Cloning, Expression, and Bioinformatics Analysis of Heavy Metal Resistance Gene afe_1862 from Acidithiobacillus ferrooxidans L1 in Escherichia coli

Biol Trace Elem Res. 2019 May;189(1):291-300. doi: 10.1007/s12011-018-1462-3. Epub 2018 Aug 16.

Abstract

Molecular studies of copper and cadmium resistances in acidophilic bacteria are significant in biomining. In this study, afe_1862, which encodes a heavy metal-binding protein in Acidithiobacillus ferrooxidans L1, was amplified using PCR, cloned into the pET32a plasmid, and sequenced. Following SDS-PAGE analysis, optimization of the expression conditions and heterologous overexpression of afe_1862 in Escherichia coli BL21 in the presence of Cu2+ and Cd2+ were studied as well. The results indicated that AFE_1862 has higher resistance to Cu2+ than Cd2+. Bioinformatics analysis illustrated that AFE_1862 has a conserved HMA domain containing heavy metal-binding sites, which may play a role in transporting or detoxifying heavy metals.

Keywords: Acidithiobacillus ferrooxidans; Bioinformatics; Expression; Metal resistance; afe_1862.

MeSH terms

  • Acidithiobacillus / drug effects
  • Acidithiobacillus / genetics
  • Acidithiobacillus / metabolism*
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism*
  • Bacterial Proteins / toxicity*
  • Computational Biology
  • Escherichia coli / drug effects
  • Escherichia coli / genetics
  • Escherichia coli / metabolism*
  • Metals, Heavy / toxicity*

Substances

  • Bacterial Proteins
  • Metals, Heavy