Bioremediation and cloning of a DNA fragment that encodes the nitrite-eliminating enzyme activity

Abstract

Bioremediation tends to be an effective technology used to remove pollutants and involving relatively little capital. This study identified a DNA fragment which encodes the nitrite-eliminating enzyme (NEE) activity. This was achieved by enriching the activity using the fast performance liquid chromatography (FPLC) and protein sequencing. The two internal peptide sequences were obtained: L P G D Y T D Q L L R and S G A I V G L G H A Q V D R. These were used to design the degenerate primers in order to pool a clone from nocardioform genome. A genomic library using Nocardioides simplex FJ2-1A total DNA was constructed. The number of clones required to ensure a 99% probability of a given DNA sequence being contained within a library composed of 10 kb inserts was 2305. The similarity with fatty acid hydroxylase from Nocardioides sp. JS6614 was detected.