Engineering Formaldehyde Dehydrogenase from Pseudomonas putida to Favor Nicotinamide Cytosine Dinucleotide
- Resource Type
- Authors
- Junting Wang; Xiaojia Guo; Li Wan; Yuxue Liu; Haizhao Xue; Zongbao K. Zhao
- Source
- ChemBioChem. 23
- Subject
- Molecular Docking Simulation
Niacinamide
Cytosine
Pseudomonas putida
Formaldehyde
Organic Chemistry
Molecular Medicine
NAD
Aldehyde Oxidoreductases
Molecular Biology
Biochemistry
- Language
- ISSN
- 1439-7633
1439-4227
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.