat/Km) with L-pipecolic acid.

公开(公告)号：US11136560B2

公开(公告)日：2021-10-05

申请号：US15947220

申请日：2018-04-06

Applicant: API CORPORATION

Inventor： Makoto Hibi ,  Jun Ogawa ,  Ryoma Miyake ,  Hiroshi Kawabata

IPC: C12N9/02 ,  C12N1/20 ,  C12P13/04 ,  C12P17/12 ,  C12P17/10

Abstract: The present invention provides a pipecolic acid 4-hydroxylase protein exemplified by the following (A), (B), and (C), having activity to react with L-pipecolic acid in the presence of 2-oxoglutaric acid and iron(II) ions to produce trans-4-hydroxy-L-pipecolic acid, and a method for producing 4-hydroxy amino acid, which method comprises reacting the pipecolic acid 4-hydroxylase protein, cells containing the protein, a treated product of the cells, and/or a culture liquid obtained by culturing the cells, with α-amino acid to produce 4-hydroxy amino acid: (A) a polypeptide comprising the amino acid sequence represented by SEQ ID NO:2, 4, 6, 8, 10, 12, 16, or 18; (B) a polypeptide comprising the amino acid sequence represented by SEQ ID NO:2, 4, 6, 8, 10, 12, 16, or 18 except that one or several amino acids are deleted, substituted, and/or added, and having pipecolic acid 4-hydroxylase activity; and (C) a polypeptide having an amino acid sequence that is not less than 80% identical to the amino acid sequence represented by SEQ ID NO:2, 4, 6, 8, 10, 12, 16, or 18, and having pipecolic acid 4-hydroxylase activity.

公开(公告)号：US09512452B2

公开(公告)日：2016-12-06

申请号：US14431193

申请日：2014-02-18

Applicant: API Corporation

Inventor： Kuniki Kino ,  Ryotaro Hara ,  Ryoma Miyake ,  Hiroshi Kawabata

IPC: C12P13/04 ,  C12P13/08 ,  C12P17/12 ,  C12N9/02

CPC classification number: C12P13/08 ,  C12N9/0071 ,  C12P13/04 ,  C12P17/12 ,  C12Y114/11 ,  C12Y114/11004 ,  Y02P20/52

Abstract: The present invention aims to provide a method for efficiently producing hydroxy-L-lysine. The present invention provides a method for producing hydroxy-L-lysine, the method comprising allowing 2-oxoglutarate-dependent L-lysine hydroxylase, a cell containing 2-oxoglutarate-dependent L-lysine hydroxylase, a processed product of the cell, and/or a culture broth obtained by culturing the cell, to act on L-lysine to produce hydroxy-L-lysine represented by the following General Formula (I) (wherein each of R1, R2 and R3 represents a hydrogen atom or hydroxyl group, with the proviso that at least one of R1, R2 and R3 represents a hydroxyl group).

公开(公告)号：US20140127762A1

公开(公告)日：2014-05-08

申请号：US14122365

申请日：2012-03-29

Applicant: API CORPORATION

Inventor： Hisatoshi Uehara ,  Ryoma Miyake ,  Keisuke Bando ,  Hiroshi Kawabata ,  Tomoko Maeda

IPC: C07D207/16 ,  C07D207/20

CPC classification number: C07D207/16 ,  C07D207/20 ,  C12P13/24 ,  C12P41/007

Abstract: The present invention aims to provide an industrial method practically suitable for producing optically active α-substituted prolines from an acyclic ketone compound by a small number of steps under mild conditions. The present invention relates to a production method of an optically active α-substituted proline (4) and/or an optically active α-substituted prolinamide (5), including (a) reacting an acyclic ketone compound (1) with at least one selected from ammonia, an ammonium salt, primary amine and a salt of primary amine, and a cyanating agent to give a cyclic nitrogen-containing compound (2), (b) hydrating the cyclic nitrogen-containing compound (2) to give an α-substituted prolinamide (3), and (c) resolving the α-substituted prolinamide (3) by one or more of (d) enzymatical hydrolysis, (e) resolution by diastereomeric salt formation, and (f) separation by column chromatography.

Abstract translation: 本发明旨在提供一种实际上适用于在温和条件下通过少量步骤从无环酮化合物制备光学活性α-取代的脯氨酸的工业方法。 本发明涉及光学活性α-取代脯氨酸（4）和/或光学活性α-取代脯氨酰胺（5）的制备方法，包括（a）使无环酮化合物（1）与至少一种选择的 由氨，铵盐，伯胺和伯胺的盐和氰化剂得到环状含氮化合物（2），（b）使环状含氮化合物（2）水合，得到α- 取代的脯氨酰胺（3），和（c）通过（d）酶水解，（e）通过非对映异构体盐形成的拆分，和（f）通过柱色谱法分离的α-取代的脯氨酰胺（3）