Biocatalysts and methods for the synthesis of substituted lactams

Patent metadata

Field	Value
Publication number	US-12163162-B2
Application number	US-202318164522-A
Country	US
Kind code	B2
Filing date	Feb 3, 2023
Priority date	Sep 8, 2011
Publication date	Dec 10, 2024
Grant date	Dec 10, 2024

Abstract

The present disclosure relates to transaminase polypeptides capable of aminating a dicarbonyl substrate, and polynucleotides, vectors, host cells, and methods of making and using the transaminase polypeptides.

First claim

What is claimed is:

1. An engineered transaminase polypeptide comprising an amino acid sequence having at least 80% sequence identity to reference sequence SEQ ID NO: 4 and at least an amino acid residue difference as compared to SEQ ID NO:4 at residue position X136 is L, and wherein the amino acid residue at residue position X284 is selected from A, G, P, S, T, or V.

2. The engineered transaminase polypeptide of claim 1 , wherein the amino acid sequence further comprises one or more residue differences as compared to SEQ ID NO: 4 selected from: X42G; X54P; X54R; X69A; X69G; X69T; X122M; X124F; X124L; X124N; X124R; X126A; X126T; X150S; X152C; X152G; X152I; X152L; X152S; X156A; X156S; X156T; X157L; X165N, X192A, X192G, X192H, X192K, X192N, X192Q, X192R, and X192S.

3. The engineered transaminase polypeptide of claim 1 , wherein the amino acid sequence further comprises a combination of residue differences selected from:
(a) X124W and X327L;
(b) X209M and X300G;
(c) X122F and X223V;
(d) XI92A, X215H and X311T;
(e) X62N, X124F, and X126A;
(f) X124W, X126A, and X192A; and
(g) X124W, X126A, X152R/X152L/X152I, and X192A.

4. The engineered transaminase polypeptide of claim 1 , in which the amino acid sequence further comprises at least one or more residue differences as compared to SEQ ID NO: 4 selected from: X54A; X54L; X56D; X56E; X61G; X61W; X62A; X62L; X62N; X62S; X64C; X68I; X122F, X122W; X122Y; X124K; X124M; X124S; X124W; X126C; X126I; X139E; X140K; X143V; X150L; X155I; X155L; X159G; X160L; X176C; X199L; X199V; X209F; X209M; X209Q; X209V; X223S; X223T; X223V; X227I; X282L, and X282V.

5. The engineered transaminase polypeptide of claim 1 , wherein said polypeptide is purified.

6. A composition comprising the engineered transaminase polypeptide of claim 1 .

7. An engineered polynucleotide encoding the engineered transaminase polypeptide of claim 1 .

8. A vector comprising the engineered polynucleotide of claim 7 .

9. A host cell comprising the vector of claim 8 .

10. The host cell of claim 9 , wherein said host cell is an Escherichia coli cell.

11. The vector of claim 8 , further comprising at least one control sequence.

12. A host cell comprising the vector of claim 11 .

13. The host cell of claim 12 , wherein said host cell is an Escherichia coli cell.

Assignees

• Codexis Inc

Inventors

• Cabirol Fabien Louis
• Chen Haibin
• Gohel Anupam
• Salim Paulina Elena
• Smith Derek J
• Janey Jacob
• Kosjek Birgit
• Tang Weng Lin
• Hsieh Helen
• Pham Son Q

Classifications

• C12P17/12

  containing a six-membered hetero ring · CPC title

• C12P13/005

  Amino acids other than alpha- or beta amino acids, e.g. gamma amino acids · CPC title

• C12P13/001

  Amines; Imines · CPC title

• C12Y206/01

  Transaminases (2.6.1) · CPC title

• C07D211/76

  attached in position 2 or 6 · CPC title