Biocatalysts and methods for the synthesis of substituted lactams

Patent metadata

Field	Value
Publication number	US-11597918-B2
Application number	US-202017087484-A
Country	US
Kind code	B2
Filing date	Nov 2, 2020
Priority date	Sep 8, 2011
Publication date	Mar 7, 2023
Grant date	Mar 7, 2023

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 polynucleotide encoding 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 two amino acid residue differences as compared to SEQ ID NO:4 at residue positions X140 and X323.

2. The engineered polynucleotide encoding an engineered polypeptide of claim 1 , in which X323 is selected from C, E, N, and R.

3. The engineered polynucleotide encoding an engineered polypeptide of claim 2 , 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, X193G, X192H, X192K, X192N, X193Q, X192R, and X192S.

4. The engineered polynucleotide encoding an engineered polypeptide of claim 2 , wherein the amino acid sequence further comprises a combination of residue differences selected from:
(a) X124W and X327L;
(b) X209M and X300G;
(c) X122F, X223V and X284A;
(d) XI92A, X215H and X311T;
(e) X62N, X124F, X126A and X136L;
(f) X124W, X126A, X136L, X192A and X284A; and
(g) X124W, X126A, X136L, X152R/X152L/X1521, and X192A.

5. The engineered polynucleotide encoding an engineered 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; X143V; X150L; X155I; X155L; X159G; X160L; X176C; X199L; X199V; X209F; X209M; X209Q; X209V; X223S; X223T; X223V; X227I; X282L, X282V; X284A; X284P; X284S; X284T; and X284V.

6. A vector comprising the engineered polynucleotide of claim 1 .

7. A host cell comprising the vector of claim 6 .

8. The vector of claim 6 , further comprising at least one control sequence.

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.

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

• C12N9/1096 Primary

  transferring nitrogenous groups (2.6) · CPC title

• C12Y206/01

  Transaminases (2.6.1) · CPC title

• C12P13/005

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

• C07D211/76

  attached in position 2 or 6 · CPC title

• C12P17/12

  containing a six-membered hetero ring · CPC title