Part:BBa_K5041001

Peroxidase-mimicking DNAzyme

Notice: Functional DNA

This part is a sequence of a functional ssDNA. It is only active as single-stranded DNA. It can not be cloned into a plasmid. For use order it as a DNA oligo.

This part forms a G-quadruplex structure when folded, and exhibits peroxidase activity when complexed with hemin. the G-quadruplex-hemin complex can oxidize chromogenic substrate, such as 3,3′,5,5′-Tetramethylbenzidine (TMB) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) in the presence of hydrogen peroxide [1].

This G-quadruplex DNAzyme was discovered by Chen et al. in an investigation into the effects of non-guanine bases on the activity of G-quadruplex DNAzyme, which exhibits the highest activity among the variants investigated in the study [1]. In our project, this DNAzyme is produced when the antibiotic resistance gene is present, thus serving as a reporter. We believe that this part could also serve as a reporter for hydrogen peroxide in other projects.

Chen et al. reported that this DNAzyme has a V0 of 369.9 nM/s [1].

In addition, we also measured the activity of BBa_K5041001 against its predecessor variants. After successfully replicating the optimum signal of BBa_K3343000 and BBa_K3343001 as presented by van den Brink [3], we also plotted the activity of BBa_K5041001 along with the other variants of G-quadruplex DNAzyme. As shown above, BBa_K5041001 exhibited a higher activity compared to the other variants.

[1] J. Chen et al., “How Proximal Nucleobases Regulate the Catalytic Activity of G‑Quadruplex/Hemin DNAzymes,” vol. 8, no. 12, pp. 11352–11361, Dec. 2018, doi: 10.1021/acscatal.8b03811.

[2] F. A. Sorgenfrei, “BBa_K1614007 HRP-mimicking DNAzyme,” Registry of Standard Biological Parts, https://parts.igem.org/Part:BBa_K1614007 (accessed Oct. 22, 2024).

[3] M. van den Brink, “BBa_K3343000 DNAzyme with peroxidase activity,” Registry of Standard Biological Parts, https://parts.igem.org/Part:BBa_K3343000 (accessed Oct. 22, 2024).

[4] M. van den Brink, “BBa_K3343001 DNAzyme with peroxidase activity,” Registry of Standard Biological Parts, https://parts.igem.org/Part:BBa_K3343001 (accessed Oct. 22, 2024).

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]