Biocatalysts and methods for synthesizing derivatives of tryptamine and tryptamine analogs

Patent metadata

Field	Value
Publication number	US-11098291-B2
Application number	US-201916707724-A
Country	US
Kind code	B2
Filing date	Dec 9, 2019
Priority date	Mar 23, 2012
Publication date	Aug 24, 2021
Grant date	Aug 24, 2021

Abstract

The present disclosure provides engineered transaminase polypeptides for the production of amines, polynucleotides encoding the engineered transaminases, host cells capable of expressing the engineered transaminases, and methods of using the engineered transaminases to prepare compounds useful in the production of active pharmaceutical agents.

First claim

What is claimed is:

1. An engineered polypeptide having transaminase activity, comprising an amino acid sequence having at least 98% sequence identity to the amino acid sequence of SEQ ID NO: 134 and residues difference as compared to SEQ ID NO: 2 at residue positions X14, X391 and X427.

2. The engineered polypeptide of claim 1 , further comprises one or more further residue differences as compared to SEQ ID NO:2 at residue positions selected from: X26, X31, X33, X57, X86, X88, X113, X148, X163, X168, X173, X203, X250, X284, X314, X315, X324, X346, X398, X400, X417, and X448.

3. The engineered polypeptide of claim 1 , wherein the residue difference at the residue position X14 is X14V.

4. The engineered polypeptide of claim 1 , wherein the amino acid sequence further comprises at least one or more residue differences selected from X26R, X31S/D, X86D, X163I/L/R/V, X315G, X398L/V/W, and X400G.

5. The engineered polypeptide of claim 1 , wherein the amino acid sequence further comprises a combination of residue differences selected from:
X163I/L/R/V,
X86D and X400G,
X57F/Y and X163I/L/R/V,
X57F/Y and X398L/V/W,
X163I/L/R/V, and X424V,
X31S, X57F/Y, X163I/L/R/V, X315G, X346L, and X398L/V/W,
X26R, X163L, and X400G,
X26R and X88L,
X57F, X163L, X168K, X314N, X315G, X346L, and X398V,
X163L, X173A, X400G, and X420N,
X26R, X163L, and X400G, and
X33T, X57F, and X163L.

6. The engineered polypeptide of claim 1 , wherein said polypeptide has at least 1.2 fold increased stability as compared to the polypeptide of SEQ ID NO:4.

7. The engineered polypeptide of claim 1 , wherein the amino acid sequence comprises SEQ ID NO: 134.

8. The engineered polypeptide of claim 1 , wherein said polypeptide is immobilized on a solid support.

Assignees

• Codexis Inc

Inventors

• Nazor Jovana
• Smith Derek J
• Crowe Michael A
• Song Shiwei
• Collier Steven J

Classifications

• C12N9/1096 Primary

  transferring nitrogenous groups (2.6) · CPC title

• C12Y206/01

  Transaminases (2.6.1) · CPC title

• C12P17/10

  Nitrogen as only ring hetero atom · CPC title

• C12Y206/01018

  Beta-alanine-pyruvate transaminase (2.6.1.18) · CPC title

• Y02P20/52

  using catalysts, e.g. selective catalysts · CPC title