Difference between revisions of "Part:BBa K4259003"
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| − | + | ''copA'' is a periplasmic Cu-oxidase in ''Cupriavidus metallidurans'' (''C. metallidurans'').<sup>1</sup>. | |
===Usage and Biology=== | ===Usage and Biology=== | ||
| − | In C. metallidurans a reductive precipitation mechanism is used where Au(I)-S complexes are reduced to Au<sup>0</sup> or Au(I)-C in a process mediated by | + | In ''C. metallidurans'' a reductive precipitation mechanism is used where Au(I)-S complexes are reduced to Au<sup>0</sup> or Au(I)-C in a process mediated by proteins associated with the cop loci. It was found that ''copA'' was strongly upregulated after exposure to Au<sup>3+</sup> , suggesting that it plays an important role in the gold detoxification mechanism in C. metallidurans.<sup>2</sup> Additionally, it was shown that CopA functions as an Au(I)-oxidase to promote formation of Au<sup>0</sup> nanoparticles from Au<sup>3+</sup> in ''C. metallidurans''.<sup>3</sup> This makes CopA an interesting candidate for the production of bimetallic gold and silver NPs. |
| + | It should be noted that due to old nomenclature, CopA refers to a different protein when mentioned for ''Escherichia coli'' (''E. coli''). The ortholog of CopA in ''C. metallidurans'' is called CueO in ''E. coli''.<sup>1</sup> | ||
| + | |||
===Design=== | ===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. | + | 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''. Two overhang sequences with the restriction enzymes ''BamH''I and ''Nde''I 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=== | ===References=== | ||
Revision as of 08:30, 11 October 2022
copA is a periplasmic Cu-oxidase in Cupriavidus metallidurans (C. metallidurans).1.
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 proteins associated with 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 Au0 nanoparticles from Au3+ in C. metallidurans.3 This makes CopA an interesting candidate for the production of bimetallic gold and silver NPs. It should be noted that due to old nomenclature, CopA refers to a different protein when mentioned for Escherichia coli (E. coli). The ortholog of CopA in C. metallidurans is called CueO in E. coli.1
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. 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.