Part:BBa_K4596004

NCD dependent formaldehyde dehydrogenase ->FalDH

This enzyme catalyzes the oxidation of formaldehyde to formic acid, and in doing so uses the non-natural coenzyme nicotinamide cytidine dinucleotide (NCD), rather than the nicotinamide adenine dinucleotide (NAD) used by conventional formaldehyde dehydrogenase.

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 837
  Illegal AgeI site found at 325
  Illegal AgeI site found at 520
  Illegal AgeI site found at 1006
  
• 1000
  COMPATIBLE WITH RFC[1000]

Profile

Name: FalDH
Base Pairs: 1218 bp
Origin: Pseudomonas putida, synthetic
Properties: Using NCD as a hydrogen acceptor, formaldehyde was catalyzed to formic acid

Characterization

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

We expressed the formaldehyde dehydrogenase in Escherichia coli BL21.The extracted formaldehyde dehydrogenase was then used for the reaction.

Figure 1.Enzymatic activity of FalDH variants.

References

Wang, J., Guo, X., Wan, L., Liu, Y., Xue, H., & Zhao, Z. K. (2022). Engineering Formaldehyde Dehydrogenase from Pseudomonas putida to Favor Nicotinamide Cytosine Dinucleotide. Chembiochem: A European Journal of Chemical Biology, 23(7), e202100697. https://doi.org/10.1002/cbic.202100697