A novel formaldehyde dismutase gene was identified in Pseudomonas putida J3. High expression levels were obtained in an Escherichia coli rhamnose-inducible system (51 U mg−1). Chaperone GroESL coexpression increased yield of activity in raw extracts 8.6 fold. First report of a functional formaldehyde dismutase with a C-terminal 6xHis-Tag allowing simple affinity purification. Formaldehyde dismutase will be useful for biomethanol production from biogas.