Part:BBa_K4596005

NCD dependent formate dehydrogenase ->FDH

This enzyme catalyzes the oxidation of formic acid to carbon dioxide and water, and in doing so uses the non-natural coenzyme nicotinamide cytidine dinucleotide (NCD), rather than the nicotinamide adenine dinucleotide (NAD) used by conventional formate dehydrogenase. It's a very orthogonality enzyme.

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
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 300
  Illegal NgoMIV site found at 942
  
• 1000
  COMPATIBLE WITH RFC[1000]

Profile

Name: FDH
Base Pairs: 1224 bp
Origin: Pseudomonas, synthetic
Properties: Using NCD as a hydrogen acceptor, formic acid is catalyzed into carbon dioxide and water

Characterization

We cloned the FDH gene into the pet-28(a) vector with a 6xHis tag for easy purification of the formate dehydrogenase.

We expressed the formate dehydrogenase in Escherichia coli BL21.The enzyme activity was verified in vitro.

Figure 1.Enzymatic activity of FDH variants.

References

Guo, X., Liu, Y., Wang, Q., Wang, X., Li, Q. X., Liu, W.-J., & Zhao, Z. K. (2020). Non‐natural Cofactor and Formate‐Driven Reductive Carboxylation of Pyruvate. Angew. Chem., 59(8), 3143–3146. https://doi.org/10.1002/anie.201915303