Immobilized transaminases and process for making and using immobilized transaminase

Patent metadata

Field	Value
Publication number	US-9587229-B2
Application number	US-201214117410-A
Country	US
Kind code	B2
Filing date	Jun 18, 2012
Priority date	Jun 24, 2011
Publication date	Mar 7, 2017
Grant date	Mar 7, 2017

Abstract

The invention is directed to immobilized transaminases and methods of making and using them.

First claim

What is claimed is:

1. An immobilized transaminase comprising:
a recombinant transaminase that is SEQ ID NO: 110; and
a resin, wherein the recombinant transaminase is attached to the resin, wherein the resin comprises polymethacrylate with epoxide functional groups, polymethacrylate with amino epoxide functional groups, styrene/DVB copolymer or polymethacrylate with octadecyl functional groups and wherein the immobilized transaminase loses less than 10% activity per hour in a solvent system that comprises at least 90% of organic solvents.

2. An immobilized transaminase of claim 1 wherein the immobilized transaminase loses less than 10% activity per hour in a solvent system that comprises at least 95% of organic solvents.

3. A process for making (2R)-4-oxo-4-[3-(trifluoromethyl)-5,6-dihydro[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl]-1-(2,4,5-trifluorophenyl)butan-2-amine comprising the steps of:
1) dissolving 4-oxo-4-[3-(trifluoromethyl)-5,6-dihydro[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl]-1-(2,4,5-trifluorophenyl)butan-2-one in an organic solvent;
2) contacting 4-oxo-4-[3-(trifluoromethyl)-5,6-dihydro[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl]-1-(2,4,5-trifluorophenyl)butan-2-one with an immobilized transaminase of claim 1 in presence of an amino group.

4. A process of making an immobilized transaminase comprising:
1) incubating a solution of transaminase with a resin and an enzyme solution to form an immobilized transaminase, wherein that transaminase is SEQ ID NO: 110 and the resin comprises polymethacrylate with epoxide functional groups, polymethacrylate with amino epoxide functional groups, styrene/DVB copolymer or polymethacrylate with octadecyl functional groups;
2) filtering and rinsing the immobilized transaminase; and
3) drying the immobilized transaminase,
wherein the immobilized transaminase loses less than 10% activity per hour in a solvent system that comprises at least 90% of organic solvents.

Assignees

• Truppo Matthew D
• Janey Jacob M
• Hughes Gregory
• Merck Sharp & Dohme

Inventors

• Truppo Matthew D
• Janey Jacob M
• Hughes Gregory

Classifications

• C12P13/001

  Amines; Imines · CPC title

• C12N9/1096 Primary

  transferring nitrogenous groups (2.6) · CPC title

• Y02P20/52

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

• C12P17/182

  Heterocyclic compounds containing nitrogen atoms as the only ring heteroatoms in the condensed system (alloxazine or isoalloxazine, e.g. riboflavine C12P25/00) · CPC title

• C12N11/08

  the carrier being a synthetic polymer · CPC title