Biocatalysts and methods for the synthesis of substituted lactams

Patent metadata

Field	Value
Publication number	US-10633640-B2
Application number	US-201715449677-A
Country	US
Kind code	B2
Filing date	Mar 3, 2017
Priority date	Sep 8, 2011
Publication date	Apr 28, 2020
Grant date	Apr 28, 2020

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 90% sequence identity to SEQ ID NO: 4, wherein the amino acid sequence comprises residue differences at position X136 and position X323 as compared to SEQ ID NO: 4.

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, X193G, X192H, X 192K, X192N, X193Q, 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, X223V, and X284A;
(d) X192A, X215H and X311T;
(e) X62N, X124F and X126A;
(f) X124W, X126A, X192A and X284A; and
(g) X124W, X126A, X152R/X152I/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, X282V; X284A; X284P; X284S; X284T; and X284V.

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

6. A vector comprising the polynucleotide of claim 5 .

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

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

• C07D211/76

  attached in position 2 or 6 · CPC title

• C12P13/005

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

• C12P17/12

  containing a six-membered hetero ring · CPC title

• C12Y206/01

  Transaminases (2.6.1) · CPC title

• C12N9/1096 Primary

  transferring nitrogenous groups (2.6) · CPC title