Part:BBa_K4596006

NCD-dependent malic enzyme -> ME

This enzyme catalyzes the synthesis of malic acid from carbon dioxide and pyruvate, and in this process uses the non-natural coenzyme nicotinamide cytidine dinucleotide (NCD), which oxidizes NCDH to NCD, rather than using nicotinamide adenine dinucleotide (NAD). 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
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

Profile

Name: ME
Base Pairs: 1743 bp
Origin: Escherichia coli, synthetic
Properties: Malic acid is synthesized by NCDH, carbon dioxide and pyruvate to ensure the balance of NCD and NCDH

Characterization

We cloned the ME gene into the pet-28(a) vector with a 6xHis tag for easy purification of the malic enzyme.

We expressed the malic enzyme in Escherichia coli BL21. And the activity of malic enzyme was verified in vitro.

Figure 1.malic enzyme catalyzes the synthesis of malic acid over time.

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