A kinetic analysis of three modified novel nitroreductases

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 [BLN...

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Bibliographic Details
Published in:Biodegradation (Dordrecht) 2011-04, Vol.22 (2), p.463-474
Main Authors: Gwenin, Christopher D., Kalaji, Maher, Williams, Peter A., Kay, Catherine M.
Format: Article
Language:English
Subjects:
Amino acids
Analysis
Animal genetic engineering
Aquatic Pollution
Bacillus - enzymology
Bacillus - genetics
Bacillus licheniformis
Bacteria
Biodegradation of pollutants
Biological and medical sciences
Biomedical and Life Sciences
Biotechnology
Environment and pollution
Enzymes
Explosives
Fundamental and applied biological sciences. Psychology
Genomics
Geochemistry
Hydrogen-Ion Concentration
Industrial applications and implications. Economical aspects
Kinetics
Life Sciences
Microbiology
Nitroreductases - genetics
Nitroreductases - metabolism
Original Paper
Pets
Polymerase Chain Reaction
Protein Binding
Soil Science & Conservation
Terrestrial Pollution
Waste Management/Waste Technology
Waste Water Technology
Water Management
Water Pollution Control
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Summary: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.
ISSN:0923-9820
1572-9729
DOI:10.1007/s10532-010-9418-0