Biocatalysts and methods for synthesizing derivatives of tryptamine and tryptamine analogs

Patent metadata

Field	Value
Publication number	US-10544402-B2
Application number	US-201715623698-A
Country	US
Kind code	B2
Filing date	Jun 15, 2017
Priority date	Mar 23, 2012
Publication date	Jan 28, 2020
Grant date	Jan 28, 2020

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 80% sequence identity to SEQ ID NO:2 and as compared to SEQ ID NO: 2 residue difference at residue X424V, X448E and X451D, and wherein said engineered polypeptide comprises an amino acid sequence having at least 90% sequence identity to SEQ ID NO:4, and wherein said engineered polypeptide comprises an amino acid sequence having one or more further residue differences as compared to SEQ ID NO:2 at residue positions selected from: X14, X26, X31, X33, X41, X47, X57, X70, X86, X88, X107, X132, X148, X168, X173, X203, X250, X284, X314, X315, X324, X346, X395, X398, X400, X417, X419 and, X423.

2. The engineered polypeptide of claim 1 , wherein the amino acid sequence further comprises one or more residue differences selected from X383A and X420N.

3. The engineered polypeptide of claim 1 , wherein the amino acid sequence further comprises at least one or more residue differences selected from X14V, X26R, X31S/D, X86D, X284A, X315G, X398L/V/W, and X400G.

4. The engineered polypeptide of claim 1 , wherein the amino acid sequence comprises a combination of residue differences selected from:
X86D and X400G,
X57F/Y,
X57F/Y and X398L/V/W,
X31S, X57F/Y, X315G, X346L, and X398L/V/W
X14V, X284A, and X424V,
X14V, X26R, and X88L,
X57F, X168K, X314N, X315G, X346L, and X398V,
X14V, X173A, X400G, and X420N,
X14V, and X284A,
X14V, X26R, X284A, and X400G, and
X14V, X33T, and X57F.

5. 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

• C12Y206/01

  Transaminases (2.6.1) · CPC title

• C12Y206/01018

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

• C12P17/10

  Nitrogen as only ring hetero atom · CPC title

• C12N9/1096 Primary

  transferring nitrogenous groups (2.6) · CPC title

• Y02P20/52

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