Difference between revisions of "Part:BBa K4259002"

Latest revision as of 08:25, 11 October 2022

NapA

napA is a periplasmic nitrate reductase and catalyzes the transformation of nitrate to nitrite in prokaryotes¹. This sequence comes from the metal resistant bacteria Cupriavidus metallidurans (C. metallidurans).

Usage and Biology

The periplasmic nitrate reductases catalyze the transformation of nitrate (NO3-) to nitrite (NO2-), meaning that NapA has a high affinity for nitrate.¹ In previous studies it was seen that it has emerged as a strong candidate for the production of silver nanoparticles (AgNP) in bacteria, especially in a nitrate-rich medium. Nitrate-rich media are known inducers of nitrate reductase. It was shown that AgNP absorbance was nearly tripled in this medium compared to a nitrate-poor media. Additionally, silver resistant E. coli strains were exposed to Ag+ and found to produce AgNPs.² These were promising results to use this gene in the Binanox project to produce bimetallic silver and gold nanoparticles.

Design

In our project we used napA from the bacterium Cupriavidus metallidurans, as it is metal resistant and a suitable candidate for NP formation.² 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. Sparacino-Watkins, C., Stolz, J. F. & Basu, P. Nitrate and periplasmic nitrate reductases. Chemical Society Reviews vol. 43 676–706 Preprint at https://doi.org/10.1039/c3cs60249d (2014).

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.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
INCOMPATIBLE WITH RFC[21]
Illegal BamHI site found at 2517
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal NgoMIV site found at 519
Illegal NgoMIV site found at 1310
Illegal NgoMIV site found at 1754
1000
COMPATIBLE WITH RFC[1000]

@@ Line 3: / Line 3: @@
 <partinfo>BBa_K4259002 short</partinfo>
-NapA is a periplasmic nitrate reductase and part of the nap operon
+''napA'' is a periplasmic nitrate reductase and catalyzes the transformation of nitrate to nitrite in prokaryotes<sup>1</sup>. This sequence comes from the metal resistant bacteria ''Cupriavidus metallidurans'' (''C. metallidurans'').
 ===Usage and Biology===
-The periplasmic nitrate reductases catalyze the transformation of nitrate to nitrite, meaning that NapA has a high affinity for nitrate. In previous studies it was seen that it has emerged as a strong candidate for the production of silver nanoparticles (AgNP) in bacteria, especially in a nitrate-rich media. A nitrate-rich media is a known inducer of nitrate reductase and it was shown that it nearly tripled the absorbance when comparing it to a nitrate-poor media. Additionally, silver resistant E. coli strains were exposed to Ag and found to produce AgNPs <sup>1</sup>. These were promising results to use this gene in the Binanox project to produce bimetallic silver and gold nanoparticles.
+The periplasmic nitrate reductases catalyze the transformation of nitrate (NO3-) to nitrite (NO2-), meaning that NapA has a high affinity for nitrate.<sup>1</sup> In previous studies it was seen that it has emerged as a strong candidate for the production of silver nanoparticles (AgNP) in bacteria, especially in a nitrate-rich medium. Nitrate-rich media are known inducers of nitrate reductase. It was shown that AgNP absorbance was nearly tripled in this medium compared to a nitrate-poor media.  Additionally, silver resistant ''E. coli'' strains were exposed to Ag+ and found to produce AgNPs.<sup>2</sup> These were promising results to use this gene in the Binanox project to produce bimetallic silver and gold nanoparticles.
 ===Design===
-In our project we used NapA 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.
+In our project we used ''napA'' from the bacterium ''Cupriavidus metallidurans'', as it is metal resistant and a suitable candidate for NP formation.<sup>2</sup> The sequence was codon-optimized for ''E. coli''. Then 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===
-. 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.
+. Sparacino-Watkins, C., Stolz, J. F. & Basu, P. Nitrate and periplasmic nitrate reductases. Chemical Society Reviews vol. 43 676–706 Preprint at https://doi.org/10.1039/c3cs60249d (2014).
+. 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.
 <!-- Add more about the biology of this part here