A 2‐Tyr‐1‐carboxylate Mononuclear Iron Center Forms the Active Site of a Paracoccus Dimethylformamidase

Abstract

N,N‐dimethyl formamide (DMF) is an extensively used organic solvent but is also a potent pollutant. Certain bacterial species from genera such as Paracoccus, Pseudomonas, and Alcaligenes have evolved to use DMF as a sole carbon and nitrogen source for growth via degradation by a dimethylformamidase (DMFase). We show that DMFase from Paracoccus sp. strain DMF is a halophilic and thermostable enzyme comprising a multimeric complex of the α2β2 or (α2β2)2 type. One of the three domains of the large subunit and the small subunit are hitherto undescribed protein folds of unknown evolutionary origin. The active site consists of a mononuclear iron coordinated by two Tyr side‐chain phenolates and one carboxylate from Glu. The Fe3+ ion in the active site catalyzes the hydrolytic cleavage of the amide bond in DMF. Kinetic characterization reveals that the enzyme shows cooperativity between subunits, and mutagenesis and structural data provide clues to the catalytic mechanism.

Thermostable enzyme research advances: a bibliometric analysis

“…Fig. 3 Annual scientific production of articles on thermostable enzymes Che Hussian and Leong Journal of Genetic Engineering and Biotechnology (2023) 21:37 [78]. The most important enzymes: Dehydrogenase, peroxidases, phosphatase and pectate lyse.…”

Genomic profiling and characteristics of a C1 degrading heterotrophic fresh-water bacterium Paracoccus sp. strain DMF

“…3A). The translational product of gene dmfA1A2 is a α 2 β 2 -type metallo-amidohydrolase enzyme, N, N-dimethylformamidase (DMFase) that converts DMF into DMA (Arya et al, 2020). dmfR belongs to a LuxR family transcriptional regulators and positively regulates the activation of the dmfRA1A2 operon when DMF is present in the bulk (Dziewit et al, 2014).…”