Part:BBa_K4259003

CopA is a periplasmic Cu-oxidase in C. metallidurans1.

Usage and Biology

In C. metallidurans a reductive precipitation mechanism is used where Au(I)-S complexes are reduced to Au⁰ or Au(I)-C in a process mediated by members of the cop loci. It was found that CopA was strongly upregulated after exposure to Au³⁺ , suggesting that it plays an important role in the gold detoxification mechanism in C. metallidurans ². Additionally, it was shown that CopA functions as an Au(I)-oxidase to promote formation of Au nanoparticles from Au(III) in C. metallidurans³. This makes CopA an interesting candidate for the production of bimetallic gold and silver NPs.

Design

In our project we used CopA from the bacterium Cupriavidus metallidurans as it is a metal resistant bacteria and a suitable candidate for NP formation. The sequence was codon-optimized for E. coli. Then two overhang sequences with the restriction enzymes BamHI and NdeI were designed and added to the sequence. This part is therefore compatible with vectors that have those two restriction sites. In the Binanox project, the vector pET-16b was used.

References

1. Wiesemann N, Bütof L, Herzberg M, Hause G, Berthold L, Etschmann B, Brugger J, Martinez-Criado G, Dobritzsch D, Baginsky S, Reith F, Nies DH. Synergistic Toxicity of Copper and Gold Compounds in Cupriavidus metallidurans. Appl Environ Microbiol. 2017 Nov 16;83(23):e01679-17. doi: 10.1128/AEM.01679-17. PMID: 28939602; PMCID: PMC5691418.

2.Chen, A., Keitz, B. K., & Contreras, L. M. (2018). Biological links between nanoparticle biosynthesis and stress responses in bacteria. Mexican journal of biotechnology, 3(4), 44-69.

3. Butof L, Wiesemann N, Herzberg M, Altzschner M, Holleitner A, Reith F, Nies DH. 2018. Synergistic gold-copper detoxification at the core of gold biomineralisation in Cupriavidus. Metallomics 10:278–286.