Immobilization of Escherichia coli containing ω‐transaminase activity in LentiKats®

Max Cárdenas‐Fernández, Watson Lima Afonso Neto, Carmen López, Gregorio Álvaro, Pär Tufvesson, John M. Woodley

Research output: Contribution to journalJournal articleResearchpeer-review

Abstract

Whole Escherichia coli cells overexpressing ω‐transaminase (ω‐TA) and immobilized cells entrapped in LentiKats® were used as biocatalysts in the asymmetric synthesis of the aromatic chiral amines 1‐phenylethylamine (PEA) and 3‐amino‐1‐phenylbutane (APB). Whole cells were permeabilized with different concentrations of cetrimonium bromide (CTAB) and ethanol; the best results were obtained with CTAB 0.1% which resulted in an increase in reωaction rate by 40% compared to the whole cells. The synthesis of PEA was carried out using isopropyl amine (IPA) and L‐alanine (Ala) as amino donors. Using whole cell biocatalysis, the reaction with IPA was one order of magnitude faster than with Ala. No reaction was detected when permeabilized E. coli cells containing ω‐TA were employed using Ala as the amino donor. Additionally, the synthesis of APB from 4‐phenyl‐2‐butanone and IPA was studied. Whole and permeabilized cells containing ω‐TA and their immobilized LentiKats® counterparts showed similar initial reactions rates and yields in the reaction systems, indicating 100% of immobilization efficiency (observed activity/activity immobilized) and absence of diffusional limitations (due to the immobilization). Immobilization of whole and permeabilized cells containing ω‐TA in LentiKats® allowed improved stability as the biocatalyst was shown to be efficiently reused for five reaction cycles, retaining around 80% of original activity. © 2012 American Institute of Chemical Engineers Biotechnol. Prog., 2012
Original languageEnglish
JournalBiotechnology Progress
Volume28
Issue number3
Pages (from-to)693-698
ISSN8756-7938
DOIs
Publication statusPublished - 2012

Keywords

  • Whole cell biocatalysis
  • ω-transaminase
  • Asymmetric synthesis
  • Chiral amines
  • Cell permeabilization
  • Immobilization by entrapment
  • LentiKats®

Cite this

Cárdenas‐Fernández, M., Lima Afonso Neto, W., López, C., Álvaro, G., Tufvesson, P., & Woodley, J. M. (2012). Immobilization of Escherichia coli containing ω‐transaminase activity in LentiKats®. Biotechnology Progress, 28(3), 693-698. https://doi.org/10.1002/btpr.1538