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

Patent metadata

Field	Value
Publication number	US-9932613-B2
Application number	US-201715792446-A
Country	US
Kind code	B2
Filing date	Oct 24, 2017
Priority date	May 11, 2012
Publication date	Apr 3, 2018
Grant date	Apr 3, 2018

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 polynucleotide encoding an engineered polypeptide having imine reductase activity, wherein said polypeptide has at least 90% sequence identity to the polypeptide of SEQ ID NO: 2, and comprises a substitution at a position corresponding to position 280 of SEQ ID NO: 2, wherein the amino acid at position 280 has been replaced with an aliphatic or non-polar residue.

2. The engineered polynucleotide of claim 1 , wherein said polypeptide has at least 90% sequence identity to the polypeptide of SEQ ID NO: 2, wherein the amino acid at the position corresponding to position 280 of SEQ ID NO: 2 has been replaced with leucine.

3. The engineered polynucleotide 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 polynucleotide 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 polynucleotide 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 polynucleotide 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 polynucleotide 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

• C12P13/001

  Amines; Imines · CPC title

• C12P17/185

  Heterocyclic compounds containing sulfur atoms as ring hetero atoms in the condensed system (cepam nucleus C12P35/00; penam nucleus C12P37/00) · 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

• C12P13/06

  Alanine; Leucine; Isoleucine; Serine; Homoserine · CPC title

• C12Y105/01023

  Tauropine dehydrogenase (1.5.1.23) · CPC title