Engineered transaminase polypeptides for industrial biocatalysis

Patent metadata

Field	Value
Publication number	US-2025250550-A1
Application number	US-202519183477-A
Country	US
Kind code	A1
Filing date	Apr 18, 2025
Priority date	Jun 14, 2017
Publication date	Aug 7, 2025
Grant date	—

Abstract

The present invention provides engineered transaminase polypeptides useful for the synthesis of chiral amine compounds under industrially relevant conditions. The invention also provides polynucleotides encoding the engineered transaminase polypeptides, host cells capable of expressing the engineered transaminases, and methods of using the engineered transaminases for the production of chiral amine compounds.

First claim

We claim:

1 . An engineered transaminase comprising a polypeptide sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to SEQ ID NOS: 4, 18, 20, 30, 48, 66, 70, 88, 110, 112, 120, 138, and/or 156, or a functional fragment thereof, wherein said engineered transaminase comprises at least one substitution or substitution set in said polypeptide sequence at one or more positions selected from 17, 17/259/347, 20, 45/48/78/81/92, 57, 57/319, 85, 86, 157, 158, 161, 203/416, 314, 316, 319, 375/416, 404, 415, 416, 417, 420, 421, 422, and 424, and wherein the amino acid positions of said polypeptide sequence are numbered with reference to SEQ ID NO: 4, 18, 20, 30, 48, 66, 70, 88, 110, 112, 120, 138, or 156.

2 . The engineered transaminase of claim 1 , wherein said polypeptide sequence has at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to SEQ ID NO: 18, wherein said engineered transaminase comprises at least one substitution or substitution set in said polypeptide sequence at one or more positions selected from 5/17/86/157/259/314/316/422, 17/86/157/203/259/314/316/422, 17/86/157/314/316/347/422, 17/86/157/316/422, 85/86, 86/157/314/316/347/416/422, 86/157/416, 86/203/316/416, 86/203/316/422, 86/316/416, 139/221/310/316, 157/316, 195/316, 207/316, 227/422, 280/381/416/445, 314, 314/316, 316, 416, 417, and 422, and wherein the amino acid positions of said polypeptide sequence are numbered with reference to SEQ ID NO: 18.

3 . The engineered transaminase of claim 1 , wherein said polypeptide sequence has at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to SEQ ID NO: 20, wherein said engineered transaminase comprises at least one substitution or substitution set in said polypeptide sequence at one or more positions selected from 17/139/314/416/417/422, 17/195/316/414/416/422, 17/314/316/414/416, 17/314/316/414/416/417/422, 20, 31, 54, 57, 83, 86, 86/194,121, 139/195/227/314/316/416, 139/202/207/227/314/316/416/422, 139/207/227/314/414/416/422, 139/227/310/414/416/417, 139/316/414/416/417/422, 146/157, 164/207/213/227/310/316/416, 166, 178/314/316/416, 190/195/207/213/244/314/414/416/417/422, 194/213/310/314/316/422, 195/227/314/316/416, 195/227/314/416/417, 195/227/314/422, 195/227/414/416, 195/227/414/416/422, 195/227/416/417, 195/316/416/417/422, 207/208/220/241/291/314/316, 226, 227, 227/416, 228, 258, 259, 271/310/316/416/422, 282, 310/314/316/414/416/417/422, 314/316/414/416/422, 314/316/416, 314/316/416/422, 316, 319, 320, 322, and 417, and wherein the amino acid positions of said polypeptide sequence are numbered with reference to SEQ ID NO: 20.

4 . The engineered transaminase of claim 1 , wherein said polypeptide sequence has at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to SEQ ID NO: 30, wherein said engineered transaminase comprises at least one substitution or substitution set in said polypeptide sequence at one or more positions selected from 21, 21/23/53/146/162/395, 21/23/53/146/162/395/432, 21/23/53/162/395/432, 21/23/53/395/432, 21/23/395, 21/53/159/395, 21/53/163/395/432, 23/53/146/159/395/431, 23/53/146/159/395/432, 23/53/146/162/395/432, 23/53/146/163, 23/146/163/432, 23/162, 53/146/159/395, 53/146/162/395, 53/146/162/432, 53/146/163/395, 53/146/395/432, 53/162/163/395/432, 53/162/395, 53/162/395/432, 53/163/395/432, 53/395, 57/86, 57/86/121/146/157, 57/86/121/157/226/259/282/332, 57/86/121/332, 57/86/157/259, 57/86/157/282/332, 57/86/332, 57/121/417/422, 57/146/226/259, 57/146/226/259/282, 57/259, 86, 86/157, 86/226,121, 121/146/157/226/332, 121/208/316/417/422, 121/226/332, 121/316/422, 121/332, 121/416/417/422, 121/417/422, 146, 146/162/395/432, 157, 162/432, 226/282/332, 227, 227/417, 259, 282, 282/332, 316/417/422, 332, 395/432, 416, 416/422, 417, and 422, and wherein the amino acid positions of said polypeptide sequence are numbered with reference to SEQ ID NO: 30.

5 . The engineered transaminase of claim 1 , wherein said polypeptide sequence has at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to SEQ ID NO: 66, wherein said engineered transaminase comprises at least one substitution or substitution set in said polypeptide sequence at one or more positions selected from 107, 141, 161, 174, 245, 281, 293, 432, and 453, and wherein the amino acid positions of said polypeptide sequence are numbered with reference to SEQ ID NO: 66.

6 . The engineered transaminase of claim 1 , wherein said polypeptide sequence has at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to SEQ ID NO: 70, wherein said engineered transaminase comprises at least one substitution or substitution set in said polypeptide sequence at one or more positions selected from 2, 4, 5, 5/38/56/346, 13, 17, 17/31, 19, 31, 57, 74, 79, 86, 121, 155, 161, 161/174/432, 163/167, 164, 190/291, 195, 195/408, 203/207/219, 205/419, 207, 207/235/419, 214/228/251, 222, 223, 226/228/230/231/311, 228, 233, 246, 247/271/293, 259, 282, 284, 309, 327, 346, 359, 362, 375, 404, 408, 419, 420, 422, and 443, and wherein the amino acid positions of said polypeptide sequence are numbered with reference to SEQ ID NO: 70.

7 . The engineered transaminase of claim 1 , wherein said polypeptide sequence has at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to SEQ ID NO: 88, wherein said engineered transaminase comprises at least one substitution or substitution set in said polypeptide sequence at one or more positions selected from 31, 86, 149, 157, 163, 163/164, 191/208/211/286, 192/294, 195/207/208/286, 208/294, 260, 284, 286, 294, 313, 332, 395/396, and 424 and wherein the amino acid positions of said polypeptide sequence are numbered with reference to SEQ ID NO: 88.

8 . The engineered transaminase of claim 1 , wherein said polypeptide sequence has at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to SEQ ID NO: 110, wherein said engineered transaminase comprises at least one substitution or substitution set in said polypeptide sequence at one or more positions selected from 14/31/208/260/286/294/424, 14/31/260/286/294/424, 31, 31/208, 31/208/241/286/294, 31/208/260/286/294, 31/208/260/286/294/313/424, 31/208/260/286/294/424, 31/208/260/286/313/424, 31/208/260/294/313/424, 31/208/260/294/424, 31/208/286/294, 31/208/286/294/313, 31/208/286/294/313/424, 31/208/286/294/424, 31/208/286/313/424, 31/208/286/424, 31/208/294, 31/208/294/313/424, 31/208/294/424, 31/260/286, 31/260/286/294/313, 31/260/286/294/313/424, 31/260/286/313/424, 31/260/286/424, 31/260/294, 31/260/294/313, 31/286/294, 31/286/294/313, 31/286/294/424, 31/294/313/424, 31/294/424, 208/260/286/294/313/424, 208/260/286/294/424, 208/260/286/424, 208/260/294, 208/260/294/313/424, 208/286/294/424, 260/286/294/424, 260/286/424, 286/294/313/424, and 286/313/424, and wherein the amino acid positions of said polypeptide sequence are numbered with reference to SEQ ID NO: 110.

9 . The engineered transaminase of claim 1 , wherein said polypeptide sequence has at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to SEQ ID NO: 112, wherein said engineered transaminase comprises at least one substitution or substitution set in said polypeptide sequence at one or more positions selected from 16/272, 17, 19/86, 19/86/321, 20, 21, 22, 22/255, 22/255/362/421, 22/294, 22/362, 24/255/362, 31, 41, 42/187/294, 66, 66/190, 66/

Assignees

• Codexis Inc

Inventors

• Novick Scott J
• Dellas Nikki
• Alivizo Oscar
• Garcia Ravi David
• Ching Charlene
• Entwistle David

Classifications

• C12Y206/01

  Transaminases (2.6.1) · CPC title

• C12N15/63

  Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression · CPC title

• C12N9/1096 Primary

  transferring nitrogenous groups (2.6) · CPC title