Engineered imine reductases and methods for the reductive amination of ketone and amine compounds

Patent metadata

Field	Value
Publication number	US-10494656-B2
Application number	US-201916391036-A
Country	US
Kind code	B2
Filing date	Apr 22, 2019
Priority date	May 11, 2012
Publication date	Dec 3, 2019
Grant date	Dec 3, 2019

Abstract

The present disclosure provides engineered polypeptides having imine reductase activity, polynucleotides encoding the engineered imine reductases, host cells capable of expressing the engineered imine reductases, and methods of using these engineered polypeptides with a range of ketone and amine substrate compounds to prepare secondary and tertiary amine product compounds.

First claim

What is claimed is:

1. An engineered polypeptide having imine reductase activity, wherein said polypeptide has at least 90% sequence identity to SEQ ID NO:2, and comprises a substitution at a position corresponding to position 292 of SEQ ID NO:2, wherein the amino acid at position 292 has been replaced with a proline or with an aromatic, non-polar, aliphatic, or polar residue.

2. The engineered polypeptide having imine reductase activity of claim 1 , wherein said polypeptide has at least 90% sequence identity to SEQ ID NO:2, wherein the amino acid at the position corresponding to position 292 of SEQ ID NO:2 has been replaced with valine.

3. The engineered polypeptide of claim 1 , wherein said polypeptide is capable of converting substrate compound (1a) pyruvate,

and substrate compound (2b) butylamine

to product compound (3b), N-2-(butylamino)propanoic acid,

under suitable reaction conditions.

4. The engineered polypeptide of claim 1 , wherein said polypeptide is capable of converting substrate compound (1b) cyclohexanone,

and substrate compound (2a) L-norvaline

to product compound (3c), (S)-2-(cyclohexylamino)pentanoic acid,

under suitable reaction conditions.

5. The engineered polypeptide of claim 1 , wherein said polypeptide is capable of converting substrate compound (1b) cyclohexanone,

and substrate compound (2b) butylamine

to product compound (3d), N-butylcyclohexanamine,

under suitable reaction conditions.

6. The engineered polypeptide of claim 1 , wherein said polypeptide is capable of converting substrate compound (1i),

and substrate compound (2b)

to product compound (3n),

under suitable reaction conditions.

7. The engineered polypeptide of claim 1 , wherein said polypeptide is capable of converting substrate compound (1j),

and substrate compound (2b)

to product compound (3o),

under suitable reaction conditions.

Assignees

• Codexis Inc

Inventors

• Chen Haibin
• Collier Steven J
• Nazor Jovana
• Sukumaran Joly
• Smith Derek
• Moore Jeffrey C
• Hughes Gregory
• Janey Jacob
• Huisman Gjalt W
• Novick Scott J
• Agard Nicholas J
• Alvizo Oscar
• Cope Gregory A
• Yeo Wan Lin
• Minor Stefanie Ng

Classifications

• C12Y105/01024

  N5-(Carboxyethyl)ornithine synthase (1.5.1.24) · CPC title

• C12P13/02 Primary

  Amides, e.g. chloramphenicol {or polyamides; Imides or polyimides; Urethanes, i.e. compounds comprising N-C=O structural element or polyurethanes (peptides C12P21/00 or C07K)} · CPC title

• C12P17/10

  Nitrogen as only ring hetero atom · CPC title

• Y02P20/52

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

• C12P13/001

  Amines; Imines · CPC title