A kinetic analysis of three modified novel nitroreductases

Christopher D. Gwenin*, Maher Kalaji, Peter A. Williams, Catherine M. Kay

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

9 Citations (Scopus)

Abstract

A kinetic comparison between three nitroreductase enzymes isolated from the genome of Bacillus licheniformis ATCC 14580 for prospective use as immobilised enzymes for explosives detection has been conducted. The genes encoding the three enzymes (yfkO [BLNfnB] encoding an NfsB-like enzyme; nfrA [BLNfrA1] and ycnD [BLNfrA2] encoding PnrA-like enzymes) have been PCR amplified from the native genome and cloned into pET-28a(+) and a modified cysteine (6)-tagged pET-28a(+) and subsequently over-expressed, purified, and biochemically characterised. The previously uncharacterised nitroreductases exhibited activity against a wide range of explosives, including cyclic nitramines. Amino acid alignments and overall structural comparisons with other nitroreductase family members suggest that the B. licheniformis enzymes are members of the NfsA-Frp/NfsB-FRase I family group. Despite the overall low amino acid identity, regions for flavin mononucleotide binding and active site residues were highly conserved.

Original languageEnglish
Pages (from-to)463-474
Number of pages12
JournalBiodegradation
Volume22
Issue number2
Early online date23 Sep 2010
DOIs
Publication statusPublished - 1 Apr 2011
Externally publishedYes

Keywords

  • Bacillus licheniformis
  • Nitroreductase
  • Explosives

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