Covalently immobilized Trp60Cys mutant of Ω-transaminase from Chromobacterium violaceum for kinetic resolution of racemic amines in batch and continuous-flow modes

Emese Abaházi, Péter Sátorhelyi, Balázs Erdélyi, B. Vértessy, Henrik Land, Csaba Paizs, Per Berglund, L. Poppe

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

Covalent immobilization of an engineered ω-transaminase mutant Trp60Cys from Chromobacterium violaceum (CvTAW60C) was performed on bisepoxide-activated aminoalkyl resins. Activity of the various CvTAW60C preparations was evaluated in kinetic resolution of four racemic amines (rac-1a–d). The most active EA-G-CvTAW60C preparation (CvTAW60C attached to polymeric resin with ethylamine function activated with glycerol diglycidyl ether—EA-G) could perform the kinetic resolution of racemic 4-phenylbutan-2-amine (rac-1a) over 49% conversion up to 19 consecutive reaction cycles or in media containing up to 50% v/v DMSO as cosolvent in batch mode reactions. The immobilization process of CvTAW60C onto the EA-G resin filled in stainless steel bioreactors was also tested in flow-through mode. Kinetic resolution of three racemic amines containing aromatic moieties (rac-1a-c) was performed in continuous-flow mode resulting in easy-to-separate mixture of the corresponding ketone (2a–c) and the non-converted (R)-amine in high enantiopurity (ee(R)-1a-c ≥ 96%).

Original languageEnglish
Pages (from-to)270-278
Number of pages9
JournalBiochemical Engineering Journal
Volume132
DOIs
Publication statusPublished - Apr 15 2018

Fingerprint

Chromobacterium
Transaminases
Amines
Kinetics
Resins
Immobilization
Stainless Steel
Bioreactors
Dimethyl Sulfoxide
Ketones
Glycerol
Stainless steel

Keywords

  • Amine
  • Continuous-flow biocatalysis
  • Covalent immobilization
  • Kinetic resolution
  • Transaminase

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Environmental Engineering
  • Biomedical Engineering

Cite this

Covalently immobilized Trp60Cys mutant of Ω-transaminase from Chromobacterium violaceum for kinetic resolution of racemic amines in batch and continuous-flow modes. / Abaházi, Emese; Sátorhelyi, Péter; Erdélyi, Balázs; Vértessy, B.; Land, Henrik; Paizs, Csaba; Berglund, Per; Poppe, L.

In: Biochemical Engineering Journal, Vol. 132, 15.04.2018, p. 270-278.

Research output: Contribution to journalArticle

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