US 12,522,810 B2Granted

Transaminase mutant and use thereof

AssigneeAsymchem Laboratories (Tianjin) Co., Ltd.

InventorsHao Hong, Gage James, Yi Xiao, Na Zhang, Xuecheng Jiao, Yulei Ma, Huiyan Mu, Shan Cao

Life of this patent

Estimating the term…

Abstract

Provided are a transaminase mutant and an application thereof. Compared with an amino acid sequence shown in SEQ ID NO:1, an amino acid sequence of the transaminase mutant includes at least one of the following mutation sites: L166, K149, K146, A168, H73, F133, H82, E24, V194, T294, A295, G235 and F236. The mutant of the present invention has the improved catalytic activity for a transammonization reaction of ketone substrates, and is suitable for industrial production of chiral amines.

Claims6

Dependent claims

A transaminase mutant, wherein the transaminase mutant comprises:

a protein having the amino acid sequence of SEQ ID NO:
1 with a mutation of one or more amino acids, wherein the mutation is selected from the group consisting of:
L166I+A168I, L166I+K146R+F133M,
L166I+A168V, L166I+K146R+F133Q,
L166I+A168M, L166I+K146R+F133D,
L166I+K149V, L166I+K149Y+A168I+H73N,
L166I+K149C, L166I+K146R+A168I+K149H,
L166I+K146R+A168I+K149L,
L166I+K146R+A168I+K149I,
L166I+K146R+A168I+K149V,
L166I+K146R+A168I+K149F,
L166I+K146R+A168I+K149M,
L166I+F133S+K149Q, L166I+K146R+A168I+K149Y,
L166I+F133S, L166I+K146R+A168I+H73T,
L166I+K146R, L166I+K146R+A168I+H73S,
L166I+K146M, L166I+K146R+A168I+H73N,
L166I+K146A, L166I+K146R+A168I+H73M,
L166I+K146R+A168I, L166I+K146R+A168I+H73Q,
L166I+K146R+A168S, L166I+K146R+A168I+K149H,
L166I+K146R+A168P, L166I+K146R+A168I+K149H+R145K,
L166I+K146R+A168F, L166I+K146R+A168I+K149H+R145S,
L166I+A168I+K149H, L166I+K146R+A168I+K149H+R145M,
L166I+A168I+K149Y, L166I+K146R+A168I+K149H+R145P
L166I+A168I+H73N, L166I+K146R+A168I+K149H+R145Y,
L166I+A168I+K149H+R145K, L166I+K146R+A168I+K149H+R145K+T294A,
L166I+A168I+K149H+K146P, L166I+K146R+A168I+K149H+R145K+T294E,
L166I+K146R+F133S, L166I+K146R+A168I+K149H+R145K+T294R,
L166I+K146R+F133R, L166I+K146R+A168I+K149H+R145K+T294Y,
L166I+K146R+F133A, L166I+K146R+A168I+K149H+R145K+T294F,
L166I+K146R+F133S+K149Q,
L166I+K146R+F133S+K149R,
L166I+K146R+F133S+K149L,
L166I+K146R+F133S+K149M,
L166I+K146R+F133S+K149Y,
L166I+K146R+F133S+K149Q+H73R,
L166I+K146R+F133S+K149Q+H73Q,
L166I+K146R+F133S+K149Q+H73K,
L166I+K146R+F133S+K149Q+H73T,
L166I+K146R+F133S+K149Q+H73S,
L166I+K146R+F133S+K149Q+H73W,
L166I+K146R+F133S+K149Q+H73R+V194I,
L166I+K146R+F133S+K149Q+H73R+V194S,
L166I+K146R+F133S+K149Q+H73R+V194A,
L166I+K146R+F133S+K149Q+H73R+V194H,
L166I+K146R+F133S+K149Q+H73R+V194N,
L166I+K146R+F133S+K149Q+H73R+V194I+H82S,
L166I+K146R+F133S+K149Q+H73R+V194I+H82Q,
L166I+K146R+F133S+K149Q+H73R+V194I+H82E,
L166I+K146R+F133S+K149Q+H73R+V194I+H82T,
L166I+K146R+F133S+K149Q+H73R+V194I+H82Y,
L166I+K146R+F133S+K149Q+H73R+V194I+E24V,
L166I+K146R+F133S+K149Q+H73R+V194I+E24S,
L166I+K146R+F133S+K149Q+H73R+V194I+E24A,
L166I+K146R+F133S+K149Q+H73R+V194I+E24F,
L166I+K146R+F133S+K149Q+H73R+V194I+L151R,
L166I+K146R+F133S+K149Q+H73R+V194I+L151M,
L166I+K146R+F133S+K149Q+H73R+V194I+L151A,
L166I+K146R+F133S+K149Q+H73R+V194I+E24V+A295S,
L166I+K146R+F133S+K149Q+H73R+V194I+E24V+A295Y,
L166I+K146R+F133S+K149Q+H73R+V194I+E24V+A295I,
L166I+K146R+F133S+K149Q+H73R+V194I+E24V+A295M,
L166I+K146R+F133S+K149Q+H73R+V194I+E24V+A295G,
L166I+K146R+F133S+K149Q+H73R+V194I+E24V+T294A,
L166I+K146R+F133S+K149Q+H73R+V194I+E24V+T294R,
L166I+K146R+F133S+K149Q+H73R+V194I+E24V+T294Q,
L166I+K146R+F133S+K149Q+H73R+V194I+E24V+T294W,
L166I+K146R+F133S+K149Q+H73R+V194I+E24V+T294V,
L166I+K146R+F133S+K149Q+H73R+V194I+E24V+G235Y,
L166I+K146R+F133S+K149Q+H73R+V194I+E24V+G235H,
L166I+K146R+F133S+K149Q+H73R+V194I+E24V+F236I,
L166I+K146R+F133S+K149Q+H73R+V194I+E24V+F236T,
L166I+K146R+F133S+K149Q+H73R+V194I+E24V+F236P,
L166I+K146R+F133S+K149Q+H73R+V194I+E24V+F236M,
L166I+K146R+F133S+K149Q+H73R+V194I+E24V+F236V,
L166I+K146R+F133S+K149Q+H73R+V194I+E24V+F236D,
L166I+K146R+F133S+K149Q+H73R+V194I+E24V+F236S,
L166I+K146P+F133S+K149Q+H73R+V194I+H82S,
L166I+K146R+F133S+K149Q+H73R+V194I+H82S+Y9N,
L166I+K146R+F133S+K149Q+H73R+V194I+H82S+E24V,
L166I+K146R+F133S+K149Q+H73R+V194I+H82S+S132K,
L166I+K146R+F133S+K149Q+H73R+V194I+H82S+T178M,
L166I+K146R+F133S+K149Q+H73R+V194I+H82S+E24A, and
L166I+K146R+F133S+K149Q+H73R+V194I+H82S+E24V.

80 more elements

Dependencies are inferred from the claim text.

Facts

Application: 17/997,419Filed 29 May 2020
Priority: CN 202010353069.829 Apr 2020
Published: US 2023/0235300 A127 Jul 2023
Granted: 13 Jan 2026
Term ends: Not estimatedThe source records don’t support an estimate.

Classification12 codes

C12N 9/1096
Transferases: transferring nitrogenous groups
C12N 15/52
DNA or RNA fragments; Modified forms thereof ; Non-coding nucleic acids having a biological activity: genes encoding for enzymes or proenzymes
C12N 15/70
Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression: vectors or expression systems…
C12P 13/001
Preparation of nitrogen-containing organic compounds: amines; Imines

Family

US 2023/0235300 A1Pre-grant publication · published 27 Jul 2023
US 12,522,810 B2This patent

From the US filing history. Foreign family members aren’t listed.

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Abstract

A transaminase mutant, wherein the transaminase mutant comprises:

a protein having the amino acid sequence of SEQ ID NO:
1 with a mutation of one or more amino acids, wherein the mutation is selected from the group consisting of:
L166I+A168I, L166I+K146R+F133M,
L166I+A168V, L166I+K146R+F133Q,
L166I+A168M, L166I+K146R+F133D,
L166I+K149V, L166I+K149Y+A168I+H73N,
L166I+K149C, L166I+K146R+A168I+K149H,
L166I+K146R+A168I+K149L,
L166I+K146R+A168I+K149I,
L166I+K146R+A168I+K149V,
L166I+K146R+A168I+K149F,
L166I+K146R+A168I+K149M,
L166I+F133S+K149Q, L166I+K146R+A168I+K149Y,
L166I+F133S, L166I+K146R+A168I+H73T,
L166I+K146R, L166I+K146R+A168I+H73S,
L166I+K146M, L166I+K146R+A168I+H73N,
L166I+K146A, L166I+K146R+A168I+H73M,
L166I+K146R+A168I, L166I+K146R+A168I+H73Q,
L166I+K146R+A168S, L166I+K146R+A168I+K149H,
L166I+K146R+A168P, L166I+K146R+A168I+K149H+R145K,
L166I+K146R+A168F, L166I+K146R+A168I+K149H+R145S,
L166I+A168I+K149H, L166I+K146R+A168I+K149H+R145M,
L166I+A168I+K149Y, L166I+K146R+A168I+K149H+R145P
L166I+A168I+H73N, L166I+K146R+A168I+K149H+R145Y,
L166I+A168I+K149H+R145K, L166I+K146R+A168I+K149H+R145K+T294A,
L166I+A168I+K149H+K146P, L166I+K146R+A168I+K149H+R145K+T294E,
L166I+K146R+F133S, L166I+K146R+A168I+K149H+R145K+T294R,
L166I+K146R+F133R, L166I+K146R+A168I+K149H+R145K+T294Y,
L166I+K146R+F133A, L166I+K146R+A168I+K149H+R145K+T294F,
L166I+K146R+F133S+K149Q,
L166I+K146R+F133S+K149R,
L166I+K146R+F133S+K149L,
L166I+K146R+F133S+K149M,
L166I+K146R+F133S+K149Y,
L166I+K146R+F133S+K149Q+H73R,
L166I+K146R+F133S+K149Q+H73Q,
L166I+K146R+F133S+K149Q+H73K,
L166I+K146R+F133S+K149Q+H73T,
L166I+K146R+F133S+K149Q+H73S,
L166I+K146R+F133S+K149Q+H73W,
L166I+K146R+F133S+K149Q+H73R+V194I,
L166I+K146R+F133S+K149Q+H73R+V194S,
L166I+K146R+F133S+K149Q+H73R+V194A,
L166I+K146R+F133S+K149Q+H73R+V194H,
L166I+K146R+F133S+K149Q+H73R+V194N,
L166I+K146R+F133S+K149Q+H73R+V194I+H82S,
L166I+K146R+F133S+K149Q+H73R+V194I+H82Q,
L166I+K146R+F133S+K149Q+H73R+V194I+H82E,
L166I+K146R+F133S+K149Q+H73R+V194I+H82T,
L166I+K146R+F133S+K149Q+H73R+V194I+H82Y,
L166I+K146R+F133S+K149Q+H73R+V194I+E24V,
L166I+K146R+F133S+K149Q+H73R+V194I+E24S,
L166I+K146R+F133S+K149Q+H73R+V194I+E24A,
L166I+K146R+F133S+K149Q+H73R+V194I+E24F,
L166I+K146R+F133S+K149Q+H73R+V194I+L151R,
L166I+K146R+F133S+K149Q+H73R+V194I+L151M,
L166I+K146R+F133S+K149Q+H73R+V194I+L151A,
L166I+K146R+F133S+K149Q+H73R+V194I+E24V+A295S,
L166I+K146R+F133S+K149Q+H73R+V194I+E24V+A295Y,
L166I+K146R+F133S+K149Q+H73R+V194I+E24V+A295I,
L166I+K146R+F133S+K149Q+H73R+V194I+E24V+A295M,
L166I+K146R+F133S+K149Q+H73R+V194I+E24V+A295G,
L166I+K146R+F133S+K149Q+H73R+V194I+E24V+T294A,
L166I+K146R+F133S+K149Q+H73R+V194I+E24V+T294R,
L166I+K146R+F133S+K149Q+H73R+V194I+E24V+T294Q,
L166I+K146R+F133S+K149Q+H73R+V194I+E24V+T294W,
L166I+K146R+F133S+K149Q+H73R+V194I+E24V+T294V,
L166I+K146R+F133S+K149Q+H73R+V194I+E24V+G235Y,
L166I+K146R+F133S+K149Q+H73R+V194I+E24V+G235H,
L166I+K146R+F133S+K149Q+H73R+V194I+E24V+F236I,
L166I+K146R+F133S+K149Q+H73R+V194I+E24V+F236T,
L166I+K146R+F133S+K149Q+H73R+V194I+E24V+F236P,
L166I+K146R+F133S+K149Q+H73R+V194I+E24V+F236M,
L166I+K146R+F133S+K149Q+H73R+V194I+E24V+F236V,
L166I+K146R+F133S+K149Q+H73R+V194I+E24V+F236D,
L166I+K146R+F133S+K149Q+H73R+V194I+E24V+F236S,
L166I+K146P+F133S+K149Q+H73R+V194I+H82S,
L166I+K146R+F133S+K149Q+H73R+V194I+H82S+Y9N,
L166I+K146R+F133S+K149Q+H73R+V194I+H82S+E24V,
L166I+K146R+F133S+K149Q+H73R+V194I+H82S+S132K,
L166I+K146R+F133S+K149Q+H73R+V194I+H82S+T178M,
L166I+K146R+F133S+K149Q+H73R+V194I+H82S+E24A, and
L166I+K146R+F133S+K149Q+H73R+V194I+H82S+E24V.

80 more elements

Facts

Application

17/997,419Filed 29 May 2020

Priority

CN 202010353069.829 Apr 2020

Published

US 2023/0235300 A127 Jul 2023

Granted

13 Jan 2026

Term ends

Not estimatedThe source records don’t support an estimate.

Classification12 codes

C12N 9/1096
Transferases: transferring nitrogenous groups
C12N 15/52
DNA or RNA fragments; Modified forms thereof ; Non-coding nucleic acids having a biological activity: genes encoding for enzymes or proenzymes
C12N 15/70
Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression: vectors or expression systems…
C12P 13/001
Preparation of nitrogen-containing organic compounds: amines; Imines