Characterization of a novel ω-transaminase from a Triassic salt mine metagenome

Stephen A. Kelly, Timofey Skvortsov, Damian Magill, Derek J. Quinn, John W. McGrath, Christopher C.R. Allen, Thomas S. Moody, Brendan F. Gilmore*

*Corresponding author for this work

Research output: Contribution to journalArticle

3 Citations (Scopus)
145 Downloads (Pure)

Abstract

Chiral amines are valuable building blocks for the pharmaceutical industry, and are increasingly synthesized by transaminase-mediated (TAm) synthesis. Currently available TAms, primarily isolated from the genomes of cultured mesophilic bacteria, often suffer from a number of drawbacks, including poor substrate range and an inability to tolerate the harsh conditions often demanded by industrial processes. These characteristics have, in part, driven the search for novel biocatalysts from both metagenomic sources and extreme environments. Herein, we report the isolation and characterization of an ω-TAm from a metagenome of a Triassic salt mine in Kilroot, N. Ireland, an extremely hypersaline environment formed circa 220–250 mya. The gene sequence was identified based on homology with existing bacterial TAms, synthesized within a pET28a(+) plasmid and expressed in E. coli BL21 DE3 cells. The resultant 49 kDa protein accepted (S)-methylbenzylamine (MBA) as amino donor and had a specific activity of 0.54 U/mg using α-ketoglutarate (ΑKG) as substrate. Molecular modeling and substrate docking indicated the presence of key residues, conserved in a number of other TAms. Despite the hypersaline environment from which it was isolated, the enzyme displayed low halotolerance, highlighting that not all biocatalysts will demonstrate the extreme characteristics associated with their source environment. This study does however reinforce the viability of mining metagenomic datasets as a means of discovering novel and functional biocatalysts, and adds to a currently scant list of such examples in the field of TAms.

Original languageEnglish
Pages (from-to)2936-2942
Number of pages7
JournalBiochemical and Biophysical Research Communications
Volume503
Issue number4
Early online date14 Aug 2018
DOIs
Publication statusPublished - 01 Sep 2018

Keywords

  • Biocatalysis
  • Chiral amine
  • Halophile
  • Hypersaline
  • Metagenome
  • Transaminase

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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