Engineering Formaldehyde Dehydrogenase from Pseudomonas putida to Favor Nicotinamide Cytosine Dinucleotide

The enzyme formaldehyde dehydrogenase (FalDH) from Pseudomonas putida is of particular interest for biotechnological applications as it catalyzes the oxidation of formaldehyde independent of glutathione. However, the consumption of a stoichiometric amount of nicotinamide adenine dinucleotide (NAD) can be challenging at the metabolic level as this may affect many other NAD‐linked processes. A potential solution is to engineer FalDH to utilize non‐natural cofactors. Here we devised FalDH variants to favor nicotinamide cytosine dinucleotide (NCD) by structure‐guided modification of the binding pocket for the adenine moiety of NAD. Several mutants were obtained and the best one FalDH 9B2 had over 150‐fold higher preference for NCD than NAD. Molecular docking analysis indicated that the cofactor binding pocket shrunk to better fit NCD, a smaller‐sized cofactor. FalDH 9B2 together with other NCD‐linked enzymes offer opportunities to assemble orthogonal pathways for biological conversion of C1 molecules.

• Publication year

  2022

• Venue

  ChemBioChem

• Publication date

  2022-02-10

• Fields of study

  Biology, Chemistry, Engineering, Environmental Science, Medicine

• Identifiers
  DOI 10.1002/cbic.202100697PMID 35146861
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• No claims are published for this paper.

• No concepts are published for this paper.

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