Haloalkane dehalogenases (HLDs), which catalyse the cleavage of the carbon-halogen bond of organohalogen compounds, are recognized as the key tools in many industrial and biotechnological processes [1]. The HLDs are composed of two domains a main α/β domain containing catalytic residues and a cap domain. According to phylogenetic and structural analyses, HLDs can be divided into three subfamilies − HLD-I, HLD-II and HLD-III, which differ in the cap domain architecture and the composition of their catalytic pentads [2]. Unlike the HLD-I and HLD-II members, which structures have been studied by X-ray crystallography, the structural information on HLD-III enzymes is missing. A major problem with structural characterization of HLD-III members is the fact that these enzymes form polydisperse high-molecular weight oligomeric states [3, 4], which hampers their structural analysis. Here, we report an optimized method for recombinant production and purification of DhmeA from the halophilic archeon Haloferax mediterranei. The strategy employs a robust recombinant expression in Escherichia coli together with protein purification through a poly-histidine affinity tag and size-exclusion chromatography. Subsequent biophysical characterization by differential scanning fluorimetry and dynamic light scattering revealed that the newly developed purification method significantly improved monodispersity of DhmeA. Our findings pave a way for probing DhmeA structure by high-resolution techniques such as X-ray crystallography and single-particle cryo-electron microscopy analysis. Cryo-electron microscopy data was collected and initial 3D model of DhmeA was inferred. Future work will focus on 3D structure reconstruction, model building and experimental validation.